CN115125259A - Dwarf 20 protein, coding gene and application thereof - Google Patents
Dwarf 20 protein, coding gene and application thereof Download PDFInfo
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- CN115125259A CN115125259A CN202110324876.1A CN202110324876A CN115125259A CN 115125259 A CN115125259 A CN 115125259A CN 202110324876 A CN202110324876 A CN 202110324876A CN 115125259 A CN115125259 A CN 115125259A
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Abstract
The invention relates to a corn gene 'dwarf 20' and a coding protein thereof. The invention also researches the application of the gene.
Description
Technical Field
The invention relates to a maize gene 'dwarf 20' and a protein coded by the maize gene. The invention also relates to a method for obtaining the 'dwarf 20' transgenic plant and the function of the gene and the protein in the growth and development of the plant.
Background
"dwarf 20" encodes an IRE/NPH/phosphoinositide-dependent/ribosomal protein S6 kinase. The gene is specifically expressed in maize anther. No relevant research report of the gene in the corn is found at present.
Homologous gene AT1G51170.1 in Arabidopsis encodes an active AGC-VIII protein kinase that interacts with the putative transcription factor ATS, AGC2-3 regulates planar growth during ovule integumentary development. Mutants exhibit ectopic growth of filaments and petals, as well as abnormal embryogenesis. Balaji et al [1] It has been demonstrated that the AGC VIII protein kinase UNICORN (UCN) of Arabidopsis maintains planar growth by inhibiting the formation of ectopic multicellular processes in several floral tissues, including integuments. UCN encodes an AGC-active kinase, controlled by direct interaction with ABERRANT TESTA SHAPE (ATS) which is one of the transcription factor KANADI (KAN) family members, and inhibition of its activity (Balaji et al) [2] ，2013)。
The homologous gene LOC _ Os08g39460.1 in rice encodes an AGC kinase, Ana et al found important roles in regulating plant growth, immunity and cell death, and a link to stress-induced mitogen-activated protein kinase signaling cascade (Ana et al) [3] ,2012). For example, Matsui et al studied the interaction of the AGC kinase Osoxi1 with OsPti1a, and showed that the interaction between the two positively regulated the basal disease resistance of rice. In eukaryotes, AGC kinase family proteins are regulated by 3-phosphoinositide-dependent protein kinase 1(Pdk 1). Therefore, the research shows that OsPdk1 can be used for carrying out cascade phosphorylation through Osoxi1-OsPti1a to positively regulate and control the basic disease resistance of rice (Matsui et al) [4] ，2010)。
Therefore, the gene is probably related to the growth and development of plant reproductive organs and is involved in plant stress resistance pathways such as disease resistance and the like, and further research and confirmation are needed for the specific function of the gene in the corn.
Disclosure of Invention
The following definitions and methods serve to better define the invention and to guide those skilled in the art in the practice of the invention. Unless otherwise indicated, terms should be understood according to conventional usage by those of ordinary skill in the art.
The invention relates to a corn gene short 20, the gene sequence source is B73, the B73_ RefGen _ v4 version code is Zm00001d031426, B73_ RefGen _ v3 version code is: GRMZM2G 050427.
The "short 20" gene contains an exon, presents a transcript, translates 442 amino acids, encodes a kinase protein, and comprises 2 domains: protein kinase (8-381), AGC-kinase C-terminal (382-442).
The amino acid sequence of "short 20" protein (SEQ ID NO:1), consists of 442 amino acid residues. The coding region CDS (coding sequence) of the "dwarf 20" gene is shown as the nucleotide sequence SEQ ID NO 2. The DNA sequence of the "short 20" gene is shown in SEQ ID NO 3.
First, in a first aspect, the present invention relates to an isolated nucleic acid molecule, characterized in that it comprises a sequence selected from the group consisting of:
1) 3 or a complementary sequence thereof;
2) a sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the nucleotide sequence set forth in SEQ ID NO. 3;
3) a nucleotide sequence that hybridizes under stringent conditions to SEQ ID NO. 3; or
4) 3 by deletion, substitution, insertion and/or addition of one or more nucleotides.
In some embodiments, the isolated nucleic acid molecule comprises the nucleotide sequence set forth in SEQ ID NO. 3; in a specific embodiment, the isolated nucleic acid molecule has the nucleotide sequence set forth in SEQ ID NO 3.
In some embodiments, the CDS sequence corresponding to the nucleic acid molecule is set forth in SEQ ID NO 2.
In another aspect, the invention also provides an isolated nucleic acid molecule comprising a sequence selected from the group consisting of:
1) a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO. 1;
2) a nucleotide sequence encoding an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence set forth in SEQ ID NO. 1; or
3) 1 by deletion, substitution, insertion and/or addition of one or more amino acid residues.
In another aspect, the present invention also provides an isolated polypeptide comprising an amino acid sequence selected from the group consisting of:
1) 1, SEQ ID NO;
2) an amino acid sequence encoded by a nucleic acid molecule as described above;
3) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence shown in SEQ ID NO. 1; or
4) 1, amino acid sequence derived by deleting, substituting, inserting and/or adding one or more amino acid residues in the sequence shown in SEQ ID NO.
In some embodiments, the polypeptide of the invention comprises the amino acid sequence set forth in SEQ ID NO 1; in a specific embodiment, the polypeptide of the invention has an amino acid sequence as shown in SEQ ID NO. 1.
In another aspect, the present invention also provides a recombinant vector comprising an isolated nucleic acid molecule as defined above.
In another aspect, the invention also provides a host cell comprising an isolated nucleic acid molecule as defined above, or comprising an isolated polypeptide as defined above, or comprising a recombinant vector as defined above.
In another aspect, the invention also provides a transgenic plant comprising an isolated nucleic acid molecule as defined above, or comprising an isolated polypeptide as defined above, or comprising a recombinant vector as defined above.
In some embodiments, the transgenic plant is a monocot or a dicot; in a particular embodiment, the transgenic plant is preferably a crop plant.
