WO2009077611A2 - Plants with increased yield and/or increased tolerance to environmental stress (iy-bm) - Google Patents
Plants with increased yield and/or increased tolerance to environmental stress (iy-bm) Download PDFInfo
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- WO2009077611A2 WO2009077611A2 PCT/EP2008/067960 EP2008067960W WO2009077611A2 WO 2009077611 A2 WO2009077611 A2 WO 2009077611A2 EP 2008067960 W EP2008067960 W EP 2008067960W WO 2009077611 A2 WO2009077611 A2 WO 2009077611A2
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- nucleic acid
- polypeptide
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- acid molecule
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants
Definitions
- This invention relates generally to plant cells and/or plants with increased tolerance to environmental stress and/or increased yield as compared to a corresponding, e.g. non-transformed, wild type plant cell by increasing or generating one or more activities of polypeptides associated with abiotic stress responses and abiotic stress tolerance in plants.
- this invention relates to plant cells and/or plants tailored to grow under conditions of environmental stress, and/or to plant cells and/or plants showing increased yield under environmental stress conditions.
- the invention also deals with methods of pro- ducing and screening for and breeding such plant cells and/or plants.
- the present invention disclosed herein provides a method for producing a plant with increased yield as compared to a corresponding wild type plant comprising increasing or generating one or more activities in a plant or a part thereof.
- the present inven- tion further relates to nucleic acids enhancing or improving one or more traits of a transgenic plant, and cells, progenies, seeds and pollen derived from such plants or parts, as well as methods of making and methods of using such plant cell(s) or plant(s), progenies, seed(s) or pollen.
- said improved trait(s) are manifested in an increased yield, preferably by improving one or more yield-related trait(s).
- plant performance for example in terms of growth, development, biomass accumulation and seed generation, depends on a plant's tolerance and acclimation ability to numerous environmental conditions, changes and stresses. Since the beginning of agriculture and horticulture, there was a need for improving plant traits in crop cultivation. Breeding strategies foster crop properties to withstand biotic and abiotic stresses, to im- prove nutrient use efficiency and to alter other intrinsic crop specific yield parameters, i.e. increasing yield by applying technical advances. Plants are sessile organisms and consequently need to cope with various environmental stresses.
- Agricultural biotechnologists have used assays in model plant systems, greenhouse studies of crop plants, and field trials in their efforts to develop transgenic plants that exhibit increased yield, either through increases in abiotic stress tolerance or through increased biomass. Agricultural biotechnologists also use measurements of other parameters that indicate the potential impact of a transgene on crop yield. For forage crops like alfalfa, silage corn, and hay, the plant biomass correlates with the total yield. For grain crops, however, other parameters have been used to estimate yield, such as plant size, as measured by total plant dry weight, above-ground dry weight, above-ground fresh weight, leaf area, stem volume, plant height, rosette diameter, leaf length, root length, root mass, tiller number, and leaf number.
- Plant size at an early developmental stage will typically correlate with plant size later in development.
- a larger plant with a greater leaf area can typically absorb more light and carbon dioxide than a smaller plant and therefore will likely gain a greater weight during the same period.
- the present invention provides a method for producing a plant with increased yield as compared to a corresponding wild type plant comprising at least the following step: increasing or generating in a plant one or more activities (in the following referred to as one or more "activities” or one or more of said activities or for one selected activity as "said activity") selected from the group consisting of 3OS ribosomal protein S1 1 , 60S ribosomal protein, Adaptin medium chain homolog APM2, B0252-protein, BRICK1 -like protein, Cav1 protein, Chloroplast chaperonin, DNA polymerase, flagellar pro- O
- the invention provides a transgenic plant that over- expresses an isolated polynucleotide identified in Table I in the sub-cellular compartment and tissue indicated herein.
- the transgenic plant of the invention demonstrates an improved yield or increased yield as compared to a wild type variety of the plant.
- improved yield or “increased yield” can be used interchangeable.
- yield generally refers to a measurable produce from a plant, particularly a crop. Yield and yield increase (in comparison to a non-transformed starting or wild-type plant) can be measured in a number of ways, and it is understood that a skilled person will be able to apply the correct meaning in view of the particular embodiments, the particular crop concerned and the specific purpose or application concerned.
- the term “improved yield” or the term “increased yield” means any improvement in the yield of any measured plant product, such as grain, fruit or fiber.
- changes in different phenotypic traits may improve yield.
- parameters such as floral organ development, root initiation, root biomass, seed number, seed weight, harvest index, tolerance to abiotic environmental stress, leaf formation, phototropism, apical dominance, and fruit development, are suitable measurements of improved yield.
- Any increase in yield is an improved yield in accordance with the invention.
- the improvement in yield can comprise a 0.1 %, 0.5%, 1 %, 3%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater increase in any measured parameter.
- an increase in the bu/acre yield of soybeans or corn derived from a crop comprising plants which are transgenic for the nucleotides and polypeptides of Table I, as compared with the bu/acre yield from untreated soybeans or corn cultivated under the same conditions, is an improved yield in accordance with the invention.
- the increased or improved yield can be achieved in the absence or presence of stress conditions.
- yield refers to one or more yield parameters selected from the group consisting of biomass yield, dry biomass yield, aerial dry biomass yield, underground dry biomass yield, fresh-weight biomass yield, aerial fresh-weight biomass yield, underground fresh-weight biomass yield; enhanced yield of harvestable parts, either dry or fresh-weight or both, either aerial or un- derground or both; enhanced yield of crop fruit, either dry or fresh-weight or both, either aerial or underground or both; and preferably enhanced yield of seeds, either dry or fresh- weight or both, either aerial or underground or both.
- yield as used herein generally refers to a measurable produce from a plant, particularly a crop.
- the present invention provides methods for producing transgenic plant cells or plants with can show an increased yield-related trait, e.g. an increased tolerance to environmental stress and/or increased intrinsic yield and/or biomass production as compared to a corresponding (e.g. non-transformed) wild type or starting plant by increasing or generating one or more of said activities mentioned above.
- an increased yield-related trait e.g. an increased tolerance to environmental stress and/or increased intrinsic yield and/or biomass production as compared to a corresponding (e.g. non-transformed) wild type or starting plant by increasing or generating one or more of said activities mentioned above.
- an increase in yield refers to increased or improved crop yield or har- vestable yield, biomass yield and/or an increased seed yield.
- Crop yield is defined herein as the number of bushels of relevant agricultural product (such as grain, forage, or seed) harvested per acre. Crop yield is impacted by abiotic stresses, such as drought, heat, salinity, and cold stress, and by the size (biomass) of the plant. Traditional plant breeding strategies are relatively slow and have in general not been success- ful in conferring increased tolerance to abiotic stresses. Grain yield improvements by conventional breeding have nearly reached a plateau in maize.
- yield refers to harvestable yield of a plant.
- the yield of a plant can depend on the specific plant/ crop of interest as well as its intended application (such as food production, feed production, processed food production, bio-fuel, biogas or alcohol production, or the like) of interest in each particular case.
- yield is calculated as harvest index (expressed as a ratio of the weight of the respective harvestable parts divided by the total biomass), harvestable parts weight per area (acre, square meter, or the like); and the like.
- the harvest index i.e., the ratio of yield biomass to the total cumulative biomass at harvest, in maize has remained essentially un- changed during selective breeding for grain yield over the last hundred years.
- biomass yield refers to biomass yield, e.g. to dry weight biomass yield and/or fresh-weight biomass yield.
- Biomass yield refers to the aerial or underground parts of a plant, depending on the specific circumstances (test conditions, specific crop of interest, application of interest, and the like). In one embodiment, biomass yield refers to the aerial and underground parts. Biomass yield may be calculated as fresh-weight, dry weight or a moisture adjusted basis. Biomass yield may be calculated on a per plant basis or in relation to a specific area (e.g. biomass yield per acre/ square meter/ or the like).
- yield refers to seed yield which can be measured by one or more of the following parameters: number of seeds or number of filled seeds (per plant or per area (acre/ square meter/ or the like)); seed filling rate (ratio between number of filled seeds and total number of seeds); number of flowers per plant; seed biomass or total seeds weight (per plant or per area (acre/square meter/ or the like); thousand kernel weight (TKW; extrapolated from the number of filled seeds counted and their total weight; an increase in TKW may be caused by an increased seed size, an increased seed weight, an increased embryo size, and/or an increased endosperm). Other parameters allowing to measure seed yield are also known in the art. Seed yield may be determined on a dry weight or on a fresh weight basis, or typically on a moisture adjusted basis, e.g. at 15.5 percent moisture.
- the term "increased yield” means that the photosynthetic active organ- ism, especially a plant, exhibits an increased growth rate, under conditions of abiotic environmental stress, compared to the corresponding wild-type photosynthetic active organism.
- An increased growth rate may be reflected inter alia by or confers an increased biomass production of the whole plant, or an increased biomass production of the aerial parts of a plant, or by an increased biomass production of the underground parts of a plant, or by an increased biomass production of parts of a plant, like stems, leaves, blossoms, fruits, and/or seeds.
- increased yield includes higher fruit yields, higher seed yields, higher fresh matter production, and/or higher dry matter production.
- the term "increased yield” means that the photosynthetic active organism, preferably plant, exhibits an prolonged growth under conditions of abiotic environmental stress, as compared to the corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- a prolonged growth comprises survival and/or continued growth of the photosynthetic active organism, preferably plant, at the moment when the non-transformed wild type photosynthetic active organism shows visual symptoms of defi- ciency and/or death.
- the plant used in the method of the invention is a corn plant.
- Increased yield for corn plants means in one embodiment, increased seed yield, in particular for corn varieties used for feed or food.
- Increased seed yield of corn refers in one embodiment to an increased kernel size or weight, an increased kernel per pod, or in- creased pods per plant.
- the cob yield is increased, this is particularly useful for corn plant varieties used for feeding.
- the length or size of the cob is increased.
- increased yield for a corn plant relates to an improved cob to kernel ratio.
- the plant used in the method of the invention is a soy plant.
- Increased yield for soy plants means in one embodiment, increased seed yield, in particular for soy varieties used for feed or food.
- Increased seed yield of soy refers in one embodiment to an increased kernel size or weight, an increased kernel per pod, or increased pods per plant.
- the plant used in the method of the invention is an oil seed rape (OSR) plant.
- Increased yield for OSR plants means in one embodiment, increased seed yield, in particular for OSR varieties used for feed or food.
- Increased seed yield of OSR refers in one embodiment to an increased kernel size or weight, an increased kernel per pod, or increased pods per plant.
- the plant used in the method of the invention is a cotton plant.
- Increased yield for cotton plants means in one embodiment, increased lint yield.
- Increased cotton yield of cotton refers in one embodiment to an increased length of lint.
- Increased seed yield of corn refers in one embodiment to an increased kernel size or weight, an increased kernel per pod, or increased pods per plant.
- Said increased yield in accordance with the present invention can typically be achieved by enhancing or improving, in comparison to an origin or wild-type plant, one or more yield- related traits of the plant.
- yield-related traits of a plant the improvement of which results in increased yield comprise, without limitation, the increase of the intrinsic yield capac- ity of a plant, improved nutrient use efficiency, and/or increased stress tolerance, in particular increased abiotic stress tolerance.
- yield can be increased by improving one or more of the yield-related traits as defined herein:
- the yield-related trait conferring an increase of the plant's yield is an increase of the intrinsic yield capacity of a plant and can be, for example, manifested by improving the specific (intrinsic) seed yield (e.g. in terms of increased seed/ grain size, increased ear number, increased seed number per ear, improvement of seed filling, improvement of seed composition, embryo and/or endosperm improvements, or the like); modification and improvement of inherent growth and development mechanisms of a plant (such as plant height, plant growth rate, pod number, pod position on the plant, number of internodes, incidence of pod shatter, efficiency of nodulation and nitrogen fixation, efficiency of carbon assimilation, improvement of seedling vigour/early vigour, enhanced efficiency of germination (under stressed or non-stressed conditions), improvement in plant architecture, cell cycle modifications, photosynthesis modifications, various signaling pathway modifications, modification of transcriptional regulation, modification of translational regulation, modification of enzyme activities, and the like); and/or the like.
- specific (intrinsic) seed yield e.
- the yield-related trait conferring an increase of the plant's yield is an improvement or increase of stress tolerance of a plant and can be for example manifested by improving or increasing a plant's tolerance against stress, particularly abiotic stress.
- abiotic stress refers generally to abiotic environmental conditions a plant is typically confronted with, including conditions which are typically referred to as "abiotic stress" conditions including, but not limited to, drought (tolerance to drought may be achieved as a result of improved water use efficiency), heat, low temperatures and cold conditions (such as freezing and chilling conditions), salinity, osmotic stress , shade, high plant density, mechanical stress, oxidative stress, and the like.
- an increased plant yield is mediated by increasing the "nutrient use efficiency of a plant", e.g. by improving the use efficiency of nutrients including, but not limited to, phosphorus, potassium, and nitrogen.
- a plant e.g. by improving the use efficiency of nutrients including, but not limited to, phosphorus, potassium, and nitrogen.
- nutrients including, but not limited to, phosphorus, potassium, and nitrogen.
- plant yield is increased by increasing nitrogen use efficiency of a plant or a part thereof.
- Enhanced nitrogen use efficency of the plant can for example be determined and quantified according to the following method: Transformed plants are grown in pots in a growth chamber (Sval ⁇ f Weibull, Sval ⁇ v, Sweden).
- the plants are Arabidopsis thaliana seeds thereof are sown in pots containing a 1 :1 (v:v) mixture of nutrient depleted soil ("Einheit- serde Typ 0", 30% clay, Tantau, Wansdorf Germany) and sand. Germination is induced by a four day period at 4 0 C, in the dark. Subsequently the plants are grown under standard growth conditions. In case the plants are Arabidopsis thaliana, the standard growth conditions are: photoperiod of 16 h light and 8 h dark, 20 0 C, 60% relative humidity, and a photon flux density of 200 ⁇ E.
- the plants are Arabidopsis thaliana they are watered every second day with a N-depleted nutrient solution. After 9 to 10 days the plants are individual- ized. After a total time of 29 to 31 days the plants are harvested and rated by the fresh weight of the aerial parts of the plants, preferably the rosettes.
- the increased yield can be determined according to the method described in the examples, e.g.: (a) measuring the nitrogen content in the soil, and (b) determining, whether the nitrogen -content in the soil is optimal or suboptimal for the growth of an origin or wild type plant, e.g. a crop, and (c1 ) growing the plant of the invention in said soil, if the nitrogen -content is suboptimal for the growth of the origin or wild type plant, or (c2) growing the plant of the invention in the soil and comparing the yield with the yield of a standard, an origin or a wild type plant, selecting and growing the plant, which shows the highest yield, if the nitrogen - content is optimal for the origin or wild type plant. Plant yield is increased by increasing the plant's stress tolerance(s).
- the term " increased tolerance to stress” can be defined as survival of plants, and/or higher yield production, under stress conditions as compared to a n on -transformed wild type or starting plant.
- the plant of the invention or produced according to the method of the invention is better adapted to the stress conditions.
- "Improved adaptation" to environmental stress like e.g. draught, heat, nutrient depletion, freezing and/or chilling temperatures refers o
- Stress condition a condition where biotic stress may generally be divided into biotic and abiotic (environmental) stresses. Unfavorable nutrient conditions are sometimes also referred to as “environmental stress”.
- the present invention does also contemplate solutions for this kind of environmental stress, e.g. referring to increased nutrient use efficiency. Plant yield can be further increased by increasing the abiotic stress tolerance(s) of a plant or a part thereof.
- the terms “enhanced tolerance to abiotic stress”, “enhanced resistance to abiotic environmental stress”, “enhanced tolerance to environmental stress”, “improved adaptation to environmental stress” and other variations and expressions similar in its meaning are used interchangeably and refer, without limitation, to an improvement in tolerance to one or more abiotic environmental stress(es) as described herein and as compared to a corresponding origin or wild type plant or a part thereof.
- abiotic stress tolerance(s) refers for example low temperature tolerance, drought tolerance or improved water use efficiency (WUE), heat tolerance, salt stress tolerance and others. Studies of a plant's response to desiccation, osmotic shock, and temperature extremes are also employed to determine the plant's tolerance or resistance to abiotic stresses.
- Stress tolerance in plants like low temperature, drought, heat and salt stress tolerance can have a common theme important for plant growth, namely the availability of water. Plants are typically exposed during their life cycle to conditions of reduced environmental water content. The protection strategies are similar to those of chilling tolerance.
- said yield-related trait relates to an increased water use efficiency of the plant of the invention and/ or an increased tolerance to drought conditions of the plant of the in- vention.
- Water use efficiency is a parameter often correlated with drought tolerance. An increase in biomass at low water availability may be due to relatively improved efficiency of growth or reduced water consumption.
- a decrease in water use, without a change in growth would have particular merit in an irrigated agricultural system where the water input costs were high.
- An increase in growth without a corre- sponding jump in water use would have applicability to all agricultural systems. In many agricultural systems where water supply is not limiting, an increase in growth, even if it came at the expense of an increase in water use also increases yield.
- Drought stress means any environmental stress which leads to a lack of water in plants or reduction of water supply to plants, including a secondary stress by low temperature and/or salt, and/or a primary stress during drought or heat, e.g. desiccation etc.
- Increased tolerance to drought conditions can for example be determined and quantified according to the following method.
- transformed plants are grown individually in pots in a growth chamber (York lndustriekalte GmbH, Mannheim, Germany). Germination is induced.
- the plants are Arabidopsis thaliana sown seeds are kept at 4 0 C, in the dark, for 3 days in order to induce germination.
- conditions are changed for 3 days to 2O 0 C/ 6 0 C day/night temperature with a 16/8h day-night cycle at 150 ⁇ E/m2s.
- the plants are grown under standard growth conditions.
- the plants are Arabidopsis thaliana
- the standard growth conditions are: photoperiod of 16 h light and 8 h dark, 20 0 C, 60% relative humidity, and a photon flux density of 200 ⁇ E. Plants are grown and cultured until they develop leaves. In case the plants are Arabidopsis thaliana they are watered daily until they were approximately 3 weeks old. Starting at that time drought was imposed by withholding water. After the non-transformed wild type plants show visual symptoms of injury, the evaluation starts and plants are scored for symptoms of drought symptoms and biomass production comparison to wild type and neighboring plants for 5 - 6 days in succession.
- the tolerance to drought e.g. the tolerance to cycling drought is determined according to the method described in the examples.
- the tolerance to drought can be a tolerance to cycling drought.
- a method for increasing the yield can comprise the following steps: (a) determining, whether the water supply in the area for planting is optimal or suboptimal for the growth of an origin or wild type plant, e.g.
- Visual symptoms of injury for example state for one or any combination of two, three or more of the following features:(a) wilting; (b) leaf browning; (c) loss of turgor, which results in drooping of leaves or needles stems, and flowers, (d) drooping and/or shedding of leaves or needles, (e) the leaves are green but leaf angled slightly toward the ground compared with controls, (f) leaf blades begun to fold (curl) inward, (g) premature senescence of leaves or needles, (h) loss of chlorophyll in leaves or needles and/or yellowing.
- Said yield-related trait of the plant of the invention can be an increased tolerance to heat conditions of said plant.
- Said yield-related trait of the plant of the invention can be an increased low temperature tolerance of said plant, e.g. comprising freezing tolerance and/or chilling tolerance.
- Low temperatures impinge on a plethora of biological processes. They retard or inhibit almost all metabolic and cellular processes
- the response of plants to low temperature is an important determinant of their ecological range. The problem of coping with low temperatures is exacerbated by the need to prolong the growing season beyond the short summer found at high latitudes or altitudes. Most plants have evolved adaptive strategies to protect themselves against low temperatures. Generally, adaptation to low temperature may be divided into chilling tolerance, and freezing tolerance. Chilling tolerance is naturally found in species from temperate or boreal zones and allows survival and an enhanced growth at low but non-freezing temperatures.
- improved or enhanced “chilling tolerance” or variations thereof refers herein to improved adaptation to low but non-freezing temperatures around 10 0 C, preferably temperatures between 1 to 18 0 C, more preferably 4-14 0 C, and most preferred 8 to 12 0 C; hereinafter called "chilling temperature”.
- Freezing tolerance allows survival at near zero to particularly subzero temperatures. It is believed to be promoted by a process termed cold-acclimation which occurs at low but non- freezing temperatures and provides increased freezing tolerance at subzero temperatures. In addition, most species from temperate regions have life cycles that are adapted to seasonal changes of the temperature. For those plants, low temperatures may also play an important role in plant development through the process of stratification and vernalisation. It becomes obvious that a clear-cut distinction between or definition of chilling tolerance and freezing tolerance is difficult and that the processes may be overlapping or interconnected.
- Improved or enhanced "freezing tolerance” or variations thereof refers herein to improved adaptation to temperatures near or below zero, namely preferably temperatures below 4 0 C, more preferably below 3 or 2 0 C, and particularly preferred at or below 0 (zero) 0 C or below -4 0 C, or even extremely low temperatures down to -10 0 C or lower; hereinafter called "freezing temperature.
- the plant may show an early seedling growth after exposure to low temperatures to an chilling-sensitive wild type or origin, improving in a further embodiment seed germination rates.
- the process of seed germination strongly depends on environmental temperature and the properties of the seeds determine the level of activity and performance during germination and seedling emergence when being exposed to low temperature.
- the method of the invention further provides in one embodiment a plant which show under chilling condition an reduced delay of leaf development.
- the method of the invention relates to a production of a tolerant major crop, e.g. corn (maize), bean, rice, soy bean, cotton, tomato, banana, cucumber or potato because most major crops are chilling- sensitive.
- a tolerant major crop e.g. corn (maize), bean, rice, soy bean, cotton, tomato, banana, cucumber or potato because most major crops are chilling- sensitive.
- Enhanced tolerance to low temperature may, for example, be determined according to the following method: Transformed plants are grown in pots in a growth chamber (e.g. York, Mannheim, Germany). In case the plants are Arabidopsis thaliana seeds thereof are sown in pots containing a 3.5:1 (v:v) mixture of nutrient rich soil (GS90, Tantau, Wansdorf, Ger- many) and sand. Plants are grown under standard growth conditions. In case the plants are Arabidopsis thaliana, the standard growth conditions are: photoperiod of 16 h light and 8 h dark, 20 0 C, 60% relative humidity, and a photon flux density of 200 ⁇ mol/m2s. Plants are grown and cultured.
- the plants are Arabidopsis thaliana they are watered every second day. After 9 to 10 days the plants are individualized. Cold (e.g. chilling at 1 1 - 12 0 C) is applied 14 days after sowing until the end of the experiment. After a total growth period of 29 to 31 days the plants are harvested and rated by the fresh weight of the aerial parts of the plants, in the case of Arabidopsis preferably the rosettes.
- a method for increasing yield may comprise the following steps: (a) determining, whether the temperature in the area for planting is optimal or suboptimal for the growth of an origin or wild type plant, e.g. a crop; and (b1) growing the plant of the invention in said soil; if the temperature is suboptimal low for the growth of an origin or wild type plant growing in the area; or (b2) growing the plant of the invention in the soil and comparing the yield with the yield of a standard, an origin or a wild type plant and selecting and growing the plant, which shows the highest yield, if the temperature is optimal for the origin or wild type plant.
- an origin or wild type plant e.g. a crop
- Yield-related trait may also be increased salinity tolerance (salt tolerance), tolerance to osmotic stress, increased shade tolerance, increased tolerance to a high plant density, increased tolerance to mechanical stresses, and/or increased tolerance to oxidative stress.
- the term "enhanced tolerance to abiotic environmental stress" in a photo- synthetic active organism can also mean that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced dry biomass yield as compared to a corresponding, e.g. non-transformed, wild type photo- synthetic active organism like a plant.
- the term "enhanced tolerance to abiotic environmental stress" in a photosyn- thetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced aerial dry biomass yield as compared to a corresponding, e.g. non-transformed, wild type photo- synthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced underground dry biomass yield as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced fresh weight biomass yield as compared to a corresponding, e.g. non-transformed, wild type pho- tosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced aerial fresh weight biomass yield as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced underground fresh weight biomass yield as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of harvestable parts of a plant as compared to a corresponding, e.g. non- transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of dry harvestable parts of a plant as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of dry aerial harvestable parts of a plant as compared to a corresponding, e.g. non- transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of underground dry harvestable parts of a plant as compared to a corresponding, e.g. non- transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of fresh weight harvestable parts of a plant as compared to a corresponding, e.g. non- transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions an enhanced yield of aerial fresh weight harvestable parts of a plant as compared to a corresponding, e.g. non- transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of underground fresh weight harvestable parts of a plant as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of the crop fruit as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of the fresh crop fruit as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of the dry crop fruit as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosynthetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced grain dry weight as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosyn- thetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of seeds as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosyn- thetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of fresh weight seeds as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the term "enhanced tolerance to abiotic environmental stress" in a photosyn- thetic active organism means that the photosynthetic active organism, preferably a plant, when confronted with abiotic environmental stress conditions exhibits an enhanced yield of dry seeds as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- the abiotic environmental stress conditions, the organism is confronted with can, however, be any of the abiotic environmental stresses mentioned herein.
- An increased nitrogen use efficiency of the produced corn relates in one embodiment to an improved protein content of the corn seed, in particular in corn seed used as feed. Increased nitrogen use efficiency relates in another embodiment to an increased kernel size or number. A increased water use efficiency of the produced corn relates in one embodiment to an increased kernel size or number. Further, an increased tolerance to low temperature relates in one embodiment to an early vigor and allows the early planting and sowing of a corn plant produced according to the method of the present invention.
- a increased nitrogen use efficiency of the produced soy plant relates in one embodiment to an improved protein content of the soy seed, in particular in soy seed used as feed.
- Increased nitrogen use efficiency relates in another embodiment to an increased kernel size or number.
- a increased water use efficiency of the produced soy plant relates in one embodiment to an increased kernel size or number.
- an increased tolerance to low temperature relates in one embodiment to an early vigor and allows the early planting and sowing of a soy plant produced according to the method of the present invention.
- a increased nitrogen use efficiency of the produced OSR plant relates in one embodiment to an improved protein content of the OSR seed, in particular in OSR seed used as feed, lncresed nitrogen use efficiency relates in another embodiment to an in- creased kernel size or number.
- a increased water use efficiency of the produced OSR plant relates in one embodiment to an increased kernel size or number.
- an increased tolerance to low temperature relates in one embodiment to an early vigor and allows the early planting and sowing of a soy plant produced according to the method of the present invention.
- the present invention relates to a method for the production of hardy oil seed rape (OSR with winter hardness) comprising using a hardy oil seed rape plant in the above mentioned method of the invention.
- a increased nitrogen use efficiency of the produced cotton plant relates in one embodiment to an improved protein content of the cotton seed, in particular in cotton seed used for feeding. Increased nitrogen use efficiency relates in another embodiment to an increased kernel size or number. A increased water use efficiency of the produced cotton plant relates in one embodiment to an increased kernel size or number. Further, an increased tolerance to low temperature relates in one embodiment to an early vigor and allows the early planting and sowing of a soy plant produced according to the method of the present invention.
