WO1999063100A1 - Metodo para la obtencion de plantas transgenicas que tienen una capacidad mejorada para la toma de nutrientes y la tolerancia a compuestos toxicos presentes en el suelo - Google Patents
Metodo para la obtencion de plantas transgenicas que tienen una capacidad mejorada para la toma de nutrientes y la tolerancia a compuestos toxicos presentes en el suelo Download PDFInfo
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- WO1999063100A1 WO1999063100A1 PCT/MX1998/000020 MX9800020W WO9963100A1 WO 1999063100 A1 WO1999063100 A1 WO 1999063100A1 MX 9800020 W MX9800020 W MX 9800020W WO 9963100 A1 WO9963100 A1 WO 9963100A1
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0012—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7)
- C12N9/0026—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on CH-NH groups of donors (1.5)
- C12N9/0028—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on CH-NH groups of donors (1.5) with NAD or NADP as acceptor (1.5.1)
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- 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
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/88—Lyases (4.)
Definitions
- the invention relates to a method for obtaining transgenic plants that have a high capacity to synthesize and exude organic acids when compared to equivalent non-transformed plants, a DNA molecule to produce said transgenic plants, transgenic plants with increased capacity to synthesis and excression of organic acids and the uses of transgenic plants.
- the elements that have been described include C, H, O, N, K, P, Mg, Ca, S, Fe, B, Mn, Cu, Zn, Mo, Cl and Ni, some of them, micronutrients, such as For example, molybdenum is required at concentrations less than one part per million, but if they are absent, the plant cannot complete its life cycle. Other nutrients such as nitrogen and phosphorus (macronutrients) are needed at high levels and reach concentrations of up to 3% of the total dry weight of the plant (Crawford, NM (1994). In Arabidopsis. Cold Spring Harbor Press pp 1 1 19-1 145).
- Plant roots excrete a wide variety of ions and organic substances that affect the availability of nutrients in the soil.
- chelating compounds organic acids and phytokelatins
- P, Fe, Mn and Zn Marschner, H. (1995) Mineral nutrition of higher plants. Academic Press, San Diego, CA.
- exudation and excresion are functionally considered the same process, which we define as the process by which plants, through their roots, send organic and inorganic compounds to the soil.
- the chemical nature of the substances that are excreted by the root depends on several factors: the species, the genetic variability, the age of the plant and the nutritional status of the same.
- the components that have been identified in root exudates can be included in two large groups: mucilaginous substances and organic solutes.
- Organic solutes include sugars, organic acids, amino acids and phenolic compounds.
- Organic acids are very versatile molecules that take part in different physiological processes in all living organisms including plants.
- the biosynthesis of these compounds seems to be a general phenomenon that is conserved in living beings, although in animals their participation is known in detail as precursors of important metabolic pathways such as the Krebs cycle or the glyoxylate cycle, their transport mechanism and the enzymes with which they interact, there is very little information in plants about it (Srere, PA (1992). Curr. Top. Cell. Reg. 33: 261-275).
- Organic acid excretion has been correlated with the capacity of some plant species, such as rapeseed (Brassica napus) and white lupine (Lupinus albus), to solubilize phosphate (and therefore, convert phosphate into biologically assimilable forms), from insoluble compounds such as aluminum or iron phosphates, or phosphoric rock.
- rapeseed Brassica napus
- white lupine Liupinus albus
- citric acid In addition to the correlation observed between the ability of certain plant species to solubilize phosphate and their ability to exude organic acids, especially citric acid, since the 1950s it is known that organic acids, such as citrate, malate, oxalate, tartrate, malonate and lactate they are capable of dissolving aluminum and iron phosphates that are normally insoluble. Of all the aforementioned organic acids, citrate seems to be the most effective for dissolving insoluble phosphate compounds.
- Another important problem that affects the intake of nutrients from the soil is the presence of toxic compounds, such as certain forms of soluble aluminum or other metals that can interfere with root growth.
- Aluminum is the most abundant metal on the earth's layer (representing approximately 7% of its mass) and is found in the soil mainly in the form of insoluble aluminum silicates and oxides. However, when solubilized in acidic soils (mainly in the form of Al + 3), Aluminum is highly toxic to many crops. Al toxicity is considered as the main factor that limits plant productivity in acidic soils. Acidification of soils occurs naturally when basic cations are washed away from the soil, but can also be significantly accelerated by certain agricultural practices and by acid rain. Acid soils comprise approximately 40% of the world's arable land and are particularly abundant in tropical and subtropical regions of the world (Foy, CD. Et al. (1978). Annu.Rev.Plant Physiol. 29: 51 1- 66). A common practice to sustain agricultural productivity in acidic soils is the application of calcium hydroxide (lime) or calcium sulfate (gypsum,
- citrate binds strongly to iron and aluminum, avoiding the precipitation of phosphate at different pH values, this interaction was more efficient compared to the effect promoted by other organic acids.