In some specific embodiments, the plant is selected from the group consisting of maize (Zea mays), oilseed rape (Brassica napus), turnip (Brassica rapa), mustard (Brassica juncea), alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cereale), Sorghum (Sorghum biocolor, Sorghum vulgare), pearl millet (Pennisetum glaucum), glutinous rice (Panicum Miliaceum), millet (Setaria italica), finger millet (Eleusines corana), sunflower (Helianthus annuus), safflower (Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), peanut (Arachis hypocoea), cotton (Gossypium barbadense), Gossypium hirsutum (Gossypium hirsutum), sweet potato (Ipomoea batatus), cassava (Manihot esculenta), beet (Beta vulgaris), sugarcane (Saccharum spp.), oat (Avena sativa), barley (Hordeum vulgare), and Arabidopsis thaliana (Arabidopsis thaliana); in a preferred embodiment, the plant of the invention is selected from arabidopsis thaliana or maize.
In some embodiments, the transgenic plant of the invention has an altered trait compared to a plant or wild type plant not having the corresponding transgene, wherein the altered trait is selected from the group consisting of yield, plant height, ear length of ear, ear thickness, bald tip length, ear row number, normal ear number, grain width, grain thickness, cell grain weight, grain moisture, hundred grain weight, internode spacing, and the like; e.g. with increased yield, reduced plant height, increased hundred grain weight, reduced internode spacing, etc.
In another aspect, the present invention also provides a method for producing a transgenic plant having an altered trait, the method comprising introducing into a plant a nucleic acid molecule as defined above, or a recombinant vector as defined above, or a host cell as defined above, to obtain a corresponding transgenic plant; the transgenic plant has altered traits as compared to a plant not transformed accordingly or a wild type plant, wherein the altered traits are selected from yield, plant height, ear length of ear, ear thickness, bald tip length, ear row number, normal ear number, grain width, grain thickness, cell grain weight, grain moisture, grain weight per hundred, internode spacing; wherein said alteration of a trait has e.g. increased yield, decreased plant height, increased weight per hundred particles, decreased internode spacing, etc.
In another aspect, the present invention also provides a method of modulating plant growth or plant stress resistance, which comprises transforming a plant with a recombinant vector as defined above, or transfecting a plant with a host cell as defined above; wherein the transgenic plant has increased yield as compared to a plant not correspondingly transformed/transfected or as compared to a wild type plant.
In another aspect, the invention relates to the use of a nucleic acid molecule as defined above or a polypeptide as defined above or a recombinant vector as defined above for the modulation of plant growth or plant stress resistance.
In some embodiments, the above-described plant or transgenic plant is a monocot or dicot; in a particular embodiment, the plant is preferably a crop plant.
In a preferred embodiment, the plant is selected from the group consisting of maize (Zea mays), oilseed rape (Brassica napus), turnip (Brassica rapa), mustard (Brassica juncea), alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cereale), Sorghum (Sorghum bicolor, Sorghum vulgare), pearl millet (Pennisetum glaucum), yellow rice (Panicum militicum), millet (Setaria italica), finger millet (Eleusines corana), sunflower (Helnthhus annuus), safflower (Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), peanut (Arachis hypocoea), cotton (Gossypium barbadense), Gossypium hirsutum (Gossypium hirsutum), sweet potato (Ipomoea batatus), cassava (Manihot esculenta), beet (Beta vulgaris), sugarcane (Saccharum spp.), oat (Avena sativa), barley (Hordeum vulgare), and Arabidopsis thaliana (Arabidopsis thaliana); more preferably, the plant of the invention is selected from arabidopsis thaliana or maize.
As used herein, the term "plant" includes whole plants, transgenic plants, meristematic tissues, shoot organs/structures (e.g., leaves, stems and tubers), roots, flowers and floral organs/structures (e.g., bracts, sepals, petals, stamens, carpels, anthers and ovules), seeds (including embryos, endosperms and seed coats) and fruits (mature ovaries), plant tissues (e.g., vascular tissue, basal tissue, etc.) and cells (e.g., guard cells, egg cells, pollen, mesophyll cells, etc.) and progeny thereof. The class of plants useful in the present invention is generally as broad as the class of higher or lower plants that can be treated by transformation and breeding techniques, including angiosperms (monocotyledonous and dicotyledonous plants), gymnosperms, ferns, equisetum, gymnosperms, lycopodium, bryophytes, and multicellular algae.
As used herein, the term "transgene" refers to a nucleotide molecule that is artificially introduced into the genome of a host cell. Such a transgene may be heterologous to the host cell. As used herein, "transgenic plant" refers to a plant whose genome has been altered by the stable integration of recombinant DNA. Transgenic plants include plants regenerated from the originally transformed plant cells and progeny transgenic plants from subsequent generation or crossing of the transgenic plants.
As used herein, "control plant" refers to a plant that does not contain recombinant DNA that alters a trait. Control plants are used to identify and screen transgenic plants having altered traits. Suitable control plants may be non-transgenic plants which are used to generate the parental lines of the transgenic plants, e.g. wild-type plants which do not contain the corresponding recombinant DNA. Suitable control plants may also be transgenic plants containing recombinant DNA conferring other traits, for example, transgenic plants with enhanced herbicide tolerance.
As used herein, a "trait" is a physiological, morphological, biochemical or physical characteristic of a plant or a particular plant material or cell. In some cases, the characteristic is visible to the human eye, such as seed or plant size, or can be measured by biochemical techniques (such as detecting protein, starch, metabolite, or oil content of seed or leaf) or by observing metabolic or physiological processes (such as by measuring tolerance to water deprivation or specific salt or sugar concentrations) or by measuring the expression level of one or more genes (by using Northern Blotting, Western Blotting, RT-PCR, microarray gene expression assays, or reporter gene expression systems) or by agricultural observations (such as hyperosmotic stress tolerance and yield). Any technique can be used to measure the amount, comparative level, or difference of any selected chemical compound or macromolecule in the transgenic plant.