- antioxidant enzymes or ROS- scavenging enzymes is one possibility to engineer tolerance, e.g. transgenic alfalfa plants expressing Mn-superoxide dismutase tend to have reduced injury after water-deficit stress (McKersie et al., 1996. Plant Physiol. 1 1 1 , 1177-1 181 ). These same transgenic plants show increased yield in field trials (McKersie et al., 1999. Plant Physiology, 119: 839-847; McKersie et al., 1996. Plant Physiol. 11 1 , 1 177-1 181 ).
- Transgenic plants that overproduce osmolytes such as mannitol, fructans, proline or glycine-betaine also show increased toler- ance to some forms of abiotic stress and it is proposed that the synthesized osmolytes act as ROS scavengers (Tarczynski. et al. 1993. Science 259, 508-510; Sheveleva,. et al. 1997. Plant Physiol.1 15, 121 1-1219).
- the transformed and stress resistant plants cited above generally exhibit slower growth and reduced biomass, due to an imbalance in development and physiology of the plant, thus having significant fitness cost (Kasuga et al., 1999; Danby and Gehring et al., 2005). Despite maintaining basic metabolic function this leads to severe biomass and yield loss.
- the present invention provides a method for producing a transgenic plant showing an increased yield- related trait as compared to a corresponding origin or the wild type plant, by increasing or generating one or more activities (in the following "activities") selected from the group consisting of 3OS ribosomal protein S1 1 , 60S ribosomal protein, Adaptin medium chain ho- molog APM2, B0252-protein, BRICK1-like protein, Cav1 protein, Chloroplast chaperonin, DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GREI -protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoenzyme cyclin-like subunit, Serine/threonine-protein kinase, Short chain dehydrogenase, Ste
- activities selected from the group consisting of 3OS ribosomal protein S1 1 , 60S ribosomal protein, Adaptin medium chain
- the present invention provides a method for producing a transgenic plant cell or plant with increased tolerance to environmental stress and/or increased yield or biomass production as compared to a corresponding (non-transformed) wild type or starting plant cell by increasing or generating one or more activities selected from the group consisting of: 3OS ribosomal protein S1 1 , 60S ribosomal protein, Adaptin medium chain homolog APM2, B0252-protein, BRICK1 -like protein, Cav1 protein, Chloroplast chaperonin, DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GREI -protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoenzyme cyclin-like subunit, Ser- ine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, transcription regulator (farR), Ykr015c-protein
- the present invention provides a method for producing a transgenic plant showing an increased yield as compared to a corresponding origin or wild type plant, by increasing or generating one or more said "activities" according to the methods disclosed below.
- enhanced or increased “yield” refers to one or more yield parameters selected from the group consisting of biomass yield, dry biomass yield, aerial dry biomass yield, underground dry biomass yield, freshweight biomass yield, aerial freshweight biomass yield, underground freshweight biomass yield; enhanced yield of har- vestable parts, either dry or freshweight or both, either aerial or underground or both; enhanced yield of crop fruit, either dry or freshweight or both, either aerial or underground or both; and preferably enhanced yield of seeds, either dry or freshweight or both, either aerial or underground or both.
- yield is, mainly, dependent on the crop of inter- est, and it is understood, that the skilled person will understand in each particular case what is meant from the circumstances of the description.
- the activity is increased by increasing the amount and/or activity of one or more proteins having an activity selected from the group consisting of: 3OS ribosomal protein S1 1 , 60S ribosomal protein, Adaptin medium chain homolog APM2, B0252-protein, BRICK1 -like protein, Cav1 protein, Chloroplast chaperonin, DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GREI -protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoenzyme cyclin-like subunit, Ser- ine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, tran- scription regulator (farR), Ykr015c-protein, and YPL167C_2-protein and the polypeptides as depicted in table II, column 5 and 7.
- said activity is increased in one or more specific compartments of a cell and confers an increased yield, e.g. the plant shows an increased or improved said yield- related trait.
- said activity is increased in the plastid of a cell as indicated in table I or Il in column 6 and increases yield in a corresponding plant.
- the specific plastidic localization of said activity which can be derived form the disclosure of table I or Il in column 6 confers an improved or increased yield-related trait as shown in table Villa to VIIId.
- said activity can be increased in mitochondria of a cell and increases yield in a corresponding plant, e.g. conferring an improved or increased yield-related trait as the relevant activities indicated in table Villa to VIIId, if applicable.
- the present invention relates to method for producing a plant with increased yield as compared to a corresponding wild type plant comprising at least one of the steps selected from the group consisting of: (i) increasing or generating the activity of a polypeptide comprising a polypeptide, a consensus sequence or at least one polypeptide motif as de- picted in column 5 or 7 of table Il or of table IV, respectively; (ii) increasing or generating the activity of an expression product of one or more nucleic acid molecule(s) comprising one or more polynucleotide(s) as depicted in column 5 or 7 of table I, and (iii) increasing or generating the activity of a functional equivalent of (i) or (ii).
- the increase or generation of one or more said activities is for example con- ferred by one or more expression products of said nucleic acid molecule, e.g. proteins. Accordingly, in the present invention described above, the increase or generation of one or more said activities is for example conferred by one or more protein(s) each comprising a polypeptide selected from the group as depicted in table II, column 5 and 7.
- the method of the invention comprises in one embodiment the following steps: (i) increas- ing or generating of the expression of; and/or (ii) increasing or generating the expression of an expression product; and/or (iii) increasing or generating one or more activities of an expression product encoded by; at least one nucleic acid molecule (in the following "Yield Related Protein (YIP)"-encoding gene or "YIP”-gene) comprising a nucleic acid molecule selected from the group consisting of: (a) a nucleic acid molecule encoding the polypeptide shown in column 5 or 7 of table II;
- YIP Yield Related Protein
- nucleic acid molecule which, as a result of the degeneracy of the genetic code, can be derived from a polypeptide sequence depicted in column 5 or 7 of table Il and confers an increased yield as compared to a corresponding, e.g. non-transformed, wild type plant cell, a transgenic plant or a part thereof ;
- nucleic acid molecule having at least 30, for example 50, 60, 70, 80, 85, 90, 95, 97, 98, or 99 % identity with the nucleic acid molecule sequence of a polynucleotide comprising the nucleic acid molecule shown in column 5 or 7 of table I and confers an increased yield as compared to a corresponding, e.g. non-transformed, wild type plant cell, a transgenic plant or a part thereof;
- nucleic acid molecule encoding a polypeptide having at least 30, for example 50, 60, 70, 80, 85, 90, 95, 97, 98, or 99 % identity with the amino acid sequence of the polypeptide encoded by the nucleic acid molecule of (a) to (c) and having the activity represented by a nucleic acid molecule comprising a polynucleotide as depicted in col- umn 5 of table I and confers an increased yield as compared to a corresponding, e.g. non-transformed, wild type plant cell, a transgenic plant or a part thereof;
- nucleic acid molecule which hybridizes with a nucleic acid molecule of (a) to (c) under stringent hybridization conditions and confers an increased yield as compared to a corresponding, e.g. non-transformed, wild type plant cell, a transgenic plant or a part thereof;
- nucleic acid molecule encoding a polypeptide which can be isolated with the aid of monoclonal or polyclonal antibodies made against a polypeptide encoded by one of the nucleic acid molecules of (a) to (e) and having the activity represented by the nucleic acid molecule comprising a polynucleotide as depicted in column 5 of table I;
- nucleic acid molecule encoding a polypeptide comprising the consensus sequence or one or more polypeptide motifs as shown in column 7 of table IV and preferably having the activity represented by a nucleic acid molecule comprising a polynucleotide as depicted in column 5 of table Il or IV;
- nucleic acid molecule encoding a polypeptide having the activity represented by a protein as depicted in column 5 of table Il and conferring increased yield as compared to a corresponding, e.g. non-transformed, wild type plant cell, a transgenic plant or a part thereof;
- nucleic acid molecule which comprises a polynucleotide, which is obtained by amplifying a cDNA library or a genomic library using the primers in column 7 of table III and preferably having the activity represented by a nucleic acid molecule comprising a polynucleotide as depicted in column 5 of table Il or IV;
- nucleic acid molecule which is obtainable by screening a suitable nucleic acid library under stringent hybridization conditions with a probe comprising a complementary sequence of a nucleic acid molecule of (a) or (b) or with a fragment thereof, having at least 15nt, preferably 20nt, 30nt, 50nt, 100nt, 200nt, or 500nt, 1000nt, 1500nt, 2000nt or 3000nt of a nucleic acid molecule complementary to a nucleic acid molecule sequence characterized in (a) to (e) and encoding a polypeptide having the activity represented by a protein comprising a polypeptide as depicted in column 5 of table II.
- the present invention provides YIP and YIP genes. Further, the present inven- tion provides YIP and YIP genes derived from plants and other organisms in column 5 as well as in column 7 of tables I or II. Accordingly, the present invention provides YIP and YIP genes derived from plants. In particular, gene from plants are described in column 5 as well as in column 7 of tables I or II.
- abiotic environmental stress As used herein, the terms “environmental stress” or “abiotic environmental stress” and variations thereof are used interchangeably and refer to any abiotic suboptimal growing condition and include, without limitation, suboptimal conditions associated with low temperature, heat, oxidative stress, drought and salinity or combinations thereof.
- the term "increased tolerance to environmental stress” relates to an increased tolerance to water stress, which is produced as a secondary stress by low temperature and/or salt, and/or as a primary stress during drought or heat.
- the term " increased tolerance to environmental stress” relates to an increased low temperature tolerance. In one embodiment of the invention the term “increased tolerance to environmental stress” relates to an increased salt tolerance. In a preferred embodiment of the invention the term “increased tolerance to environmental stress” relates to an increased drought tolerance.
- the term "increased tolerance to environmental stress” relates to an increased low temperature tolerance, comprising freezing tolerance and/or chilling tolerance.
- the term "increased tolerance to environmental stress” is defined as survival of plants, and/or higher yield or biomass production, under stress conditions as compared to non-transformed wild type or starting plants.
- these traits are achieved by a process for an enhanced tolerance to abiotic environmental stress in a photosynthetic active organism, preferably a plant, as compared to a corresponding (non-transformed) wild type or starting photosynthetic active organism.
- the terms “enhanced tolerance to environmental stress”, “enhanced resistance to abiotic environmental stress”, “enhanced tolerance to environmental stress”, “improved adaptation to environmental stress” and variations thereof are used interchangeably and refer preferably, without limitation, to an improvement in tolerance to one or more abiotic environmental stress(es) selected from the group consisting of salt stress, heat stress, drought stress and low temperature stress, while low temperature stress refers to chilling tolerance and/or freezing tolerance of a pho- tosynthetic active organism as compared to a corresponding (non-transformed) wild type (or starting) photosynthetic active organism.
- Improved adaptation or “enhanced tolerance” to environmental stress like refers to an improved plant performance, while improved plant performance may also be achieved by im- proving the intrinsic plant properties resulting to an increase in yield and/or biomass production.
- the present invention refers to a process of developing or obtaining a plant or plant cell that exhibits increased yield, a plant or plant cell that exhib- its enhanced tolerance to environmental stress, or a plant or plant cell that exhibits increased yield in the presence of abiotic environmental stress conditions, as compared to a corresponding, e.g. non-transformed, wild type plant.
- This is generally referred to as a "method for producing a transgenic plant or plant cell with increased yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass".
- these traits are achieved by a process for increased yield in the presence and/or absence of environmental stress, particularly abiotic environmental stress, in a photosynthetic active organism, preferably a plant, as compared to a corresponding (non-transformed) wild type or starting photosynthetic active organism.
- the term "increased yield” means that the photosynthetic active organism, especially a plant, exhibits an increased growth rate, compared to the corresponding wild-type photosynthetic active organism.
- An increased growth rate may be re- fleeted inter alia by an increased biomass production of the whole plant, or by an increased biomass production of the aerial parts of a plant, or by an increased biomass production of the underground parts of a plant, or by an increased biomass production of parts of a plant, like stems, leaves, blossoms, fruits, and/or seeds.
- increased yield includes higher fruit yields, higher seed yields, higher fresh matter production, and/or higher dry matter production.
- the term "increased yield” means that the photosynthetic active organism, preferably plant, exhibits a prolonged growth, as compared to the corresponding, e.g. non-transformed, wild type photosynthetic active organism.
- a prolonged growth comprises survival and/or continued growth of the photosynthetic active organism, preferably plant, at the moment when the non-transformed wild type photosynthetic active organism shows visual symptoms of deficiency and/or death, confronted or in the absence of environmental stress.
- the present invention relates to a process for the production of a plant (or plant cell) exhibiting increased yield as defined above in the absence of stress conditions, as compared to the corresponding, e.g. non-transformed, wild type plant (cell).
- the present invention provides a method for producing a transgenic plant cell with increased yield, e.g an improved yield- related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g.
- 3OS ribosomal protein S1 1 60S ribosomal protein
- Adaptin medium chain homolog APM2, B0252-protein BRICK1 -like protein
- Cav1 protein Chloroplast chaperonin
- DNA polymerase DNA polymerase
- flagellar protein G2/mitotic-specific cyclin
- GREI -protein(Hydrophilin) Membrane protein
- ORF YPL249c-a RNA polymerase Il holoenzyme cyclin-like subunit, Ser- ine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, transcription regulator (farR), Ykr015c-protein, and YPL167C_2-protein.
- the present invention provides a method for producing a transgenic plant cell with increased yield, in the absence of environmental stress, as compared to a corresponding, e.g. non-transformed, wild type plant cell, by increasing or generating one or more activities selected from the group consisting of 3OS ribosomal protein S1 1 , 60S ribosomal protein, Adaptin medium chain homolog APM2, B0252-protein, BRICK1-like protein, Cav1 protein, Chloroplast chaperonin, DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GREI -protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoenzyme cyclin-like subunit, Serine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, transcription regulator (farR), Ykr015c-protein, and
- YIP Yield Increase Protein
- a photosynthetic active organism particularly a plant, shows an enhanced tolerance to abiotic environmental stress.
- a photosynthetic active organism, especially a plant shows increased yield under conditions of abiotic environmental stress.
- this invention fulfills the need to identify new, unique genes capable of conferring enhanced tolerance to abiotic environmental stress to photosynthetic active organism, preferably plants, upon expression or over-expression of endogenous and/or ex- ogenous genes. In another embodiment thereof this invention fulfills the need to identify new, unique genes capable of conferring enhanced tolerance to abiotic environmental stress to photosynthetic active organism, preferably plants, upon expression or over-expression of endogenous genes. In another embodiment thereof this invention fulfills the need to identify new, unique genes capable of conferring enhanced tolerance to abiotic environmental stress to photosynthetic active organism, preferably plants, upon expression or over-expression of exogenous genes.
- this invention fulfills the need to identify new, unique genes capable of conferring an increase of yield to photosynthetic active organism, preferably plants, upon expression or over-expression of endogenous and/or exogenous genes.
- this invention fulfills the need to identify new, unique genes capable of conferring an increase of yield to photosynthetic active organism, preferably plants, upon expression or over-expression of endogenous genes.
- this invention fulfills the need to identify new, unique genes capable of conferring an increase of yield to photosynthetic active organism, preferably plants, upon expression or over-expression of exogenous genes.
- this invention fulfills the need to identify new, unique genes capable of conferring an enhanced tolerance to abiotic environmental stress in combination with an increase of yield to photosynthetic active organism, preferably plants, upon expression or over-expression of endogenous and/or exogenous genes.
- this invention fulfills the need to identify new, unique genes capable of conferring an enhanced tolerance to abiotic environmental stress in combination with an increase of yield to photosynthetic active organism, preferably plants, upon expression or over-expression of endogenous genes.
- this invention fulfills the need to identify new, unique genes capable of conferring an enhanced tolerance to abiotic environmental stress in combination with an increase of yield to photosynthetic active organisms, preferably plants, upon expression or over-expression of exogenous genes.
- the present invention relates to a method for producing a transgenic photosynthetic active organism or a part thereof, preferably a plant cell, a plant or a part thereof, with enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g.
- non-transformed, wild type photosynthetic active organism or a part thereof preferably a plant cell, a plant or a part thereof, which comprises (a) increasing or generating one or more activities selected from the group consisting of 3OS ribosomal protein S11 , 60S ribosomal protein, Adaptin medium chain homolog APM2, B0252-protein, BRICK1 -like protein, Cav1 protein, Chloroplast chaperonin, DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GRE1 - protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoen- zyme cyclin-like subunit, Serine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, transcription regulator (farR), Ykr015c-protein, and YPL167C_2-protein in a photosynthetic active
- the present invention relates to a method for producing a transgenic photosynthetic active organism or a part thereof, preferably a plant cell, a plant or a part thereof with enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g.
- non-transformed, wild type photosynthetic active organism or a part thereof preferably a plant cell, a plant or a part thereof, which comprises (a) increasing or generating one or more activities selected from the group consisting of: 3OS ribosomal protein S1 1 , 60S ribosomal protein, Adaptin medium chain homolog APM2, B0252-protein, BRICK1 -like protein, Cav1 protein, Chloroplast chaperonin, DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GRE1 - protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoen- zyme cyclin-like subunit, Serine/threonine-protein kinase, Short chain dehydrogenase,
- Sterol-C-methyltransferase a photosynthetic active organism or a part thereof, preferably a plant cell, a plant or a part thereof,
- thetic active organism or a part thereof preferably a plant cell, a plant or a part thereof, after the non-transformed wild type photosynthetic active organism or a part thereof, preferably a plant cell, a plant or a part thereof, show visual symptoms of deficiency and/or death.
- Enhanced yield, particularly biomass production may, for example and preferably, be determined according to the following method:
- Transformed plants are grown in pots in a growth chamber (e.g. Sval ⁇ f Weibull, Sval ⁇ v, Sweden).
- a growth chamber e.g. Sval ⁇ f Weibull, Sval ⁇ v, Sweden.
- the plants are Arabidopsis thaliana seeds thereof are sown in pots containing a 3.5:1 (v:v) mixture of nutrient rich soil (GS90, Tantau, Wansdorf, Germany) and sand.
- Plants are grown under standard growth conditions.
- the plants are A. thaliana
- the standard growth conditions are: photoperiod of 16 h light and 8 h dark, 20 0 C, 60% relative humidity, and a photon flux density of 200 ⁇ mol/m 2 s.
- Plants are grown and cultured.
- the plants are Arabidopsis thaliana they are watered every second day. After 9 to 10 days the plants are individualized. After a total growth period of 29 to 30 days the plants are harvested and rated by the
- the present invention relates to a method for producing a transgenic photosynthetic active organism or a part thereof, preferably a plant cell, a plant or a part thereof with enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type photosynthetic active organism or a part thereof, preferably a plant cell, a plant or a part thereof, which comprises
- the present invention relates to a method for producing a transgenic plant cell, a plant or a part thereof with enhanced yield, e.g an improved yield- related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g.
- non-transformed, wild type plant cell a plant or a part thereof, which comprises (a) increasing or generating one or more activities selected from the group consisting of 3OS ribosomal protein S1 1 , 60S ribosomal protein, Adaptin medium chain homolog APM2, B0252-protein, BRICK1 -like protein, Cav1 protein, Chloroplast chaperonin, DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GRE1 - protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoen- zyme cyclin-like subunit, Serine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, transcription regulator (farR), Ykr015c-protein, and
- the present invention relates to a method for producing a transgenic plant cell, a plant or a part thereof with enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, a plant or a part thereof, which comprises
- DNA polymerase DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GRE1 - protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoen- zyme cyclin-like subunit, Serine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, transcription regulator (farR), Ykr015c-protein, and YPL167C_2-protein in the cytoplasm of a plant cell, and
- the present invention relates to a method for producing a transgenic plant cell, a plant or a part thereof with enhanced yield, e.g an improved yield- related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, a plant or a part thereof, which comprises
- the present invention relates to a method for producing a transgenic plant cell, a plant or a part thereof with enhanced yield, e.g an improved yield- related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, a plant or a part thereof, which comprises
- the present invention is related to a method for producing a transgenic plant cell, a plant or a part thereof with enhanced yield, e.g an im- proved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, a plant or a part thereof, which comprises
- DNA polymerase DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GRE1 - protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoen- zyme cyclin-like subunit, Serine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, transcription regulator (farR), Ykr015c-protein, and YPL167C_2-protein in an organelle of a plant cell or
- the present invention relates to a method for producing a transgenic plant cell, a plant or a part thereof with enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant.
- a method for producing a transgenic plant cell, a plant or a part thereof with enhanced yield e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, a plant or a part thereof, which comprises
- the nucleic acid sequence encoding a transit peptide can be isolated from every organism such as microorganisms such as algae or plants containing plastids preferably chloroplasts.
- a "transit peptide” is an amino acid sequence, whose encoding nucleic acid sequence is translated together with the corresponding structural gene. That means the transit peptide is an integral part of the translated protein and forms an amino terminal extension of the protein. Both are translated as so called “preprotein”.
- preprotein the transit peptide is cleaved off from the preprotein during or just after import of the protein into the correct cell organelle such as a plastid to yield the mature protein.
- the transit peptide ensures correct localization of the mature protein by facilitating the transport of proteins through intracellular membranes.
- Preferred nucleic acid sequences encoding a transit peptide are derived from a nucleic acid sequence encoding a protein finally resided in the plastid and stemming from an organism selected from the group consisting of the genera Acetabularia, Arabidopsis, Brassica, Capsicum, Chlamydomonas, Cururbita, Dunaliella, Euglena, Flaveria, Glycine, Helianthus, Hor- deum, Lemna, Lolium, Lycopersion, Malus, Medicago, Mesembryanthemum, Nicotiana, Oenotherea, Oryza, Petunia, Phaseolus, Physcomitrella, Pinus, Pisum, Raphanus, Silene, Sinapis, Solanum, Spinacea, Stevia, Synechococcus, Triticum and Zea.
- Transit peptides which are beneficially used in the inventive process, are derived from the nucleic acid sequence encoding a protein selected from the group consisting of ribulose bisphosphate carboxylase/oxygenase, 5-enolpyruvyl- shikimate-3-phosphate synthase, acetolactate synthase, chloroplast ribosomal protein CS17, Cs protein, ferredoxin, plastocyanin, ribulose bisphosphate carboxylase activase, -.
- a protein selected from the group consisting of ribulose bisphosphate carboxylase/oxygenase, 5-enolpyruvyl- shikimate-3-phosphate synthase, acetolactate synthase, chloroplast ribosomal protein CS17, Cs protein, ferredoxin, plastocyanin, ribulose bisphosphate carboxylase activase, -.
- nucleic acid sequence encoding a transit peptide is derived from a nucleic acid sequence encoding a protein finally resided in the plastid and stemming from an organism selected from the group consisting of the species Acetabularia mediterranea, Arabidopsis thaliana, Brassica campestris, Brassica napus, Capsicum an- nuum, Chlamydomonas reinhardtii, Cururbita moschata, Dunaliella salina, Dunaliella tertio- lecta, Euglena gracilis, Flaveria trinervia, Glycine max, Helianthus annuus, Hordeum vul- gare, Lemna gibba, Lolium perenne, Lycopersion esculentum, Malus domestica, Medicago falcata, Medicago sativa, Mesembryanthemum crystallinum, Nicotiana plumbaginifoli
- nucleic acid sequences are encoding transit peptides as disclosed by von Heijne et al. (Plant Molecular Biology Reporter, 9 (2), 104, (1991 )), which are hereby incorparated by reference. Table V shows some examples of the transit peptide sequences disclosed by von Heijne et al. According to the disclosure of the invention especially in the examples the skilled worker is able to link other nucleic acid se- o
- transit peptides can easely isolated from plastid- localized proteins, which are expressed from nuclear genes as precursors and are then targeted to plastids.
- Such transit peptides encoding sequences can be used for the construc- tion of other expression constructs.
- the transit peptides advantageously used in the inventive process and which are part of the inventive nucleic acid sequences and proteins are typically 20 to 120 amino acids, preferably 25 to 1 10, 30 to 100 or 35 to 90 amino acids, more preferably 40 to 85 amino acids and most preferably 45 to 80 amino acids in length and functions post-translationally to direct the protein to the plastid preferably to the chloro- plast.
- nucleic acid sequences encoding such transit peptides are localized upstream of nucleic acid sequence encoding the mature protein.
- nucleic acid sequence encoding the mature protein For the correct molecular joining of the transit peptide encoding nucleic acid and the nucleic acid encoding the protein to be targeted it is sometimes necessary to introduce additional base pairs at the joining position, which forms restriction enzyme recognition sequences useful for the molecular joining of the different nucleic acid molecules. This procedure might lead to very few additional amino acids at the N-terminal of the mature imported protein, which usually and preferably do not interfer with the protein function.
- the additional base pairs at the joining position which forms restriction enzyme recognition sequences have to be choosen with care, in order to avoid the formation of stop codons or codons which encode amino acids with a strong influence on protein folding, like e.g. proline. It is preferred that such additional codons encode small structural flexible amino acids such as glycine or alanine.
- nucleic acid sequences coding for the proteins as shown in table II, column 3 and its homologs as disclosed in table I, columns 5 and 7 can be joined to a nucleic acid sequence encoding a transit peptide.
- This nucleic acid sequence encoding a transit peptide ensures transport of the protein to the plastid.
- the nucleic acid sequence of the gene to be expressed and the nucleic acid sequence encoding the transit peptide are operably linked. Therefore the transit peptide is fused in frame to the nucleic acid sequence coding for proteins as shown in table II, column 3 and its homologs as dis- closed in table I, columns 5 and 7.
- organelle shall mean for example “mitochondria” or preferably “plastid” (throughout the specification the "plural” shall comprise the “singular” and vice versa).
- plastid according to the invention are in- tended to include various forms of plastids including proplastids, chloroplasts, chromo- plasts, gerontoplasts, leucoplasts, amyloplasts, elaioplasts and etioplasts, preferably chloroplasts. They all have as a common ancestor the aforementioned proplasts.
- Other transit peptides are disclosed by Schmidt et al. (J.
- Transit peptide sequences which are used in the inventive process and which form part of the inventive nucleic acid sequences are generally enriched in hydroxylated amino acid residues (serine and threonine), with these two residues generally constituting 20 to 35 % of the total. They often have an amino-terminal region empty of GIy, Pro, and charged residues. Furthermore they have a number of small hydrophobic amino acids such as valine and alanine and generally acidic amino acids are lacking. In addition they generally have a middle region rich in Ser, Thr, Lys and Arg. Overall they have very often a net positive charge.
- nucleic acid sequences coding for the transit peptides may be chemically synthesized either in part or wholly according to structure of transit peptide sequences disclosed in the prior art. Said natural or chemically synthesized sequences can be directly linked to the sequences encoding the mature protein or via a linker nucleic acid sequence, which may be typically less than 500 base pairs, preferably less than 450, 400, 350, 300, 250 or 200 base pairs, more preferably less than 150, 100, 90, 80, 70, 60, 50, 40 or 30 base pairs and most preferably less than 25, 20, 15, 12, 9, 6 or 3 base pairs in length and are in frame to the coding sequence.
