Ways to increase corn yields

Abstract

The present invention relates to a method for increasing corn yield, belonging to the field of molecular genetics, and discloses a corn yield regulating gene and a method for increasing corn yield and / or biomass.

Description

Technical Field

[0001] The invention relates to a method for increasing corn yield and belongs to the field of molecular genetics. Background Art

[0002] Corn is an important food and feed crop in my country. Furthermore, it serves as an indispensable raw material for numerous industries, including medicine, chemicals, and brewing, playing a crucial role in my country's national economy. High corn yields are the result of the coordinated development of three key yield factors: number of ears per unit area, number of kernels per ear, and kernel weight. In recent years, numerous yield-related genes have been cloned. However, due to the complexity of corn yield and biomass regulation mechanisms, a greater number of yield / biomass regulatory genes are needed for practical applications in yield / biomass improvement and the development of new corn germplasm with higher yields and biomass.

[0003] During previous research, the inventors discovered that the GRMZM2G005732 gene affects the flowering period of corn. Overexpressing this gene will delay the flowering time of corn, while mutating this gene using gene editing technology will advance the flowering time of corn. However, these operations will not change the yield traits of corn (see patent CN 112646820A for details).

[0004] The present invention unexpectedly discovered that another transcript, T03, of the GRMZM2G005732 gene is associated with both maize yield and biomass traits. Expressing the T03 transcript in maize can increase maize yield and / or biomass, thereby enabling the development of new high-yield / high-biomass maize germplasm. This provides a new approach for developing new high-yield / high-biomass maize germplasm. Summary of the Invention

[0005] One of the objectives of the present invention is to provide a nucleic acid molecule that affects corn yield and / or biomass, as well as an expression cassette, an expression vector and a host cell containing the nucleic acid molecule.

[0006] A second object of the present invention is to provide a method for increasing corn yield and / or biomass.

[0007] To achieve the above object, the present invention adopts the following technical solutions:

[0008] The present invention provides an application of a nucleic acid molecule in regulating corn yield and / or biomass, characterized in that the nucleotide sequence of the nucleic acid molecule is shown as SEQ ID NO.2.

[0009] The present invention also provides an application of an expression cassette in regulating corn yield and / or biomass, characterized in that the expression cassette contains the above-mentioned nucleic acid molecule.

[0010] The present invention also provides an application of an expression vector in regulating corn yield and / or biomass, characterized in that the expression vector contains the above-mentioned expression cassette.

[0011] The present invention also provides an application of a host cell in regulating corn yield and / or biomass, characterized in that the host cell contains the above-mentioned expression vector.

[0012] In some embodiments, the host cell is a prokaryotic cell.

[0013] In some embodiments, the host cell is an Escherichia coli or Agrobacterium cell.

[0014] The present invention also provides a method for increasing corn yield and / or biomass, characterized by expressing the above nucleic acid molecule in corn and selecting plants with increased corn yield and / or biomass.

[0015] Compared to existing technologies, the present invention has the following beneficial effects: It unexpectedly discovered a new function of the T03 transcript of GRMZM2G005732—influencing maize yield and / or biomass. Nucleic acid molecules expressing the T03 transcript in maize can increase maize yield and / or biomass. These nucleic acid molecules, as well as expression cassettes, expression vectors, or host cells containing them, can be used to improve maize yield and / or biomass traits. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Yield / biomass-related traits of three maize lines expressing the T01 transcript. Plant height: Plant height; Ear height: Ear height; Aboveground biomass fresh weight: Aboveground biomass fresh weight; Aboveground biomass dry weight: Aboveground biomass dry weight. "+" indicates positive transgenic plants; "-" indicates negative control; "ns" indicates no significant difference.

[0017] Figure 2 Diagram of the expression vector containing the T03 nucleic acid molecule.

[0018] Figure 3 Phenotype of maize expressing the T03 transcript. “+” positive transgenic plants; “-” negative control; long lines represent bar = 10 cm.

