WO2013093637A2 - Procédés de traitements des plantes et moyens associés - Google Patents

Procédés de traitements des plantes et moyens associés Download PDF

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WO2013093637A2
WO2013093637A2 PCT/IB2012/003045 IB2012003045W WO2013093637A2 WO 2013093637 A2 WO2013093637 A2 WO 2013093637A2 IB 2012003045 W IB2012003045 W IB 2012003045W WO 2013093637 A2 WO2013093637 A2 WO 2013093637A2
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plant
seq
pad4
lsd1
eds1
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WO2013093637A3 (fr
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Stanislaw Karpinski
Magdalena Szechynska-Hebda
Ireneusz SLESAK
Weronika WITUSZYNSKA
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Szkola Glowna Gospodarstwa Weijskeigo W Warszawie
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    • C—CHEMISTRY; METALLURGY
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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
    • 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
    • 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
    • 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
    • C12N15/8273—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 for drought, cold, salt resistance
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • 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

  • the invention relates to the field of plant and agricultural technology. More specifically, the present invention relates to a method for regulation (increasing) of tree growth, development, biomass production, and stress tolerance and means therefor. In particular, the invention relates to a method for altering the biomass production, in optimal conditions and under water stress by genetically engineering plants to specific expression of PAD4, EDS1, LSD1 genes, alone or in combinations. BACKGROUND
  • biomass is the fourth largest energy source in the world after coal, petroleum and natural gas, accounting for 14% of the world's primary energy consumption.
  • the object of the present invention is to obtain transgenic trees and crop plants with improved water use efficiency and biomass production in stress and semi-stress field (and laboratory) conditions.
  • Increase in stress tolerance opens opportunities for successful cultivation of crop and other plant species under conditions that were not previously possible, e.g. in areas with poor irrigation.
  • increase tolerance toward water stress it is meant that under water deficiency conditions, the plant exhibits the ability to grow in a more successful manner than the control plant that is not genetically engineered as described herein.
  • the genetically engineered plant is able to develop better traits with respect to growth rate, biomass, color, maturation, time of flowering, fruit production, etc. under stress conditions.
  • an isolated nucleic acid molecule including a nucleotide sequence includes at least two amino acid sequences chosen from among amino acid sequences having at least 60% identity with SEQ ID NO. 4, at least 60% identity with SEQ ID NO. 5, and at least 60% identity with SEQ ID NO. 6; or chosen from among amino acid sequences having at least 60% identity with SEQ ID NO. 7, at least 60% identity with SEQ ID NO. 8, and at least 60% identity with SEQ ID NO. 9; wherein SEQ ID NO. 4 and SEQ ID NO. 7 encode phytoalexin deficient 4 (PAD4) protein in a plant; SEQ ID NO. 5 and SEQ ID NO. 8 encode lesion simulating disease 1 (LSD1 ) protein in a plant; and SEQ ID NO. 6 and SEQ ID NO. 9 encode enhanced disease susceptibility 1 (EDS1 ) protein in a plant.
  • PAD4 phytoalexin deficient 4
  • allelic variant or a homolog of the nucleotide sequence or a DNA fragment of the nucleotide sequence is also provided, wherein one or more functional characteristics of the protein are retained.
  • a vector construct including the isolated nucleic acid molecule is provided wherein the nucleotide sequence is operably linked to an expression control sequence.
  • a host cell including the isolated nucleic acid molecule is provided, wherein the nucleotide sequence is flanked by exogenous sequence.
  • a host cell Including the vector construct or a progeny of the cell is provided, wherein the cell is operably linked to an expression control sequence and expresses the polypeptide.
  • a plant, plant material, plant cell, or a seed of a plant which includes the isolated nucleic acid molecule, wherein the nucleotide sequence is exogenous or heterologous to the plant or the plant cell.
  • a plant, plant material, plant cell, or a seed of a plant further includes the host cell.
  • a plant regenerated from a plant cell or seed that includes the isolated nucleic acid molecule is also provided.
  • a method of regulating growth and increasing biomass production in a plant further includes over expressing at least two of phytoalexin deficient 4 (PAD4) gene, lesion simulating disease 1 (LSD1 ) gene, and enhanced disease susceptibility 1 (EDS1 ) gene in the plant, wherein the overexpression step further includes contacting the genome of the plant with an expression cassette comprising an isolated nucleic acid molecule comprising the nucleotide sequence encoding at least two of PAD4, LSD1 , or EDS1 proteins, wherein the nucleotide sequence is operably linked to an expression control sequence that is functional in plants under conditions that permit integration of the expression cassette into the genome of the plant; integrating the expression cassette into the genome of the plant; and overexpressing the nucleotide sequence in the plant.
  • PAD4 phytoalexin deficient 4
  • LSD1 lesion simulating disease 1
  • EDS1 enhanced disease susceptibility 1
  • a method of regulating growth and increasing biomass production in a plant including attenuating the expression of at least two of phytoalexin deficient 4 (PAD4) gene, lesion simulating disease 1 (LSD1 ) gene, and enhanced disease susceptibility 1 (EDS1 ) gene in the plant according to claim 1 , wherein the attenuating step further includes contacting the genome of the plant with an expression cassette comprising an isolated nucleic acid molecule comprising a nucleotide sequence encoding at least two of PAD4, LSD1 , or EDS1 proteins, wherein the nucleotide sequence is operably linked to an expression control sequence that is functional in plants under conditions that permit integration of the expression cassette into the genome of the plant; integrating the expression cassette into the genome of the plant; and attenuating the expression of the nucleotide sequence in the plant.