- a linker nucleic acid sequence which may be typically less than 500 base pairs, preferably less than 450, 400, 350, 300, 250 or 200 base pairs, more preferably less than 150, 100, 90, 80, 70, 60, 50, 40 or 30 base pairs and most preferably less than 25, 20, 15, 12, 9, 6 or 3 base pairs in length and are in frame to the coding sequence.
- nucleic acid sequences encoding transit peptides may comprise sequences derived from more than one biological and/or chemical source and may include a nucleic acid sequence derived from the amino- terminal region of the mature protein, which in its native state is linked to the transit peptide.
- said amino-terminal region of the mature protein is typically less than 150 amino acids, preferably less than 140, 130, 120, 110, 100 or 90 amino acids, more preferably less than 80, 70, 60, 50, 40, 35, 30, 25 or 20 amino acids and most preferably less than 19, 18, 17, 16, 15, 14, 13, 12, 1 1 or 10 amino acids in length. But even shorter or longer stretches are also possible.
- target sequences which facilitate the transport of proteins to other cell compartments such as the vacuole, endoplasmic reticulum, golgi complex, glyoxysomes, peroxisomes or mitochondria may be also part of the inventive nucleic acid sequence.
- the proteins translated from said inventive nucleic acid sequences are a kind of fusion proteins that means the nucleic acid sequences encoding the transit peptide for example the ones shown in table V, preferably the last one of the table are joint to the nucleic acid sequences shown in table I, columns 5 and 7. The person skilled in the art is able to join said sequences in a functional manner.
- the transit peptide part is cleaved off from the protein part shown in table II, columns 5 and 7 during the transport preferably into the plastids.
- All products of the cleavage of the preferred transit peptide shown in the last line of table V have preferably the N-terminal amino acid sequences QIA CSS or QIA EFQLTT in front of the start methionine of the protein metioned in table II, columns 5 and 7.
- Other short amino acid sequences of an range of 1 to 20 amino acids preferable 2 to 15 amino acids, more preferable 3 to 10 amino acids most preferably 4 to 8 amino acids are also possible in front of the start methionine of the protein metioned in table II, columns 5 and 7.
- Said short amino acid sequence is preferred in the case of the expression of Escherichia coli genes.
- the amino acid sequence QIA EFQLTT the six amino acids in front of the start methionine are stemming from the LIC cassette.
- Said short amino acid sequence is preferred in the case of the expression of Saccharomyces cerevisiae genes.
- the skilled worker knows that other short sequences are also useful in the expression of the genes metioned in table I, columns 5 and 7. Furthermore the skilled worker is aware of the fact that there is not a need for such short sequences in the expression of the genes.
- nucleic acids of the invention can directly be introduced into the plastidal genome. Therefore in a preferred embodiment the nucleic acid sequences shown in table I, columns 5 and 7 are directly introduced and expressed in plastids.
- a plastid such as a chloroplast
- a plastid has been "transformed” by an exogenous (preferably foreign) nucleic acid sequence if nucleic acid sequence has been introduced into the plastid that means that this sequence has crossed the membrane or the membranes of the plastid.
- the foreign DNA may be integrated (covalently linked) into plastid DNA making up the genome of the plastid, or it may remain unintegrated (e.g., by including a chloroplast origin of replication).
- "Stably" integrated DNA sequences are those, which are inherited through plastid replication, thereby transferring new plastids, with the features of the integrated DNA sequence to the progeny.
- microspore-derived hypocotyl or cotyledonary tissue which are green and thus contain numerous plastids
- a method for the transformation bombarding of the plant material or the use of independently replicating shuttle vectors are well known by the skilled worker. But also a PEG-mediated transformation of the plastids or Agrobacterium transformation with binary vectors is possible.
- Useful markers for the transformation of plastids are positive selection markers for example the chloramphenicol-, streptomycin-, kanamycin-, neomycin-, amikamycin-, spectinomycin-, triazine- and/or lincomycin-tolerance genes.
- reporter genes are for example ⁇ -galactosidase-, ⁇ -glucuronidase-(GUS), al- kaline phosphatase- and/or green-fluorescent protein-genes (GFP).
- a further preferred embodiment of the invention relates to the use of so called "chloroplast localization sequences", in which a first RNA sequence or molecule is capable of transporting or “chaperoning" a second RNA sequence, such as a RNA sequence transcribed from the sequences depicted in table 1 , columns 5 and 7 or a sequence encoding a protein, as depicted in table II, columns 5 and 7, from an external environment inside a cell or outside a plastid into a chloroplast.
- the chloroplast localization signal is substantially similar or complementary to a complete or intact viroid sequence.
- the chloroplast localization signal may be encoded by a DNA sequence, which is transcribed into the chloro- plast localization RNA.
- the term "viroid" refers to a naturally occurring single stranded RNA molecule (Flores, C. R. Acad Sci III. 324 (10), 943 (2001 )). Viroids usually contain about OD
- viroids that contain chloroplast localization signals include but are not limited to ASBVd, PLMVd, CChMVd and ELVd.
- the viroid sequence or a functional part of it can be fused to the sequences depicted in table I, columns 5 and 7 or a sequence encoding a protein, as depicted in table II, columns 5 and 7 in such a manner that the viroid sequence transports a sequence transcribed from a sequence as depicted in table 1 , columns 5 and 7 or a sequence encoding a protein as depicted in table II, columns 5 and 7 into the chloroplasts.
- a preferred embodiment uses a modified ASBVd (Navarro et al., Virology. 268 (1), 218 (2000)).
- the protein to be expressed in the plastids such as the pro- teins depicted in table II, columns 5 and 7 are encoded by different nucleic acids.
- WO 2004/040973 teaches a method, which relates to the translocation of an RNA corresponding to a gene or gene fragment into the chloroplast by means of a chloroplast localization sequence.
- the genes, which should be expressed in the plant or plants cells, are split into nucleic acid fragments, which are introduced into different compartments in the plant e.g. the nucleus, the plastids and/or mitochondria.
- the chloroplast contains a ribozyme fused at one end to an RNA encoding a fragment of a protein used in the inventive process such that the ribozyme can trans-splice the translocated fusion RNA to the RNA encoding the gene fragment to form and as the case may be reunite the nucleic acid fragments to an intact mRNA encoding a functional protein for example as disclosed in table II, columns 5 and 7.
- nucleic acid sequences as shown in table I, columns 5 and 7 used in the inventive process are transformed into plas- tids, which are metabolically active.
- Those plastids should preferably maintain at a high copy number in the plant or plant tissue of interest, most preferably the chloroplasts found in green plant tissues, such as leaves or cotyledons or in seeds.
- nucleic acid sequences as shown in table I, columns 5 and 7 are introduced into an expression cassette using preferably a promoter and terminator, which are active in plastids preferably a chloroplast promoter.
- promoters include the psbA promoter from the gene from spinach or pea, the rbcL promoter, and the atpB promoter from corn.
- plant cell or the term “organism” as understood herein relates always to a plant cell or a organelle thereof, preferably a plastid, more preferably chloroplast.
- plant is meant to include not only a whole plant but also a part thereof i.e., one or more cells, and tissues, including for example, leaves, stems, shoots, roots, flowers, fruits and seeds.
- transgenic expression of the Saccaro- myces cerevisiae protein as shown in table II, column 3 and/or the transgenic expression of the E. coli protein as shown in table II, column 3 in a plant such as A. thaliana for example conferred enhanced tolerance to abiotic environmental stress, particularly low temperature, and/or increased yield to the transgenic plant cell, plant or a part thereof as compared to a corresponding, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 39 or polypeptide SEQ ID NO. 40, respectively, is increased or generated or if the activity "Chloroplast chaperonin" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 40 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 40, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 39, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Arabidopsis thaliana nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 39 or polypeptide SEQ ID NO. 40, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 39 or polypeptide SEQ ID NO. 40, respectively, is increased or generated or if the activity "Chloroplast chaperonin" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a corresponding non-modified e.g. a non-transformed, wild type plant cell, a plant or a part thereof
- an increase of yield from 1.1-fold to 1.365-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 40 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 39, or a homolog of said nucleic acid molecule or polypeptide, e.g.
- Chloroplast chaperonin is increased or generated in a plant cell, plant or part thereof, es- pecially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.419-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 40 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 40, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 39, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Arabidopsis thaliana nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 39 or polypeptide SEQ ID NO. 40, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 39 or polypeptide SEQ ID NO. 40, respectively, is increased or generated or if the activity
- Chloroplast chaperonin is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased intrinsic yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased yield under standard conditions e.g. in the absence of nutrient deficiency as well as stress conditions, is conferred.
- an increase of yield from 1.05-fold to 1.075-fold, for example plus at least 100% thereof, under conditions of low temperature is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 138 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 137, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Azotobacter vinelandii nucleic acid molecule or a polypeptide, re- spectively, comprising the nucleic acid SEQ ID NO. 137 or polypeptide SEQ ID NO. 138, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 137 or polypeptide SEQ ID NO. 138, respectively, is in- creased or generated or if the activity "3OS ribosomal protein S1 1 " is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 138 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 137, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Azotobacter vinelandii nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 137 or polypeptide SEQ ID NO. 138, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nu- cleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 137 or polypeptide SEQ ID NO. 138, respectively, is increased or generated or if the activity "3OS ribosomal protein S1 1 " is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, com- pared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.231 -fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 138 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 137, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Azotobacter vinelandii nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 137 or polypeptide SEQ ID NO. 138, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 137 or polypeptide SEQ ID NO. 138, respectively, is increased or generated or if the activity "3OS ribosomal protein S1 1 " is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.872-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 138 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 137, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Azotobacter vinelandii nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 137 or polypeptide SEQ ID NO. 138, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 137 or polypeptide SEQ ID NO. 138, respectively, is increased or generated or if the activity "3OS ribosomal protein S1 1 " is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased intrinsic yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased yield under standard conditions e.g. in the absence of nutrient deficiency as well as stress conditions, is conferred.
- an increase of yield from 1.05-fold to 1.083-fold, for example plus at least 100% thereof, under conditions of low temperature is conferred corn-pared to a correspond- ing non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 983 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 982, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Azotobacter vinelandii nucleic acid molecule or a polypeptide, re- spectively, comprising the nucleic acid SEQ ID NO. 982 or polypeptide SEQ ID NO. 983, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 982 or polypeptide SEQ ID NO. 983, respectively, is in- creased or generated or if the activity "Short chain dehydrogenase" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 982 or polypeptide SEQ ID NO. 983, respectively, is increased or generated or if the activity "Short chain dehydrogenase" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.429-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 983 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 982, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Azotobacter vinelandii nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 982 or polypeptide SEQ ID NO. 983, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 982 or polypeptide SEQ ID NO. 983, respectively, is increased or generated or if the activity "Short chain dehydrogenase" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.590-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 1226, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1225, or a homolog of said nucleic acid molecule or polypeptide e.g.
- a polypeptide according to the polypeptide SEQ ID NO. 1226 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1225, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Escherichia coli nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 1225 or polypeptide SEQ ID NO. 1226, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in ta- ble I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 1225 or polypeptide SEQ ID NO. 1226, respectively, is increased or generated or if the activity "B0252-protein" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is Cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.489-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 1226, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1225, , or a homolog of said nucleic acid molecule or polypeptide e.g.
- an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.617-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the poly- peptide SEQ ID NO. 1328 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1327, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Escherichia coli nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 1327 or polypeptide SEQ ID NO. 1328, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypep- tide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 1327 or polypeptide SEQ ID NO. 1328, respectively, is increased or generated or if the activity "transcription regulator (farR)" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- farR transcription regulator
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 1327 or polypeptide SEQ ID NO. 1328, respectively, is increased or generated or if the activity "transcription regulator (farR)" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- farR transcription regulator
- an increase of yield from 1.1-fold to 1.593-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in ta- ble I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 1327 or polypeptide SEQ ID NO. 1328, respectively, is increased or generated or if the activity "transcription regulator (farR)" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use effi- ciency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.408-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corre- sponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 1328 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1327, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Escherichia coli nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 1327 or polypeptide SEQ ID NO. 1328, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 1327 or polypeptide SEQ ID NO. 1328, respectively, is increased or generated or if the ac- tivity "transcription regulator (farR)" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased intrinsic yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased yield under standard conditions e.g. in the absence of nutrient deficiency as well as stress conditions, is con- ferred.
- an increase of yield from 1.05-fold to 1.221 -fold, for example plus at least 100% thereof, under conditions of low temperature is conferred corn-pared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the poly- peptide SEQ ID NO. 1426 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1425, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Escherichia coli nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 1425 or polypeptide SEQ ID NO. 1426, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypep- tide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 1425 or polypeptide SEQ ID NO. 1426, respectively, is increased or generated or if the activity "flagellar protein" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as com- pared to a corresponding non-modified, e.g. a non -transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 1426 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1425, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Escherichia coli nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 1425 or polypeptide SEQ ID NO. 1426, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in ta- ble I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 1425 or polypeptide SEQ ID NO. 1426, respectively, is increased or generated or if the activity "flagellar protein" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is Cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.325-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 1426, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1425, , or a homolog of said nucleic acid molecule or polypeptide e.g.
- an increase of yield from 1.05-fold to 1.741 -fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 1426 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1425, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Escherichia coli nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 1425 or polypeptide SEQ ID NO. 1426, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in ta- ble I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 1425 or polypeptide SEQ ID NO. 1426, respectively, is increased or generated or if the activity "flagellar protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased intrinsic yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased yield under standard conditions e.g. in the absence of nutrient deficiency as well as stress conditions, is conferred.
- an increase of yield from 1.05-fold to 1.192-fold, for example plus at least 100% thereof, under conditions of low temperature is conferred corn-pared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 1450 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1449, or a homolog of said nucleic acid molecule or polypeptide, e.g.
- a polypeptide according to the polypeptide SEQ ID NO. 1450 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1449, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Escherichia coli nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 1449 or polypeptide SEQ ID NO. 1450, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in ta- ble I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 1449 or polypeptide SEQ ID NO. 1450, respectively, is increased or generated or if the activity "Short chain dehydrogenase" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.246-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 1450 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 1449, , or a homolog of said nucleic acid molecule or polypeptide, e.g.
- an increase of yield from 1.05-fold to 1.808-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2173 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2172, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Glycine max nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2172 or polypeptide SEQ ID NO. 2173, respectively, is increased or generated, e.g.
- a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2172 or polypeptide SEQ ID NO. 2173, respectively, is increased or generated or if the activity "BRICK1 -like protein" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2173 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2172, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Glycine max nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2172 or polypeptide SEQ ID NO. 2173, respectively, is increased or gen- erated, e.g.
- a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2172 or polypeptide SEQ ID NO. 2173, respectively, is increased or generated or if the activity "BRICK1 -like protein" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is Cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.373-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2173 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 2173, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2172, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Glycine max nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2172 or polypeptide SEQ ID NO. 2173, respectively, is increased or generated, e.g.
- a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2172 or polypeptide SEQ ID NO. 2173, respectively, is increased or generated or if the activity "BRICK1 -like protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.994-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2173 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 2173, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2172, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Glycine max nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2172 or polypeptide SEQ ID NO. 2173, respectively, is increased or generated, e.g.
- a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2172 or polypeptide SEQ ID NO. 2173, respectively, is increased or generated or if the activity "BRICK1 -like protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased intrinsic yield as com- pared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased yield under standard conditions e.g. in the absence of nutrient deficiency as well as stress conditions, is conferred.
- an increase of yield from 1.05-fold to 1.126-fold, for example plus at least 100% thereof, under conditions of low temperature is conferred corn-pared to a correspond- ing non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2215 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 2215, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2214, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Synechocystis sp. nucleic acid molecule or a polypeptide, respec- tively, comprising the nucleic acid SEQ ID NO. 2214 or polypeptide SEQ ID NO. 2215, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2214 or polypeptide SEQ ID NO. 2215, respectively, is increased or generated or if the activity "Sterol-C-methyltransferase" is increased or generated in a plant cell, plant or part thereof, especially if localized mitochondrial, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2215 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 2215, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2214, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Synechocystis sp. nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2214 or polypeptide SEQ ID NO. 2215, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nu- cleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2214 or polypeptide SEQ ID NO. 2215, respectively, is increased or generated or if the activity "Sterol-C-methyltransferase" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is mitochondrial localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.235-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2215 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 2215, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2214, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Synechocystis sp. nucleic acid molecule or a polypeptide comprising the nu- cleic acid SEQ ID NO. 2214 or polypeptide SEQ ID NO. 2215, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2214 or polypeptide SEQ ID NO. 2215, respectively, is increased or generated or if the activity "Sterol-C-methyltransferase" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is mitochondrial localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.443-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2342 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 2342, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2341 , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2341 or polypeptide SEQ ID NO. 2342, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2341 or polypeptide SEQ ID NO. 2342, respectively, is increased or generated or if the activity "Cav1 protein" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2342 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 2342, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2341 , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2341 or polypeptide SEQ ID NO. 2342, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2341 or polypeptide SEQ ID NO. 2342, respectively, is increased or generated or if the activity "Cav1 protein" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.391 -fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2342 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 2342, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2341 , , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2341 or polypeptide SEQ ID NO. 2342, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2341 or polypeptide SEQ ID NO. 2342, respectively, is increased or generated or if the activity "Cav1 protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is Cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.620-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2342 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2341 , , or a homolog of said nucleic acid molecule or polypeptide, e.g.
- the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2345 or polypeptide SEQ ID NO. 2346, respectively, is increased or generated, e.g. if the activity of a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2345 or polypeptide SEQ ID NO.
- G2/mitotic-specific cyclin is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2346 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2345, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2345 or polypeptide SEQ ID NO. 2346, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2345 or polypeptide SEQ ID NO. 2346, respectively, is increased or generated or if the activity "G2/mitotic-specific cyclin" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.312-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2346 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2345, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2345 or polypeptide SEQ ID NO. 2346, respectively, is increased or generated, e.g.
- an increase of yield from 1.05-fold to 1.291 -fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corre- sponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2464 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2463, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2463 or polypeptide SEQ ID NO. 2464, respectively, is increased or generated, e.g.
- Adaptin medium chain homolog APM2 is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2464 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2463, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2463 or polypeptide SEQ ID NO. 2464, respectively, is increased or generated, e.g.
- Adaptin medium chain homolog APM2 is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is Cytoplasmic local- ized, an increased biomass, compared to a corresponding non-modified, e.g. a non- transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.367-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2464 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2463, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2463 or polypeptide SEQ ID NO. 2464, respectively, is in- creased or generated, e.g.
- Adaptin medium chain homolog APM2 is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is Cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.343-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 251 1 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2510, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2510 or polypeptide SEQ ID NO. 251 1 , respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2510 or polypeptide SEQ ID NO. 251 1 , respectively, is increased or generated or if the activity "Serine/threonine-protein kinase" is increased or generated in a plant cell, plant or part thereof, especially if localized Cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 251 1 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2510, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2510 or polypeptide SEQ ID NO. 251 1 , respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2510 or polypeptide SEQ ID NO. 251 1 , respectively, is increased or generated or if the activity "Serine/threonine-protein kinase" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g.
- a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.522-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 251 1 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO.
- nucleic acid molecule or polypeptide e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2510 or polypeptide SEQ ID NO. 251 1 , respectively, is in- creased or generated, e.g. if the activity of a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2510 or polypeptide SEQ ID NO.
- 251 1 is increased or generated or if the activity "Serine/threonine-protein kinase" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non- transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.389-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 251 1 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2510, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2510 or polypeptide SEQ ID NO. 251 1 , respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2510 or polypeptide SEQ ID NO. 251 1 , respectively, is increased or generated or if the activity "Serine/threonine-protein kinase" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased intrinsic yield as compared to a corresponding non-modified, e.g. a non- transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased yield under standard conditions e.g. in the absence of nutrient deficiency as well as stress conditions, is conferred.
- an increase of yield from 1.05-fold to 1.227-fold, for example plus at least 100% thereof, under conditions of low temperature is conferred corn-pared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2596 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2595, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2595 or polypeptide SEQ ID NO. 2596, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2595 or polypeptide SEQ ID NO. 2596, respectively, is increased or generated or if the activity "Ykr015c-protein" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2596 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2595, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2595 or polypeptide SEQ ID NO. 2596, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide com- prising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2595 or polypeptide SEQ ID NO. 2596, respectively, is increased or generated or if the activity "Ykr015c-protein" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.347-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2596 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2595, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2595 or polypeptide SEQ ID NO. 2596, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2595 or polypeptide SEQ ID NO. 2596, respectively, is increased or generated or if the activity "Ykr015c-protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.160-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2600 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2599, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2599 or polypeptide SEQ ID NO. 2600, respectively, is increased or generated, e.g.
- RNA polymerase Il holoenzyme cyclin-like sub- unit is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non- transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2600 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2599, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2599 or polypeptide SEQ ID NO. 2600, respectively, is in- creased or generated, e.g.
- RNA polymerase Il holoenzyme cyclin-like subunit is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non- transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.376-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2600 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2599, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising OO
- nucleic acid SEQ ID NO. 2599 or polypeptide SEQ ID NO. 2600 is increased or generated, e.g. if the activity of a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2599 or polypeptide SEQ ID NO.
- RNA polymerase Il holoenzyme cyclin-like subunit is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is Cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non- modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.649-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the poly- peptide SEQ ID NO. 2646 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2645, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2645 or polypeptide SEQ ID NO. 2646, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2645 or polypeptide SEQ ID NO. 2646, respectively, is increased or generated or if the activity "Membrane protein" is increased or generated in a plant cell, plant or part thereof, especially if localized plastidic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2646 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2645, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2645 or polypeptide SEQ ID NO. 2646, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2645 or polypeptide SEQ ID NO. 2646, respectively, is increased or generated or if the activity "Membrane protein" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is plastidic localized, an increased bio- mass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.261 -fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2646, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2645, , or a homolog of said nucleic acid molecule or polypeptide e.g.
- 2646 is increased or generated or if the activity "Membrane protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is Plastidic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.778-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2662 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2661 , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2661 or polypeptide SEQ ID NO. 2662, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2661 or polypeptide SEQ ID NO. 2662, respectively, is increased or generated or if the activity "DNA polymerase" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2662 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2661 , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2661 or polypeptide SEQ ID NO. 2662, respectively, is in- creased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2661 or polypeptide SEQ ID NO. 2662, respectively, is increased or generated or if the activity "DNA polymerase" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.373-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2662 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2661 , , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2661 or polypeptide SEQ ID NO. 2662, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2661 or polypeptide SEQ ID NO. 2662, respectively, is increased or generated or if the activity "DNA polymerase" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.642-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2662 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2661 , or a homolog of said nucleic acid molecule or polypeptide, e.g.
- a polypeptide according to the polypeptide SEQ ID NO. 2737 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2736, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2736 or polypeptide SEQ ID NO. 2737, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2736 or polypeptide SEQ ID NO. 2737, respectively, is increased or generated or if the activity "GREI -protein(Hydrophilin)" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2737 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2736, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2736 or polypeptide SEQ ID NO. 2737, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2736 or polypeptide SEQ ID NO. 2737, respectively, is increased or generated or if the activity "GREI -protein(Hydrophilin)" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.326-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2736 or polypeptide SEQ ID NO. 2737, respectively, is increased or generated, e.g. if the activity of a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2736 or polypeptide SEQ ID NO.
- GREI -protein(Hydrophilin) is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non- transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.268-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2743, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2742, or a homolog of said nucleic acid molecule or polypeptide e.g.
- a polypeptide according to the polypeptide SEQ ID NO. 2743 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2742, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2742 or polypeptide SEQ ID NO. 2743, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2742 or polypeptide SEQ ID NO. 2743, respectively, is increased or generated or if the activity "60S ribosomal protein" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.546-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a correspond-ing non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a correspond-ing non-modified e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2743, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2742, , or a homolog of said nucleic acid molecule or polypeptide e.g.
- 2743 is increased or generated or if the activity "60S ribosomal protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.223-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2743 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 2743, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2742, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2742 or polypeptide SEQ ID NO. 2743, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2742 or polypeptide SEQ ID NO. 2743, respectively, is increased or generated or if the activity "60S ribosomal protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased intrinsic yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased yield under standard conditions e.g. in the absence of nutrient deficiency as well as stress conditions, is conferred.
- an increase of yield from 1.05-fold to 1.230-fold, for example plus at least 100% thereof, under conditions of low temperature is conferred corn-pared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2908 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2907, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Escherichia coli nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 2907 or polypeptide SEQ ID NO. 2908, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2907 or polypeptide SEQ ID NO. 2908, respectively, is in- creased or generated or if the activity "Short chain dehydrogenase" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 2908 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2907, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Escherichia coli nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2907 or polypeptide SEQ ID NO. 2908, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2907 or polypeptide SEQ ID NO. 2908, respectively, is increased or generated or if the activity "Short chain dehydrogenase" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is cytoplasmic localized, an increased biomass, com- pared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.246-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 2908 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 2907, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Escherichia coli nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 2907 or polypeptide SEQ ID NO. 2908, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 2907 or polypeptide SEQ ID NO. 2908, respectively, is increased or generated or if the ac- tivity "Short chain dehydrogenase" is increased or generated in a plant cell, plant or part DO
- an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.808-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 3633 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 3632, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Glycine max nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 3632 or polypeptide SEQ ID NO. 3633, respectively, is increased or generated, e.g.
- a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypep- tide motif as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3632 or polypeptide SEQ ID NO. 3633, respectively, is increased or generated or if the activity "BRICK1 -like protein" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3632 or polypeptide SEQ ID NO. 3633, respectively, is increased or generated or if the ac- tivity "BRICK1 -like protein" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is Cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1-fold to 1.373-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- the Glycine max nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 3632 or polypeptide SEQ ID NO. 3633, respectively, is increased or generated, e.g. if the activity of a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in ta- ble I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3632 or polypeptide SEQ ID NO.
- BRICK1 -like protein is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.994-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 3633 in case the activity of a polypeptide according to the polypeptide SEQ ID NO. 3633, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 3632, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Glycine max nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 3632 or polypeptide SEQ ID NO. 3633, respectively, is increased or generated, e.g.