A trait of particular economic interest is increased yield. Yield is generally defined as the measurable production of economic value from a crop. This may be defined in terms of quantity and/or quality. Yield is directly dependent on several factors, e.g., the number and size of organs, plant architecture (e.g., number of branches), seed production, leaf senescence, etc. Root development, nutrient uptake, stress tolerance, etc. may also be important factors in determining yield. In the case of crop plants, such as corn, rice, etc., yield means the amount of grain harvested, and particle size and grain weight traits are critical in determining yield. Therefore, increasing the particle size and the particle weight are important for increasing the yield of crops.
As used herein, the term "nucleic acid" refers to any polymer comprising deoxyribonucleotides or ribonucleotides, including but not limited to modified or unmodified DNA, RNA, and the length thereof is not limited in any way. For the nucleic acid used to construct the recombinant construct, preferably DNA, it is more stable and easier to manipulate than RNA.
The term "DNA" refers to a double-stranded DNA molecule of genomic or synthetic origin, i.e., a polymer of deoxyribonucleotide bases or a nucleotide molecule, reading from the 5 'end (upstream) to the 3' end (downstream). The term "nucleotide sequence" refers to the sequence of nucleotides of a DNA or RNA molecule that is typically shown from the 5 '(upstream) end to the 3' (downstream) end.
Methods known to those skilled in the art can be used to isolate and describe the DNA molecules or fragments thereof described herein. For example, PCR (polymerase chain reaction) techniques can be used to amplify a particular starting DNA molecule and/or to generate variants of the original molecule. The DNA molecule or fragment thereof may also be obtained by other techniques, such as direct synthesis of the fragment by chemical means (e.g., an automated oligonucleotide synthesizer).
As used herein, the term "isolated" refers to a molecule that is at least partially separated from other molecules with which it is normally associated in its natural or native state. In some embodiments, the term "isolated nucleic acid molecule" or "isolated DNA molecule" refers to a nucleic acid molecule (e.g., a DNA molecule) that is at least partially separated from the nucleic acid that normally flanks the gene or DNA molecule in its natural or native state. Thus, a nucleic acid molecule fused by recombinant techniques to regulatory or coding sequences normally unrelated thereto is considered herein to be isolated. Even when integrated into the chromosome of the host cell or present in nucleic acid solution with other DNA molecules, the molecules are considered isolated.
As used herein, a "polypeptide" comprises a plurality of consecutive polymerized amino acid residues, for example, at least about 15 consecutive polymerized amino acid residues. Typically a polypeptide comprises a series of polymeric amino acid residues that are transcriptional modulators or domains or portions or fragments thereof. Further, the polypeptide may comprise: (i) a localization domain; (ii) an activation domain; (iii) an inhibitory domain; (iv) an oligomerization domain; (v) a protein-protein interaction domain; (vi) a DNA binding domain; or other parts. The polypeptide optionally comprises modified amino acid residues, naturally occurring amino acid residues not encoded by codons, non-naturally occurring amino acid residues. As used herein, "protein" refers to a series of amino acids, oligopeptides, peptides, polypeptides, or portions thereof, whether naturally occurring or synthetic.
The term "isolated polypeptide", whether naturally occurring or recombinant, refers to a polypeptide that is present in the cell (or extracellularly) in an amount that is higher than the polypeptide in its native state in the wild-type cell, e.g., in an amount of greater than about 5% or greater than about 10% or greater than about 20% or greater than about 50% or greater, i.e., alternatively expressed as: the content is 105%, 110%, 120%, 150% or more with respect to the wild-type polypeptide normalized at 100%. This is not the result of the natural response of wild type plants. In addition, the isolated polypeptide is separated from other normally associated cellular components, for example, using various protein purification methods.
As used herein, the term "stringent conditions" are those described by Sambrook et al (1989) and Haymes, et al: Nucleic Acid Hybridization, A Practical Approach, IRL Press, Washington, DC (1985). Suitable stringency conditions for promoting DNA hybridization, for example, 6.0 Xsodium chloride/sodium citrate (SSC), about 45 ℃, followed by a wash with 2.0 XSSC at 50 ℃, are known to those skilled in the art, or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. For example, the salt concentration in the washing step can be selected from a low stringency of about 2.0 XSSC at 50 ℃ to a high stringency of about 0.2 XSSC at 50 ℃. In addition, the temperature in the washing step can be increased from room temperature, low stringency conditions of about 22 ℃ to high stringency conditions of about 65 ℃. Both temperature and salt can be varied, or temperature or salt concentration can be held constant while the other variable is changed. For example, moderately stringent conditions are about 2.0 XSSC and about 65 ℃.
In one aspect of the invention, an isolated nucleic acid molecule of the invention comprises the nucleotide sequence set forth in SEQ ID NO. 3. In another aspect of the invention, an isolated nucleic acid molecule of the invention has 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 98.5%, 99% and 99.5% sequence identity to the nucleotide sequence set forth in SEQ ID NO. 3. In some embodiments, the isolated nucleic acid molecule of the invention is 95% 96%, 97%, 98%, 98.5%, 99% and 99.5% identical to the nucleotide sequence set forth in SEQ ID No. 3.
The term "percent identity" or "% identity" is a comparison between amino acid sequences or nucleotides, as determined by a comparison between two sequences that are optimally matched over a comparison window. The person skilled in the art knows how to calculate the percentage identity between two sequences and there are many tools available (e.g.Clustal, Bestfit, Blast, Fasta, etc. software). One of the two sequences may have insertions, substitutions and/or deletions of one or more amino acids or nucleotides relative to the other sequence.
In the vector, the gene of the invention is typically operably linked to a promoter, a terminator and/or any other sequence necessary for its expression in yeast.
The terms "operably linked" and "operably linked" are used interchangeably and refer to a functional linkage between elements that enable expression of a gene, and optionally, the regulatory (5 'and 3' regulatory sequences) and sequences of a reporter gene that these elements control. The skilled person knows how to select promoters, terminators and other regulatory sequences required for expression of a gene.