  • PAD4 phytoalexin deficient 4
  • LSD1 lesion simulating disease 1
  • EDS1 enhanced disease susceptibility 1
  • the method further comprises an amino acid sequence set forth in SEQ ID NO. 1 or SEQ ID NO. 10 that encodes PAD4 protein; an amino acid sequence set forth in SEQ ID NO. 2 or SEQ ID NO. 1 1 that encodes LSD1 protein; and an amino acid sequence set forth in SEQ ID NO. 3 or SEQ ID NO. 12 that encodes EDS1 protein.
  • the method may further include a nucleotide sequence with at least 60% identity to the nucleotide sequence encoding at least two of PAD4, LSD1 , or EDS1 .
  • a transgenic plant having a gene construct including a nucleotide sequence encoding at least two of PAD4, LSD1 , or EDS1 proteins is provided.
  • the nucleotide sequence is operably linked to a promoter such that the nucleotide sequence is overexpressed or attenuated, thereby causing the transgenic plant to exhibit increased biomass, increased stress tolerance, efficient growth development, changes in tissue density, modified cell division to programmed cell death ratio, greater fruit or seed yield, changes in chemical composition, physical, mechanical, and technological properties, , efficient use of water or early appearance of fruit or seed
  • transgenic plant and the non-transgenic plant are cultivated under identical growth conditions and identical stress conditions.
  • the transgenic plant is an annual plant or a perennial plant, wherein the annual plant is selected from a group consisting of Arabidopsis, lettuce, tobacco, soybeans, potato, tomato, canola, rice, corn, and wheat and wherein the perennial plant is a woody plant.
  • the woody plant is a hardwood plant selected from a group consisting of acacia, eucalyptus, hornbeam, beech, mahogany, walnut, oak, ash, willow, hickory, birch, chestnut, poplar, alder, maple, and sycamore, wherein the hardwood plant is a plant of the Populus or Salicaceae groups.
  • biomass is a renewable, potentially sustainable and relatively environmentally benign source of energy;
  • biomass fuels have negligible sulfur content and therefore do not contribute to sulfur dioxide emissions that cause acid rain;
  • combustion of biomass produces less ash than coal combustion and the ash produced can be used as a soil fertilizer;
  • the combustion of agricultural and forestry residues, and municipal solid wastes (MSW) for energy production is an effective use of waste products that reduces the significant problem of waste disposal, particularly in municipal areas;
  • MSW municipal solid wastes
  • the higher growth rate means that the transgenic plant increases its fresh and/or dry biomass at a rate that is more rapid than that of a control plant that has not been genetically engineered in the manner described herein. In other words, the plant with accelerated growth rate needs less time than the control plant to accumulate a particular amount of biomass.
  • the yield (fruit, seeds) production may also be accelerated and/or appear sooner.
  • Perennial plants are plants that have a life cycle which takes longer than 2 years and involves a long juvenile period with vegetative stage only. In contrast, annual plants such as Arabidopsis thaliana have life cycle which is completed within one year. Biomass accumulation in perennial woody plant species begin with cell divisions in the primary and secondary meristems.
  • the primary apical meristems provide cells for root and shoot tip growth.
  • a circumferential secondary meristem called the vascular cambium (VC)
  • VC vascular cambium
  • Wood is produced by cells proliferation in the VC.
  • Cells can divide in 2 ways: (i) periclinal division - cells divide along the longitudinal axis of the stem (responsible for diameter growth of cambium), (ii) anticlinal division - cells divide in radial direction (responsible for circumference growth of tree).
  • plants need to coordinate metabolism and growth between various cells, tissues and organs. Because of their sessile nature, plants are equipped in different defense mechanisms against both biotic and abiotic stresses. In the course of evolution, plants have developed discrete, dynamic and emerging physiological responses that are able to process simultaneously various environmental stimuli and thereby optimize Darwinian fitness under short- and long- term changes in the natural environment (S. Karpinski et al., (1999) Science 284, 654- 657; D. Peak, J. D. West, S. M. Messinger, K.A. Mott, (2004) Proc. Natl. Acad. Sci. USA 101 , 918-22; M. Szechynska-Hebda, J. Kruk, M. Gorecka, B.
  • This biological hardware includes quantum-redox sensing and modifications in photosystem II, e.g.
  • PHYTOALEXIN DEFICIENT 4 encodes a protein that operate upstream of pathogen-induced SA accumulation (Glazebrook et al., 1997; Zhou et al., 1998) and therefore pad4 (phytoalexin deficient 4) mutant demonstrates blocked biosynthesis of salicylic acid triggered by infection with avirulent pathogens (Feys, B. J., Moisan, L. J., Newman, M. A., and Parker, J. E. (2001 ). EMBO J. 20, 5400-541 1 ; Jirage, D., Tootle, T. L, Reuber, T.
  • PAD4 expression can be enhanced by exogenous applications of SA, suggesting that it is regulated by SA-dependent positive feedback (Falk et al., 1999; Jirage et al., 1999; Feys et al., 2001 ).
  • PAD4 was mainly characterized to be involved in defense response to bacteria, insects and other pathogens, its implication in many other processes e.g.
  • PAD4 acts also as a positive regulator of cell death in response to abiotic stresses such as high light or water stress.