- a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3632 or polypeptide SEQ ID NO. 3633, respectively, is increased or generated or if the ac- tivity "BRICK1 -like protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is Cytoplasmic localized, an increased intrinsic yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased yield under standard conditions e.g. in the absence of nutrient deficiency as well as stress conditions, is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 3677 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 3676, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 3676 or polypeptide SEQ ID NO. 3677, respectively, is increased or generated, e.g. if the activity of a nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as o
- nucleic acid molecule SEQ ID NO. 3676 or polypeptide SEQ ID NO. 3677, respectively is increased or generated or if the activity "YPL167C_2-protein" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 3677 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 3676, or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 3676 or polypeptide SEQ ID NO. 3677, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3676 or polypeptide SEQ ID NO. 3677, respectively, is increased or generated or if the activity "YPL167C_2-protein" is increased or generated in a plant cell, plant or part thereof, especially, if the polypeptide is Cytoplasmic localized, an increased biomass, compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increase of yield from 1.1 -fold to 1.373-fold, for example plus at least 100% thereof, under standard conditions, e.g. in the absence of nutrient deficiency as well as stress conditions is conferred compared to a corresponding non-modified, e.g. non- transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 3677 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 3676, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 3676 or polypeptide SEQ ID NO. 3677, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3676 or polypeptide SEQ ID NO. 3677, respectively, is increased or generated or if the activity "YPL167C_2-protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is Cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non- transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.642-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 3677 or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 3676, , or a homolog of said nucleic acid molecule or polypeptide, e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 3676 or polypeptide SEQ ID NO. 3677, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3676 or polypeptide SEQ ID NO. 3677, respectively, is increased or generated or if the activity "YPL167C_2-protein" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is Cytoplasmic localized, an increased intrinsic yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased yield under standard conditions e.g. in the absence of nutrient deficiency as well as stress condi- tions, is conferred.
- an increase of yield from 1.05-fold to 1.255-fold, for example plus at least 100% thereof, under conditions of low temperature is conferred corn-pared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the poly- peptide SEQ ID NO. 3699, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 3698, or a homolog of said nucleic acid molecule or polypeptide e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide, respectively, comprising the nucleic acid SEQ ID NO. 3698 or polypeptide SEQ ID NO. 3699, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3698 or polypeptide SEQ ID NO. 3699, respectively, is increased or generated or if the activity "ORF YPL249c-a" is increased or generated in a plant cell, plant or part thereof, especially if localized cytoplasmic, an increased yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- a polypeptide according to the polypeptide SEQ ID NO. 3699, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 3698, or a homolog of said nucleic acid molecule or polypeptide e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 3698 or polypeptide SEQ ID NO. 3699, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3698 or polypeptide SEQ ID NO. 3699, respectively, is increased or o
- a polypeptide according to the polypeptide SEQ ID NO. 3699, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 3698, , or a homolog of said nucleic acid molecule or polypeptide e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 3698 or polypeptide SEQ ID NO. 3699, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide com- prising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3698 or polypeptide SEQ ID NO. 3699, respectively, is increased or generated or if the activity "ORF YPL249c-a" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased nutrient use efficiency as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred. In one embodiment an increased nitrogen use efficiency is conferred.
- an increase of yield from 1.05-fold to 1.223-fold, for example plus at least 100% thereof, under conditions of nitrogen deficiency is conferred compared to a corre- sponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- a polypeptide according to the polypeptide SEQ ID NO. 3699, or encoded by a nucleic acid molecule comprising the nucleic acid SEQ ID NO. 3698, , or a homolog of said nucleic acid molecule or polypeptide e.g. in case the activity of the Saccharomyces cerevisiae nucleic acid molecule or a polypeptide comprising the nucleic acid SEQ ID NO. 3698 or polypeptide SEQ ID NO. 3699, respectively, is increased or generated, e.g.
- nucleic acid molecule or a polypeptide comprising the nucleic acid or polypeptide or the consensus sequence or the polypeptide motif, as depicted in table I, Il or IV, column 7 in the respective same line as the nucleic acid molecule SEQ ID NO. 3698 or polypeptide SEQ ID NO. 3699, respectively, is increased or generated or if the activity "ORF YPL249c-a" is increased or generated in a plant cell, plant or part thereof, especially if the polypeptide is cytoplasmic localized, an increased intrinsic yield as compared to a corresponding non-modified, e.g. a non-transformed, wild type plant cell, a plant or a part thereof is conferred.
- an increased yield under standard conditions e.g. in the absence of nutrient deficiency as well as stress conditions.
- an increase of yield from 1.05-fold to 1.230-fold, for example plus at least 100% thereof, under conditions of low temperature is conferred corn-pared to a corresponding non-modified, e.g. non-transformed, wild type plant cell, a plant or a part thereof.
- ratios indicated above particularly refer to an increased yield actually measured as increase of biomass, especially as fresh weight biomass of aerial parts.
- sequence may relate to polynucleotides, nucleic acids, nucleic acid molecules, peptides, polypeptides and proteins, depending on the context in which the term “sequence” is used.
- nucleic acid molecule(s) refers to a polymeric form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. The terms refer only to the primary structure of the molecule.
- the terms "gene(s)", “polynucleotide”, “nucleic acid sequence”, “nucleotide sequence”, or “nucleic acid molecule(s)” as used herein include double- and single-stranded DNA and/or RNA. They also include known types of modifications, for example, methylation, "caps", substitutions of one or more of the naturally occurring nucleotides with an analogue.
- the DNA or RNA sequence comprises a coding sequence encoding the herein defined polypeptide.
- a "coding sequence” is a nucleotide sequence, which is transcribed into an RNA, e.g.
- a regulatory RNA such as a miRNA, a ta-siRNA, co-suppression molecule, an RNAi, a h- bozyme, etc. or into a mRNA which is translated into a polypeptide when placed under the control of appropriate regulatory sequences.
- the boundaries of the coding sequence are determined by a translation start codon at the 5'-terminus and a translation stop codon at the 3'-terminus.
- a coding sequence can include, but is not limited to mRNA, cDNA, recombinant nucleotide sequences or genomic DNA, while introns may be present as well under certain circumstances.
- nucleic acid molecule may also encompass the untranslated sequence located at the 3' and at the 5' end of the coding gene region, for example at least 500, preferably 200, especially preferably 100, nucleotides of the sequence upstream of the 5' end of the coding region and at least 100, preferably 50, especially preferably 20, nucleotides of the sequence downstream of the 3' end of the coding gene region.
- the antisense, RNAi, snRNA, dsRNA, siRNA, miRNA, ta-siRNA, co- suppression molecule, ribozyme etc. technology is used coding regions as well as the 5'- and/or 3'-regions can advantageously be used. However, it is often advantageous only to choose the coding region for cloning and expression purposes.
- Polypeptide refers to a polymer of amino acid (amino acid sequence) and does not refer to a specific length of the molecule. Thus, peptides and oligopeptides are included within the definition of polypeptide. This term does also refer to or include post-translational modifications of the polypeptide, for example, glycosylations, acetylations, phosphorylations and the like. Included within the definition are, for example, polypeptides containing one or more analogs of an amino acid (including, for example, unnatural amino acids, etc.), polypeptides with substituted linkages, as well as other modifications known in the art, both naturally oc- curring and non-naturally occurring.
- tablette I used in this specification is to be taken to specify the content of table I A and table I B.
- table II used in this specification is to be taken to specify the content of table Il A and table Il B.
- table I A used in this specification is to be taken to specify the content of table I A.
- table I B used in this specification is to be taken to specify the content of table I B.
- table Il A used in this specification is to be taken to specify the content of table Il A.
- table Il B used in this specification is to be taken to specify the content of table Il B.
- the term "table I" means table I B.
- the term “table II” means table Il B.
- a protein or polypeptide has the "activity of an protein as shown in table II, column 3" if its de novo activity, or its increased expression directly or indirectly leads to and confers an enhanced yield, e.g an improved yield- related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof and the protein has the above mentioned activities of a protein as shown in table II, column 3.
- the activity or preferably the biological activity of such a protein or polypeptide or an nucleic acid molecule or sequence encoding such protein or polypeptide is identical or similar if it still has the biological or enzymatic activity of a protein as shown in table II, column 3, or which has at least 10% of the original enzymatic activity, preferably 20%, 30%, 40%, 50%, particularly preferably 60%, 70%, 80% most particularly preferably 90%, 95 %, 98%, 99% in comparison to a protein as shown in table II, column 3 of S. cerevisiae or E. coli or Synechocystis sp. or A. thaliana.
- the biological or enzymatic activity of a protein as shown in table II, column 3 has at least 101 % of the original enzymatic activity, preferably 1 10%, 120%, %, 150%, particularly preferably 150%, 200%, 300% in comparison to a protein as shown in table II, column 3 of S. cerevisiae or E. coli or Synechocystis sp. or A. thaliana [0053.1.1.1]
- the terms “increased”, “rised”, “extended”, “enhanced”, “improved” or “amplified” relate to a corresponding change of a property in a plant, an organism, a part of an organism such as a tissue, seed, root, leave, flower etc.
- the overall activity in the volume is increased or enhanced in cases if the increase or enhancement is related to the increase or enhancement of an activity of a gene product, independent whether the amount of gene product or the specific activity of the gene product or both is increased or enhanced or whether the amount, stability or translation efficacy of the nucleic acid sequence or gene encoding for the gene product is in- creased or enhanced.
- the terms "increase” relate to a corresponding change of a property an organism or in a part of a plant, an organism, such as a tissue, seed, root, leave, flower etc. or in a cell.
- the overall activity in the volume is increased in cases the increase relates to the increase of an activity of a gene product, independent whether the amount of gene product or the specific activity of the gene product or both is increased or generated or whether the amount, stability or translation efficacy of the nucleic acid sequence or gene encoding for the gene product is increased.
- the terms "increase” include the change of said property in only parts of the subject of the present invention, for example, the modification can be found in compartment of a cell, like a organelle, or in a part of a plant, like tissue, seed, root, leave, flower etc. but is not detect- able if the overall subject, i.e. complete cell or plant, is tested.
- the term "increase” means that the specific activity of an enzyme as well as the amount of a compound or metabolite, e.g. of a polypeptide, a nucleic acid molecule of the invention or an encoding mRNA or DNA, can be increased in a volume.
- wild type can be a cell or a part of organisms such as an organelle like a chloroplast or a tissue, or an organism, in particular a plant, which was not modified or treated according to the herein described process according to the invention.
- the cell or a part of organisms such as an organelle like a chloroplast or a tissue, or an organism, in particular a plant used as wild type, control or reference corresponds to the cell, organism, plant or part thereof as much as possible and is in any other property but in the result of the process of the invention as identical to the subject matter of the invention as possible.
- the wild type, control or reference is treated identically or as identical as possible, saying that only conditions or properties might be different which do not influence the quality of the tested property.
- analogous conditions means that all conditions such as, for example, culture or growing conditions, soil, nutrient, water content of the soil, temperature, humidity or surrounding air or soil, assay conditions (such as buffer composition, temperature, substrates, pathogen strain, con- centrations and the like) are kept identical between the experiments to be compared.
- the "reference”, "control”, or “wild type” is preferably a subject, e.g. an organelle, a cell, a tissue, an organism, in particular a plant, which was not modified or treated according to the herein described process of the invention and is in any other property as similar to the subject matter of the invention as possible.
- the reference, control or wild type is in its genome, transcriptome, proteome or metabolome as similar as possible to the subject of the present invention.
- the term "reference-" "control-” or “wild type-”-organelle, -cell, -tissue or -organism, in particular plant relates to an organelle, cell, tissue or organism, in particular plant, which is nearly genetically identical to the organelle, cell, tissue or organism, in particular plant, of the present invention or a part thereof preferably 95%, more preferred are 98%, even more preferred are 99,00%, in particular 99,10%, 99,30%, 99,50%, 99,70%, 99,90%, 99,99%, 99,999% or more.
- the "reference", "control”, or “wild type” is a subject, e.g.
- a control, reference or wild type differing from the subject of the present invention only by not being subject of the process of the invention can not be pro- vided
- a control, reference or wild type can be an organism in which the cause for the modulation of an activity conferring the enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof or expression of the nucleic acid molecule of the invention as described herein has been switched back or off, e.g. by knocking out the expression of responsible gene product, e.g.
- a gene production can for example be knocked out by introducing inactivating point mutations, which lead to an enzymatic activity inhibition or a destabilization or an inhibition of the ability to bind to cofactors etc.
- preferred reference subject is the starting subject of the present process of the invention.
- the reference and the subject matter of the inven- tion are compared after standardization and normalization, e.g. to the amount of total RNA, DNA, or protein or activity or expression of reference genes, like housekeeping genes, such as ubiquitin, actin or ribosomal proteins.
- the increase or modulation according to this invention can be constitutive, e.g. due to a stable permanent transgenic expression or to a stable mutation in the corresponding endogenous gene encoding the nucleic acid molecule of the invention or to a modulation of the expression or of the behavior of a gene conferring the expression of the polypeptide of the invention, or transient, e.g. due to an transient transformation or temporary addition of a modulator such as a agonist or antagonist or inducible, e.g. after trans- formation with a inducible construct carrying the nucleic acid molecule of the invention under control of a inducible promoter and adding the inducer, e.g. tetracycline or as described herein below.
- a modulator such as a agonist or antagonist or inducible
- the increase in activity of the polypeptide amounts in a cell, a tissue, an organelle, an organ or an organism, preferably a plant, or a part thereof preferably to at least 5%, preferably to at least 20% or at to least 50%, especially preferably to at least 70%, 80%, 90% or more, very especially preferably are to at least 100%, 150 % or 200%, most preferably are to at least 250% or more in comparison to the control, reference or wild type.
- the term increase means the increase in amount in relation to the weight of the organism or part thereof (w/w).
- the increase in activity of the polypeptide amounts in an organelle such as a plastid.
- the increase in activity of the polypeptide amounts in the cytoplasm.
- the term "increase” includes, that a compound or an activity, especially an activity, is introduced into a cell, the cytoplasm or a subcellular compartment or organelle de novo or that the compound or the activity, especially an activity, has not been detected before, in other words it is "generated”. Accordingly, in the following, the term “increasing” also comprises the term “generating” or “stimulating”.
- the increased activity manifests itself in an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding e.g. non-transformed wild type plant cell, plant or part thereof.
- AT3G60210 from Arabidopsis thaliana, e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as Chloroplast chaperonin.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "Chloroplast chaperonin” from Arabidopsis thaliana or its functional equivalent or its homolog, e.g.
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "Chloroplast chaperonin", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule, which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "Chloroplast chaperonin” is increased Cytoplasmic.
- AVINDRAFT_2382 from Azotobacter vinelandii, e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as 3OS ri- bosomal protein S1 1.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "3OS ribosomal protein S1 1" from Azotobacter vinelandii or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said AVINDRAFT_2382 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said AVINDRAFT_2382, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic envi- ronmental stress and increased yield.
- an enhanced yield e.
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "3OS ribosomal protein S1 1 ", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "3OS ribosomal protein S11 ", is increased Cytoplasmic.
- AVINDRAFT_2913 from Azotobacter vinelandii, e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331), 1453 (1997)), and/or its activity is described as Short chain dehydrogenase.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "Short chain dehydrogenase" from Azotobacter vinelandii or its functional equivalent or its homolog, e.g. the in- crease of
- DRAFT_2913 or (b) a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said AVINDRAFT_2913 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said AVINDRAFT_2913, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g.
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "Short chain dehydrogenase", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "Short chain de- hydrogenase", is increased Cytoplasmic.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "B0252-protein" from Escherichia coli or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said B0252 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said B0252, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. en- hanced tolerance to abiotic environmental stress and/or increased yield like biomass as o
- a corresponding, e.g. non-transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "B0252-protein", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "B0252-protein", is increased Cytoplasmic.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "transcription regulator (farR)" from Escherichia coli or its functional equivalent or its homolog, e.g. the increase of (a) a gene product of a gene comprising the nucleic acid molecule as shown in column 5 of table I, and being depicted in the same respective line as said B0730 or a functional equivalent or a homologue thereof as shown depicted in column 7 of table I, preferably a homologue or functional equivalent as shown depicted in column 7 of table I B, and being depicted in the same respective line as said B0730; or (b) a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said B0730 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV,
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "transcription regulator (farR)", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule, which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "transcription regulator (farR)” is increased Cytoplasmic.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "flagellar protein" from Escherichia coli or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said B1926 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said B1926, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an enhanced yield e.g an improved yield-related trait, e.g. enhanced tolerance to
- the molecule which activity is to be increased in the proc- ess of the invention is the gene product with an activity of described as a "flagellar protein", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "flagellar protein", is increased Cytoplasmic.
- the sequence of B2426 from Escherichia coli e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as Short chain dehydrogenase. o
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "Short chain dehydrogenase" from Escherichia coli or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said B2426 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said B2426, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic envi- ronmental stress and increased yield.
- an enhanced yield e.g an improved yield-related trait, e.
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "Short chain dehydrogenase", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "Short chain dehydrogenase", is increased Cytoplasmic.
- G M 02 LC 13630 from Glycine max e.g. as shown in column 5 of table I
- sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996)
- sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)
- BRICK1 -like protein e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996)
- sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)
- its activity is described as BRICK1 -like protein.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "BRICK1 -like protein" from Glycine max or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said G M 02 LC 13630 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said G M 02 LC 13630, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic envi- ronmental stress and increased yield.
- an enhanced yield e.g an
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "BRICK1 -like protein", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "BRICK1 -like protein", is increased Cytoplasmic.
- sequence of SLL0418 from Synechocystis sp. e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as Sterol-C- methyltransferase.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "Sterol-C- methyltransferase" from Synechocystis sp. or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said SLL0418 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said SLL0418, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof .
- an enhanced yield e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part
- the molecule which activity is to be increased in the proc- ess of the invention is the gene product with an activity of described as a "Sterol-C- methyltransferase", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "Sterol-C- methyltransferase", is increased Mitochondrial.
- the sequence of YCR085W from Saccharomyces cerevisiae e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as Cav1 protein.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "Cav1 protein" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YCR085W or a functional equivalent or a homo- logue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YCR085W, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. n on -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an enhanced yield e.g an improved yield-related trait
- the molecule which activity is to be increased in the proc- ess of the invention is the gene product with an activity of described as a "Cav1 protein", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "Cav1 protein", is increased Cytoplasmic.
- sequence of YDL155W from Saccharomyces cerevisiae e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as G2/mitotic- specific cyclin.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "G2/mitotic-specific cyclin" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g.
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "G2/mitotic- specific cyclin", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule, which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "G2/mitotic- specific cyclin” is increased Cytoplasmic.
- sequence of YHL019C from Saccharomyces cerevisiae e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as Adaptin medium chain homolog APM2.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "Adaptin medium chain homolog APM2" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YHL019C or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YHL019C, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. en- hanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g.
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "Adaptin medium chain homolog APM2", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "Adaptin medium chain homolog APM2", is increased Cytoplasmic.
- sequence of YJR066w from Saccharomyces cerevisiae e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as Serine/threonine- protein kinase.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "Serine/threonine-protein kinase" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g. the increase of (a) a gene product of a gene comprising the nucleic acid molecule as shown in column 5 of table I, and being depicted in the same respective line as said YJR066w or a functional equivalent or a homologue thereof as shown depicted in column 7 of table I, preferably a homologue or functional equivalent as shown depicted in column 7 of table I B, and being depicted in the same respective line as said YJR066w; or OO
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YJR066w or a functional equivalent or a homo- logue thereof as depicted in column 7 of table Il or IV, preferably a homologue or func- tional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YJR066w, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an enhanced yield e.g an
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "Serine/threonine- protein kinase", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "Ser- ine/threonine-protein kinase", is increased Cytoplasmic.
- sequence of YKR015C from Saccharomyces cerevisiae e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as Ykr015c-protein.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "Ykr015c-protein" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YKR015C or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or func- tional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YKR015C, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to OD
- abiotic environmental stress or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "Ykr015c-protein", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "YkrO15c- protein", is increased Cytoplasmic.
- RNA polymerase Il holoenzyme cyclin-like subunit The sequence of YNL025C from Saccharomyces cerevisiae, e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as RNA polymerase Il holoenzyme cyclin-like subunit.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "RNA polymerase Il holoenzyme cyclin-like subunit" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YNL025C or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YNL025C, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. en- hanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an enhanced yield e.g an improved yield-
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "RNA polymerase Il holoenzyme cyclin-like subunit", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "RNA polymerase Il holoenzyme cyclin-like subunit", is increased Cytoplasmic.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "Membrane protein" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YOL073C or a functional equivalent or a homo- logue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YOL073C, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an enhanced yield e.g an improved yield-related trait,
- the molecule which activity is to be increased in the proc- ess of the invention is the gene product with an activity of described as a "Membrane protein", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "Membrane protein", is increased Plastidic.
- sequence of YPL167C from Saccharomyces cerevisiae e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as DNA poly- merase.
- sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996)
- sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)
- its activity is described as DNA poly- merase.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "DNA polymerase" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YPL167C or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YPL167C, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic envi- ronmental stress and increased yield.
- an enhanced yield e.g an improved yield-related
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "DNA polymerase", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "DNA polymerase", is increased Cytoplasmic.
- sequence of YPL223C from Saccharomyces cerevisiae e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as GRE1 - protein(Hydrophilin).
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "GRE1 - protein(Hydrophilin)" from Saccharomyces cerevisiae or its functional equivalent or its ho- molog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YPL223C or a functional equivalent or a homo- logue thereof as depicted in column 7 of table Il or IV, preferably a homologue or func- tional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YPL223C, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an enhanced yield e.g an improved yield
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "GRE1 - protein(Hydrophilin)", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "GRE1 - protein(Hydrophilin)", is increased Cytoplasmic.
- sequence of YPL249C-A from Saccharomyces cerevisiae e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as 60S ri- bosomal protein.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "60S ribosomal protein" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YPL249C-A or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YPL249C-A, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an enhanced yield e.g an improved yield-related trait
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "60S ribosomal protein", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule, which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "60S ribosomal protein” is increased Cytoplasmic.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "Short chain dehydro- genase" from Escherichia coli or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said B2426_2 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said B2426_2, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an enhanced yield e.g an improved yield-related trait, e.g. enhanced tolerance
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "Short chain de- hydrogenase", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule, which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "Short chain dehydrogenase" is increased Cytoplasmic.
- GM02LC13630_2 from Glycine max e.g. as shown in column 5 of table I
- sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996)
- sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)
- BRICK1 -like protein e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996)
- sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)
- its activity is described as BRICK1 -like protein.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "BRICK1 -like protein" from Glycine max or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said GM02LC13630_2 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said GM02LC13630_2, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an enhanced yield e.g an improved yield-related trait,
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "BRICK1 -like pro- tein", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "BRICK1 -like protein", is increased Cytoplasmic.
- YPL167C_2 from Saccharomyces cerevisiae, e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331 ), 1453 (1997)), and/or its activity is described as YPL167C_2-protein.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "YPL167C_2-protein" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g. the increase of
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YPL167C_2 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YPL167C_2, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic envi- ronmental stress and increased yield.
- an enhanced yield e.g an improved
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "YPL167C_2- protein", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule, which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "YPL167C_2- protein” is increased Cytoplasmic.
- sequence of YPL249C-A_2 from Saccharomyces cerevisiae e.g. as shown in column 5 of table I, is published (e.g. sequences from S. cerevisiae have been published in Goffeau et al., Science 274 (5287), 546 (1996), sequences from E. coli have been published in Blattner et al., Science 277 (5331), 1453 (1997)), and/or its activity is described as ORF YPL249c-a.
- the process of the present invention comprises increasing or generating the activity of a gene product with the activity of a "ORF YPL249c-a" from Saccharomyces cerevisiae or its functional equivalent or its homolog, e.g.
- a polypeptide comprising a polypeptide, a consensus sequence or a polypeptide motif as shown depicted in column 5 of table Il or column 7 of table IV, and being depicted in the same respective line as said YPL249C-A_2 or a functional equivalent or a homologue thereof as depicted in column 7 of table Il or IV, preferably a homologue or functional equivalent as depicted in column 7 of table Il B, and being depicted in the same respective line as said YPL249C-A_2, as mentioned herein, for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non -transformed wild type plant cell, plant or part thereof in plant cell, plant or part thereof, as mentioned, especially for an enhanced tolerance to abiotic environmental stress, or increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an enhanced yield e.g an improved yield
- the molecule which activity is to be increased in the process of the invention is the gene product with an activity of described as a "ORF YPL249c-a", preferably it is the molecule of section (a) or (b) of this paragraph.
- said molecule which activity is to be increased in the process of the invention and which is the gene product with an activity as described as a "ORF YPL249c- a", is increased Cytoplasmic.
- thaliana conferred an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass in the transformed plants as compared to a corresponding, e.g. non-transformed, wild type plant, especially an enhanced tolerance to abiotic environmental stress, or an increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- an improved yield-related trait e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass in the transformed plants as compared to a corresponding, e.g. non-transformed, wild type plant, especially an enhanced tolerance to abiotic environmental stress, or an increased yield, or an enhanced tolerance to abiotic environmental stress and increased yield.
- Adaptin medium chain homolog APM2 encoded by a gene comprising the nucleic acid sequence SEQ ID NO.: 2463 in A. thaliana conferred an increased yield, e.g. an increased yield-related trait, for example enhanced tolerance to abiotic environmental stress, for example an increased drought tolerance and/or low temperature tolerance and/or an increased nutrient use efficiency, intrinsic yield and/or another mentioned yield-related trait as compared to wild type control.
- an increased yield-related trait for example enhanced tolerance to abiotic environmental stress, for example an increased drought tolerance and/or low tem- perature tolerance and/or an increased nutrient use efficiency, intrinsic yield and/or another mentioned yield-related trait as compared to wild type control.
- RNA polymerase Il holoenzyme cyclin-like subunit encoded by a gene comprising the nucleic acid sequence SEQ ID NO.: 2599 in A. thaliana conferred an increased biomass compared to the wild type control.
- increasing or generating the activity of a gene product being encoded by a gene comprising the nucleic acid sequence SEQ ID NO.: 2599 localized as indicated in table I, column 6, e.g. Cytoplasmic in A. thaliana, for example with the activity of a "RNA polymerase Il holoenzyme cyclin- like subunit” conferred an increased yield, for example biomass increase.
- RNA polymerase Il holoenzyme cyclin-like subunit encoded by a gene comprising the nucleic acid sequence SEQ ID NO. 2599 in A. thaliana conferred an increased tolerance to abiotic environmental stress and an increased yield compared with the wild type control.
- increasing or generating the activity of a gene product with the activity of a "Membrane protein” encoded by a gene comprising the nucleic acid sequence SEQ ID NO.: 2645 in A. thaliana conferred an increased yield, e.g.
- an increased yield-related trait for example enhanced tolerance to abiotic environmental stress, for example an increased drought tolerance and/or low temperature tolerance and/or an in- creased nutrient use efficiency, intrinsic yield and/or another mentioned yield-related trait as compared to wild type control.
- an increased yield-related trait for example enhanced tolerance to abiotic environmental stress, for example an increased drought tolerance and/or low temperature tolerance and/or an in- creased nutrient use efficiency, intrinsic yield and/or another mentioned yield-related trait as compared to wild type control.
- an increased yield-related trait for example enhanced tolerance to abiotic environmental stress, for example an increased drought tolerance and/or low temperature tolerance and/or an in- creased nutrient use efficiency, intrinsic yield and/or another mentioned yield-related trait as compared to wild type control.