As used herein, the term "promoter" generally refers to a DNA molecule that is involved in the recognition and binding of RNA polymerase II and other proteins (trans-acting transcription factors) to initiate transcription. In some embodiments, the promoter may be initially isolated from the 5 'untranslated region (5' UTR) of the genomic copy of the gene; alternatively, the promoter may be a synthetically produced or manipulated DNA molecule. In some embodiments, the promoter may also be chimeric, i.e., a promoter produced by the fusion of two or more heterologous DNA molecules. Plant promoters include promoter DNA obtained from plants, plant viruses, fungi, and bacteria (e.g., Agrobacterium). In some embodiments, the promoter is a developmentally-regulated, organelle-specific, tissue-specific, inducible, constitutive, or cell-specific promoter. In some specific embodiments, the expression of a gene is controlled by a so-called "strong" promoter (i.e., a promoter with high transcription potential such that the gene is strongly expressed).
The isolated nucleic acid molecules of the invention also comprise variant sequences which are derived from the sequence shown in SEQ ID No. 3 by deletion, substitution, insertion and/or addition of one or more nucleotides.
As described above, one or more nucleotides or amino acids may be "inserted", "deleted", "substituted" or "added", wherein the insertion, deletion, substitution and/or addition does not impair the function of the original sequence (e.g., as used herein, it is intended to refer to a function which still maintains the function of regulating plant growth or regulating plant stress resistance). The skilled person is aware of methods for performing one or more nucleotide/amino acid insertions, deletions and/or substitutions in the original sequence while preserving the biological function of the original sequence. For example, selecting for such insertions, deletions, additions, substitutions and/or in non-conserved regions; or modifying a nucleotide by "silent variation" based on the degeneracy of the genetic code without altering the polypeptide encoded by the nucleotide; alternatively, one amino acid in a protein may be replaced by another amino acid of similar nature by a "conservative substitution" without affecting the biological function of the protein.
Conservative substitutions may occur within the following groups: 1) acidic (negatively charged) amino acids such as aspartic acid and glutamic acid; 2) basic (positively charged) amino acids, such as arginine, histidine and lysine; 3) neutral polar amino acids such as glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine; and 4) neutral nonpolar (hydrophobic) amino acids such as alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine. Conservative substitutions for amino acids within a native protein or polypeptide may be selected from other members of the group to which the naturally occurring amino acid belongs. For example, the group of amino acids having aliphatic side chains is glycine, alanine, valine, leucine, and isoleucine; the amino acid group with aliphatic-hydroxyl side chains is serine and threonine; the amino acid group having amide-containing side chains is asparagine and glutamine; the group of amino acids with aromatic side chains is phenylalanine, tyrosine and tryptophan; the amino acid group with basic side chains is lysine, arginine and histidine; and the group of amino acids with sulfur-containing side chains are cysteine and methionine. The natural conservative amino acid substitution set is: valine-leucine, valine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine valine, aspartic acid-glutamic acid, and asparagine-glutamine.
As used herein, "mutation" refers to a sudden and heritable variation in a genomic DNA molecule. At the molecular level, genetic mutation means a change in the sequence of the arrangement of base pair composition or structure. The generation of the gene mutation may be spontaneous or induced. Methods for artificially inducing gene mutations include physical factors (e.g., gamma rays, x-rays, UV, neutron beam, etc.), chemical factors (e.g., alkylating agents, base analogs, antibiotics, etc.), and biological factors (e.g., certain viruses, bacteria, etc.). In addition, recombinant DNA techniques can be used to introduce specific variations at a given site in a DNA molecule for site-directed mutagenesis. Any of these well-known mutagenesis methods may be used by those skilled in the art to obtain variant sequences of the sequence shown in SEQ ID NO. 3 comprising deletions, substitutions, insertions and/or additions of one or more nucleotides.
As used herein, the term "recombinant" refers to a form of DNA and/or protein and/or organism that does not normally occur in nature and is therefore produced by human intervention. Such human intervention may result in recombinant DNA molecules and/or recombinant plants. As used herein, a "recombinant DNA molecule" is a DNA molecule that comprises a combination of DNA molecules that do not naturally occur together and that are the result of human intervention, e.g., a DNA molecule comprising a combination of: at least two DNA molecules heterologous to each other, and/or DNA molecules which are artificially synthesized and comprise a polynucleotide sequence derived from a polynucleotide sequence which is usually present in nature, and/or DNA molecules which comprise a transgene which is artificially introduced into the genomic DNA of a host cell and the associated flanking DNA of the genome of the host cell. An example of a recombinant DNA molecule is a DNA molecule described herein that results from the insertion of a gene into the genome of maize, which can ultimately result in the transcription of a recombinant RNA in an organism and/or the expression of a polypeptide or protein in that organism.
As used herein, "host cell" refers to a cell that contains a recombinant vector and supports replication and/or expression of the expression vector. The host cell may be a prokaryotic cell (e.g., an E.coli cell, an Agrobacterium tumefaciens cell) or a eukaryotic cell (e.g., a yeast, insect, plant or animal cell).
In some embodiments, the host cell is preferably a monocot or dicot plant cell, including but not limited to cells from maize, oilseed rape, turnip, mustard, alfalfa, rice, rye, sorghum, pearl millet, finger millet, sunflower, safflower, wheat, soybean, tobacco, potato, peanut, cotton (gossypium barbadense, gossypium hirsutum), sweet potato, cassava, sugar beet, sugarcane, oat, barley, and arabidopsis thaliana. Preferably, the host cell is a maize cell or a rice cell; more preferably, the host cell is a maize cell.
As used herein, the "introduction" of a nucleic acid molecule or expression vector into a plant or plant cell refers to the transfection, transformation, transduction, or incorporation of the nucleic acid molecule or recombinant expression vector into a host cell such that the nucleic acid molecule is capable of autonomous replication or expression in the host cell.
In some embodiments, the introduced nucleic acid molecule is integrated into the host cell genomic DNA (e.g., chromosomal, plasmid, plastid, or mitochondrial DNA) and expression of the nucleotide sequence is controlled by the regulatory promoter region. In other embodiments, the introduced nucleic acid molecule is not integrated into the genome of the cell.