  • EDS1 ENHANCED DISEASE SUSCEPTIBILITY1
  • PAD4 PAD4
  • Both proteins share homology to triglyceride lipases and both act upstream of SID2 and EDS5 in the regulation of SA accumulation (Glazebrook, J., Rogers, E. E., and Ausubel, F. M. (1997). Annu. Rev. Genet.
  • Knock-out mutant of LESION SIMULATING DISEASE1 gene belongs to one of the best characterized mutants deregulated in terms of programmed cell death (PCD) in Arabidopsis thaliana (Dietrich et al., 1994; Jabs et al., 1996, Hunt et al. 1997, Rusterucci et al. 2001 , Epple et al. 2003, Mateo et al., 2004; Torres et al. 2005; Muhlenbock et al., 2007; 2008).
  • PCD programmed cell death
  • the Isd1 mutant was initially characterized for its ROS- and SA-dependent uncontrolled spread of PCD that develops under long (>16 h) or continuous photoperiods, supply of superoxide ion (O 2 ⁇ ) or after infection with avirulent pathogen.
  • LSD1 in light acclimation and in restricting pathogen-induced cell death are functionally linked (Mateo A, Muhlenbock P, Rusterucci C, Chi-Chen Ch, Miszalski Z, Karpinska B, Parker JE, Mullineaux PM, Karpinski S (2004). Plant Phys. 136: 2818-2830).
  • the Isd1 phenotype is indicative for failure to stop both the initiation and propagation of PCD, therefore it was named runaway cell death (red).
  • LSD1 function was proposed as a negative regulator of PCD, acting as a ROS rheostat (Dietrich et al., 1994; Jabs et al., 1996) and preventing the pro-death pathway below certain ROS level (Dietrich et al., 1997; Klingenstein et al., 1999).
  • LSD1 , EDS1 and PAD4 constitute a molecular hub, which coordinates the signaling of programmed cell death, light acclimation, and holistic defense responses that are initiated, at least in part, by redox changes of the plastoquinone pool (Muhlenbock P, Szechynska-Hebda M, Plaszczyca M, Baudo M, Mateo A, Mullineaux PM, Parker JE, Karpinska B, Karpinski S., 2008, Plant Cell. 20(9): 2339-56).
  • the LSD1/EDS1/PAD4 hub regulates many aspects of plant defense and acclimation by affecting SA, jasmonate, ethylene, and other yet undefined signal intermediates. All of these features point to an intrinsic activity of LSD1 , EDS1 , and PAD4 in the processing and transduction of redox-derived signals from different subcellular compartments and different plant organs and tissues during a variety of environmental stresses. Laboratory versus field experiments
  • the gene function study based on the mutant phenotype is conventionally performed under highly controlled laboratory conditions. These conditions do not represent the multivariable signals that plants perceive in their natural environment. The phenotype of particular mutant may differ depending on condition. Thus, the study of specific gene function should be studied under multiple environments. So far only a few studies focused on the role of plant genes in natural, field conditions (Kulheim et al. 2002, Ganeteg et al. 2004, and Athanasiou et al. 2010).
  • Arabidopsis thaliana wild type (Ws-0) and five different mutants of the same accession: Isd1, edsl, pad4, eds1/lsd1 and pad4/lsd1 were grown in standard laboratory conditions (9 or 16 h photoperiod, PPFD: 100 ⁇ 25 ⁇ m " V, 50% relative air humidity and temperature day/night: 22/18 °C).
  • Arabidopsis thaliana Ws-0 and mutant plants were also grown in the field at two different locations (Cracow, 50°03'41 "N 19°56'18"E and Warsaw, 52°09'38"N 21 °02'52”E) in four independent experiments during several seasons (June-September, 2006-2010). The smallest experimental field unit was approximately 100 cm 2 where all the six representative genotypes were grown together. For measurements of physiological and morphological parameters, 3- to 4-week-old plants were used, whereas for determination of seed yield, 8-week-old plants were harvested
  • Poplar (Populus tremula x tremuloides) transgenic lines were generated based on the stable plant transformation using Agrobacterium tumefaciens strains containing proper binary plasmids - pCAMBIA or pH7GWIWG2(l). This method is widely used in woody plant species transformation as discussed in Toriyama, et al. (1988) Bio/Technology 6, 1072-1074; Zhang, et al . (1988) Plant Cell Rep. 7, 379- 384; Zhang, et al. (1988) Theor Appl Genet 16, 835-840; Shimamoto, et al. (1989) Nature 338, 274-276; Datta, et al.
  • pH7GWIWG2( ⁇ ) vector based on Gateway technology was a kind gift of the Department of Plant System Biology, Gent University, Belgium (M., Inze, D., Depicker, A. Trends Plant Sci. 2002 May;7(5): 193-195).
  • AtPAD4 T-DNA containing the antisense sequences of poplar LSD1, EDS1 or PAD4 was inserted into binary vectors pCAMBIA and pH7GWIWG2(l).
  • T-DNA regions from vectors were incorporated into plant cell genome together with selective markers conferring resistance to antibiotics (kanamycin and hygromycin).
  • antibiotics kanamycin and hygromycin
  • plants were regenerated from leaf discs via callus formation (Vasil et al., Cell Culture and Somatic Cell Genetics of Plants, Vol I, II and III, Laboratory Procedures and Their Applications, Academic Press, 1984, and Weissbach and Weissbach, Methods for Plant Molecular Biology, Academic Press, 1989).