- an increased yield-related trait for example enhanced tolerance to abiotic environmental stress, for example an increased drought tolerance and/or low temperature tolerance and/or an in- creased nutrient use efficiency, intrinsic yield and/or another mentioned yield-related trait as compared to wild type control.
- an increased yield-related trait for example enhanced tolerance to abiotic environmental stress, for example an increased drought tolerance and/or low temperature tolerance and/or an in- creased nutrient use efficiency, intrinsic yield and/or another mentioned yield-related trait as compared to wild type control.
- nucleic acid molecule indicated in Table Villa or its homolog as indicated in Table I or the expression product is used in the method of the present invention to increased nutrient use efficiency, e.g. to increased the nitrogen use efficiency, of the a plant compared with the wild type control. It was further observed that increasing or generating the activity of a nucleic acid molecule derived from the nucleic acid molecule shown in Table VIIIb in A.
- thaliana conferred increased stress tolerance, e.g. increased low temperature tolerance, compared with the wild type control.
- a nucleic acid molecule indicated in Table VIIIb or its homolog as indicated in Table I or the expression product is used in the method of the present invention to increase stress tolerance, e.g. increase low temperature, of a plant compared with the wild type control.
- nucleic acid molecule indicated in Table VIIIc or its homolog as indicated in Table I or the expression product is used in the method of the present invention to increase stress tolerance, e.g. increase cycling drought tolerance, of a plant compared with the wild type control.
- nucleic acid molecule indicated in Table VIIId or its homolog as indicated in Table I or the expression product is used in the method of the present invention to increase intrinsic yield, e.g. to increase yield under standard conditions, e.g. increase biomass under non-deficiency or non-stress conditions, of a plant compared with the wild type control.
- an enhanced yield e.g an improved yield- related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass in a plant cell, plant or a part thereof compared to a control or wild type can be achieved.
- expression refers to the transcription and/or translation of a codogenic gene segment or gene.
- the resulting product is an mRNA or a protein.
- expression products can also include functional RNA such as, for example, an- tisense, nucleic acids, tRNA, snRNA, rRNA, RNAi, siRNA, ribozymes etc.
- Expression may be systemic, local or temporal, for example limited to certain cell types, tissues organs or organelles or time periods.
- the process of the present invention comprises one or more of the following steps
- Chloroplast chaperonin DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GREI -protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoenzyme cyclin-like subunit, Serine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, transcription regulator (farR), Ykr015c-protein, and YPL167C_2-protein and confering an enhanced yield, e.g an improved yield- related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof;
- YPL167C_2-protein and conferring an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield o
- a transgenic gene encoding a protein conferring the increased expression of a polypeptide encoded by the nucleic acid molecule of the present invention or a polypeptide of the present invention, having the herein-mentioned activity selected from the group consisting of 3OS ribosomal protein S1 1 , 60S ribosomal protein, Adaptin medium chain homolog APM2, B0252-protein, BRICK1-like protein, Cav1 protein, Chloroplast chaperonin, DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GREI -protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoenzyme cyclin-like subunit, Serine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, transcription regulator (farR), YkrO15c- protein, and YPL167
- Further gene conversion methods can be used to disrupt repressor elements or to enhance to activity of positive elements- positive elements can be randomly introduced in plants by T-DNA or transpo- son mutagenesis and lines can be identified in which the positive elements have been integrated near to a gene of the invention, the expression of which is thereby enhanced; and/or (i) modulating growth conditions of the plant in such a manner, that the expression or activity of the gene encoding the protein of the invention or the protein itself is enhanced;
- said mRNA is the nucleic acid molecule of the present invention and/or the protein conferring the increased expression of a protein encoded by the nu- cleic acid molecule of the present invention alone or linked to a transit nucleic acid sequence or transit peptide encoding nucleic acid sequence or the polypeptide having the herein mentioned activity, e.g. conferring an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof after increasing the expression or activity of the encoded polypeptide or having the activity of a polypeptide having an activity as the protein as shown in table Il column 3 or its homolog.
- the amount of mRNA or polypeptide in a cell or a compartment of an organism correlates with the amount of encoded protein and thus with the overall activity of the encoded protein in said volume. Said correlation is not always linear, the activity in the volume is dependent on the stability of the molecules or the presence of activating or inhibiting co-factors. Further, product and educt inhibition of enzymes are well known and described in textbooks, e.g. Stryer, Biochemistry.
- the amount of mRNA, polynucleotide or nucleic acid molecule in a cell or a compartment of an organism correlates with the amount of encoded protein and thus with the overall activity of the encoded protein in said volume. Said correlation is not always linear, the activity in the volume is dependent on the stability of the molecules, the degradation of the molecules or the presence of activating or inhibiting co-factors. Further, product and educt inhibition of enzymes are well known, e.g. Zinser et al. "Enzymin- hibitoren'V ⁇ nzyme inhibitors".
- the activity of the abovementioned proteins and/or polypeptides encoded by the nucleic acid molecule of the present invention can be increased in various ways.
- the activity in an organism or in a part thereof, like a cell is increased via increasing the gene product number, e.g. by increasing the expression rate, like introducing a stronger promoter, or by increasing the stability of the mRNA expressed, thus increasing the translation rate, and/or increasing the stability of the gene product, thus reducing the proteins decayed.
- the activity or turnover of enzymes can be influenced in such a way that a reduction or increase of the reaction rate or a modification (reduction or increase) of the affinity to the substrate results, is reached.
- a mutation in the catalytic centre of an polypeptide of the invention can modulate the turn over rate of the enzyme, e.g. a knock out of an essential amino acid can lead to a reduced or completely knock out activity of the enzyme, or the deletion or mutation of regulator binding sites can reduce a negative regulation like a feedback inhibition (or a substrate inhibition, if the sub- strate level is also increased).
- the specific activity of an enzyme of the present invention can be increased such that the turn over rate is increased or the binding of a co-factor is improved. Improving the stability of the encoding mRNA or the protein can also increase the activity of a gene product.
- the stimulation of the activity is also under the scope of the term "increased activity".
- the regulation of the abovementioned nucleic acid sequences may be modified so that gene expression is increased. This can be achieved advantageously by means of heterologous regulatory sequences or by modifying, for example mutating, the natural regulatory sequences which are present. The advantageous methods may also be combined with each other.
- an activity of a gene product in an organism or part thereof, in particular in a plant cell or organelle of a plant cell, a plant, or a plant tissue or a part thereof or in a microorganism can be increased by increasing the amount of the specific encoding mRNA or the corresponding protein in said organism or part thereof.
- “Amount of protein or mRNA” is understood as meaning the molecule number of polypeptides or mRNA molecules in an organism, especially a plant, a tissue, a cell or a cell compartment.
- Increase in the amount of a protein means the quantitative increase of the molecule number of said protein in an organism, especially a plant, a tissue, a cell or a cell compartment such as an organelle like a plastid or mitochondria or part thereof - for example by one of the methods described herein below - in comparison to a wild type, control or reference.
- the increase in molecule number amounts preferably to at least 1 %, preferably to more than 10%, more preferably to 30% or more, especially preferably to 50%, 70% or more, very especially preferably to 100%, most preferably to 500% or more.
- a de novo expression is also regarded as subject of the present invention.
- a modification i.e. an increase
- an increase in activity in an organism or a part thereof can be caused by adding a gene product or a precursor or an activator or an agonist to the media or nutrition or can be caused by introducing said subjects into a organism, transient or stable.
- an increase can be reached by the introduction of the inventive nucleic acid sequence or the encoded protein in the correct cell compartment for example into the nucleus or cytoplasm respectively or into plastids either by transformation and/or target- ing.
- cytoplasmic shall indicate, that the nucleic acid of the invention is expressed without the addition of an non-natural transit peptide encoding sequence.
- a non-natural transient peptide encoding sequence is a sequence which is not a natural part of a nucleic acid of the invention but is rather added by molecular manipulation steps as for example described in the example under "plastid targeted expression". Therefore the term “cytoplasmic” shall not exclude a targeted localisation to any cell compartment for the products of the inventive nucleic acid se- quences by their naturally occurring sequence properties.
- the enhancement of the yield e.g an improved yield- related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell in the plant or a part thereof, e.g. in a cell, a tissue, a organ, an organelle, the cytoplasm etc.
- the present invention relates to a process wherein the gene copy number of a gene encoding the polynucleotide or nucleic acid molecule of the invention is increased.
- the endogenous level of the polypeptide of the invention can for example be increased by modifying the transcriptional or translational regulation of the polypeptide.
- the enhanced yield e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass of the plant or part thereof can be altered by targeted or random mutagenesis of the endogenous genes of the invention.
- homologous recombination can be used to either introduce positive regulatory elements like for plants the 35S enhancer into the promoter or to remove repressor elements form regulatory regions.
- gene conversion like methods described by Kochevenko and Willmitzer (Plant Physiol. 132 (1), 174 (2003)) and citations therein can be used to disrupt repressor elements or to enhance to activity of positive regulatory elements.
- positive elements can be randomly introduced in (plant) genomes by T-DNA or transposon mutagenesis and lines can be screened for, in which the positive elements have been integrated near to a gene of the invention, the expression of which is thereby en- hanced.
- the activation of plant genes by random integrations of enhancer elements has been described by Hayashi et al. (Science 258,1350 (1992)) or Weigel et al. (Plant Physiol. 122, 1003 (2000)) and others cited therein.
- the enhancement of positive regulatory elements or the disruption or weakening of negative regulatory elements can also be achieved through common mutagenesis techniques:
- the production of chemically or radiation mutated populations is a common technique and known to the skilled worker. Methods for plants are described by Koorneef et al. (Mutat Res. Mar. 93 (1 ) (1982)) and the citations therein and by Lightner and Caspar in "Methods in Molecular Biology” Vol. 82. These techniques usually induce point mutations that can be identified in any known gene using methods such as TILLING (Colbert et al., Plant Physiol, 126, (2001 )).
- the expression level can be increased if the endogenous genes encoding a polypeptide conferring an increased expression of the polypeptide of the present invention, in particular genes comprising the nucleic acid molecule of the present invention, are modified via homologous recombination, Tilling approaches or gene conver- sion. It also possible to add as mentioned herein targeting sequences to the inventive nucleic acid sequences.
- Regulatory sequences in addition to a target sequence or part thereof can be operatively linked to the coding region of an endogenous protein and control its transcription and translation or the stability or decay of the encoding mRNA or the expressed protein.
- promoter, UTRs, splicing sites, processing signals, polyadenylation sites, terminators, enhancers, repressors, post transcriptional or posttranslational modification sites can be changed, added or amended.
- enhancer elements has been described by Hayashi et al. (Science 258, 1350(1992)) or Weigel et al. (Plant Physiol.
- the expression level of the endogenous protein can be modulated by replacing the endogenous promoter with a stronger transgenic promoter or by replacing the endogenous 3'UTR with a 3'UTR, which provides more stability without amending the coding region.
- the transcriptional regulation can be modulated by introduction of an artificial transcription factor as described in the examples. Alternative promoters, terminators and UTR are described below.
- n on -transformed, wild type plant cell, plant or part thereof after increase of expression or activity in the cytoplasm and/or in an organelle like a plastid can also be increased by introducing a synthetic transcription factor, which binds close to the coding region of the gene encoding the protein as shown in table II, column 3 and activates its transcription.
- a chimeric zinc finger protein can be constructed, which comprises a specific DNA-binding domain and an activation domain as e.g. the VP16 domain of Herpes Simplex virus. The specific binding domain can bind to the regulatory region of the gene encoding the protein as shown in table II, column 3.
- the methods thereto a known to a skilled person and/or disclosed e.g. in WO01/52620, Oriz, Proc. Natl. Acad. Sci. USA, 99, 13290 (2002) or Guan, Proc. Natl. Acad. Sci. USA 99, 13296 (2002).
- or- ganisms are used in which one of the abovementioned genes, or one of the abovemen- tioned nucleic acids, is mutated in a way that the activity of the encoded gene products is less influenced by cellular factors, or not at all, in comparison with the unmutated proteins.
- well known regulation mechanism of enzymic activity are substrate inhibition or feed back regulation mechanisms. Ways and techniques for the introduction of substitu- tion, deletions and additions of one or more bases, nucleotides or amino acids of a corresponding sequence are described herein below in the corresponding paragraphs and the references listed there, e.g.
- nucleic acid molecule of the invention or a polypeptide of the invention derived from a evolutionary distantly related organism as e.g. using a prokaryotic gene in a eukaryotic host, as in these cases the regulation mechanism of the host cell may not weaken the activity (cellular or specific) of the gene or its expression product.
- the mutation is introduced in such a way that the enhanced tolerance to abiotic environmental stress and/or yield increase are not adversely affected.
- the invention provides that the above methods can be performed such that yield of a plant is increased, or the tolerance to abiotic environmental stress is increased, or both, wherein particularly the yield of a plant (e.g. biomass yield) is increased.
- the invention provides that the above methods can be performed such that yield-related traits are increased.
- the invention provides that the above methods can be performed such that the yield is increased in the absence of nutrient deficiencies as well as the absence of stress conditions.
- the inven- tion provides that the above methods can be performed such that the nutrient use efficiency, particularly the nitrogen use efficiency, and the yield, in the absence of nutrient deficiencies as well as the absence of stress conditions, is increased.
- the invention provides that the above methods can be performed such that the tolerance to abiotic stress, particularly the tolerance to low temperature and/or water use effi- ciency, and at the same time, the nutrient use efficiency, particularly the nitrogen use efficiency, and the yield in the absence of nutrient deficiencies as well as the absence of stress conditions, is increased.
- the present invention also relates to isolated nucleic acids comprising a nucleic acid molecule selected from the group consisting of:
- nucleic acid molecule shown in column 7 of table I B, application no.1 (b) a nucleic acid molecule shown in column 7 of table I B, application no.1 ; (c) a nucleic acid molecule, which, as a result of the degeneracy of the genetic code, can be derived from a polypeptide sequence depicted in column 5 or 7 of table II, application no.1 , and confers an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, a plant or a part thereof;
- an enhanced yield e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, a plant or a part thereof;
- nucleic acid molecule having at least 30% identity, preferably at least 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99,5%, with the nu- oleic acid molecule sequence of a polynucleotide comprising the nucleic acid molecule shown in column 5 or 7 of table I, application no.1 , and confers an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non- transformed, wild type plant cell, a plant or a part thereof ;
- nucleic acid molecule encoding a polypeptide having at least 30% identity, preferably at least 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99,5%, with the amino acid sequence of the polypeptide encoded by the nucleic acid molecule of (a), (b), (c) or (d) and having the activity represented by a nucleic acid molecule comprising a polynucleotide as depicted in column 5 of table I, application no.1 , and confers an enhanced yield, e.g an improved yield-related trait, e.g.
- nucleic acid molecule which hybridizes with a nucleic acid molecule of (a), (b), (c), (d) or (e) under stringent hybridization conditions and confers an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g.
- nucleic acid molecule encoding a polypeptide which can be isolated with the aid of monoclonal or polyclonal antibodies made against a polypeptide encoded by one of the nucleic acid molecules of (a), (b), (c), (d), (e) or (f) and having the activity represented by the nucleic acid molecule comprising a polynucleotide as depicted in column 5 of table I, application no.1 ;
- nucleic acid molecule which comprises a polynucleotide, which is obtained by amplify- ing a cDNA library or a genomic library using the primers in column 7 of table III, application no.1 , which do not start at their 5'-end with the nucleotides ATA and preferably having the activity represented by a nucleic acid molecule comprising a polynucleotide as depicted in column 5 of table Il or IV, application no.1 ; and (k) a nucleic acid molecule which is obtainable by screening a suitable nucleic acid library, especially a cDNA library and/or a genomic library, under stringent hybridization conditions with a probe comprising a complementary sequence of a nucleic acid molecule of (a) or (b) or with a fragment thereof,
- the invention relates to homologs of the aforementioned sequences, which can be isolated advantageously from yeast, fungi, viruses, algae, bacteria, such as Acetobacter (subgenus Acetobacter) aceti; Acidithiobacillus ferrooxidans; Acinetobacter sp.; Actinobacillus sp; Aeromonas salmonicida; Agrobacterium tumefaciens; Aquifex aeolicus; Area no bacterium pyogenes; Aster yellows phytoplasma; Bacillus sp.; Bifidobacterium sp.; Borrelia burgdorferi; Brevi bacterium linens; Brucella melitensis; Buchnera sp.; Butyrivibrio fibrisolvens; Campylobacter jejuni; Caulobacter crescentus; Chlamydia sp.; Chlamydophila sp.; Chlorobi
- TA144 Mycobacterium sp.; Mycoplasma sp.; Neisseria sp.; Nitrosomonas sp.; Nostoc sp. PCC 7120; Novosphingobium aromaticivorans; Oenococcus oeni; Pantoea citrea; Pasteurella multocida; Pediococcus pentosaceus; Phor- midium foveolarum; Phytoplasma sp.; Plectonema boryanum; Prevotella ruminicola; Propi- onibacterium sp.; Proteus vulgaris; Pseudomonas sp.; Ralstonia sp.; Rhizobium sp.;
- Rhodococcus equi Rhodothermus marinus; Rickettsia sp.; Riemerella anatipestifer; Rumi- nococcus flavefaciens; Salmonella sp.; Selenomonas ruminantium; Serratia entomophila; Shigella sp.; Sinorhizobium meliloti; Staphylococcus sp.; Streptococcus sp.; Streptomyces sp.; Synechococcus sp.; Synechocystis sp.
- PCC 6803 Thermotoga maritima; Treponema sp.; Ureaplasma urealyticum; Vibrio cholerae; Vibrio parahaemolyticus; XyIeIIa fastidiosa; Yersinia sp.; Zymomonas mobilis, preferably Salmonella sp. or E. coli or plants, preferably from yeasts such as from the genera Saccharomyces, Pichia, Candida, Hansenula, Toru- lopsis or Schizosaccharomyces or plants such as A.
- thaliana maize, wheat, rye, oat, triticale, rice, barley, soybean, peanut, cotton, borage, sunflower, linseed, primrose, rapeseed, canola and turnip rape, manihot, pepper, sunflower, tagetes, solanaceous plant such as potato, tobacco, eggplant and to- mato, Vicia species, pea, alfalfa, bushy plants such as coffee, cacao, tea, Salix species, trees such as oil palm, coconut, perennial grass, such as ryegrass and fescue, and forage crops, such as alfalfa and clover and from spruce, pine or fir for example. More preferably homologs of aforementioned sequences can be isolated from S. cerevisiae, E. coli or Synechocystis sp. or plants, preferably Brassica napus, Glycine max, Zea mays, cotton or Oryza sativa.
- the proteins of the present invention are preferably produced by recombinant DNA techniques.
- a nucleic acid molecule encoding the protein is cloned into an expression vector, for example in to a binary vector, the expression vector is introduced into a host cell, for example the A. thaliana wild type NASC N906 or any other plant cell as described in the examples see below, and the protein is expressed in said host cell.
- binary vectors are pBIN19, pBI101 , pBinAR, pGPTV, pCAMBIA, pBIB-HYG, pBecks, pGreen or pPZP (Hajukiewicz, P. et al., Plant MoI. Biol. 25, 989 (1994), and HeI- lens et al, Trends in Plant Science 5, 446 (2000)).
- the protein of the present invention is preferably produced in a compartment of the cell, more preferably in the plastids. Ways of introducing nucleic acids into plastids and producing proteins in this compartment are known to the person skilled in the art have been also described in this application.
- the polypeptide of the invention is a protein localized after expression as indicated in column 6 of table II, e.g. non- targeted, mitochondrial or plastidic, for example it is fused to a transit peptide as decribed above for plastidic localisation.
- the protein of the present invention is preferably produced in the cytoplasm of the cell.
- Ways of producing proteins in the cytoplasm are known to the person skilled in the art.
- Ways of producing proteins without artificial targeting are known to the person skilled in the art.
- the nucleic acid sequences according to the invention or the gene construct together with at least one reporter gene are cloned into an expression cassette, which is introduced into the organism via a vector or directly into the genome.
- This reporter gene should allow easy detection via a growth, fluorescence, chemical, biolumi- nescence or tolerance assay or via a photometric measurement.
- antibiotic- or herbicide-tolerance genes hydrolase genes, fluorescence protein genes,
- a nucleic acid construct for example an ex- pression cassette, comprises upstream, i.e. at the 5' end of the encoding sequence, a promoter and downstream, i.e. at the 3' end, a polyadenylation signal and optionally other regulatory elements which are operably linked to the intervening encoding sequence with one of the nucleic acids of SEQ ID NO as depicted in table I, column 5 and 7.
- an operable linkage is meant the sequential arrangement of promoter, encoding sequence, termi- nator and optionally other regulatory elements in such a way that each of the regulatory elements can fulfill its function in the expression of the encoding sequence in due manner.
- sequences preferred for operable linkage are targeting sequences for ensuring subcellular localization in plastids.
- a nucleic acid construct for example an expression cassette may, for ex- ample, contain a constitutive promoter or a tissue-specific promoter (preferably the USP or napin promoter) the gene to be expressed and the ER retention signal.
- a constitutive promoter or a tissue-specific promoter preferably the USP or napin promoter
- the ER retention signal the KDEL amino acid sequence (lysine, aspartic acid, glutamic acid, leucine) or the KKX amino acid sequence (lysine-lysine-X-stop, wherein X means every other known amino acid) is preferably employed.
- the expression cassette is advantageously inserted into a vector such as by way of example a plasmid, a phage or other DNA which allows optimal expression of the genes in the host organism.
- a vector such as by way of example a plasmid, a phage or other DNA which allows optimal expression of the genes in the host organism.
- suitable plasmids are: in E. coli pLG338, pACYC184, pBR series such as e.g.
- pBR322 pUC series such as pUC18 or pUC19, M 113mp series, pKC30, pRep4, pHS1 , pHS2, pPLc236, pMBL24, pLG200, pUR290, plN-IM 113 -B1 , ⁇ gt1 1 or pBdCI; in Streptomyces plJ101 , plJ364, plJ702 or plJ361 ; in Bacillus pUB1 10, pC194 or pBD214; in Corynebacte- rium pSA77 or pAJ667; in fungi pALS1 , plL2 or pBB1 16; other advantageous fungal vectors are described by Romanos M.A.
- yeast promoters examples include 2 ⁇ M, pAG-1 , YEp6, YEpI 3 or pEMBLYe23.
- algal or plant promoters examples include pLGV23, pGHIac + , pBIN19, pAK2004, pVKH or pDH51 (see Schmidt, R. and Willmitzer, L., Plant Cell Rep. 7, 583 (1988))).
- the vectors identified above or derivatives of the vectors identified above are a small selection of the possible plasmids.
- vectors is meant with the exception of plasmids all other vectors known to those skilled in the art such as by way of example phages, viruses such as SV40, CMV, baculovirus, adenovirus, transposons, IS elements, phasmids, phagemids, cosmids, linear or circular DNA.
- phages viruses such as SV40, CMV, baculovirus, adenovirus, transposons, IS elements, phasmids, phagemids, cosmids, linear or circular DNA.
- viruses such as SV40, CMV, baculovirus, adenovirus, transposons, IS elements, phasmids, phagemids, cosmids, linear or circular DNA.
- viruses such as SV40, CMV, baculovirus, adenovirus, transposons, IS elements, phasmids, phagemids, cosmids, linear or circular DNA.
- the expression cassette according to the invention may also advantageously be introduced into the organisms in the form of a linear DNA and be integrated into the genome of the host organism by way of heterologous or homologous recombination.
- This linear DNA may be composed of a linearized plasmid or only of the expression cassette as vector or the nucleic acid sequences according to the invention.
- nucleic acid sequence according to the invention can also be introduced into an organism on its own.
- nucleic acid sequence according to the invention further genes are to be introduced into the organism, all together with a reporter gene in a single vector or each single gene with a reporter gene in a vector in each case can be introduced into the organism, whereby the different vectors can be introduced simultaneously or successively.
- the invention further provides an isolated recombinant expression vector comprising a nucleic acid encoding a polypeptide as depicted in table II, column 5 or 7, wherein expression of the vector in a host cell results in enhanced tolerance to abiotic environmental stress and/or yield as compared to a wild type variety of the host cell.
- vector refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.
- plasmid refers to a circular double stranded DNA loop into which additional DNA segments can be ligated.
- viral vector Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome.
- Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g. bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g.
- non-episomal mammalian vectors are integrated into the genome of a host cell or a organelle upon introduction into the host cell, and thereby are replicated along with the host or organelle genome.
- certain vectors are capable of directing the expression of genes to which they are opera- tively linked.
- Such vectors are referred to herein as "expression vectors.”
- expression vectors of utility in recombinant DNA techniques are often in the form of plasmids.
- plasmid and vector can be used interchangeably as the plasmid is the most commonly used form of vector.
- the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses, and adeno-associated viruses), which serve equivalent functions.
- the recombinant expression vectors of the invention comprise a nucleic acid of the invention in a form suitable for expression of the nucleic acid in a host cell, which means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis of the host cells to be used for expression, which is operatively linked to the nucleic acid sequence to be expressed.
- "operatively linked" is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner which allows for expression of the nucleotide sequence (e.g. in an in vitro transcription/translation system or in a host cell when the vector is introduced into the host cell).
- regulatory sequence is intended to include promoters, enhancers, and other expression control elements (e.g. polyadenyla- tion signals). Such regulatory sequences are described, for example, in Goeddel, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, CA (1990), and Gruber and Crosby, in: Methods in Plant Molecular Biology and Biotechnology, eds. Glick and Thompson, Chapter 7, 89-108, CRC Press; Boca Raton, Florida, including the references therein. Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cells and those that direct expression of the nucleotide sequence only in certain host cells or under certain conditions.
- the expression vectors of the invention can be introduced into host cells to thereby produce polypeptides or peptides, including fusion polypeptides or peptides, encoded by nucleic acids as described herein (e.g., YIPs, mutant forms of YIPs, fusion polypeptides, "Yield Related Proteins" or "YIPs” etc.).
- the recombinant expression vectors of the invention can be designed for expression of the polypeptide of the invention in plant cells.
- YIP genes can be expressed in plant cells (see Schmidt R., and Willmitzer L., Plant Cell Rep. 7 (1988); Plant Molecular Biology and Biotechnology, C Press, Boca Raton, Florida, Chapter 6/7, p. 71 -1 19 (1993); White F. F., Jenes B. et al., Techniques for Gene Transfer, in: Transgenic Plants, Vol. 1 , Engineering and Utilization, eds. Kung und Wu R., 128-43, Academic Press: 1993; Potrykus, Annu. Rev. Plant Physiol. Plant Molec. Biol.
- the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.
- Fusion vectors add a number of amino acids to a polypeptide encoded therein, usually to the amino terminus of the recombinant polypeptide but also to the C-terminus or fused within suitable regions in the polypeptides.