According to the invention, through overexpression of a gene 'dwarf 20' in corn, plants with significant changes in the traits of spike length, spike thickness, bald tip length, spike row number, line grain number, normal spike grain number, grain width, grain thickness, cell grain weight, grain moisture, hundred grain weight and the like are obtained; the suggestion that the gene is possibly related to the growth and development of plants and participates in the stress-resistant way of the plants.
A more detailed description of some preferred embodiments of the invention follows.
Drawings
FIG. 1: "short 20" gene structure and protein functional domain
FIG. 2: pBCXUN vector structure schematic diagram
FIG. 3: schematic structure of gene overexpression vector
FIG. 4: expression analysis of dwarf 20 in leaves of maize inbred lines B73-329(WT) and T3 transgenic inbred lines
FIG. 5: comparison of seedling vigor of transgenic plants before 2018 years of harvest
FIG. 6: pre-harvest SPAD (chlorophyll content) value comparison of transgenic plants 2018
FIG. 7: comparison of the days to produce silks before harvesting transgenic plants in 2018 years
FIG. 8: comparison of days of pollen dispersal before harvest in 2018 years for transgenic plants
FIG. 9: plant height data comparison of transgenic plants before 2018 year harvest
FIG. 10: grain weight comparison of transgenic plants in cell after 2018 years of harvest
FIG. 11: comparison of ear length data of transgenic plants harvested in 2018 years
FIG. 12: comparison of ear width data of transgenic plants harvested in 2018 years
FIG. 13: comparison of ear row number of transgenic plants harvested in 2018 years
FIG. 14 is a schematic view of: comparison of the number of grains in transgenic plants harvested in 2018 years
FIG. 15 is a schematic view of: comparison of grain number of normal ears of transgenic plants harvested in 2018 years
FIG. 16: grain width contrast of transgenic plants harvested in 2018 years
FIG. 17: grain thickness comparison of transgenic plants harvested in 2018 years
FIG. 18 is a schematic view of: comparison of hundred grain weight of transgenic plants harvested in 2018 years
FIG. 19: comparison of spinning days before harvest in 2020 for transgenic plants
FIG. 20: comparison of days of pollen dispersal before harvest in 2020 years for transgenic plants
FIG. 21: transgenic plant 2020 Pre-harvest plant height contrast
FIG. 22: comparison of ear height before harvest of transgenic plants in 2020
FIG. 23: pre-harvest internode distance comparison of transgenic plants in 2020
FIG. 24: comparative plant height photographs of control plants and transgenic plants
FIG. 25: photograph comparing the internodal distance between control and transgenic plants (left transgenic plant, right control plant)
Detailed description of the preferred embodiments
The following examples describe some specific embodiments of the invention. It is to be understood, however, that the examples and drawings are given by way of illustration only and do not limit the scope of the invention.
pBCXUN: the HYG gene is transformed on the basis of a vector pCXUN (NCBI GenBank: FJ905215), and Bar is replaced by the HYG gene through an Xho I site; bar gene comes from pCAMBIA3301, the promoter for starting the expression of target gene in pCXUN vector is maize Ubiquitin-1, and the terminator is T-nos.
Example 1: construction of overexpression vector for "dwarf 20" Gene
1. Obtaining the maize "dwarf 20" gene
RNA was extracted from anthers of corn variety B73 using the magnetic bead method plant Total RNA extraction Kit (Beijing Baitach Biotechnology Co., Ltd., cat # AU3402) according to the manufacturer's instructions, and then the RNA was Reverse transcribed into cDNA using the High-Capacity cDNA Reverse Transcription Kit (Thermo Scientific Co., cat # 4368814) to create a cDNA template.
The cDNA of corn is used as a template, a primer is designed according to the sequence analysis of a short 20 coding region, the coding region of the gene is amplified by utilizing an upstream primer 'short 20' -F, a downstream primer 'short 20' -R and high fidelity enzyme to obtain a target gene sequence, and the target gene sequence is connected to a pBCXUN vector with a promoter of Ubiquitin. Primer sequences are shown in table 1:
TABLE 1 "dwarf 20" amplification primers
| Primer numbering | Primer sequences |
| "short 20" -F (SEQ ID NO:4) | 5’-GCCAGCAGACATGGACATCGAC-3’ |
| "short 20" -R (SEQ ID NO:5) | 5’-TAGCCCGAGCTCAGAGTCAGAAC-3’ |
2. Preparation of an overexpression vector
The DNA molecule of the "dwarf 20" insertion sequence shown in SEQ ID NO. 6 is connected to the pBCXUN vector by a TA cloning method to obtain the recombinant expression vector pBCXUN- "dwarf 20" (sequencing verification is carried out), and the method specifically comprises the following steps:
the PCR product was purified using 1% agarose gel and gel recovery kit, and the purified "short 20" cDNA sequence was ligated to the vector pBCXUN (NCBI GenBank: FJ905215, see, e.g., Plant Physiol.150(3), 1111-. The recombinant expression vector is used for transforming escherichia coli, positive clones are screened, and cloning construction is completed by a one-step method.
And (3) sequencing the obtained plasmid by a sequencing company, comparing and analyzing a sequencing result with a target sequence to ensure that the vector contains a complete target gene sequence, indicating that the target gene is transferred into the pBCXUN, and completing cloning construction.
Example 2: preparation of "dwarf 20" overexpression transgenic maize
The recombinant expression vector pBCXUN- "dwarf 20" is introduced into the agrobacterium EHA105 strain to obtain recombinant bacteria. Then, the recombinant strain is introduced into a maize inbred line B73-329 by an agrobacterium-mediated method to obtain a T0 generation transgenic plant.
Taking the seedling leaves of T0 generation transgenic plants, and extracting genome DNA. Using genome DNA as template, PCR amplification is carried out by using primer Ubip-F (aiming at the 5 'end of Ubi1P promoter of recombinant expression vector pBCXUN- "dwarf 20") and primer Nos-R (aiming at the 3' end of Nos terminator of recombinant expression vector pBCXUN-dwarf 20) as well as self primer "dwarf 20" -F and "dwarf 20" -R of target gene for combination pairing.