  • Transgenic lines of P. tremula x tremuloides with significantly diminished expression of poplar orthologues of AtLSDI, AtEDSI and AtPAD4 were selected (3-4 independent lines for each transgen).
  • transgenic poplar lines were grown together with the wild-type control (T89) in a greenhouse under a photoperiod of 18 h and a temperature of 22°C/15°C (day/night). The humidity was ca. 70%. The plants were grown for 4-9 months before harvest. During this time their positions within the glasshouse were altered every 2-3 days and their heights and diameters were measured as described in the results. The transgenic lines were also grown in natural field conditions near Warsaw.
  • Poplar plants exhibited an exponential growth pattern (plant height) up to an approximate height of 220-250 cm or a maximum of up to month 9 in the greenhouse. Height measurements were taken starting from apical part of plant (0.01 m) and subsequently on the levels corresponding to length measured at 0.1 m, 0.2m, 0.3m, 0.5m, 0.7m, 0.9m, 1 m, 1 .5m, 2.0m, 2.5m. Under the above defined growth conditions, stem width exhibited a comparatively linear increase over height. Morphology of main stems (length (cm), volume (cm 3 ) and internodes (cm)), morphology of secondary stems (number and length (cm)) and leaves area (cm 2 ) were determined.
  • leaf area (cm 2 ), main stem volume (cm 3 ) and internodes (cm) were determined for 4-9-month-old poplar wild type plants (t89) and three different transgenic lines: pad4, Isd1, edsl in l-lll greenhouse experiments from two different localizations (Stockholm and Warsaw). Values represent mean from different sublines (four for pad4, one for Isd1 and two for edsl) and replications (3-6 plant within sublines). leaf area main stem volume internodes length exp.
  • Diameter of main stems was determined for 4 to 9-month-old poplar wild type plants (t89) and three different transgenic lines: pad4, Isd1, edsl in three (l-lll) greenhouse experiments. Diameter of main stems were measured starting from apical part of plant (0.01 m) and subsequently on the levels corresponding to length measured at 0.1 m, 0.2m, 0.3m, 0.5m, 0.7m, 0.9m, 1 m, 1 .5m, 2.0m and 2.5m. Values represent mean from different sublines (four for pad4, one for Isd1 and two for edsl) and replications (3-6 plant within sublines). exp 0.01 m 0.1 m 0.2m 0.3m 0.5m 0.7m 0.9m 1 m 1.5m 2.0m 2.5m t89 I 1.27 1.53 2.40 2.87 3.73 5.00 5.00 - - - - -
  • Water use was determined as water used during 4 weeks of plant growth in closed system. Plants were grown in 50 ml tubes filled with perlite and soil in 1 :1 proportion and 35 ml of water. Seeds were placed in a hole (0 ca. 1 .5 mm) made in the cap. After germination, the system was weighed. The water lost corresponds to water transpired by plants.
  • Poplar leaves were cut from plants and immediately transferred into 1 ml Falcon tubes filled with water. Leaves were kept in greenhouse conditions (experiment I) or in laboratory conditions (experiments ll-IV; in 22 °C, ca. 60% humidity, light intensity - 100 ⁇ m 2 s "1 ). Tubes were weighted (or water volume changes were measured) at 0 and 24 hours of incubation. Leaves area was measured after experiment with ImageJ 3.0 software. Stomatal density
  • Water use and stomata number was calculated for fully developed leaves of 8-month-old poplar stem of wild type plants (t89) and three different transgenic lines: pad4, Isd1, edsl were measured. Leaves were collected from part of plant about 1 m above ground. Values represent means from replications (3-6 plant within sublines) and 3-5 places o leaf surface.
  • LSD1 poplar gene modifications resulted in the changes of water use similarly to Isd1 Arabidopsis plants. Moreover, transgenic line pad4 and edsl had also reduced water consumption. Although lower transpiration rate changes were observed in comparison to wild type for all transgenic lines, the relation between stomata number and water use was determined only in case of Isd1. Therefore, the internal control of stomata opening seems to be more important factor than total stomata number.
  • Percentage of wood tissue in cross section of 8-month-old poplar stem was determined via microscopic analysis.
  • Cross sections were prepared by stem cutting at 0.1 m 0.2m, 0.5m and 1 m starting from apical part of the stem. Calculation of cross section area of wood and other tissue was preformed with ImageJ 3.0 software.
  • the three points bending flexural test preformed with hydraulic power supply provides values for the characteristic of elasticity of wood: maximal force (N) required to wood rupture and maximal bending of wood.
  • the maximum force capacity was 0.6 kN and the speed of loading was 1 mm/min in the flexural test.
  • Wood sample were cut from air-dried stem in the form 5 (radial) mm ⁇ 5 (tangential) mm x 30 (longitudinal) mm pieces.
  • Thermal analysis techniques include: simultaneous differential scanning calorimetry (DSC), thermogravimetry (TG) combined with quadruple mass spectrometry (QMS). It is a useful tool to obtain complete information on physical and chemical properties of plant cell wall.
  • Wood density (g/cm 3 ), wood abilities to water binding (weight changes (%) and wood swelling (%) during tissue hydratation, dimensional changes under heating (wood shrinkage)) and mechanical properties of wood were determined for 8-month- old poplar wild type plants (t89) and three different transgenic lines: pad4, Isd1, eds1. Values represent means from different sublines (four for pad4, one for Isd1 and two for edsl) and replications (3-6 plant within sublines). wood maximal dilatometrical maximal weight wood
  • the density of poplar lines varied from 0.420 g/cm 3 for Isd1 to 0.441 g/cm 3 for pad4 (Table 10).