- Such fusion vectors typically serve three purposes: 1 ) to increase expression of a recombinant polypeptide; 2) to increase the solubility of a recombinant polypeptide; and 3) to aid in the purification of a recombinant polypeptide by acting as a ligand in affinity purification.
- a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant polypeptide to enable separation of the recombinant polypeptide from the fusion moiety subsequent to purification of the fusion polypeptide.
- enzymes, and their cognate recognition sequences include Factor Xa, thrombin, and enterokinase.
- the plant expression cassette can be installed in the pRT transformation vector ((a) Toepfer et al., Methods Enzymol. 217, 66 (1993), (b) Toepfer et al., Nucl. Acids. Res. 15, 5890 (1987)).
- Fusion vectors employed in prokaryotes frequently make use of induc- ible systems with and without fusion proteins or fusion oligopeptides, wherein these fusions can ensue in both N-terminal and C-terminal manner or in other useful domains of a protein.
- Such fusion vectors usually have the following purposes: 1 ) to increase the RNA expression rate; 2) to increase the achievable protein synthesis rate; 3) to increase the solubility of the protein; 4) or to simplify purification by means of a binding sequence usable for affinity chromatography.
- Proteolytic cleavage points are also frequently introduced via fusion proteins, which allow cleavage of a portion of the fusion protein and purification.
- recognition sequences for proteases are recognized, e.g. factor Xa, thrombin and enterokinase.
- Typical advantageous fusion and expression vectors are pGEX (Phar- macia Biotech Inc; Smith D. B. and Johnson K.S., Gene 67, 31 (1988)), pMAL (New Eng- land Biolabs, Beverly, MA) and pRIT5 (Pharmacia, Piscataway, NJ) which contains glutathione S-transferase (GST), maltose binding protein or protein A.
- GST glutathione S-transferase
- the coding sequence of the polypeptide of the inven- tion is cloned into a pGEX expression vector to create a vector encoding a fusion polypeptide comprising, from the N-terminus to the C-terminus, GST-thrombin cleavage site-X polypeptide.
- the fusion polypeptide can be purified by affinity chromatography using glutathione-agarose resin. Recombinant YIP unfused to GST can be recovered by cleavage of the fusion polypeptide with thrombin.
- E. coli expression vectors are pTrc (Amann et al., Gene 69, 301 (1988)) and pET vectors (Studier et al., Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, California (1990) 60-89; Stratagene, Amsterdam, The Netherlands).
- Target gene expression from the pTrc vector relies on host RNA polymerase transcription from a hybrid trp-lac fusion promoter.
- Target gene expression from the pET 1 1 d vector relies on transcription from a T7 gn10-lac fusion promoter mediated by a co-expressed viral RNA polymerase (T7 gn1 ).
- This viral polymerase is supplied by host strains BL21 (DE3) or HMS174(DE3) from a resident I prophage harboring a T7 gn1 gene under the transcriptional control of the lacUV 5 promoter.
- the YIPs are expressed in plants and plants cells such as unicellular plant cells (e.g. algae) (see Falciatore et al., Marine Biotechnology 1 (3), 239 (1999) and references therein) and plant cells from higher plants (e.g., the spermatophytes, such as crop plants).
- a nucleic acid molecule coding for YIP as depicted in table II, column 5 or 7 may be "introduced" into a plant cell by any means, including transfection, transformation or transduction, electroporation, particle bombardment, agroinfection, and the like.
- One transformation method known to those of skill in the art is the dipping of a flowering plant into an Agrobacteria solution, wherein the Agro- bacteria contains the nucleic acid of the invention, followed by breeding of the transformed gametes.
- Forage crops include, but are not limited to Wheatgrass, Canarygrass, Bromegrass, Wildrye Grass, Bluegrass, Or- chardgrass, Alfalfa, Salfoin, Birdsfoot Trefoil, Alsike Clover, Red Clover and Sweet Clover.
- transfection of a nucleic acid molecule coding for YIP as depicted in table II, column 5 or 7 into a plant is achieved by Agrobacterium mediated gene transfer.
- Agrobacterium mediated plant transformation can be performed using for example the GV3101 (pMP90) (Koncz and Schell, MoI. Gen. Genet. 204, 383 (1986)) or LBA4404 (Clontech) Agrobacterium tumefaciens strain. Transformation can be performed by standard transformation and regeneration techniques (Deblaere et al., Nucl. Acids Res. 13, 4777 (1994), Gelvin, Stanton B.
- rapeseed can be transformed via cotyledon or hy- pocotyl transformation (Moloney et al., Plant Cell Report 8, 238 (1989); De Block et al., Plant Physiol. 91 , 694 (1989)).
- Agrobacterium and plant selection de- pends on the binary vector and the Agrobacterium strain used for transformation. Rapeseed selection is normally performed using kanamycin as selectable plant marker.
- Agrobacterium mediated gene transfer to flax can be performed using, for example, a technique described by Mlynarova et al., Plant Cell Report 13, 282 (1994). Additionally, transformation of soybean can be performed using for example a technique described in European Patent No. 424 047, U.S. Patent No. 5,322,783, European Patent No. 397 687, U.S. Patent No.
- Transformation of maize can be achieved by particle bombardment, polyethylene glycol mediated DNA uptake or via the silicon carbide fiber technique. (See, for example, Freeling and Walbot "The maize handbook” Springer Verlag: New York (1993) ISBN 3-540-97826-7).
- a specific example of maize transformation is found in U.S. Patent No. 5,990,387, and a specific example of wheat transformation can be found in PCT Application No. WO 93/07256.
- the introduced nucleic acid molecule coding for YIP as depicted in table II, column 5 or 7 may be maintained in the plant cell stably if it is incorporated into a non-chromosomal autonomous replicon or integrated into the plant chromosomes or organelle genome.
- the introduced YIP may be present on an extra-chromosomal non-replicating vector and be transiently expressed or transiently active.
- a homologous recombinant microorganism can be created wherein the YIP is integrated into a chromosome, a vector is prepared which contains at least a portion of a nucleic acid molecule coding for YIP as depicted in table II, col- umn 5 or 7 into which a deletion, addition, or substitution has been introduced to thereby alter, e.g., functionally disrupt, the YIP gene.
- the YIP gene is a yeast, E. coli gene, but it can be a homolog from a related plant or even from a mammalian or insect source.
- the vector can be designed such that, upon homologous recombination, the en- dogenous nucleic acid molecule coding for YIP as depicted in table II, column 5 or 7 is mutated or otherwise altered but still encodes a functional polypeptide (e.g., the upstream regulatory region can be altered to thereby alter the expression of the endogenous YIP).
- a functional polypeptide e.g., the upstream regulatory region can be altered to thereby alter the expression of the endogenous YIP.
- the biological activity of the protein of the invention is increased upon homologous recombination.
- DNA-RNA hybrids can be used in a technique known as chimeraplasty (Cole-Strauss et al., Nucleic Acids Research 27 (5), 1323 (1999) and Kmiec, Gene Therapy American Scientist. 87 (3), 240 (1999)).
- Homologous recombination procedures in Physcomitrella patens are also well known in the art and are contemplated for use herein.
- the altered portion of the nucleic acid molecule coding for YIP as depicted in table II, column 5 or 7 is flanked at its 5' and 3' ends by an additional nucleic acid molecule of the YIP gene to allow for homologous recombination to occur between the exogenous YIP gene carried by the vector and an endogenous YIP gene, in a microorganism or plant.
- the additional flanking YIP nu- cleic acid molecule is of sufficient length for successful homologous recombination with the endogenous gene.
- flanking DNA typically, several hundreds of base pairs up to kilobases of flanking DNA (both at the 5' and 3' ends) are included in the vector. See, e.g., Thomas K.R., and Capecchi M. R., Cell 51 , 503 (1987) for a description of homologous recombination vectors or Strepp et al., PNAS, 95 (8), 4368 (1998) for cDNA based recombination in Physcomi- trella patens.
- the vector is introduced into a microorganism or plant cell (e.g. via polyethylene glycol mediated DNA), and cells in which the introduced YIP gene has homologously recombined with the endogenous YIP gene are selected using art-known techniques.
- nucleic acid molecule coding for YIP as depicted in table II, column 5 or 7 preferably resides in a plant expression cassette.
- a plant expression cassette preferably contains regulatory sequences capable of driving gene expression in plant cells that are operatively linked so that each sequence can fulfill its function, for example, termination of transcription by polyadenylation signals.
- Preferred polyadenylation signals are those originating from Agrobacterium tumefaciens t-DNA such as the gene 3 known as octopine synthase of the Ti-plasmid pTiACH ⁇ (Gielen et al., EMBO J. 3, 835 (1984)) or functional equivalents thereof but also all other terminators functionally active in plants are suitable.
- a plant expression cassette preferably contains other operatively linked se- quences like translational enhancers such as the overdrive-sequence containing the 5 ' - untranslated leader sequence from tobacco mosaic virus enhancing the polypeptide per RNA ratio (GaIMe et al., Nucl.
- plant expres- sion vectors include those detailed in: Becker D. et al., Plant MoI. Biol. 20, 1195 (1992); and Bevan M.W., Nucl. Acid. Res. 12, 871 1 (1984); and "Vectors for Gene Transfer in Higher Plants” in: Transgenic Plants, Vol. 1 , Engineering and Utilization, eds. Kung and Wu R., Academic Press, 1993, S. 15-38.
- Transformation is defined herein as a process for introducing heterologous DNA into a plant cell, plant tissue, or plant. It may occur under natural or artificial conditions using various methods well known in the art. Transformation may rely on any known method for the insertion of foreign nucleic acid sequences into aprokaryotic or eukaryotic host cell. The method is selected based on the host cell being transformed and may include, but is not limited to, viral infection, electroporation, lipofection, and particle bombardment. Such "transformed” cells include stably transformed cells in which the inserted DNA is capable of replication either as an autonomously replicating plasmid or as part of the host chromosome. They also include cells which transiently express the inserted DNA or RNA for limited periods of time. Transformed plant cells, plant tissue, or plants are understood to encompass not only the end product of a transformation process, but also transgenic progeny thereof.
- transformed refers to a host organism such as a bacterium or a plant into which a heterologous nucleic acid molecule has been intro- prised.
- the nucleic acid molecule can be stably integrated into the genome of the host or the nucleic acid molecule can also be present as an extrachromosomal molecule. Such an extrachromosomal molecule can be auto-replicating.
- Transformed cells, tissues, or plants are understood to encompass not only the end product of a transformation process, but also transgenic progeny thereof.
- a “non-transformed”, “non-transgenic” or “non- recombinant” host refers to a wild-type organism, e.g. a bacterium or plant, which does not contain the heterologous nucleic acid molecule.
- transgenic plant refers to a plant which contains a foreign nucleotide sequence inserted into either its nuclear genome or organelle genome. It encompasses further the offspring generations i.e. the T1 -, T2- and consecutively generations or BC1 -, BC2- and consecutively generation as well as crossbreeds thereof with non-transgenic or other transgenic plants.
- transgenic plants are, for example, selected from the families Aceraceae, Anacardiaceae, Apiaceae, Asteraceae, Brassicaceae, Cactaceae, Cucurbitaceae, Euphorbiaceae, Fabaceae, Malvaceae, Nym- phaeaceae, Papaveraceae, Rosaceae, Salicaceae, Solanaceae, Arecaceae, Bromeliaceae, Cyperaceae, Iridaceae, Liliaceae, Orchidaceae, Gentianaceae, Labiaceae, Magnoliaceae, -.D
- Ranunculaceae Carifolaceae, Rubiaceae, Scrophulariaceae, Caryophyllaceae, Ericaceae, Polygonaceae, Violaceae, Juncaceae or Poaceae and preferably from a plant selected from the group of the families Apiaceae, Asteraceae, Brassicaceae, Cucurbitaceae, Fabaceae, Papaveraceae, Rosaceae, Solanaceae, Liliaceae or Poaceae.
- crop plants such as plants advantageously selected from the group of the genus peanut, oilseed rape, canola, sunflower, safflower, olive, sesame, hazelnut, almond, avocado, bay, pumpkin/squash, linseed, soya, pistachio, borage, maize, wheat, rye, oats, sorghum and millet, triticale, rice, barley, cassava, potato, sugarbeet, egg plant, alfalfa, and perennial grasses and forage plants, oil palm, vegetables (brassicas, root vegetables, tuber vegetables, pod vegetables, fruiting vegetables, onion vegetables, leafy vegetables and stem vegetables), buckwheat, Jerusalem artichoke, broad bean, vetches, lentil, dwarf bean, lupin, clover and Lucerne for mentioning only some of them.
- transgenic plants are selected from the group comprising cereals, soybean, rapeseed (including oil seed rape, especially canola and winter oil seed rape), cotton sugarcane and potato, especially corn, soy, rapeseed (including oil seed rape, especially canola and winter oil seed rape), cotton, wheat and rice.
- the transgenic plant is a gymnosperm plant, especially a spruce, pine or fir.
- the host plant is selected from the families Aceraceae, Ana- cardiaceae, Apiaceae, Asteraceae, Brassicaceae, Cactaceae, Cucurbitaceae, Euphor- biaceae, Fabaceae, Malvaceae, Nymphaeaceae, Papaveraceae, Rosaceae, Salicaceae, Solanaceae, Arecaceae, Bromeliaceae, Cyperaceae, Iridaceae, Liliaceae, Orchidaceae, Gentianaceae, Labiaceae, Magnoliaceae, Ranunculaceae, Carifolaceae, Rubiaceae, Scrophulariaceae, Caryophyllaceae, Ericaceae, Polygonaceae, Violaceae, Juncaceae or Poaceae and preferably from a plant selected from the group of the families Apiaceae, Asteraceae, Brassicaceae, Cucurbitaceae, Euphor- bi
- foliosa Brassica nigra, Brassica sinapioides, Melanosinapis communis, Brassica oleracea, Arabidopsis thaliana, Anana comosus, Ananas ananas, Bromelia comosa, Carica papaya, Cannabis sative, lpomoea batatus, lpomoea pandurata, Convolvulus batatas, Convolvulus tiliaceus, lpomoea fas- tigiata, lpomoea tiliacea, lpomoea triloba, Convolvulus panduratus, Beta vulgaris, Beta vulgaris var. altissima, Beta vulgaris var.
- Anacardiaceae such as the genera Pistacia, Mangifera, Anacardium e.g. the species Pis- tacia vera [pistachios, Pistazie], Mangifer indica [Mango] or Anacardium occidentale [Cashew]; Asteraceae such as the genera Calendula, Carthamus, Centaurea, Cichorium, Cynara, Helianthus, Lactuca, Locusta, Tagetes, Valeriana e.g.
- foliosa Brassica nigra, Brassica sinapioides, Melanosinapis communis [mustard], Brassica oleracea [fodder beet] or Arabidopsis thaliana
- Bromeliaceae such as the genera Anana, Bromelia e.g. the species Anana comosus, Ananas ananas or Bromelia comosa [pineapple]
- Caricaceae such as the genera Carica e.g. the species Carica papaya [papaya]
- Cannabaceae such as the genera Cannabis e.g.
- Convolvulaceae such as the genera Ipomea, Convolvulus e.g. the species lpomoea batatus, lpomoea pandurata, Convolvulus batatas, Convolvulus tiliaceus, lpomoea fastigiata, lpomoea tiliacea, lpomoea triloba or Convolvulus panduratus [sweet potato, Man of the Earth, wild potato], Chenopodiaceae such as the genera Beta, i.e. the species Beta vulgaris, Beta vulgaris var. altissima, Beta vulgaris var. Vulgaris, Beta mari- tima, Beta vulgaris var.
- Convolvulaceae such as the genera Ipomea, Convolvulus e.g. the species lpomoea batatus, lpomoea pandurata, Convolvulus batatas, Convolvulus tiliaceus, lpomoea fastigiata,
- Beta vulgaris var. conditiva or Beta vulgaris var. esculenta [sugar beet]; Cucurbitaceae such as the genera Cucubita e.g. the species Cucurbita maxima, Cucurbita mixta, Cucurbita pepo or Cucurbita moschata [pumpkin, squash]; Elaeagnaceae such as the genera Elaeagnus e.g. the species Olea europaea [olive]; Ericaceae such as the genera Kalmia e.g.
- Kalmia latifolia Kalmia angustifolia, KaI- mia microphylla, Kalmia polifolia, Kalmia occidentalis, Cistus chamaerhodendros or Kalmia lucida [American laurel, broad-leafed laurel, calico bush, spoon wood, sheep laurel, alpine laurel, bog laurel, western bog-laurel, swamp-laurel]
- Euphorbiaceae such as the genera Manihot, Janipha, Jatropha, Ricinus e.g.
- Manihot utilissima Janipha manihot,, Jatropha manihot.
- Manihot aipil Manihot dulcis, Manihot manihot, Manihot melanobasis, Manihot esculenta [manihot, arrowroot, tapioca, cassava] or Ricinus communis [castor bean, Castor Oil Bush, Castor Oil Plant, Palma Christi, Wonder Tree];
- Fabaceae such as the genera Pisum, Albizia, Cathormion, Feuillea, Inga, Pithecolobium, Acacia, Mimosa, Medicajo, Glycine, Dolichos, Phaseolus, Soja e.g.
- Linaceae such as the genera Linum, Adenolinum e.g. the species Linum usitatissimum, Linum humile, Linum austriacum, Linum bienne, Linum angustifolium, Linum catharticum, Linum flavum, Linum grandiflorum, Adenolinum grandiflorum, Linum lewisii, Linum narbonense, Linum perenne, Linum perenne var. lewisii, Linum pratense or Linum trigynum [flax, linseed]; Lythrarieae such as the genera Punica e.g.
- Palmae such as the genera Elacis e.g. the species Elaeis guineensis [oil plam]
- Papaveraceae such as the genera Papaver e.g. the species Papaver orientale, Papaver rhoeas, Papaver dubium [poppy, oriental poppy, corn poppy, field poppy, shirley poppies, field poppy, long-headed poppy, long-pod poppy]
- Ped- aliaceae such as the genera Sesamum e.g.
- Piperaceae such as the genera Piper, Artanthe, Peperomia, Steffensia e.g. the species Piper disposecum, Piper amalago, Piper angustifolium, Piper auritum, Piper betel, Piper cu- beba, Piper longum, Piper nigrum, Piper retrofractum, Artanthe disposeca, Artanthe elongata, Peperomia elongata, Piper elongatum, Steffensia elongata.
- Macadamia intergrifolia [macadamia]
- Rubiaceae such as the genera Cof- fea e.g. the species Cofea spp., Coffea arabica, Coffea canephora or Coffea liberica [coffee]
- Scrophulariaceae such as the genera Verbascum e.g.
- Verbascum blat- taria Verbascum chaixii, Verbascum densiflorum, Verbascum lagurus, Verbascum longi- folium, Verbascum lychnitis, Verbascum nigrum, Verbascum olympicum, Verbascum phlo- moides, Verbascum phoenicum, Verbascum pulverulentum or Verbascum thapsus
- mullein white moth mullein, nettle-leaved mullein, dense-flowered mullein, silver mullein, long- leaved mullein, white mullein, dark mullein, greek mullein, orange mullein, purple mullein, hoary mullein, great mullein]
- Solanaceae such as the genera Capsicum, Nicotiana, So- lanum, Lycopersicon e.g.
- nucleic acids according to the invention can in principle be done by all of the methods known to those skilled in the art.
- the introduction of the nucleic acid sequences gives rise to recombinant or transgenic organisms.
- sequence may relate to polynucleotides, nucleic acids, nucleic acid molecules, peptides, polypeptides and proteins, depending on the context in which the term “sequence” is used.
- nucleic acid molecule(s) refers to a polymeric form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. The terms refer only to the primary structure of the molecule.
- the terms "gene(s)”, “polynucleotide”, “nucleic acid sequence”, “nucleotide sequence”, or “nucleic acid molecule(s)” as used herein include double- and single- stranded DNA and RNA. They also include known types of modifications, for example, me- thylation, "caps", substitutions of one or more of the naturally occurring nucleotides with an analog.
- the DNA or RNA sequence of the invention comprises a coding sequence encoding the herein defined polypeptide.
- genes of the invention coding for an activity selected from the group consisting of 3OS ribosomal protein S1 1 , 60S ribosomal protein, Adaptin medium chain ho- molog APM2, B0252-protein, BRICK1-like protein, Cav1 protein, Chloroplast chaperonin, DNA polymerase, flagellar protein, G2/mitotic-specific cyclin, GREI -protein(Hydrophilin), Membrane protein, ORF YPL249c-a, RNA polymerase Il holoenzyme cyclin-like subunit, Serine/threonine-protein kinase, Short chain dehydrogenase, Sterol-C-methyltransferase, transcription regulator (farR), Ykr015c-protein, and YPL167C_2-protein are also called "YIP gene”.
- a "coding sequence” is a nucleotide sequence, which is transcribed into mRNA and/or translated into a polypeptide when placed under the control of appropriate regulatory sequences. The boundaries of the coding sequence are determined by a translation start codon at the 5'-terminus and a translation stop codon at the 3'-terminus.
- a coding sequence can include, but is not limited to mRNA, cDNA, recombinant nucleotide sequences or genomic DNA, while introns may be present as well under certain circumstances.
- transfor- mation The transfer of foreign genes into the genome of a plant is called transfor- mation.
- methods described for the transformation and regeneration of plants from plant tissues or plant cells are utilized for transient or stable transformation. Suitable methods are protoplast transformation by poly(ethylene glycol)-induced DNA uptake, the ,,biolistic" method using the gene cannon - referred to as the particle bombardment method, electroporation, the incubation of dry embryos in DNA solution, microinjection and gene transfer mediated by Agrobacterium. Said methods are described by way of example in Jenes B. et al., Techniques for Gene Transfer, in: Transgenic Plants, Vol. 1 , Engineering and Utilization, eds.. Kung S.
- nucleic acids or the construct to be expressed is preferably cloned into a vector which is suitable for trans- forming Agrobacterium tumefaciens, for example pBini 9 (Bevan et al., Nucl. Acids Res. 12, 871 1 (1984)).
- Agrobacteria transformed by such a vector can then be used in known manner for the transformation of plants, in particular of crop plants such as by way of example tobacco plants, for example by bathing bruised leaves or chopped leaves in an agrobacte- rial solution and then culturing them in suitable media.
- the transformation of plants by means of Agrobacterium tumefaciens is described, for example, by H ⁇ fgen and Willmitzer in Nucl. Acid Res. 16, 9877 (1988) or is known inter alia from White F. F., Vectors for Gene Transfer in Higher Plants; in Transgenic Plants, Vol. 1 , Engineering and Utilization, eds. Kung S. D. and Wu R., Academic Press, 1993, pp. 15-38.
- Agrobacteria transformed by an expression vector according to the invention may likewise be used in known manner for the transformation of plants such as test plants like Arabidopsis or crop plants such as cereal crops, corn, oats, rye, barley, wheat, soybean, rice, cotton, sugar beet, canola, sunflower, flax, hemp, potatoes, tobacco, tomatoes, carrots, paprika, oilseed rape, tapioca, cassava, arrowroot, tagetes, alfalfa, lettuce and the various tree, nut and vine species, in particular oil-containing crop plants such as soybean, peanut, castor oil plant, sunflower, corn, cotton, flax, oilseed rape, coconut, oil palm, safflower (Carthamus tinctorius) or cocoa bean, or in particular corn, wheat, soybean, rice, ⁇ ⁇ -
- cotton and canola e.g. by bathing bruised leaves or chopped leaves in an agrobacterial solution and then culturing them in suitable media.
- the genetically modified plant cells may be regenerated by all of the methods known to those skilled in the art. Appropriate methods can be found in the publications referred to above by Kung S. D. and Wu R., Potrykus or H ⁇ fgen and Willmitzer.
- a further aspect of the invention relates to transgenic organisms transformed by at least one nucleic acid sequence, expression cassette or vector according to the invention as well as cells, cell cultures, tissue, parts - such as, for example, leaves, roots, etc. in the case of plant organisms - or reproductive material derived from such organisms.
- the terms "host organism”, “host cell”, “recombinant (host) organism” and “transgenic (host) cell” are used here interchangeably. Of course these terms relate not only to the particular host organism or the particular target cell but also to the descendants or potential descendants of these organisms or cells. Since, due to mutation or environmental effects certain modifications may arise in successive generations, these descendants need not necessarily be identical with the parental cell but nevertheless are still encompassed by the term as used here.
- (C) (a) and (b); are not found in their natural, genetic environment or have been modified by genetic engineering methods, wherein the modification may by way of example be a substitution, addition, deletion, inversion or insertion of one or more nucleotide residues.
- Natural genetic environment means the natural genomic or chromosomal locus in the organism of origin or inside the host organism or presence in a genomic library. In the case of a genomic library the natural genetic environment of the nucleic acid sequence is preferably retained at least in part. The environment borders the nucleic acid sequence at least on one side and has a sequence length of at least 50 bp, preferably at least 500 bp, particularly preferably at least 1 ,000 bp, most particularly preferably at least 5,000 bp.
- a naturally occurring expression cassette - for example the naturally occurring combination of the natural promoter of the nucleic acid sequence according to the invention with the corresponding gene - turns into a transgenic expression cassette when the latter is modified by unnatural, synthetic ("artificial") methods such as by way of example a mutagenesis.
- Appropriate methods are described by way of example in US 5,565,350 or WO 00/15815.
- Suitable organisms or host organisms for the nucleic acid, expression cassette or vector according to the invention are advantageously in principle all organisms, which are suitable for the expression of recombinant genes as described above.
- plants such as Arabidopsis, Asteraceae such as Calendula or crop plants such as soybean, peanut, castor oil plant, sunflower, flax, corn, cotton, flax, oilseed rape, coconut, oil palm, safflower (Carthamus tinctorius) or cocoa bean.
- host plants for the nucleic acid, expression cassette or vector according to the invention are selected from the group comprising corn, soy, oil seed rape (including canola and winter oil seed rape), cotton, wheat and rice.
- a further object of the invention relates to the use of a nucleic acid construct, e.g. an expression cassette, containing DNA sequences encoding polypeptides shown in table Il or DNA sequences hybridizing therewith for the transformation of plant cells, tissues or parts of plants.
- a nucleic acid construct e.g. an expression cassette, containing DNA sequences encoding polypeptides shown in table Il or DNA sequences hybridizing therewith for the transformation of plant cells, tissues or parts of plants.
- sequences shown in table I can be expressed specifically in the leaves, in the seeds, the nodules, in roots, in the stem or other parts of the plant.
- Those transgenic plants overproducing sequences as depicted in table I, the reproductive material thereof, together with the plant cells, tissues or parts thereof are a further object of the present invention.
- the expression cassette or the nucleic acid sequences or construct according to the inven- tion containing sequences according to table I can, moreover, also be employed for the transformation of the organisms identified by way of example above such as bacteria, yeasts, filamentous fungi and plants.
- enhanced tolerance to abiotic environmental stress and/or yield means, for example, the artificially acquired trait of enhanced tolerance to abiotic environmental stress and/or yield due to functional over expression of polypeptide sequences of table Il encoded by the corresponding nucleic acid molecules as depicted in table I, column 5 or 7 and/or homologs in the organisms according to the invention, advantageously in the transgenic plants according to the invention, by comparison with the nongenetically modified initial plants at least for the duration of at least one plant generation.