The genome DNA of the seedling leaves of the maize inbred line B73-329 is used as a negative control, and the plasmid of the recombinant expression vector pBCXUN- "dwarf 20" is used as a positive control.
Table 2: identification primer for 'short 20' T0 transgenic material
The PCR amplification products were detected by agarose gel electrophoresis. The result shows that the single band of the target gene can be amplified by the transgenic plant and the plasmid: the combined amplified fragment size of Ubip-F (SEQ ID NO:7) and "dwarf 20" -R (SEQ ID NO:5) is 1464 bp; the combined amplified fragment size of "short 20" -F (SEQ ID NO:4) and Nos-R (SEQ ID NO:8) was 1436bp, while the parental B73-329 did not amplify a corresponding band, indicating that the "short 20" gene was successfully transferred into the transgenic plant.
Selfing the identified T0 transgenic plants to obtain T1 transgenic plant progeny; selfing the T1 transgenic plant filial generation to obtain T2 transgenic plant filial generation; selfing the T2 transgenic plant filial generation to obtain T3 transgenic plant filial generation; the positive transgenic plants are identified by adopting the PCR amplification method in each generation and then selfed. 5 representative T3 generation homozygous transgenic lines (i.e., dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4, dwarf 20-5) were selected for subsequent functional analysis experiments.
Example 3: detection of Gene expression level of dwarf 20
This example uses 5 representative T3 generation homozygous transgenic lines (i.e., dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4, dwarf 20-5) and maize inbred line B73-329(WT) as plants to be tested.
RNA of leaves at stage V4 of a plant to be tested was extracted using a magnetic bead method plant total RNA extraction Kit (Beijing Baitach Biotechnology Co., Ltd., cat. No. AU3402) according to the manufacturer's instructions, and then the RNA was Reverse-transcribed into cDNA using a High-Capacity cDNA Reverse Transcription Kit (Thermo Scientific Co., cat. No. 4368814).
SYBR Premix Ex TaqTM II (Tli RNaseH Plus) kit (Takara, Inc., cat No. RR820A) was used to perform real-time fluorescent quantitative PCR amplification using GTY-rF (SEQ ID NO: 9: AGTATTGGGGATCCGAATTTC) and GTY-rR (SEQ ID NO: 10: TAATCATAAAAACCCATCTCATAA) using cDNA as a template to detect the expression level of the transferred "short 20" gene (refer to the patent of Wang Xiqing et al, university of agriculture, China, application No. CN201810636371.7, publication No. CN 108624709B). The cDNA of corn inbred line B73-329(WT) is used as a control, the corn Actin gene is used as an internal reference gene, and the detection primers are as follows: ZmActin-rF (SEQ ID NO: 11: GAGCTCCGTGTTTCGCCTGA) and ZmActin-rR (SEQ ID NO: 12: CAGTTGTTCGCCCACTAGCG). The reaction procedure for the fluorescent quantitative PCR amplification is shown in Table 3 below.
Table 3: reaction procedure for fluorescent quantitative PCR amplification
The results of the fluorescent quantitative PCR are shown in FIG. 4. As can be seen from FIG. 4, the expression level of "dwarf 20" in the transgenic lines dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5 is significantly higher than that of the maize inbred line B73-329 (WT). These results indicate that the "dwarf 20" gene was successfully overexpressed in T3-generation homozygous transgenic lines dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4, dwarf 20-5.
Example 4: performance in the field
5 over-expressing transgenic lines were subjected to a multi-point test for years with control plants. Dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4, dwarf 20-5 and WT (wild type) were planted in 2018 at 2 trials (princess ridge, 28095; state, 6 repeats per trial) and 2020 at 2 trials (princess ridge, Kaifeng, 3 repeats per trial), respectively.
And (4) investigating the field properties of each strain from the seedling stage to the milk stage, wherein the field properties comprise seedling vigor, SPAD (chlorophyll content) value, silking period, powder scattering period, plant height and spike height. After harvesting, the field properties of each strain, including yield, ear length of the ear, ear thickness, bald tip length, ear row number, row grain number, normal ear grain number, grain width, grain thickness, cell grain weight, grain moisture, hundred grain weight, etc., were determined.
The field character comparison result of the transgenic corn and the control plant measured before 2018 harvest shows that the seedling potentials of 5 transgenic lines, namely dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5, are respectively reduced by 15.4%, 25.4%, 20.4% and 17.9% to reach the extremely significant level (figure 5); the SPAD values of 3 transgenic lines, namely dwarf 20-1, dwarf 20-3 and dwarf 20-5, are respectively reduced by 6.1%, 11.3% and 13.2% to reach an extremely significant level (figure 6); 1 transgenic line dwarf 20-3 delays the silking stage by 5 percent and reaches a remarkable level (figure 7); 1 transgenic line dwarf 20-3 is delayed by 5.7% in the pollen scattering stage, and reaches a significant level (figure 8); the plant heights of 5 transgenic lines, namely dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5, are respectively reduced by 43.9 percent, 42.0 percent, 42.6 percent, 34.1 percent and 41.6 percent to reach extremely remarkable levels (figure 9); the heights of 5 transgenic lines, namely dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5, of spikes are respectively reduced by 70.8%, 66.2%, 63.1%, 57.6% and 63.1%, and the levels are extremely obvious.