  • the highest wood density of pad4 confirms the compact wood structure consisting higher number of small cells per volume unit. Presence of tracheids in Isd1 results in similar to wild type wood density. Higher tissues compaction determines structure stability e.g. lower volumetric swelling during wood tissue hydratation and higher parameters of dilatometrical test (wood shrinking). The small dimensional changes during the shrinking and swelling indicate that transgenic line pad4 was able to retain less "free" (non-chemically bounded water) water in wood tissue than wild-type t89.
  • the fiber saturation point is defined as that moisture content at which all of the liquid water, essentially sap, has been removed from the cell cavities but the cell walls are still saturated with adsorbed water. Therefore small pad4 cells form compact wood, what prevent filling tissue with water. However this process occurred in more loose structure of tissue of wild type plant.
  • Hemicelluloses exhibit lower molecular weights than cellulose and decompose at temperatures of 200-260°C. They are a mixture of various polymerized monosaccharides such as glucose, mannose, galactose, xylose, arabinose, 4-O- methyl glucuronic acid and galacturonic acid residues. Different thermal behaviors (higher mass loss and higher energetic effects, indicate different hemicellulose composition and the higher amount of easily depredated components for transgenic lines, when they are compared to wild type (with exception of edsl).
  • Cellulose degradation occurs at 240-350°C and the reaction is complete at 360°C.
  • Cellulose forms long chains that are bonded to each other by a long network of hydrogen bonds.
  • Groups of cellulose chains twist in space to make up ribbon like microfibril sheets, which are the basic construction units for a variety of complex fibers.
  • These microfibrils form composite tubular structures that run along a longitudinal tree axis.
  • the crystalline structure resists thermal decomposition better than hemicelluloses.
  • Amorphous regions in cellulose exist that contain waters of hydration, and free water is present within the wood. This water, when rapidly heated, disrupts the structure by a steam explosion-like process prior to chemical dehydration of the cellulose molecules.
  • Lignin is more difficult to dehydrate than cellulose or hemicelluloses and decomposes when heated at 300-600°C.
  • the mass loss of lignins suggest their slightly higher level in pad4 and Isd2, however their decomposition was the most exothermic for pad4 (area below DSC curve). It results also in the highest total mass loss.
  • a method of producing a perennial plant comprising the incorporation of a heterologous nucleic acid encoding a PAD4, LSD1 , EDS1 polypeptides alone or in combinations, into a perennial plant cell by means of transformation and; regenerating a perennial plant from one or more transformed cells.
  • a woody plants produced by such a method may accumulate biomass more effectively than control plants.
  • the nucleic acid recombines with the cell genome nucleic acid such that it is stably incorporated therein.
  • a PAD4, LSD1 , EDS1 polypeptide may have the amino acid sequence of SEQ ID NO: 1 to 3 (respectively) or may be a fragment or variant of the SEQ ID NO: 1 to 3 sequences, which retains PAD4, LSD1 , EDS1 activity (respectively), including for example induction of biomass accumulation, and regulation of cell division and programmed cell death.
  • a PAD4, LSD1 , EDS1 polypeptides which are variants of SEQ ID NO: 1 to 3 may comprise an amino acid sequences which shares greater than 60% sequences identity with the amino acid sequences of SEQ ID NO: 1 to 3 (respectively), preferably greater than 65%, greater than 70%, greater than 80%, greater than 90% or greater than 95%.
  • a PAD4, LSD1 , EDS1 polypeptides which are variants of SEQ ID NO: 1 to 3 may share greater than 60% sequences similarity with the amino acid sequences of SEQ ID NO: 1 to 3 (respectively), preferably greater than 65%, greater than 70%, greater than 80%, greater than 90% or greater than 95%. Similarity allows for "conservative variation", i.e. substitution of one hydrophobic residue such as isoleucine, valine, leucine or methionine for another or the substitution of one polar residue for another, such as arginine for lysine, glutamic for aspartic acid, or glutamine for asparagine.
  • Particular amino acid sequences variants may differ from a known PAD4, LSD1 , EDS1 polypeptides sequences as described herein by insertion, addition, substitution or deletion of 1 amino acid, 2, 3, 4, 5-10, 10-20 20-30, 30-50, or more than 50 amino acids.
  • the Arabidopsis PAD4, LSD1 , EDS1 polypeptides sequences have the database accession numbers NP_19081 1 .1 , and NP_001 154257.1 and NP_190392.1 , respectively.
  • Nucleic acids as described herein may be wholly or partially synthetic. In particular, they may be recombinant in that nucleic acid sequences which are not found together in nature (do not run contiguously) have been ligated or otherwise combined artificially. Alternatively they may have been synthesized directly e.g. using an automated synthesizer.
  • a nucleic acid encoding a PAD4, LSD1 , EDS1 polypeptides may comprise or consist of the nucleotide sequences of SEQ ID NO: 4-6 (respectively) or may be a variant or fragment of the SEQ ID NO: 4-6 sequences (respectively).
  • a variant sequences may be a mutant, homologue, or allele of the SEQ ID NO: 4-6 sequences and may differ from the sequences of SEQ ID NO: 4-6 by one or more of addition, insertion, deletion or substitution of one or more nucleotides in the nucleic acid, leading to the addition, insertion, deletion or substitution of one or more amino acids in the encoded polypeptide.