- a constitutive expression of the polypeptide sequences of table II, encoded by the corresponding nucleic acid molecule as depicted in table I, column 5 or 7 and/or ho- mologs is, moreover, advantageous. On the other hand, however, an inducible expression may also appear desirable.
- Expression of the polypeptide sequences of the invention can be either direct to the cytoplasm or the organelles, preferably the plastids of the host cells, preferably the plant cells.
- the efficiency of the expression of the sequences of the of table II, encoded by the corre- sponding nucleic acid molecule as depicted in table I, column 5 or 7 and/or homologs can be determined, for example, in vitro by shoot meristem propagation.
- an expression of the sequences of table II, encoded by the corresponding nucleic acid molecule as depicted in table I, column 5 or 7 and/or homologs modified in nature and level and its effect on the metabolic pathways performance can be tested on test plants in greenhouse trials.
- An additional object of the invention comprises transgenic organisms such as transgenic plants transformed by an expression cassette containing sequences of as depicted in table I, column 5 or 7 according to the invention or DNA sequences hybridizing therewith, as well as transgenic cells, tissue, parts and reproduction material of such plants.
- transgenic crop plants such as by way of example barley, wheat, rye, oats, corn, soybean, rice, cotton, sugar beet, oilseed rape and canola, sunflower, flax, hemp, thistle, potatoes, tobacco, tomatoes, tapioca, cassava, arrowroot, alfalfa, lettuce and the various tree, nut and vine species.
- transgenic plants transformed by an expression cas- sette containing sequences of as depicted in table I, column 5 or 7 according to the invention or DNA sequences hybridizing therewith are selected from the group comprising corn, soy, oil seed rape (including canola and winter oil seed rape), cotton, wheat and rice.
- plants are mono- and dicotyledonous plants, mosses or algae, especially plants, for example in one embodiment monocotyledon- ous plants, or for example in another embodiment dicotyledonous plants.
- a further refinement according to the invention are transgenic plants as described above which contain a nucleic acid sequence or construct according to the invention or a expression cassette according to the invention.
- transgenic also means that the nucleic acids according to the invention are located at their natural position in the genome of an organism, but that the sequence, e.g. the coding sequence or a regulatory sequence, for example the promoter sequence, has been modified in comparison with the natural sequence.
- trans- genic/recombinant is to be understood as meaning the transcription of one or more nucleic acids or molecules of the invention and being shown in table I, occurs at a non-natural position in the genome.
- the expression of the nucleic acids or molecules is homologous.
- the expression of the nucleic acids or molecules is heterologous. This expression can be transiently or of a sequence integrated stably into the genome. oo
- transgenic plants used in accordance with the invention also refers to the progeny of a transgenic plant, for example the Ti, T 2 , T 3 and subsequent plant generations or the BC-I, BC2, BC3 and subsequent plant generations.
- the transgenic plants according to the invention can be raised and selfed or crossed with other individuals in order to obtain further transgenic plants according to the invention.
- Transgenic plants may also be obtained by propagating transgenic plant cells vegetatively.
- the present invention also relates to transgenic plant material, which can be derived from a transgenic plant population according to the invention.
- Such material includes plant cells and certain tissues, organs and parts of plants in all their manifestations, such as seeds, leaves, anthers, fibers, tubers, roots, root hairs, stems, embryo, calli, cotyledons, petioles, harvested material, plant tissue, reproductive tissue and cell cultures, which are derived from the actual transgenic plant and/or can be used for bringing about the transgenic plant.
- Any transformed plant obtained according to the invention can be used in a conventional breeding scheme or in vitro plant propagation to produce more transformed plants with the same characteristics and/or can be used to introduce the same characteristic in other varieties of the same or related species. Such plants are also part of the invention. Seeds obtained from the transformed plants genetically also contain the same characteristic and are part of the invention. As mentioned before, the present invention is in principle applicable to any plant and crop that can be transformed with any of the transformation method known to those skilled in the art.
- Advantageous inducible plant promoters are by way of example the PRP1 promoter (Ward et al., Plant.Mol. Biol. 22361 (1993)), a promoter inducible by benzenesul- fonamide (EP 388 186), a promoter inducible by tetracycline (Gatz et al., Plant J. 2, 397 (1992)), a promoter inducible by salicylic acid (WO 95/19443), a promoter inducible by ab- scisic acid (EP 335 528) and a promoter inducible by ethanol or cyclohexanone (WO 93/21334).
- PRP1 promoter Ward et al., Plant.Mol. Biol. 22361 (1993)
- a promoter inducible by benzenesul- fonamide EP 388 186
- a promoter inducible by tetracycline Gaatz et al., Plant J
- plant promoters which can advantageously be used are the promoter of cytoplasmic FBPase from potato, the ST-LSI promoter from potato (Stockhaus et al., EMBO J. 8, 2445 (1989)), the promoter of phosphoribosyl pyrophosphate amidotrans- ferase from Glycine max (see also gene bank accession number U87999) or a nodiene- specific promoter as described in EP 249 676.
- promoters which ensure expression upon onset of abiotic stress conditions Particular advantageous are those promoters which ensure expression upon onset of low temperature conditions, e.g. at the onset of chilling and/or freez- ing temperatures as defined hereinabove, e.g. for the expression of nucleic acid molecules as shown in table VIIIb.
- promoters which ensure expression upon conditions of limited nutrient availability e.g. the onset of limited nitrogen sources in case the nitrogen of the soil or nutrient is exhausted, e.g. for the expression of the nucleic acid molecules or their gene products as shown in table Villa.
- promoters which ensure expression upon onset of water deficiency as defined herein- above, e.g. for the expression of the nucleic acid molecules or their gene products as o
- promoters which ensure expression upon onset of standard growth conditions, e.g. under condition without stress and deficient nutrient provision, e.g. for the expression of the nucleic acid molecules or their gene products as shown in table VIIId.
- Such promoters are known to the person skilled in the art or can be isolated from genes which are induced under the conditions mentioned above.
- seed-specific promoters may be used for monocotyledonous or dicotyledonous plants.
- DNA fragments can be manipulated in order to obtain a nucleotide sequence, which usefully reads in the correct direction and is equipped with a correct reading frame.
- DNA fragments nucleic acids according to the invention
- adaptors or linkers may be attached to the fragments.
- the promoter and the terminator regions can usefully be provided in the transcription direc- tion with a linker or polylinker containing one or more restriction points for the insertion of this sequence.
- the linker has 1 to 10, mostly 1 to 8, preferably 2 to 6, restriction points.
- the size of the linker inside the regulatory region is less than 100 bp, frequently less than 60 bp, but at least 5 bp.
- the promoter may be both native or homologous as well as foreign or heterologous to the host organism, for example to the host plant.
- the expression cassette contains the promoter, a DNA sequence which shown in table I and a region for transcription termination. Different termination regions can be exchanged for one another in any desired fashion.
- nucleic acid and “nucleic acid molecule” are intended to include DNA molecules (e.g. cDNA or genomic DNA) and RNA molecules (e.g. mRNA) and analogs of the DNA or RNA generated using nucleotide analogs. This term also encompasses untranslated sequence located at both the 3' and 5' ends of the coding region of the gene - at least about 1000 nucleotides of sequence upstream from the 5' end of the coding region and at least about 200 nucleotides of sequence downstream from the 3' end of the coding region of the gene.
- the nucleic acid molecule can be single- stranded or double-stranded, but preferably is double-stranded DNA.
- nucleic acid molecule is one that is substantially separated from other nucleic acid molecules, which are present in the natural source of the nucleic acid. That means other nucleic acid molecules are present in an amount less than 5% based on weight of the amount of the desired nucleic acid, preferably less than 2% by weight, more preferably less ⁇ I
- an "isolated" nucleic acid is free of some of the sequences that naturally flank the nucleic acid (i.e., sequences located at the 5' and 3' ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived.
- the iso- lated Yield Increase Protein (YIP) encoding nucleic acid molecule can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of nucleotide sequences which naturally flank the nucleic acid molecule in genomic DNA of the cell from which the nucleic acid is derived.
- an "isolated" nucleic acid molecule such as a cDNA molecule, can be free from some of the other cellular material with which it is naturally associated, or culture medium when produced by recombinant techniques, or chemical precursors or other chemicals when chemically synthesized.
- a nucleic acid molecule of the present invention e.g., a nucleic acid molecule encoding an YIP or a portion thereof which confers enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass in plants, can be isolated using standard molecular biological techniques and the sequence information provided herein.
- an A. thaliana YIP encoding cDNA can be isolated from a A.
- thaliana c-DNA library or a Synechocystis sp., Brassica napus, Glycine max, Zea mays or Oryza sativa YIP encoding cDNA can be isolated from a Synechocystis sp., Brassica napus, Glycine max, Zea mays or Oryza sativa c-DNA library respectively using all or portion of one of the sequences shown in table I.
- a nucleic acid molecule encompassing all or a portion of one of the sequences of table I can be isolated by the polymerase chain reaction using oligonucleotide primers designed based upon this sequence.
- mRNA can be isolated from plant cells (e.g., by the gua- nidinium-thiocyanate extraction procedure of Chirgwin et al., Biochemistry 18, 5294 (1979)) and cDNA can be prepared using reverse transcriptase (e.g., Moloney MLV reverse transcriptase, available from Gibco/BRL, Bethesda, MD; or AMV reverse transcriptase, available from Seikagaku America, Inc., St. Russia, FL).
- reverse transcriptase e.g., Moloney MLV reverse transcriptase, available from Gibco/BRL, Bethesda, MD; or AMV reverse transcriptase, available from Seikagaku America, Inc., St. Russia, FL.
- Synthetic oligonucleotide primers for polymerase chain reaction amplification can be designed based upon one of the nucleo- tide sequences shown in table I.
- a nucleic acid molecule of the invention can be amplified using cDNA or, alternatively, genomic DNA, as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques.
- the nucleic acid molecule so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis.
- oligonucleotides corresponding to a YIP encoding nucleotide se- quence can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer.
- an isolated nucleic acid molecule of the invention comprises one of the nucleotide sequences shown in table I encoding the YIP (i.e., the "coding region"), as well as 5' untranslated sequences and 3' untranslated sequences.
- the nucleic acid molecule of the invention can comprise only a portion of the coding region of one of the sequences of the nucleic acid of table I, for example, a fragment oo
- portions of proteins encoded by the YIP encoding nucleic acid molecules of the invention are preferably biologically active portions described herein.
- biologically active portion of a YIP is intended to include a portion, e.g. a domain/motif, of Yield Increase Protein that participates in an enhanced tolerance to abiotic stress and/or increased yield in a plant.
- an analysis of a plant comprising the YIP may be performed. Such analysis methods are well known to those skilled in the art, as detailed in the Examples.
- nucleic acid fragments encoding biologically active portions of a YIP can be prepared by isolating a portion of one of the sequences of the nucleic acid of table I expressing the encoded portion of the YIP or peptide (e.g., by recombinant expression in vitro) and assessing the activity of the encoded portion of the YIP or peptide.
- Biologically active portions of a YIP are encompassed by the present invention and include peptides comprising amino acid sequences derived from the amino acid sequence of a YIP encoding gene, or the amino acid sequence of a protein homologous to a YIP, which include fewer amino acids than a full length YIP or the full length protein which is homologous to a YIP, and exhibits at least some enzymatic or biological activity of a YIP.
- biologically active portions e.g., peptides which are, for example, 5, 10, 15, 20, 30, 35, 36, 37, 38, 39, 40, 50, 100 or more amino acids in length
- biologically active portions comprise a domain or motif with at least one activity of a YIP.
- biologically active portions in which other regions of the protein are deleted can be prepared by recombinant techniques and evaluated for one or more of the activities described herein.
- the biologically active portions of a YIP include one or more selected domains/motifs or portions thereof having biological activity.
- biological active portion or “biological activity” means a polypeptide as depicted in table II, column 3 or a portion of said polypeptide which still has at least 10 % or 20 %, preferably 30 %, 40 %, 50 % or 60 %, especially preferably 70 %, 75 %, 80 %, 90 % or 95 % of the enzymatic or biological activity of the natural or starting enzyme or protein.
- nucleic acid sequences can be used, which, if appropriate, contain synthetic, non-natural or modified nucleotide bases, which can be incorporated into DNA or RNA.
- Said synthetic, non-natural or modified bases can for example increase the stability of the nucleic acid molecule outside or inside a cell.
- the nucleic acid molecules of the invention can contain the same modifications as aforementioned.
- nucleic acid molecule may also encompass the untranslated sequence located at the 3' and at the 5' end of the coding o
- gene region for example at least 500, preferably 200, especially preferably 100, nucleotides of the sequence upstream of the 5' end of the coding region and at least 100, preferably 50, especially preferably 20, nucleotides of the sequence downstream of the 3' end of the coding gene region. It is often advantageous only to choose the coding region for clon- ing and expression purposes.
- the nucleic acid molecule used in the process according to the invention or the nucleic acid molecule of the invention is an isolated nucleic acid molecule.
- An "isolated" polynucleotide or nucleic acid molecule is separated from other polynucleotides or nucleic acid molecules, which are present in the natural source of the nucleic acid molecule.
- An isolated nucleic acid molecule may be a chromosomal fragment of several kb, or preferably, a molecule only comprising the coding region of the gene.
- an isolated nucleic acid molecule of the invention may comprise chromosomal regions, which are adjacent 5' and 3' or further adjacent chromosomal regions, but preferably comprises no such sequences which naturally flank the nucleic acid molecule sequence in the genomic or chromosomal context in the organism from which the nucleic acid molecule originates (for example sequences which are adjacent to the regions encoding the 5'- and 3'-UTRs of the nucleic acid molecule).
- the isolated nucleic acid molecule used in the process according to the invention may, for example comprise less than approximately 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb nucleotide sequences which naturally flank the nucleic acid molecule in the genomic DNA of the cell from which the nucleic acid molecule originates.
- nucleic acid molecules used in the process for example the polynucleotide of the invention or of a part thereof can be isolated using molecular-biological standard techniques and the sequence information provided herein. Also, for example a homologous sequence or homologous, conserved sequence regions at the DNA or amino acid level can be identified with the aid of comparison algorithms.
- the former can be used as hybridization probes under standard hybridization techniques (for example those described in Sambrook et al., Molecular Cloning: A Laboratory Manual. 2nd Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989) for isolating further nucleic acid sequences useful in this process.
- a nucleic acid molecule encompassing a complete sequence of the nucleic acid molecules used in the process, for example the polynucleotide of the invention, or a part thereof may additionally be isolated by polymerase chain reaction, oligonucleotide primers based on this sequence or on parts thereof being used.
- a nucleic acid molecule comprising the complete sequence or part thereof can be isolated by polymerase chain reaction using oligonucleotide primers which have been generated on the basis of this very sequence.
- mRNA can be isolated from cells (for example by means of the guanidinium thiocyanate extraction method of Chirgwin et al., Biochemistry 18, 5294(1979)) and cDNA can be generated by means of reverse transcriptase (for example Moloney MLV reverse transcriptase, available from Gibco/BRL, Bethesda, MD, or AMV reverse transcriptase, obtainable from Seikagaku America, Inc., St. Russia, FL).
- reverse transcriptase for example Moloney MLV reverse transcriptase, available from Gibco/BRL, Bethesda, MD, or AMV reverse transcriptase, obtainable from Seikagaku America, Inc., St. Russia, FL.
- Synthetic oligonucleotide primers for the amplification e.g. as shown in table III, column 7, by means of polymerase chain reaction can be generated on the basis of a sequence shown herein, for example the sequence shown in table I, columns 5 and 7 or the sequences derived from table II, columns 5 and 7.
- the activity of a polypeptide is increased comprising or consisting of a consensus sequence or a polypep- tide motif shown in table IV, column 7 and in one another embodiment, the present invention relates to a polypeptide comprising or consisting of a consensus sequence or a polypeptide motif shown in table IV, column 7 whereby less than 20, preferably less than 15 or 10, preferably less than 9, 8, 7, or 6, more preferred less than 5 or 4, even more preferred less then 3, even more preferred less then 2, even more preferred 0 of the amino acids po- sitions indicated can be replaced by any amino acid. In one embodiment not more than
- amino acid position indicated by a letter are/is replaced another amino acid.
- less than 20, preferably less than 15 or 10, preferably less than 9, 8, 7, or 6, more preferred less than 5 or 4, even more preferred less than 3, even more preferred less than 2, even more preferred 0 amino acids are inserted into a consensus sequence or protein motif.
- the consensus sequence was derived from a multiple alignment of the sequences as listed in table II.
- the letters represent the one letter amino acid code and indicate that the amino acids are conserved in at least 80% of the aligned proteins.
- the letter X stands for amino acids, which are not conserved in at least 80% of the sequences. In one example, in the cases where only a small selected subset of amino acids are possible at a certain position these amino acids are given in brackets.
- the number of given X indicates the distances between conserved amino acid residues, e.g. Y-x(21 ,23)-F means that conserved tyrosine and phenylalanine residues are separated from each other by minimum 21 and maximum 23 amino acid residues in all investigated sequences.
- conserved domains were identified from all sequences and are described using a subset of the standard Prosite notation, e.g. the pattern Y-x(21 ,23)-[FW] means that a conserved tyrosine is separated by minimum 21 and maximum 23 amino acid residues from either a phenylalanine or tryptophane. conserveed patterns were identified with the software tool MEME version 3.5.1 or manually.
- MEME was developed by Timothy L. Bailey and Charles Elkan, Dept. of Computer Science and Engeneering, University of California, San Diego, USA and is described by Timothy L. Bailey and Charles Elkan (Fitting a mixture model by expectation maximization to discover motifs in biopolymers, Proceedings of the Second International Conference on Intelligent Systems for Molecular Biology, pp. 28-36, AAAI Press, Menlo Park, California, 1994).
- the source code for the stand-alone program is public available from the San Diego Supercomputer center (http://me.sdsc.edu).
- Pratt Prosite patterns for conserved domains were generated with the software tool Pratt version 2.1 or manually.
- Pratt was developed by Inge Jonassen, Dept. of Informatics, University of Bergen, Norway and is described by Jonassen et al. (I. Jonassen, J.F.Collins and
- the Prosite patterns of the conserved domains can be used to search for protein sequences matching this pattern.
- Various established Bioinformatics centres provide public internet portals for using those patterns in database searches (e.g. PIR (Protein Information Resource, located at Georgetown University Medical Center) or ExPASy (Expert Protein Analysis System)).
- PIR Protein Information Resource, located at Georgetown University Medical Center
- ExPASy Expert Protein Analysis System
- stand-alone software is available, like the program Fuzz- pro, which is part of the EMBOSS software package.
- the program Fuzzpro not only allows to search for an exact pattern-protein match but also allows to set various ambiguities in the performed search.
- the alignment was performed with the software ClustalW (version 1.83) and is described by Thompson et al. (Nucleic Acids Research 22, 4673 (1994)).
- the source code for the standalone program is public available from the European Molecular Biology Laboratory; Heidelberg, Germany.
- the analysis was performed using the default parameters of ClustalW v1.83 (gap open penalty: 10.0; gap extension penalty: 0.2; protein matrix: Gonnet; pro- tein/DNA endgap: -1 ; protein/DNA gapdist: 4).
- Degenerated primers can then be utilized by PCR for the amplification of fragments of novel proteins having above-mentioned activity, e.g. conferring the enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non- transformed, wild type plant cell, plant or part thereof after increasing the expression or activity or having the activity of a protein as shown in table II, column 3 or further functional homologs of the polypeptide of the invention from other organisms.
- a nucleic acid molecule according to the invention can be amplified using cDNA or, as an alternative, genomic DNA as template and suitable oligonucleotide primers, following standard PCR amplification techniques.
- the nucleic acid molecule amplified thus can be cloned into a suitable vector and characterized by means of DNA sequence analysis.
- Oligonucleotides, which correspond to one of the nucleic acid molecules used in the process can be generated by standard synthesis methods, for example using an automatic DNA synthesizer.
- nucleic acid molecules which are advantageously for the process accord- ing to the invention can be isolated based on their homology to the nucleic acid molecules disclosed herein using the sequences or part thereof as hybridization probe and following standard hybridization techniques under stringent hybridization conditions.
- nucleic acid molecules of at least 15, 20, 25, 30, 35, 40, 50, 60 or more nucleotides, preferably of at least 15, 20 or 25 nucleotides in length which hybridize under stringent conditions with the above-described nucleic acid molecules, in particular with those which encompass a nucleotide sequence of the nucleic acid mole- cule used in the process of the invention or encoding a protein used in the invention or of the nucleic acid molecule of the invention. Nucleic acid molecules with 30, 50, 100, 250 or more nucleotides may also be used.
- the term "homology” means that the respective nucleic acid molecules or encoded proteins are functionally and/or structurally equivalent.
- the nucleic acid molecules that are homologous to the nucleic acid molecules described above and that are derivatives of said nucleic acid molecules are, for example, variations of said nucleic acid molecules which represent modifications having the same biological function, in particular encoding proteins with the same or substantially the same biological function. They may be naturally occurring variations, such as sequences from other plant varieties or species, or mutations. These mutations may occur naturally or may be obtained by mutagenesis techniques.
- the allelic variations may be naturally occurring allelic variants as well as synthetically produced or genetically engineered variants. Structurally equivalents can, for example, be identified by testing the binding of said polypeptide to antibodies or computer based predictions. Structurally equivalent have the similar immunological characteristic, e.g. comprise similar epitopes.
- hybridizing it is meant that such nucleic acid molecules hybridize un- der conventional hybridization conditions, preferably under stringent conditions such as described by, e.g., Sambrook (Molecular Cloning; A Laboratory Manual, 2 nd Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989)) or in Current Protocols in Molecular Biology, John Wiley & Sons, N. Y. (1989), 6.3.1 -6.3.6.
- DNA as well as RNA molecules of the nucleic acid of the invention can be used as probes. Further, as template for the identification of functional homologues Northern blot assays as well as Southern blot assays can be performed.
- the Northern blot assay advantageously provides further information about the expressed gene product: e.g. expression pattern, occurrence of processing steps, like splicing and capping, etc.
- the Southern blot assay provides additional information about the chromosomal localization and organization of the gene encoding the nucleic acid molecule of the invention.
- SSC 6x sodium chloride/sodium citrate
- 0.1 % SDS at 50 to 65 0 C, for example at 5O 0 C, 55 0 C or 6O 0 C.
- these hybridization conditions differ as a function of the type of the nucleic acid and, for example when organic solvents are present, with regard to the temperature and concentration of the buffer.
- the temperature under "standard hybridization conditions” differs for example as a function of the type of the nucleic acid between 42 0 C and 58 0 C, preferably between 45 0 C and 5O 0 C in an aqueous buffer with a concentration of 0.1 x, 0.5 x, 1 x, 2 x, 3 x, 4 x or 5 x SSC (pH 7.2). If organic solvent(s) is/are present in the abovementioned buffer, for example 50% formamide, the temperature under standard conditions is approximately 4O 0 C, 42 0 C or 45 0 C.
- the hybridization conditions for DNA:DNA hybrids are preferably for example 0.1 x SSC and 2O 0 C, 25 0 C, 3O 0 C, 35 0 C, 4O 0 C or 45 0 C, preferably between 3O 0 C and 45 0 C.
- the hybridization conditions for DNA:RNA hybrids are preferably for example 0.1 x SSC and 3O 0 C, 35 0 C, 4O 0 C, 45 0 C, 5O 0 C or 55 0 C, preferably between 45 0 C and 55 0 C.
- a further example of one such stringent hybridization condition is hybridization at 4 x SSC at 65 0 C, followed by a washing in 0.1 x SSC at 65 0 C for one hour.
- an exemplary stringent hybridization condition is in 50 % formamide, 4 x SSC at 42 0 C.
- the conditions during the wash step can be selected from the range of condi- tions delimited by low-stringency conditions (approximately 2 x SSC at 5O 0 C) and high- stringency conditions (approximately 0.2 x SSC at 5O 0 C, preferably at 65 0 C) (20 x SSC : 0.3 M sodium citrate, 3 M NaCI, pH 7.0).
- the temperature during the wash step can be raised from low-stringency conditions at room temperature, approximately 22 0 C, to higher-stringency conditions at approximately 65 0 C.
- Both of the parameters salt concentra- tion and temperature can be varied simultaneously, or else one of the two parameters can be kept constant while only the other is varied.
- Denaturants for example formamide or SDS, may also be employed during the hybridization. In the presence of 50% formamide, hybridization is preferably effected at 42 0 C. Relevant factors like 1 ) length of treatment, 2) salt conditions, 3) detergent conditions, 4) competitor DNA, 5) temperature and 6) probe selection can be combined case by case so that not all possibilities can be mentioned herein.
- Northern blots are prehybridized with Rothi-Hybri-Quick buffer (Roth, Düsseldorf) at 68 0 C for 2h.
- Hybridzation with radioactive labelled probe is done overnight at 68 0 C. Subsequent washing steps are performed at 68 0 C with 1 x SSC.
- the membrane is prehybridized with Rothi-Hybri-Quick buffer (Roth, Düsseldorf) at 68 0 C for 2h.
- the hybridzation with radioactive labelled probe is conducted over night at 68 0 C. Subsequently the hybridization buffer is discarded and the filter shortly washed using 2 x SSC; 0,1 % SDS.
- Hybridization conditions can be selected, for example, from the following conditions: (a) 4 x SSC at 65 0 C,
- Wash steps can be selected, for example, from the following conditions: (a) 0.015 M NaCI/0.0015 M sodium citrate/0.1 % SDS at 5O 0 C. (b) 0.1 x SSC at 65 0 C.
- Polypeptides having above-mentioned activity i.e. conferring increased yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non- transformed, wild type plant cell, plant or part thereof, derived from other organisms, can be encoded by other DNA sequences which hybridize to the sequences shown in table I, columns 5 and 7 under relaxed hybridization conditions and which code on expression for peptides conferring the enhanced cold tolerance, and/or increased yield, as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof.
- a further example of such low-stringent hybridization conditions is 4 x SSC at 5O 0 C or hybridization with 30 to 40% formamide at 42 0 C.
- Such molecules comprise those which are fragments, analogues or derivatives of the polypeptide of the invention or used in the process of the invention and differ, for example, by way of amino acid and/or nucleotide deletion(s), insertion(s), substitution (s), addition(s) and/or recombination (s) or any other modification(s) known in the art either alone or in combination from the above-described amino acid sequences or their underlying nucleotide sequence(s). However, it is preferred to use high stringency hybridization conditions.
- Hybridization should advantageously be carried out with fragments of at least 5, 10, 15, 20, 25, 30, 35 or 40 bp, advantageously at least 50, 60, 70 or 80 bp, preferably at least 90, 100 or 110 bp. Most preferably are fragments of at least 15, 20, 25 or 30 bp. Preferably are also hybridizations with at least 100 bp or 200, very especially preferably at least 400 bp in length. In an especially preferred embodiment, the hybridization should be carried out with the entire nucleic acid sequence with conditions described above.