The field character comparison result of the transgenic corn and the control plant measured after the harvest in 2018 shows that the yield of 5 transgenic lines, namely dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5, is respectively reduced by 50.6%, 40.7%, 72.4%, 35.2% and 36.5% to reach the extremely significant level (figure 10); the ear length of 4 transgenic lines dwarf 20-1, dwarf 20-3, dwarf 20-4 and dwarf 20-5 ears is respectively reduced by 16.5%, 7.3%, 8.3% and 13.4% to reach an extremely significant level (figure 11); the thicknesses of 5 transgenic lines, namely dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5, are respectively reduced by 3.9%, 2.4%, 0.7%, 0.4% and 3.1% and the differences are not significant (figure 12); the bald tip length of 5 transgenic lines, namely dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5, is respectively increased by 25.1 percent, 6.6 percent, 20.7 percent, 15.6 percent and 23.4 percent, and reaches an extremely obvious level; the row number of 1 transgenic line dwarf 20-5 ears is reduced by 6.2 percent, which reaches a very significant level, and the difference of other lines is not significant (figure 13); the grain numbers of 4 transgenic lines of dwarf 20-1, dwarf 20-3, dwarf 20-4 and dwarf 20-5 are respectively reduced by 24.3%, 12.3%, 13.5% and 12.7% to reach the extremely significant level (figure 14); the normal grain number of dwarf strains of 5 transgenic lines, namely dwarf strains 20-1, dwarf strains 20-2, dwarf strains 20-3, dwarf strains 20-4 and dwarf strains 20-5, is respectively reduced by 29.2%, 6.4%, 6.5%, 15.5% and 20.5% to reach the extremely remarkable level (figure 15); the width of 20-2 grains of 1 transgenic line dwarf is increased by 5.4 percent, which reaches a very significant level, and the difference of other transgenic lines is not significant (figure 16); the thicknesses of 20-1 dwarf and 20-3 dwarf of 2 transgenic lines are respectively reduced by 6.4 percent and increased by 5.6 percent, the extremely significant level is reached, and the differences of other transgenic lines are not significant (figure 17); the weight of each hundred grains of 5 transgenic lines, namely dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5, is respectively reduced by 6.1%, 4.2%, 7.6%, 6.0% and 6.7% to reach a significant level (figure 18).
The field character comparison result of the transgenic corn and the control plant measured before the harvest in 2020 shows that the spinning period of 1 transgenic line of dwarf 20-3, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5 in 5 transgenic lines is delayed by 2.7 percent and reaches a significant level (figure 19); 1 transgenic line dwarf 20-3 is delayed for 1.6 percent in the powder scattering period, and reaches a remarkable level (figure 20); the plant heights of 5 transgenic lines, namely dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5, are respectively reduced by 42.8 percent, 33.8 percent, 40.6 percent, 39.5 percent and 43.2 percent to reach extremely remarkable levels (figure 21); the height of ears of 5 transgenic lines, namely dwarf 20-1, dwarf 20-2, dwarf 20-3, dwarf 20-4 and dwarf 20-5, is respectively reduced by 52.9 percent, 47.3 percent, 56.8 percent, 57.0 percent and 55.8 percent to reach the extremely remarkable level (figure 22). The 1 st internode distance under the 5 transgenic lines, namely 20-1 dwarf, 20-2 dwarf, 20-3 dwarf, 20-4 dwarf and 20-5 dwarf, is respectively reduced by 69.6 percent, 66.7 percent, 71.2 percent, 70.6 percent and 65.3 percent to reach the extremely remarkable level; the distance between the 2 nd nodes under the spike is respectively reduced by 64.5 percent, 63.7 percent, 69.4 percent, 68.7 percent and 62.0 percent, and extremely remarkable level is achieved; the distance between the 3 rd nodes under the spike is respectively reduced by 58.3%, 59.1%, 66.5%, 64.1% and 54.6%, and extremely remarkable level is achieved; the 4 th node space under the spike is respectively reduced by 52.1%, 54.0%, 63.4%, 57.8% and 46.8% to reach an extremely significant level; the distance between the 5 th nodes under the spike is respectively reduced by 51.7%, 53.3%, 65.1%, 58.4% and 42.4%, and extremely remarkable level is achieved; the 6 th internode spacing under the spike is respectively reduced by 53.6%, 58.4%, 68.5%, 62.7% and 44.2%, which reaches the extremely significant level; the space between the 7 th node under the spike is respectively reduced by 56.5 percent, 57.7 percent, 68.0 percent, 65.5 percent and 48.1 percent, and reaches a very significant level; the spacing between the ear nodes is respectively reduced by 67.0 percent, 65.1 percent, 70.3 percent, 67.9 percent and 63.5 percent, and extremely remarkable level is achieved; the 1 st internode spacing on the spike is respectively reduced by 47.9%, 44.9%, 47.1%, 48.6% and 34.7%, which reaches a very significant level; the distance between the 2 nd internode on the spike is respectively reduced by 43.7 percent, 39.6 percent, 34.2 percent, 41.2 percent and 27.9 percent, and extremely remarkable level is achieved; the distance between the 3 rd nodes on the spike is respectively reduced by 46.1%, 41.5%, 37.2%, 42.1% and 30.2%, and extremely remarkable level is achieved; the 4 th internode spacing on the spike is respectively reduced by 48.7%, 44.4%, 42.5%, 43.9% and 33.6% to reach an extremely significant level; the distance between the 5 th internode on the spike is respectively reduced by 47.3 percent, 42.2 percent, 42.4 percent, 41.7 percent and 33.1 percent, and extremely remarkable level is achieved; the 6 th internode spacing on the ear is respectively reduced by 40.3%, 36.5%, 34.8%, 35.8% and 29.2%, reaching a very significant level (fig. 23).
Small knot
The dwarf 20 gene can obviously reduce the plant height and the ear position of the corn, and can keep the leaf number and the flowering phase of the corn unchanged under the condition of reducing the plant height, so that the photosynthesis efficiency of the corn is not reduced under the condition of reducing the plant height, the accumulation requirement of grain filling is ensured, and the yield can not be obviously reduced under the normal density.
The "short 20" can significantly shorten the internode distance, and since the substutural internode distance shortening is more significant than the suprapanicular internode distance shortening, the reduction in ear position is more significant than the reduction in plant height. The plant height and the ear position are reduced, the ear position is reduced more obviously, the plant type has strong lodging resistance, the plant type can improve the planting density of the corn without causing lodging, and the yield is improved by increasing the density. The dwarf corn can be lower than the lodging rate of the high-stalk corn under the condition of the same infection of the stem rot, thereby avoiding yield loss and reducing the harvesting cost.