  • addition, insertion, deletion or substitution of one or more nucleotides in the nucleic acid leading to the addition, insertion, deletion or substitution of one or more amino acids in the encoded polypeptide.
  • changes to the nucleic acid that make no difference to the encoded amino acid sequences are included.
  • a nucleic acid encoding a PAD4, LSD1 , EDS1 polypeptides which has a nucleotide sequences which is a variant of the SEQ ID NO: 4-6 sequences (respectively) may comprise a sequences having at least 60% sequences identity with the nucleic acid sequences of SEQ ID NO: 3 to 6, than 65%, greater than 70%, greater than 80%, greater than 90% or greater than 95%. Sequences identity is described above.
  • a fragment or variant may comprise a sequences which encodes a functional PAD4, LSD1 , EDS1 polypeptides i.e. a polypeptides which retains one or more functional characteristics of the polypeptide encoded by the wild-type PAD4, LSD1 , EDS1 genes, for example, the ability to stimulate biomass accumulation in a perennial plant (woody plant).
  • a nucleic acid encoding a PAD4, LSD1 , EDS1 polypeptides which have a nucleotide sequences which is a variant of the SEQ ID NO: 4-6 sequences (respectively) may selectively hybridize under stringent conditions with the nucleic acid sequences of SEQ ID NO: 4-6 (respectively) or the complement thereof.
  • an isolated nucleic acid including a nucleotide sequence which encodes an amino acid sequence that has at least 80% amino acid sequence identity with the amino acid sequence shown in SEQ ID NO. 1 is provided.
  • the isolated nucleic acid encodes the amino acid sequence shown in SEQ ID NO: 1 to 3 and 10a to 12.
  • the isolated nucleic acid further includes a sequence having at least 80% sequence identity with the nucleic acid sequence of SEQ ID NO: 4 to 6 and 7 to 9
  • a method for increasing the growth, photosynthesis, development, biomass production and water use efficiency of a plant including the steps of genetically engineering said plant to contain and over-express or silence at least one preferably two or all three functional gene product of Phytoalexin Deficient 4 (PAD4), Enhanced Disease Susceptibilityl (EDS1 ) and Lesion Simulating Diseasel (LSD1 ) is provided.
  • PAD4 Phytoalexin Deficient 4
  • EDS1 Enhanced Disease Susceptibilityl
  • LSD1 Lesion Simulating Diseasel
  • the method further provides PAD4, EDS1 and LSD1 polypeptides, which alone or in combinations includes an amino acid sequence that shares greater than 60% sequence identity with the amino acid sequence of SEQ ID NO: 1 to 3 and 10 to 12.
  • the method further provides a nucleic acid molecule that includes a sequence having at least 60% sequence identity with the nucleic acid sequence of SEQ ID NO: 4-6 and 7 to 9.
  • the biomass of a plant is increased such that either wet biomass or dry biomass, or production of fruit, seeds, or all, of the plant is increased or may appear sooner.
  • the water use efficiency of the plant is increased such that the plant exhibits the ability to grow in a manner that is more successful than comparable control plants that are not genetically engineered, even under circumstances or conditions of stress for the plant with respect to growth rate, wet and dry biomass, color, maturation, fruit production.
  • the plant may be selected from the group consisting of annual plant Arabidopsis, lettuce, tobacco, soybeans, potato, tomato, canola, rice, corn, wheat and perennial plant, wherein the perennial plant may be a woody plant such as a hardwood plant, conifer or a fruit bearing plant or selected from the group consisting of cotton, bamboo and rubber plants.
  • the hardwood plant may be selected from the group consisting of acacia, eucalyptus, hornbeam, beech, mahogany, walnut, oak, ash, willow, hickory, birch, chestnut, poplar, alder, maple and sycamore.
  • the hardwood plant may be a plant of the Populus or Salicaceae groups.
  • the conifer may be selected from the group consisting of cypress, Douglas fir, fir, sequoia, hemlock, cedar, juniper, larch, pine, redwood, spruce and yew.
  • the fruit bearing plant may be selected from the group consisting of apple, plum, pear, banana, orange, kiwi, lemon, cherry, grapevine and fig.
  • the genetically engineered plant may cause an increase in a level of hydrogen peroxide and/or superoxide ion and/or salicylic acid and/or auxins and/or ethylene and changes the stomata density in the plant.
  • the genetically engineered plant may not cause an increase in a level of hydrogen peroxide and/or superoxide ion and/or salicylic acid and/or auxins and/or ethylene and changes the stomata density in said plant.
  • the genetically engineered plant may cause a change tissue density and compaction via modification of cell division/programmed cell death processes in the plant, and/or a change in chemical composition, physical, mechanical and technological properties of cell walls in the plant.
  • a transgenic plant wherein the transgenic plant is genetically engineered to contain and over-express or silence at least one, preferably two or three functional gene product of Phytoalexin Deficient 4 (PAD4), Enhanced Disease Susceptibilityl (EDS1 ) and Lesion Simulating Diseasel (LSD1 ).
  • PAD4 Phytoalexin Deficient 4
  • EDS1 Enhanced Disease Susceptibilityl
  • LSD1 Lesion Simulating Diseasel
  • the transgenic plant of claim 8 at least one, preferably two or three functional gene product is selected from the group consisting of Phytoalexin Deficient 4 (PAD4), Enhanced Disease Susceptibilityl (EDS1 ) and Lesion Simulating Diseasel (LSD1 ).