- a millimol of citrate is combined with a millimol of Aluminum, reducing phosphate precipitation by up to 100% in a range of 4.0-9.0 pH units.
- Enzymes that synthesize organic acids and the genes that encode them have been extensively studied in bacteria and animals. Among the most studied are citrate synthase and malate dehydrogenase, which have been characterized in great detail and the genes that encode them for a significant number of bacteria, animals and some plants have been isolated and sequenced. In the case of Citrate Syntase, more than 300 genes have been cloned and characterized, while for Malato Dehydrogenase more than 500 genes have been cloned and sequenced. Citrate synthase catalyzes the synthesis of citrate by condensation of the methyl group of acetyl coenzyme A and the carbonyl group of oxaloacetate. This enzyme known amino acids that form the active site, which are conserved in all the enzymes analyzed including those of Rickettsia and
- the invention provides a method for obtaining transgenic plants with an increased ability to synthesize and exude organic acids, which comprises the following steps. a) The preparation of a genetic construct comprising one or more enzymes that synthesize organic acids, functionally linked to a promoter sequence of active transcription in plants and a terminator sequence of active transcription in plant cells, b) The transformation of plant cells with the previous genetic construction, and c) The regeneration of transgenic plants from the transformed cells Description of the invention
- the invention relates to the overproduction of citric acid, however, the present invention is not limited to the overproduction of citric acid, since overproduction of other organic acids, such as malic acid and oxalic acid, is also possible. among others.
- One of the aspects of this invention describes the construction of a recombinant DNA molecule that encodes the enzyme citrate synthase, functionally linked to a promoter sequence of functional transcription in plants and a terminator sequence of plant transcription.
- the present invention relates, but is not limited, to the use of the gene encoding Citrate.
- citric acid occurs primarily in the mitochondria, where it is the first reaction of the Krebs cycle and is carried out by citrate synthase. Since this reaction is part of a complex cycle, where the flow of carbon skeletons does not necessarily accumulate in one of its components and is subject to complex regulatory mechanisms, it is highly desirable to carry out the invention, compartmentalize the citrate enzyme synthase in a subcellular compartment other than mitochondria, to prevent synthesized citric acid molecules from becoming other components of the Krebs cycle. Therefore, an important aspect of this invention is the description of a method for obtaining transgenic plants where Citrate Syntase is located in a subcellular compartment other than mitochondria, such as cytoplasm and chloroplast.
- the recombinant DNA molecule described in this invention may contain a signal or transit peptide that directs Citrate Syntase to a discrete compartment of the cell.
- the present invention has advantages over the prior art, since the transgenic plants obtainable by this method have a better ability to take nutrients or tolerate toxic compounds without the need to use chemical soil treatments or the use of nutrients associated with chelating compounds. Industry or farmers can use transgenic seeds to establish agricultural crops, reducing their production costs, in terms of soil treatments or fertilizer addition, or increasing their productivity in acidic soils or in those that have nutrients not available. .
- the present invention is directed to obtaining transgenic plants with an increased production capacity and exudation of organic acids.
- recombinant molecules and methods are described that allow obtaining transgenic plants with an increased capacity for synthesis and exudation of organic acids.
- the preferred organic acid form in this invention is citric acid, as it has been shown as one of the most effective in solubilizing and facilitating the absorption of nutrients from the soil.
- Citrate Syntase any enzyme capable of synthesizing citric acid.
- the present invention relates, but is not limited to Citrate Syntase, since it is possible to use other genes encoding enzymes capable of synthesizing other organic acids.
- Citrate Syntase or any other enzyme that synthesizes organic acids must have kinetic parameters compatible with the biochemical and physiological systems of the plant of interest.
- the gene or the coding part of a gene that encodes enzymes that synthesize organic acids can be derived from a complementary DNA molecule, from genomic DNA or can be chemically synthesized totally or partially.
- the desired gene can be obtained from any microorganism, from any plant or from any animal.
- the gene or part thereof will be derived from native sequences of some organism.
- enzymes capable of synthesizing organic acids and in particular Citrate Syntase
- a large number of genes have been previously identified and characterized, including the determination of their nucleotide sequence.