- fragment means a truncated sequence of the original sequence referred to.
- the truncated sequence can vary widely in length; the minimum size being a se- quence of sufficient size to provide a sequence with at least a comparable function and/or activity of the original sequence referred to or hybidizing with the nucleic acid molecule of the invention or used in the process of the invention under stringend conditions, while the maximum size is not critical. In some applications, the maximum size usually is not substantially greater than that required to provide the desired activity and/or function(s) of the origi- nal sequence.
- the truncated amino acid sequence will range from about 5 to about 310 amino acids in length. More typically, however, the sequence will be a maximum of about 250 amino acids in length, preferably a maximum of about 200 or 100 amino acids. It is usually desirable to select sequences of at least about 10, 12 or 15 amino acids, up to a maximum of about 20 or 25 amino acids.
- epitope relates to specific immunoreactive sites within an antigen, also known as antigenic determinates. These epitopes can be a linear array of mono- mers in a polymeric composition - such as amino acids in a protein - or consist of or comprise a more complex secondary or tertiary structure.
- im- munogens i.e., substances capable of eliciting an immune response
- some antigen, such as haptens are not immunogens but may be made immunogenic by coupling to a carrier molecule.
- antigen includes references to a substance to which an antibody can be generated and/or to which the antibody is specifically immunoreactive.
- the present invention relates to a epitope of the polypeptide of the present invention or used in the process of the present invention and confers an enhanced tolerance to abiotic environmental stress and/or an increased yield as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof.
- amino acids relates to at least one amino acid but not more than that number of amino acids, which would result in a homology of below 50% identity.
- identity is more than 70% or 80%, more preferred are 85%, 90%, 91 %, 92%, 93%, 94% or 95%, even more preferred are 96%, 97%, 98%, or 99% iden- tity.
- the nucleic acid molecule of the invention comprises a nucleic acid molecule, which is a complement of one of the nucleotide sequences of above mentioned nucleic acid molecules or a portion thereof.
- a nucleic acid molecule which is complemen- tary to one of the nucleotide sequences shown in table I, columns 5 and 7 is one which is sufficiently complementary to one of the nucleotide sequences shown in table I, columns 5 and 7 such that it can hybridize to one of the nucleotide sequences shown in table I, columns 5 and 7, thereby forming a stable duplex.
- the hybridization is performed under stringent hybrization conditions.
- a complement of one of the herein dis- closed sequences is preferably a sequence complement thereto according to the base pairing of nucleic acid molecules well known to the skilled person.
- the bases A and G undergo base pairing with the bases T and U or C, resp. and visa versa. Modifications of the bases can influence the base-pairing partner.
- the nucleic acid molecule of the invention comprises a nucleotide sequence which is at least about 30%, 35%, 40% or 45%, preferably at least about 50%, 55%, 60% or 65%, more preferably at least about 70%, 80%, or 90%, and even more preferably at least about 95%, 97%, 98%, 99% or more homologous to a nucleotide sequence shown in table I, columns 5 and 7, or a portion thereof and preferably has above mentioned activ- ity, in particular having a tolerance to abiotic environmental stress enhancing activity and/or yield increasing activity after increasing the acitivity or an activity of a gene product as shown in table II, column 3 by for example expression either in the cytsol or in an organelle such as a plastid or mitochondria or both, preferably in plastids.
- the nucleic acid molecule of the invention comprises a nucleotide sequence which hybridizes, preferably hybridizes under stringent conditions as defined herein, to one of the nucleotide sequences shown in table I, columns 5 and 7, or a portion thereof and encodes a protein having above-mentioned activity, e.g. conferring an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, corresponding, e.g.
- nucleic acid molecule of the invention can comprise only a portion of the coding region of one of the sequences shown in table I, columns 5 and 7, for example a fragment which can be used as a probe or primer or a fragment encoding a biologically active portion of the polypeptide of the present invention or of a polypeptide used in the process of the present invention, i.e. having above-mentioned activity, e.g. conferring an enhanced tolerance to abiotic environmental stress and/or yield as compared to a corre- sponding, e.g.
- non-transformed, wild type plant cell, plant or part thereof f its activity is increased by for example expression either in the cytsol or in an organelle such as a plastid or mitochondria or both, preferably in plastids.
- the nucleotide sequences determined from the cloning of the present protein-according-to-the-invention-encoding gene allows for the generation of probes and primers designed for use in identifying and/or cloning its homo- logues in other cell types and organisms.
- the probe/primer typically comprises substantially purified oligonucleotide.
- the oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, 15 preferably about 20 or 25, more preferably about 40, 50 or 75 consecutive nucleotides of a sense strand of one of the sequences set forth, e.g., in table I, columns 5 and 7, an anti-sense sequence of one of the sequences, e.g., set forth in table I, columns 5 and 7, or naturally occurring mutants thereof.
- Primers based on a nucleotide of invention can be used in PCR reactions to clone homologues of the polypeptide of the invention or of the polypeptide used in the process of the invention, e.g. as the primers described in the examples of the present invention, e.g. as shown in the examples.
- a PCR with the primers shown in table III, column 7 will re- suit in a fragment of the gene product as shown in table II, column 3.
- Primer sets are interchangable.
- the person skilled in the art knows to combine said primers to result in the desired product, e.g. in a full length clone or a partial sequence.
- Probes based on the sequences of the nucleic acid molecule of the invention or used in the process of the present invention can be used to detect transcripts or genomic sequences encoding the same or homologous proteins.
- the probe can further comprise a label group attached thereto, e.g. the label group can be a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor.
- Such probes can be used as a part of a genomic marker test kit for identifying cells which express an polypepetide of the invention or used in the process of the present invention, such as by measuring a level of an encoding nucleic acid molecule in a sample of cells, e.g., detecting mRNA levels or determining, whether a genomic gene comprising the sequence of the polynucleotide of the invention or used in the processs of the present invention has been mutated or deleted.
- the nucleic acid molecule of the invention encodes a polypeptide or por- tion thereof which includes an amino acid sequence which is sufficiently homologous to the amino acid sequence shown in table II, columns 5 and 7 such that the protein or portion thereof maintains the ability to participate in the enhancement of tolerance to abiotic environmental stress and/or increase of yield as compared to a corresponding, e.g. non- transformed, wild type plant cell, plant or part thereof, in particular increasing the activity as mentioned above or as described in the examples in plants is comprised.
- the language "sufficiently homologous” refers to proteins or portions thereof which have amino acid sequences which include a minimum number of identical or equivalent amino acid residues (e.g., an amino acid residue which has a similar side chain as an amino acid residue in one of the sequences of the polypeptide of the present invention) to an amino acid sequence shown in table II, columns 5 and 7 such that the protein or portion thereof is able to participate in the enhanced tolerance to abiotic environmental stress and/or increase of yield as compared to a corresponding, e.g. non- transformed, wild type plant cell, plant or part thereof.
- a corresponding, e.g. non- transformed, wild type plant cell, plant or part thereof For examples having the activity of a protein as shown in table II, column 3 and as described herein.
- the nucleic acid molecule of the present invention comprises a nucleic acid that encodes a portion of the protein of the present invention.
- the protein is at least about 30%, 35%, 40%, 45% or 50%, preferably at least about 55%, 60%, 65% or 70%, and more preferably at least about 75%, 80%, 85%, 90%, 91 %, 92%, 93% or 94% and most preferably at least about 95%, 97%, 98%, 99% or more homologous to an entire amino acid sequence of table II, columns 5 and 7 and having above-mentioned activity, e.g. conferring an enhanced yield, e.g an improved yield-related trait, e.g.
- Portions of proteins encoded by the nucleic acid molecule of the invention are preferably biologically active, preferably having above-mentioned annotated activity, e.g. conferring an enhanced tolerance to abiotic environmental stress and/or increase in yield as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof after increase of activity.
- biologically active portion is intended to include a portion, e.g., a domain/motif, that confers an enhanced tolerance to abiotic environmental stress and/or increase in yield as compared to a corresponding, e.g. non- transformed, wild type plant cell, plant or part thereof or has an immunological activity such that it is binds to an antibody binding specifially to the polypeptide of the present invention or a polypeptide used in the process of the present invention for an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof.
- the invention further relates to nucleic acid molecules that differ from one of the nucleotide sequences shown in table I A, columns 5 and 7 (and portions thereof) due to degeneracy of the genetic code and thus encode a polypeptide of the present invention, in particular a polypeptide having above mentioned activity, e.g. as that polypeptides depicted by the sequence shown in table II, columns 5 and 7 or the functional homologues.
- the nucleic acid molecule of the invention comprises, or in an other embodiment has, a nucleotide sequence encoding a protein comprising, or in an other em- bodiment having, an amino acid sequence shown in table II, columns 5 and 7 or the functional homologues.
- the nucleic acid molecule of the invention encodes a full length protein which is substantially homologous to an amino acid sequence shown in table II, columns 5 and 7 or the functional homologues.
- the nucleic acid molecule of the present invention does not consist of the se- quence shown in table I, preferably table IA, columns 5 and 7.
- DNA sequence polymorphisms that lead to changes in the amino acid sequences may exist within a population.
- Such genetic polymorphism in the gene encoding the polypeptide of the in- vention or comprising the nucleic acid molecule of the invention may exist among individuals within a population due to natural variation.
- the terms “gene” and “recombinant gene” refer to nucleic acid molecules comprising an open reading frame encoding the polypeptide of the invention or comprising the nucleic acid molecule of the invention or encoding the polypeptide used in the process of the present invention, preferably from a crop plant or from a microorgansim useful for the method of the invention. Such natural variations can typically result in 1 to 5% variance in the nucleotide sequence of the gene.
- nucleic acid molecules corresponding to natural variants homologues of a nucleic acid molecule of the invention can be isolated based on their homology to the nucleic acid molecules disclosed herein using the nucleic acid mole- cule of the invention, or a portion thereof, as a hybridization probe according to standard hybridization techniques under stringent hybridization conditions.
- a nucleic acid molecule of the inven- tion is at least 15, 20, 25 or 30 nucleotides in length. Preferably, it hybridizes under stringent conditions to a nucleic acid molecule comprising a nucleotide sequence of the nucleic acid molecule of the present invention or used in the process of the present invention, e.g. comprising the sequence shown in table I, columns 5 and 7.
- the nucleic acid molecule is preferably at least 20, 30, 50, 100, 250 or more nucleotides in length.
- hybridizes under stringent conditions is defined above.
- the term “hybridizes under stringent conditions” is intended to describe conditions for hybridization and washing under which nucleotide sequences at least 30 %, 40 %, 50 % or 65% identical to each other typically remain hybridized to each other.
- the conditions are such that sequences at least about 70%, more preferably at least about 75% or 80%, and even more preferably at least about 85%, 90% or 95% or more identical to each other typically remain hybridized to each other.
- nucleic acid molecule of the invention that hybridizes under stringent conditions to a sequence shown in table I, columns 5 and 7 corresponds to a naturally-occurring nucleic acid molecule of the invention.
- a "naturally-occurring" nucleic acid molecule refers to an RNA or DNA molecule having a nucleotide sequence that occurs in nature (e.g., encodes a natural protein).
- the nucleic acid molecule encodes a natural protein having above-mentioned activity, e.g. conferring increased yield, e.g an improved yield-related trait, e.g.
- nucleotide substitutions leading to amino acid substitutions at "non-essential" amino acid residues can be made in a sequence of the nucleic acid mole- -.
- A"non-essential" amino acid residue is a residue that can be altered from the wild-type sequence of one without altering the activity of said polypeptide, whereas an "essential" amino acid residue is required for an activity as mentioned above, e.g. leading to an enhancement of tolerance to abiotic environmental stress and/or increase of yield as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof in an organism after an increase of activity of the polypeptide.
- Other amino acid residues may not be essential for activity and thus are likely to be amenable to alteration without altering said activity.
- codon usage between organisms can differ. Therefore, he may adapt the codon usage in the nucleic acid molecule of the present invention to the usage of the organism or the cell compartment for example of the plastid or mitochondria in which the polynucleotide or polypeptide is expressed.
- the invention relates to nucleic acid molecules encoding a polypeptide having above-mentioned activity, in an organisms or parts thereof by for example expression either in the cytsol or in an organelle such as a plastid or mitochondria or both, preferably in plastids that contain changes in amino acid residues that are not essential for said activity.
- polypeptides differ in amino acid sequence from a sequence con- tained in the sequences shown in table II, columns 5 and 7 yet retain said activity described herein.
- the nucleic acid molecule can comprise a nucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence at least about 50% identical to an amino acid sequence shown in table II, columns 5 and 7 and is capable of participation in the enhancement of tolerance to abiotic environmental stress and/or increase of yield as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof after increasing its activity, e.g. its expression by for example expression either in the cytsol or in an organelle such as a plastid or mitochondria or both, preferably in plastids.
- the protein encoded by the nucleic acid molecule is at least about 60% identical to the sequence shown in table II, columns 5 and 7, more preferably at least about 70% identical to one of the sequences shown in table II, columns 5 and 7, even more preferably at least about 80%, 90%, 95% homologous to the sequence shown in table II, columns 5 and 7, and most preferably at least about 96%, 97%, 98%, or 99% identical to the sequence shown in table II, columns 5 and 7.
- amino acid residues or nucleic acid molecules at the corresponding amino acid positions or nucleotide positions are then compared. If a position in one sequence is occupied by the same amino acid residue or the same nucleic acid molecule as the corresponding position in the other sequence, the molecules are homologous at this position (i.e. amino acid or nucleic acid "homology” as used in the present context corresponds to amino acid or nucleic acid "identity”.
- Gap and “BestFit” are part of the GCG software-package (Genetics Computer Group, 575 Science Drive, Madison, Wisconsin, USA 5371 1 (1991 ); Altschul et al., (Nucleic Acids Res. 25, 3389 (1997)), "Needle” is part of the The European Molecular Biology Open Software Suite (EMBOSS) (Trends in Genetics 16 (6), 276 (2000)). Therefore preferably the calculations to determine the percentages of sequence homology are done with the programs "Gap” or “Needle” over the whole range of the sequences.
- EMBOSS European Molecular Biology Open Software Suite
- nucleic acid sequences were used for "Needle”: matrix: EDNAFULL, Gap_penalty: 10.0, Extend_penalty: 0.5.
- Gap gap weight: 50, length weight: 3, average match: 10.000, average mismatch: 0.000.
- a sequence, which has 80% homology with sequence SEQ ID NO: 39 at the nucleic acid level is understood as meaning a sequence which, upon comparison with the sequence SEQ ID NO: 39 by the above program "Needle" with the above parameter set, has a 80% identity.
- sequence which has a 80% homology with sequence SEQ ID NO: 40 at the protein level is understood as meaning a sequence which, upon comparison with the sequence SEQ ID NO: 40 by the above program "Needle" with the above parameter set, has a 80% identity.
- Functional equivalents derived from one of the polypeptides as shown in table II, columns 5 and 7 according to the invention by substitution, insertion or deletion have at least 30%, 35%, 40%, 45% or 50%, preferably at least 55%, 60%, 65% or 70% by preference at least 80%, especially preferably at least 85% or 90%, 91 %, 92%, 93% or 94%, very especially preferably at least 95%, 97%, 98% or 99% homology with one of the polypeptides as shown in table II, columns 5 and 7 according to the invention and having essentially the same properties as the polypeptide as shown in table II, columns 5 and 7.
- acitivty by for example expression either in the cytsol or in an organelle such as a plastid or mitochondria or both, preferably in plastids while increasing the amount of protein, activity or function of said functional equivalent in an organism, e.g. a microorgansim, a plant or plant tissue or animal tissue, plant or animal cells or a part of the same.
- a nucleic acid molecule encoding an homologous to a protein sequence of table II, columns 5 and 7 can be created by introducing one or more nucleotide substitutions, additions or deletions into a nucleotide sequence of the nucleic acid molecule of the present invention, in particular of table I, columns 5 and 7 such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein. Mutations can be introduced into the encoding sequences of table I, columns 5 and 7 by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis.
- conservative amino acid substitutions are made at one or more predicted non-essential amino acid residues.
- a "conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art.
- amino acids with basic side chains e.g., lysine, arginine, histidine
- acidic side chains e.g., aspartic acid, glutamic acid
- uncharged polar side chains e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine
- nonpolar side chains e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophane
- beta-branched side chains e.g., threonine, valine, isoleucine
- aromatic side chains e.g., tyrosine, phenylalanine, tryptophane, histidine
- a predicted nonessential amino acid residue in a polypeptide of the invention or a polypeptide used in the process of the invention is preferably replaced with another amino acid residue from the same family.
- mutations can be introduced randomly along all or part of a coding sequence of a nucleic acid molecule of the invention or used in the process of the invention, such as by saturation mutagenesis, and the resultant mutants can be screened for activity described herein to identify mutants that retain or even have increased above mentioned activity, e.g. conferring an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof.
- the encoded protein can be expressed recombinantly and the activity of the protein can be deter- mined using, for example, assays described herein (see Examples).
- Homologues of the nucleic acid sequences used, with the sequence shown in table I, columns 5 and 7, comprise also allelic variants with at least approximately 30%, 35%, 40% or 45% homology, by preference at least approximately 50%, 60% or 70%, more preferably at least approximately 90%, 91 %, 92%, 93%, 94% or 95% and even more preferably at least approximately 96%, 97%, 98%, 99% or more homology with one of the nu- cleotide sequences shown or the abovementioned derived nucleic acid sequences or their homologues, derivatives or analogues or parts of these.
- Allelic variants encompass in particular functional variants which can be obtained by deletion, insertion or substitution of nu- cleotides from the sequences shown, preferably from table I, columns 5 and 7, or from the derived nucleic acid sequences, the intention being, however, that the enzyme activity or the biological activity of the resulting proteins synthesized is advantageously retained or increased.
- the nucleic acid molecule of the invention or used in the process of the invention comprises the sequences shown in any of the table I, columns 5 and 7. It is preferred that the nucleic acid molecule comprises as little as possible other nucleotides not shown in any one of table I, columns 5 and 7. In one embodiment, the nucleic acid molecule comprises less than 500, 400, 300, 200, 100, 90, 80, 70, 60, 50 or 40 further nucleotides. In a further embodiment, the nucleic acid molecule comprises less than 30, 20 or 10 further nucleotides. In one embodiment, the nucleic acid molecule use in the process of the invention is identical to the sequences shown in table I, columns 5 and 7.
- nucleic acid molecule used in the process of the invention encodes a polypeptide comprising the sequence shown in table II, columns 5 and 7.
- the nucleic acid molecule encodes less than 150, 130, 100, 80, 60, 50, 40 or 30 further amino acids.
- the encoded polypeptide com- prises less than 20, 15, 10, 9, 8, 7, 6 or 5 further amino acids.
- the encoded polypeptide is identical to the sequences shown in table II, columns 5 and 7.
- the nucleic acid molecule of the invention or used in the process encodes a polypeptide comprising the sequence shown in table II, columns 5 and 7 comprises less than 100 further nucleotides. In a further embodiment, said nucleic acid molecule comprises less than 30 further nucleotides. In one embodiment, the nucleic acid molecule used in the process is identical to a coding sequence of the sequences shown in table I, columns 5 and 7.
- Polypeptides ( proteins), which still have the essential biological or enzymatic activity of the polypeptide of the present invention conferring an enhanced yield, e.g an improved yield-related trait, e.g. enhanced tolerance to abiotic environmental stress and/or increased yield like biomass as compared to a corresponding, e.g. non-transformed, wild type plant cell, plant or part thereof i.e.
- polypeptides with at least 10% or 20%, by preference 30% or 40%, especially preferably 50% or 60%, very especially preferably 80% or 90 or more of the wild type biological activity or enzyme activity, advantageously, the activity is essentially not reduced in comparison with the activity of a polypeptide shown in table II, columns 5 and 7 expressed under identi- cal conditions.
- Homologues of table I, columns 5 and 7 or of the derived sequences of table II, columns 5 and 7 also mean truncated sequences, cDNA, single-stranded DNA or RNA of the coding and noncoding DNA sequence.
- Homologues of said sequences are also understood as meaning derivatives, which comprise noncoding regions such as, for exam- pie, UTRs, terminators, enhancers or promoter variants.
- the activity of the promoters is increased by modification of their sequence, or that they are replaced completely by more active promoters, even promoters from heterologous organisms.
- Appropriate promoters are known to the person skilled in the art and are mentioned herein below.
- nucleic acid molecules encoding the YIPs described above another aspect of the invention pertains to negative regulators of the activity of a nucleic acid molecules selected from the group according to table I, column 5 and/or 7, preferably column 7.
- Antisense polynucleotides thereto are thought to inhibit the downregu- lating activity of those negative regulators by specifically binding the target polynucleotide and interfering with transcription, splicing, transport, translation, and/or stability of the target polynucleotide. Methods are described in the prior art for targeting the antisense polynucleotide to the chromosomal DNA, to a primary RNA transcript, or to a processed mRNA.
- the target regions include splice sites, translation initiation codons, translation termination codons, and other sequences within the open reading frame.
- antisense refers to a nucleic acid comprising a polynucleotide that is sufficiently complementary to all or a portion of a gene, primary transcript, or processed mRNA, so as to interfere with expression of the endogenous gene.
- “Complementary" polynucleotides are those that are capable of base pair- ing according to the standard Watson-Crick complementarity rules, bpecifically, purines will base pair with pyrimidines to form a combination of guanine paired with cytosine (G:C) and adenine paired with either thymine (A:T) in the case of DNA, or adenine paired with uracil (A:U) in the case of RNA. It is understood that two polynucleotides may hybridize to each other even if they are not completely complementary to each other, provided that each has at least one region that is substantially complementary to the other.
- antisense nucleic acid includes single stranded RNA as well as double-stranded DNA expression cassettes that can be transcribed to produce an antisense RNA.
- "Active" antisense nucleic acids are antisense RNA molecules that are capable of selectively hybridizing with a negative regulator of the activity of a nucleic acid molecules encoding a polypeptide having at least 80% sequence identity with the polypeptide selected from the group according to table II, column 5 and/or 7, preferably column 7.
- the antisense nucleic acid can be complementary to an entire negative regulator strand, or to only a portion thereof.
- the antisense nucleic acid molecule is antisense to a "noncoding region" of the coding strand of a nucleotide sequence encoding a YIP.
- the term "noncoding region" refers to 5' and 3' sequences that flank the coding region that are not translated into amino acids (i.e., also referred to as 5' and 3' untranslated regions).
- the antisense nucleic acid molecule can be complementary to only a portion of the noncoding region of YIP mRNA.
- the antisense oligonucleotide can be complementary to the region surrounding the translation start site of YIP mRNA.
- an antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 nucleo- tides in length.
- the antisense molecules of the present invention comprise an RNA having 60-100% sequence identity with at least 14 consecutive nucleotides of a non- coding region of one of the nucleic acid of table I.
- the sequence identity will be at least 70%, more preferably at least 75%, 80%, 85%, 90%, 95%, 98% and most preferably 99%.
- An antisense nucleic acid of the invention can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art.
- an antisense nucleic acid e.g., an antisense oligonucleotide
- an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and ac- ridine substituted nucleotides can be used.
Abstract
Description
Claims
Priority Applications (7)
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EP08861472A EP2222857A2 (en) | 2007-12-19 | 2008-12-19 | Plants with increased yield and/or increased tolerance to environmental stress (iy-bm) |
CN200880127041.XA CN101952305B (en) | 2007-12-19 | 2008-12-19 | Plants with increased yield and/or increased tolerance to environmental stress (IV-BM) |
DE112008003318T DE112008003318T5 (en) | 2007-12-19 | 2008-12-19 | Plants with increased yield and increased tolerance to environmental stress (IY-B) |
US12/809,142 US20110098183A1 (en) | 2007-12-19 | 2008-12-19 | Plants with increased yield and/or increased tolerance to environmental stress (iy-bm) |
BRPI0821748A BRPI0821748A2 (en) | 2007-12-19 | 2008-12-19 | method for producing a plant with increased yield, isolated nucleic acid molecule, nucleic acid construction, vector, process for producing a polypeptide, polypeptide, antibody, plant cell nucleus, plant cell, plant tissue, propagation material, seed, pollen, progeny, or a plant part, or a high yielding plant, process for the identification of a compound, method for producing an agricultural composition, composition, polypeptide or nucleic acid molecule, use of nucleic acids, and method for the identification of a plant with increased yield |
CA2708094A CA2708094A1 (en) | 2007-12-19 | 2008-12-19 | Plants with increased yield and/or increased tolerance to environmental stress (iy-bm) |
AU2008337398A AU2008337398B2 (en) | 2007-12-19 | 2008-12-19 | Plants with increased yield and/or increased tolerance to environmental stress (IY-BM) |
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EP07150175.3 | 2007-12-19 | ||
EP07150175 | 2007-12-19 |
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US (1) | US20110098183A1 (en) |
EP (2) | EP2222857A2 (en) |
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AU (1) | AU2008337398B2 (en) |
BR (1) | BRPI0821748A2 (en) |
CA (1) | CA2708094A1 (en) |
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DATABASE UniProt [Online] 3 April 2007 (2007-04-03), Mathee K. et al.: "Dynamics of Pseudomonas aeruginosa genome evolution" XP002580713 retrieved from EBI Database accession no. A3LFX1 * |
HIROHASHI T ET AL: "cDNA sequence and overexpression of chloroplast chaperonin 21 from Arabidopsis thaliana" BIOCHIMICA ET BIOPHYSICA ACTA. PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, ELSEVIER, AMSTERDAM; NL LNKD- DOI:10.1016/S0167-4838(98)00268-4, vol. 1429, no. 2, 11 January 1999 (1999-01-11), pages 512-515, XP004278612 ISSN: 0167-4838 * |
VAN CAMP W: "Yield enhancement genes: seeds for growth" CURRENT OPINION IN BIOTECHNOLOGY, LONDON, GB LNKD- DOI:10.1016/J.COPBIO.2005.03.002, vol. 16, no. 2, 1 April 2005 (2005-04-01), pages 147-153, XP004849204 ISSN: 0958-1669 * |
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EP2604622A2 (en) | 2013-06-19 |
CN101952305B (en) | 2014-12-24 |
EP2604622A3 (en) | 2013-10-09 |
CA2708094A1 (en) | 2009-06-25 |
AU2008337398B2 (en) | 2014-04-03 |
CN101952305A (en) | 2011-01-19 |
BRPI0821748A2 (en) | 2019-09-24 |
US20110098183A1 (en) | 2011-04-28 |
AR069893A1 (en) | 2010-02-24 |
DE112008003318T5 (en) | 2011-04-21 |
AU2008337398A1 (en) | 2009-06-25 |
WO2009077611A3 (en) | 2009-10-08 |
EP2222857A2 (en) | 2010-09-01 |
WO2009077611A9 (en) | 2010-08-05 |
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