  • the transgenic plant may exhibit increased biomass, wherein either wet biomass or dry biomass, either wet biomass or dry biomass, or production of fruit, seeds, or all, of said plant is increased or may appear sooner, or both is increased.
  • the transgenic plant may wherein exhibit increased stress tolerance and increased water use efficiency.
  • the transgenic plant may also exhibit the ability to grow in a manner that is more successful than comparable control plants that are not genetically engineered, even under circumstances or conditions of stress for the plant with respect to growth rate, wet and dry biomass, color, maturation, fruit production.
  • the transgenic plant may be of a type selected from the group consisting of annual plant Arabidopsis, lettuce, tobacco, soybeans, potato, tomato, canola, rice, corn, wheat and perennial plant, wherein the perennial plant is a woody plant.
  • the woody plant may be a hardwood plant, a conifer, a fruit bearing plant, or may be selected from a group consisting of cotton, bamboo and rubber plants.
  • the hardwood plant may be selected from the group consisting of acacia, eucalyptus, hornbeam, beech, mahogany, walnut, oak, ash, willow, hickory, birch, chestnut, poplar, alder, maple and sycamore.
  • the hardwood plant may be a plant of the Populus or Salicaceae groups.
  • the conifer may be selected from the group consisting of cypress, Douglas fir, fir, sequoia, hemlock, cedar, juniper, larch, pine, redwood, spruce and yew.
  • the fruit bearing plant may be selected from the group consisting of apple, plum, pear, banana, orange, kiwi, lemon, cherry, grapevine and fig.
  • the transgenic plant exhibits an increase in a level of hydrogen peroxide and/or superoxide ion and/or salicylic acid and/or auxins and/or ethylene acid and changes the stomata density or that it does not exhibit an increase in a level of hydrogen peroxide and/or superoxide ion and/or salicylic acid and/or auxins and/or ethylene and changes the stomata density.
  • the transgenic plant may include changed tissue density and modified cell division/programmed cell death ratio processes and the transgenic plant may include a changed the chemical composition, physical, mechanical and technological properties of cell wall.

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  • Genetics & Genomics (AREA)
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  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
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  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
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  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
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  • Medicinal Chemistry (AREA)
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  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
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Abstract

L'invention concerne une plante transgénique ayant une construction de gène incluant une séquence de nucléotides incluant au moins deux parmi le gène 4 déficient en phytoalexine (PAD4), le gène 1 de la maladie simulant une lésion (LSD1), et le gène 1 de sensibilité accrue à la maladie (EDS1) et un procédé de régulation de la croissance et d'augmentation de la production de la biomasse chez la plante par surexpression ou atténuation de l'expression d'au moins deux parmi le gène PAD4, le gène LSD1, et le gène EDS1 dans la plante. La plante transgénique montre une biomasse accrue, une tolérance au stress accrue, un développement de croissance efficace, un rendement en fruit et graines plus grand et/ou une utilisation efficace de l'eau ou une apparition précoce des fruits ou des graines par rapport à une plante non transgénique qui ne surexprime pas ou n'atténue pas au moins deux parmi les gènes PAD4, LSD1, et EDS1, quand la plante transgénique et la plante non transgénique sont cultivées sous des conditions de croissance identiques et des conditions de stress identiques.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110923241A (zh) * 2019-11-05 2020-03-27 南京农业大学 水稻控制通气组织形成的关键基因OsLSD1.1在减少甲烷排放上的应用

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992014828A1 (fr) 1991-02-14 1992-09-03 Svalöf Ab Procede de transformation genetique d'organes tissulaires provenant de plantes monocotyledones
WO2008125983A2 (fr) 2007-04-12 2008-10-23 Swetree Technologies Ab Procédés d'augmentation de la croissance de plantes
US20090048312A1 (en) 2007-08-16 2009-02-19 The University Of Chicago Plant pathogen resistance