The short 20 gene can ensure that farmers can enter the corn field to operate in the whole growing season after the corn is dwarfed, and the time window of fertilizing and preventing and treating plant diseases and insect pests is enlarged, thereby improving the yield and reducing the capital investment of large-scale overhead pesticide sprayers. After the dwarf 20 gene dwarfs the corn, a seed company can more easily perform emasculation on large-scale seed production, thereby remarkably reducing the labor cost of seed production.
The invention has been described in detail with reference to specific embodiments and examples, but modifications and improvements can be made on the basis of the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Reference to the literature：
[1]Balaji Enugutti,Charlotte Kirchhelle,Maxi Oelschner,Ramón Angel Torres Ruiz,Ivo Schliebner,Dario Leister,Kay Schneitz.Regulation of planar growth by the Arabidopsis AGC protein kinase UNICORN[J].Proceedings of the National Academy of Sciences of the United States of America,2012,109(37).
[2]Balaji Enugutti,Kay Schneitz.Genetic analysis of ectopic growth suppression during planar growth of integuments mediated by the Arabidopsis AGC protein kinase UNICORN[J].Kay Schneitz,2013,13(1).
[3]Ana Victoria Garcia,Mohamed Al-Yousif,Heribert Hirt.Role of AGC kinases in plant growth and stress responses[J].Cellular and Molecular Life Sciences,2012,69(19).
[4]Matsui Hidenori,Miyao Akio,Takahashi Akira,Hirochika Hirohiko.Pdk1 kinase regulates basal disease resistance through the OsOxi1-OsPti1a phosphorylation cascade in rice[J].Plant&cell physiology,2010,51(12).
Claims (17)
1. An isolated nucleic acid molecule comprising a sequence selected from the group consisting of:
1) 3 or a complementary sequence thereof;
2) a sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the nucleotide sequence set forth in SEQ ID NO. 3;
3) a nucleotide sequence that hybridizes under stringent conditions to SEQ ID NO. 3; or
4) 3 by deletion, substitution, insertion and/or addition of one or more nucleotides.
2. The nucleic acid molecule of claim 1, having the nucleotide sequence set forth in SEQ ID NO 3.
3. The nucleic acid molecule of claim 1 or 2, wherein the CDS sequence is as set forth in SEQ ID NO 2.
4. An isolated nucleic acid molecule comprising a sequence selected from the group consisting of:
1) a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO. 1;
2) a nucleotide sequence encoding an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence set forth in SEQ ID NO. 1; or
3) 1 by deletion, substitution, insertion and/or addition of one or more amino acid residues.
5. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:
1) 1, SEQ ID NO;
2) an amino acid sequence encoded by the nucleic acid molecule of any one of claims 1-4;
3) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence shown in SEQ ID NO. 1; or
4) 1, amino acid sequence derived by deleting, substituting, inserting and/or adding one or more amino acid residues in the sequence shown in SEQ ID NO.
6. The isolated polypeptide of claim 5, having an amino acid sequence as set forth in SEQ ID NO 1.
7. A recombinant vector comprising the isolated nucleic acid molecule of any one of claims 1-4.
8. A host cell comprising the isolated nucleic acid molecule of any one of claims 1-4, or comprising the isolated polypeptide of claim 5 or 6, or comprising the recombinant vector of claim 7.
9. A transgenic plant comprising the isolated nucleic acid molecule of any one of claims 1-4, or comprising the isolated polypeptide of claim 5 or 6, or comprising the recombinant vector of claim 7.
10. The transgenic plant of claim 9, wherein the transgenic plant is a monocotyledonous or dicotyledonous plant, preferably a crop plant.
11. The transgenic plant of claim 10, wherein the plant is selected from the group consisting of maize, oilseed rape, turnip, mustard, alfalfa, rice, rye, sorghum, pearl millet, finger millet, sunflower, safflower, wheat, soybean, tobacco, potato, peanut, cotton (gossypium barbadense, gossypium hirsutum), sweet potato, cassava, sugar beet, sugarcane, oat, barley, and arabidopsis, preferably arabidopsis thaliana or maize.
12. The transgenic plant of any one of claims 9-11, wherein the plant has an altered trait as compared to a plant or wild type plant not having the corresponding transgene, wherein the altered trait is selected from the group consisting of yield, plant height, ear length of ear, ear thickness, bald tip length, ear row number, row grain number, normal ear grain number, grain width, grain thickness, plot grain weight, grain moisture, hundred grain weight, internode distance.
13. A method for producing a transgenic plant with an altered trait, comprising introducing into a plant the nucleic acid molecule of claims 1-4, the recombinant vector of claim 7, or the host cell of claim 8, to obtain a corresponding transgenic plant; the transgenic plant has altered traits compared to a plant not transgenic for the corresponding or a wild type plant, wherein the altered traits are selected from the group consisting of yield, plant height, ear length of ear, ear thickness, bald tip length, ear row number, row grain number, normal ear grain number, grain width, grain thickness, cell grain weight, grain moisture, grain weight per hundred, and internode spacing.
14. A method for modulating plant growth or plant stress resistance, wherein said method comprises transforming/transfecting a plant with the recombinant vector of claim 7 or the host cell of claim 8 to obtain a transgenic plant with increased yield as compared to a plant not correspondingly transformed/transfected or a wild type plant.
15. Use of the nucleic acid molecule of any one of claims 1 to 4 or the polypeptide of claim 5 or 6 or the recombinant vector of claim 7 for modulating plant growth or plant stress resistance.
16. The method or use according to any one of claims 13 to 15, wherein the transgenic plant is a monocotyledonous or dicotyledonous plant, preferably a crop plant.
17. The method or use according to claim 16, wherein the plant is selected from the group consisting of maize, oilseed rape, turnip, mustard, alfalfa, rice, rye, sorghum, pearl millet, finger millet, sunflower, safflower, wheat, soybean, tobacco, potato, peanut, cotton (gossypium barbadense, gossypium hirsutum), sweet potato, cassava, sugar beet, sugar cane, oat, barley and arabidopsis, preferably arabidopsis thaliana or maize.
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| CN101460611A (en) * | 2006-06-08 | 2009-06-17 | 巴斯福植物科学有限公司 | Plants having improved growth characteristics and method for making the same |
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