- the gene contained in the recombinant DNA molecule is expressed in the appropriate levels and tissues, it is desirable, but not necessary for genes of plant origin, that it is contained in an expression cassette that includes a promoter sequence of the functional transcription in plants, the coding part of the enzyme gene that synthesizes organic acids and a functional transcription terminator in plants. Additionally, a sequence encoding a transit peptide that directs the enzyme to a specific cell compartment can be included.
- the transit peptide and the corresponding processing signals can be derived from any plant protein that is synthesized in the cytoplasm and translocated to the subcellular compartment of interest, be it the plastid or the mitochondria.
- sequences derived from genes encoding the small subunit of ribulose bisphosphate carboxylase or CAB proteins that can be used to locate plastids are those that bind with chlorophylls a and b to formalize light harvesting antennas (Van den Broek et al Nature (London) 313, 358-363
- a promoter In the case of genes that are not of plant origin, a promoter must be included in the transcription initiation sequence and an optimal translation start signal for plants or one of the so-called translation enhancers can also be used. which may be used include promoters of the plant to be transformed, promoters of other plants or of any origin that are functional in the white plant and direct a constitutive, inducible or specific tissue expression. For example, promoters derived from the Ti plasmid can be used.
- a robactertum tumefac ⁇ ais such as the promoters of Octopina Sintetasa, Nopahna Sintetasa or Agropina Synthetase
- promoters such as the 35S promotoi of the cauliflower mosaic virus or those derived from geminivirus
- 35S promotoi of the cauliflower mosaic virus or those derived from geminivirus may be included
- Temporary, inducible or tissue expression specific can be achieved through the use of promoters or regulatory sequences that have the specificity of expression desired
- gene promoters which encode phosphate transporters which have been shown to express themselves specifically in root epithelial cells (Muchhal US, et al Proc Nati Acad. Sci 93 10519-10523)
- the transcription terminator sequencer can be derived from the same gene from which the transcription promoter sequence was obtained or from a different gene.
- the terminator sequence can be derived from genes contained in the T-DNA of the Ti plasmid of Agrobacterium, of the cauliflower mosaic virus or of genes of plant origin. For example, the terminating sequence of the nopaline synthetase gene is frequently used in Plant Genetic Engineering.
- nucleic acid sequences that are part of the recombinant DNA molecule can be linked by conventional methods described in the literature.
- the sequences must be cloned and linked in the correct orientation and order to achieve functional expression in plant cells.
- the plants of interest are transformed with a recombinant DNA molecule that encodes at least one enzyme capable of synthesizing organic acids.
- a recombinant DNA molecule that encodes at least one enzyme capable of synthesizing organic acids.
- the different nucleotide sequences that compose it and that comprise the regulatory sequences of the transcription and the coding sequence of interest may have been subjected to different types of processing, such as ligaments, digestion with restriction enzymes , in vitro mutagenesis, oligonucleotide addition or modifications by means of the chain polymerization reaction (PCR). Therefore, the components of the recombinant molecule before being bound could be subject to deletions, insertions or internal modifications. Since the recombinant molecule is derived from components that originate from different organisms and that have been isolated, purified or synthesized, it is not a molecule that exists as such in nature.
- the inclusion of other DNA sequences may be necessary.
- a dominant selection gene is also generally included to identify and select the cells that stably incorporated the recombinant molecule into their genome.
- the recombinant molecule can be introduced into the desired plant cell by any method of genetic transformation of plants, which may include, but is not limited to, the following transformation methods: the transformation system mediated by the Ti plasmid of Agrobacterium tumefaciens , electroporation, microinjection and biobalistics or bombardment with microparticles.
- the transformation system mediated by the Ti plasmid of Agrobacterium tumefaciens electroporation, microinjection and biobalistics or bombardment with microparticles.
- Agrobacterium Ti to introduce the recombinant molecule into the genome of the plant cell, it is necessary to include the T-DNA border sequences, so that they are present at both ends of the gene that encodes the enzyme that synthesizes organic acids and in some cases the dominant selection gene.
- the use of unarmed Agrobacterium strains that is, those that have been removed from the genes responsible for tumor formation, but that maintain the ability to transfer DNA to plant cells, allows the regeneration of transgenic plants that contain the recombinant molecule of interest .
- the recombinant DNA molecule that contains the gene that codes for the synthesis of organic acids can be introduced into any plant species, including both monocot and dicotyledons.