US20090138981A1 (en) 1998-09-22 2009-05-28 Mendel Biotechnology, Inc. Biotic and abiotic stress tolerance in plants
US20100115658A1 (en) 2007-02-01 2010-05-06 Enza Zaden Beheer B.V. Disease Resistant Plants
US20100223690A1 (en) 2009-01-02 2010-09-02 Washington State University Compositions and methods for modulating plant disease resistance and immunity

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009114733A2 (fr) * 2008-03-13 2009-09-17 Ceres, Inc. Séquences nucléotidiques et polypeptides correspondants conférant un taux de croissance et une biomasse modulés dans des plantes cultivées dans des conditions salines et oxydantes

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992014828A1 (fr) 1991-02-14 1992-09-03 Svalöf Ab Procede de transformation genetique d'organes tissulaires provenant de plantes monocotyledones
US20090138981A1 (en) 1998-09-22 2009-05-28 Mendel Biotechnology, Inc. Biotic and abiotic stress tolerance in plants
US20100115658A1 (en) 2007-02-01 2010-05-06 Enza Zaden Beheer B.V. Disease Resistant Plants
WO2008125983A2 (fr) 2007-04-12 2008-10-23 Swetree Technologies Ab Procédés d'augmentation de la croissance de plantes
US20090048312A1 (en) 2007-08-16 2009-02-19 The University Of Chicago Plant pathogen resistance
US20100223690A1 (en) 2009-01-02 2010-09-02 Washington State University Compositions and methods for modulating plant disease resistance and immunity

Non-Patent Citations (35)

* Cited by examiner, † Cited by third party
Title
"Laboratory Procedures and Their Applications", 1984, ACADEMIC PRESS
CAO ET AL., PLANT CEH REP., vol. 11, 1992, pages 585 - 591
CHEILA G.; MOTHE C; LARA C. DE MIRANDA, THERM ANAL CALORIM, vol. 97, 2009, pages 661 - 665
CHRISTOU ET AL., BIO/TECHNOLOGY, vol. 9, 1991, pages 957 - 962
D. PEAK; J. D. WEST; S. M. MESSINGER; K.A. MOTT, PROC. NATL. ACAD. SCI. USA, vol. 101, 2004, pages 918 - 22
DATTA ET AL., BIO/TECHNOLOGY, vol. 8, 1990, pages 736 - 740
D'HALLUIN ET AL., PLANT CELL, vol. 4, 1992, pages 1495 - 1505
FEYS, B. J.; MOISAN, L. J.; NEWMAN, M. A.; PARKER, J. E., EMBO J., vol. 20, 2001, pages 5400 - 5411
FROMM ET AL., BIO/TECHNOLOGY, vol. 8, 1990, pages 833 - 839
GLAZEBROOK, J.; ROGERS, E. E.; AUSUBEL, F. M., ANNU. REV. GENET, vol. 31, 1997, pages 547 - 569
GORDON-KAMM ET AL., CELL, vol. 2, 1990, pages 603 - 618
JIRAGE, D.; TOOTLE, T. L.; REUBER, T. L.; FROST, L. N.; FEYS, B. J.; PARKER, J. E.; AUSUBEL, F. M.; GLAZEBROOK, J., PROC. NATL. ACAD. SCI. USA, vol. 96, 1999, pages 13583 - 13588
KOZIEL ET AL., BIOTECHNOLOGY, vol. 11, 1993, pages 194 - 200
LI ET AL., PLANT CELL REP., vol. 12, 1993, pages 250 - 255
M. SZECHYNSKA-HEBDA; J. KRUK; M. G6RECKA; B. KARPINSKA; S. KARPINSKI, PLANT CELL, vol. 22, 2010, pages 2200 - 2218
M., INZE; D., DEPICKER, A. TRENDS PLANT SCI, vol. 7, no. 5, May 2002 (2002-05-01), pages 193 - 195
MATEO A; MOHLENBOCK P; RUSTERUCCI C; CHI-CHEN CH; MISZALSKI Z; KARPINSKA B; PARKER JE; MULLINEAUX PM; KARPINSKI S, PLANT PHYS, vol. 136, 2004, pages 2818 - 2830
MUHIENBOCK P; SZECHYNSKA-HEBDA M; PLASZCZYCA M; BAUDO M; MATEO A; MULLINEAUX PM; PARKER JE; KARPINSKA B; KARPINSKI S., PLANT CELL, vol. 20, no. 9, 2008, pages 2339 - 56
NILSSON, O. ET AL., TRANSGENIC RESEARCH, vol. 1, 1992, pages 209 - 220
PENG ET AL., INTERNATIONAL RICE RESEARCH INSTITUTE, MANILA, 1991, pages 563 - 574
RAGAUSKAS ET AL., SCIENCE, vol. 113, 2006, pages 484 - 489
RATHORE ET AL., PLANT MOLECULAR BIOLOGY, vol. 21, 1993, pages 871 - 884
S. KARPINSKI ET AL., SCIENCE, vol. 284, 1999, pages 654 - 657
S. KARPINSKI; M. SZECHYNSKA-HEBDA, PLANT SIGNAL. BEHAV, vol. 11, 2010, pages 1391 - 1394
SHIMAMOTO ET AL., NATURE, vol. 338, 1989, pages 274 - 276
SOMERS ET AL., BIO/TECHNOLOGY, vol. 10, 1992, pages 1589 - 1594
STRAUS MR; RIETZ S; VER LOREN VAN THEMAAT E; BARTSCH M; PARKER JE, PLANT J, vol. 62, no. 4, 2010, pages 628 - 40
TORIYAMA ET AL., BIO/TECHNOLOGY, vol. 6, 1988, pages 1072 - 1074
VASIL ET AL., CELL CULTURE AND SOMATIC CELL GENETICS OF PLANTS, vol. II
VASIL, I. K., PLANT MOLECULAR BIOLOGY, vol. 25, 1994, pages 925 - 937
WALTERS ET AL., PLANT MOLECULAR BIOLOGY, vol. 18, 1992, pages 189 - 200
WEEKS ET AL., PLANT PHYSIOLOGY, vol. 102, 1993, pages 1077 - 1084
WEISSBACH; WEISSBACH: "Methods for Plant Molecular Biology", 1989, ACADEMIC PRESS
ZHANG ET AL., PLANT CELL REP, vol. 7, 1988, pages 379 - 384
ZHANG ET AL., THEOR APPL GENET, vol. 16, 1988, pages 835 - 840

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110923241A (zh) * 2019-11-05 2020-03-27 南京农业大学 水稻控制通气组织形成的关键基因OsLSD1.1在减少甲烷排放上的应用
CN110923241B (zh) * 2019-11-05 2022-10-14 南京农业大学 水稻控制通气组织形成的关键基因OsLSD1.1在减少甲烷排放上的应用

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