- plant species that by nature have a low capacity to solubilize phosphorus and iron from the soil or that are susceptible to the toxicity caused by aluminum.
- plant species include but are not limited to corn (Zea mays), rice (Oryza sativa), wheat (Triticum sp), sorghum (Sorghum bicolor), soybeans (Glycine max L.), tobacco (Nicotiana tabacum), tomato (Lycopersicum sculetum), papaya (Carica papaya) and potato (Solanum tuberosum).
- transgenic plants After the transformation of the tissues, the plant cells or the protoplasts derived from the plant of interest, transgenic plants that contain in their genome the recombinant molecule are regenerated. These transgenic plants are capable of stably transferring the recombinant molecule to its progeny, be it derived from seeds, cuttings, tubers or any other reproductive structure.
- Example 1 The present invention will be illustrated in the examples described below, which in no way are intended to limit the present invention.
- Example 1
- genes to be used To obtain plants that overproduce organic acids, genes that code for enzymes that have the ability to synthesize organic acids were selected.
- genes that code for enzymes that have the ability to synthesize organic acids were selected.
- the Pseudomonas aeruginosa gene that encodes the Citrate Synthase enzyme was selected (Donald et al (1989), J. Bacteriol. 171: 5542-5550). This enzyme synthesizes citric acid from oxaloacetate and acetyl-Coenzyme A.
- the cytoplasm was selected as the subcellular compartment where said enzyme would be located.
- the pB2 expression vector was first constructed.
- the region of the photoinducible promoter (pea cab 80) of the vector pGV151 1 (Jofre-Garfias et al (1997), Plant Cell Reports 16: 847-852) delimited by the restriction sites Hind III and Bam HI was replaced by the Hind III-Bam HI fragment of the vector pBI 525 (Datla et al. (1993), Plant Science 94: 139-149.), which contains the double 35S promoter of the cauliflower mosaic virus, the virus-enhancing region of the Alfalfa mosaic and the translation start site (as part of the Neo I site).
- the substitution of the promoter region of the pGV1 51 1 vector with that of pBI 525 was verified by double restriction analysis using the Xba I and Hind III sites.
- the orientation of the cloned fragment was determined by analysis of the fragment released by a double restriction using the enzymes Xba I and Bam
- Plasmid p35SCSb was conjugated from E. coli to the Agrobacterium strain LB4404 (Hoekema A. et al. (1983), Nature 303, 179-180) using the triparental conjugation system using the assistant plasmid (heper plasmid) pRK2013 ( Lam ST et al. (1985), Plasmid 13, 200-204).
- the recombinant molecule was introduced into the Tobacco genome (Nicotiana tabacum L. var. Xanthi) using the method of transformation of leaf discs (Horsh R. B. et al (1985), Science 227, 1229-1232).
- the regenerated plants, called CSb plants were selected for their growth in selective medium containing 50 micrograms of kanamycin per milliliter of culture medium.
- genomic DNA was extracted from the leaves of said plants by conventional methods, digested with the restriction enzymes Hind III and Bam HI, subjected to electrophoresis in a 1% agarose gel in TBE buffer , transferred to nylon membranes and hybridized with a specific probe for the Pseudomonas aeruginosa Citrate Syntase gene. It was found that most of the kanamycin resistant plants contained the expression cassette of the P. aeuroginosa Citrate Syntase. To verify that the expression cassette was functionally transcribed in CSb plants, total RNA from said plants was extracted using techniques.
- Enzymol, 13: 3-22) showed that several of them had high levels of Citrate Syntase activity with respect to the non-transformed control plants.
- Four lines with levels of Citrato Sintasa activity were selected between two and three times greater than the control (see figure 2).
- the presence of bacterial CS in the CSb lines was confirmed by Western-type analysis using an antiserum (Donald, J.L, et al. (1989.). Journal of Bacteriology. 5542-5550) that does not recognize the plant enzyme.
- the densitometric analysis of the detection by Western type analysis of the bacterial CS showed a good correlation between the level of bacterial CS and the increase detected in the Citrate Syntase activity in the different CSb lines, demonstrating that the increased levels of Citrate Syntase in the plants CSb is due to the expression of the recombinant molecule encoding P. aeruginosa Citrate Syntase.
- the plant material used consisted of two lines of transgenic plants called CSb 5-4 and CSb 5-18, which overexpress the P. aeuroginosa citrate synthase gene and a wild plant (1522) that was used as a control.
- the plants were established in polyethylene bags containing 2 kilograms of soil with the respective Phosphorus treatment.
- a random arrangement of the plants in the greenhouse was designed forming random blocks consisting of each treatment of 24 repetitions.
- samples were carried out in 3 stages of the development of the plants: vegetative growth, flowering and fruiting. From each stage, 8 plants were analyzed to measure the following agronomic variables: plant height, leaf area, fresh frond weight, dry frond weight, dry root weight. The number of flowers (for the flowering stage) and number and total dry weight of capsules or fruits (for the fruiting stage) were also determined.
- the capsules were collected in full senescence stage of the plant, once the grain filling had concluded. The harvest was carefully done by splitting the capsules from the base of the peduncle and depositing them in paper bags of dextrase. The bags were properly labeled and included for drying the plant material in a Heraeus Bau Herbert 6000 stove at a temperature of 70 degrees Celsius for 72 hours. The dry material was weighed on a Mettler PE 360 analytical balance and the data obtained were subjected to a statistical analysis of variance (anova) and comparison of means with the Tukey method. b) Dry weight of the frond (Includes total stem and leaf biomass).
- the leaves were harvested from the base of the petiole and were included together in paper bags of dextrase.
- the stem was split from the crown (the area of the stem closest to the ground). All material from the same plant was included in a single paper bag properly labeled.
- the samples were dried in a Heraeus Bausch 6000 oven at a temperature of 70 degrees Celsius for 72 hours. The dry material was weighed on a Mettler PE 360 analytical balance and the data obtained were subjected to a statistical analysis of variance (anova) and comparison of means with the Tukey method.
- c) Total dry weight of the plant (includes frond weight and capsules). This parameter was determined by adding the total weight of the frond and the total weight of the capsules.
- transgenic line B5 18 accumulated greater biomass than transgenic CSb 5-4 and the control plant (1522).
- the difference in the dry weight of the capsules between the CSb line 5-18 and the control 1522 was statistically significant, with a 95% confidence interval (see figure 5).
- the aluminum tolerance segregation pattern was examined in the IT progeny of the selected lines. A segregation of phenotypes was observed tolerant / susceptible with a 3: 1 pattern in the lines that contain simple insertions, and that the tolerance to Aluminum congregates with the kanamycin resistance gene present in the T-DNA of the binary vector used for the production of CSb lines, confirming that The resistance phenotype is due to the presence of the 35S-CSb construct in the genome of the Tobacco transgenic lines analyzed.
- the T-DNA contained in the recombinant molecule p35S-CSb was introduced to the papaya plant genome by the bombardment transformation system of particles (Cabrera-Ponce et al. (1995), Plant Cell Reports 15: 1-7).
- Papaya lines were found to have levels of Citrate Syntase activity 2 to 3 times higher than controls transformed with the vector without the coding sequence for CS.
- 20 regenerated plants of each line were transferred to rooting medium containing different concentrations of aluminum.
- FIG. 1 Shows the construction of p35SCSb.
- the nucleotide and amino acid sequence is also indicated at the translation initiation site of the original Pseudomonas aeruginosa (pPKB) CS gene, that of the pB2 expression vector and the resulting sequence in p35SCSb, result of fragment cloning Bel I-Bam HI containing most of the coding sequence of the CS in the expression vector pB2.
- Figure 2. Determination of citrate synthase activity present in extracts of transgenic CSb plants and 1522 control plants. It can be observed that transgenic plants have an increased citrate synthase activity with respect to the control.
- FIG. 3 Determination of citrate levels present in root extracts of transgenic CSb plants and control plants. The level of citrate exuded by CSb transgenic plants and control plants is also presented. It can be seen that transgenic plants have an increased capacity to accumulate and exude citric acid with respect to control plants.
- FIG. 4 Determination of the biomass accumulated by transgenic tobacco plants that overexpress the Pseudomonas aeuroginosa citrate synthase gene (CSb 5-4 and CSb 5-18) and control plants (1522) grown in the presence of 22 parts per million match.
- the accumulated biomass is presented as dry weight of capsules, frond and capsules + frond.
- the results presented are the average and standard error of eight repetitions per treatment.
- Statistical analysis (ANOVA and Tukey test), revealed that there is an increase in yield in transgenic plants compared to control plants in the phosphorus concentrations that were tested.
- FIG. 5 Determination of the biomass accumulated by transgenic tobacco plants that overexpress the Pseudomonas aeuroginosa citrate synthase gene (CSb 5-4 and CSb 5-18) and control plants (1522) grown in the presence of 44 parts per million match.
- the accumulated biomass is presented as dry weight of capsules, frond and capsules + frond.
- the results presented are the average and standard error of eight repetitions per treatment.
- Statistical analysis (ANOVA and Tukey test), revealed that there is an increase in yield in transgenic plants compared to control plants in the phosphorus concentrations that were tested.
- Figure 7 Determination of the effect of increasing concentrations of aluminum at pH 4.5 on the root growth of transgenic CSb plants and control plants. The effect is plotted as the percentage of root growth inhibition in the presence of aluminum with respect to the growth of the same plants in the absence of aluminum. It can be seen that the roots of the transgenic CSb plants grow better in the presence of toxic concentrations of aluminum.
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Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
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AU75533/98A AU772220B2 (en) | 1998-05-29 | 1998-05-29 | Process for obtaining transgenic plants which have an improved capacity for the uptake of nutrients and tolerance to toxic compounds which are present in the soil |
PCT/MX1998/000020 WO1999063100A1 (es) | 1998-05-29 | 1998-05-29 | Metodo para la obtencion de plantas transgenicas que tienen una capacidad mejorada para la toma de nutrientes y la tolerancia a compuestos toxicos presentes en el suelo |
EP98923194A EP1122316A4 (en) | 1998-05-29 | 1998-05-29 | PROCESS FOR OBTAINING TRANSGENIC PLANTS HAVING IMPROVED NUTRITIONAL TOLERANCE AND TOLERANCE CAPACITY FOR TOXIC COMPOUNDS IN THE SOIL |
BR9815878-3A BR9815878A (pt) | 1998-05-29 | 1998-05-29 | Método para obtenção de plantas transgênicas que apresentam capacidade aperfeiçoada de absorção de nutrientes e tolerância a compostos tóxicos presentes em solos |
US10/898,322 US20050137386A1 (en) | 1998-05-29 | 2004-07-26 | Process for obtaining transgenic plants which have improved capacity for the uptake of nutrients and tolerance to toxic compounds which are present in the soil |
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PCT/MX1998/000020 WO1999063100A1 (es) | 1998-05-29 | 1998-05-29 | Metodo para la obtencion de plantas transgenicas que tienen una capacidad mejorada para la toma de nutrientes y la tolerancia a compuestos toxicos presentes en el suelo |
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NZ542731A (en) * | 2003-04-14 | 2008-04-30 | Agriculture Victoria Serv Pty | Manipulation of organic acid biosynthesis and secretion |
US20120216315A1 (en) * | 2008-06-13 | 2012-08-23 | Basf Plant Science | Methods in Increasing Grain Value by Improving Grain Yield and Quality |
CN107058115A (zh) * | 2008-11-19 | 2017-08-18 | 国家政治研究所高级研究中心(高级研究中心) | 能够代谢亚磷酸盐作为磷源的转基因植物和真菌 |
-
1998
- 1998-05-29 EP EP98923194A patent/EP1122316A4/en not_active Withdrawn
- 1998-05-29 AU AU75533/98A patent/AU772220B2/en not_active Ceased
- 1998-05-29 WO PCT/MX1998/000020 patent/WO1999063100A1/es active IP Right Grant
-
2004
- 2004-07-26 US US10/898,322 patent/US20050137386A1/en not_active Abandoned
Non-Patent Citations (3)
Title |
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DONALD L. J., MOLGAT G. F., DUCKWORTH H. W.: "CLONING, SEQUENCING, AND EXPRESSION OF THE GENE FOR NADH-SENSITIVE CITRATE SYNTHASE OF PSEUDOMONAS AERUGINOSA.", JOURNAL OF BACTERIOLOGY, AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 171., no. 10., 1 October 1989 (1989-10-01), US, pages 5542 - 5550., XP002918866, ISSN: 0021-9193 * |
FUENTE DE LA J. M., ET AL.: "ALUMINUM TOLERANCE IN TRANSGENIC PLANTS BY ALTERATION OF CITRATE SYNTHESIS.", SCIENCE, AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE, US, vol. 276., 6 June 1997 (1997-06-06), US, pages 1566 - 1568., XP002918865, ISSN: 0036-8075, DOI: 10.1126/science.276.5318.1566 * |
See also references of EP1122316A4 * |
Also Published As
Publication number | Publication date |
---|---|
US20050137386A1 (en) | 2005-06-23 |
AU772220B2 (en) | 2004-04-22 |
EP1122316A4 (en) | 2002-08-28 |
AU7553398A (en) | 1999-12-20 |
EP1122316A1 (en) | 2001-08-08 |
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