[0019] Figure 4Yield / biomass-related traits of four maize lines expressing the T03 transcript. Plant height: Plant height; Ear height: Ear height; Aboveground biomass fresh weight: Aboveground biomass fresh weight; Aboveground biomass dry weight: Aboveground biomass dry weight. "+" indicates positive transgenic plants; "-" indicates negative control; "*" indicates a significant difference; "**" indicates a very significant difference. DETAILED DESCRIPTION

[0020] The following definitions and methods are provided to better define this application and to guide those skilled in the art in practicing this application. Unless otherwise noted, terms are to be understood according to conventional usage by those skilled in the relevant art. All patent documents, academic papers, industry standards, and other publications cited herein are hereby incorporated by reference in their entirety.

[0021] As used herein, "corn" refers to any corn plant and includes all plant varieties that can be bred with corn, including whole plants, plant cells, plant organs, plant protoplasts, plant cell tissue cultures from which plants can be regenerated, plant callus, intact plant cells in plants or plant parts, such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruits, stems, roots, root tips, anthers, etc. Unless otherwise indicated, nucleic acids are written from left to right in a 5' to 3' orientation; amino acid sequences are written from left to right in an amino to carboxyl orientation. Amino acids can be represented herein by their commonly known three-letter symbols or by the single-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Similarly, nucleotides can be represented by commonly accepted single-letter codes. Numerical ranges include numbers that define the range. As used herein, "nucleic acid" includes deoxyribonucleotides or ribonucleotide polymers in single-stranded or double-stranded form, and unless otherwise limited, includes known analogs (e.g., peptide nucleic acids) having the basic properties of natural nucleotides that hybridize to single-stranded nucleic acids in a manner similar to naturally occurring nucleotides. As used herein, the terms "encoding" or "encoded" are used in the context of a specific nucleic acid to refer to a nucleic acid that contains the necessary information to direct the translation of the nucleotide sequence into a specific protein. Codons are used to represent information encoding proteins. As used herein, "full-length sequence" relating to a specific polynucleotide or its encoded protein refers to the entire nucleic acid sequence or the entire amino acid sequence with a natural (non-synthetic) endogenous sequence. A full-length polynucleotide encodes the full-length, catalytically active form of the specific protein. The terms "polypeptide," "polypeptide," and "protein" are used interchangeably herein to refer to polymers of amino acid residues. The term is used for amino acid polymers in which one or more amino acid residues are artificial chemical analogs of the corresponding naturally occurring amino acids. The term is also used for naturally occurring amino acid polymers. The terms "residue" or "amino acid residue" or "amino acid" are used interchangeably herein to refer to an amino acid that is incorporated into a protein, polypeptide, or peptide (collectively, "protein"). The amino acid can be a naturally occurring amino acid and, unless otherwise limited, can include known analogs of a naturally occurring amino acid that can function in a manner similar to the naturally occurring amino acids.

[0022] The term "trait" refers to a physiological, morphological, biochemical or physical characteristic of a plant or a specific plant material or cell. In some cases, this characteristic is visible to the human eye, such as seed or plant size, or can be measured by biochemical techniques, such as detecting the protein, starch or oil content of seeds or leaves, or by observing metabolic or physiological processes, for example, by measuring tolerance to water deprivation or specific salt or sugar or nitrogen concentrations, or by observing the expression level of one or more genes, or by agronomic observations such as osmotic stress tolerance or yield.

[0023] "Transgenic" refers to any cell, cell line, callus, tissue, plant part, or plant whose genome is altered by the presence of a heterologous nucleic acid, such as a recombinant DNA construct. As used herein, the term "transgenic" includes those original transgenic events and those generated from the original transgenic events by sexual crosses or asexual propagation, and does not encompass genomic (chromosomal or extrachromosomal) alterations made by conventional plant breeding methods or by naturally occurring events, such as random cross-fertilization, non-recombinant viral infection, non-recombinant bacterial transformation, non-recombinant transposition, or spontaneous mutation.

[0024] "Plant" includes reference to whole plants, plant organs, plant tissues, seeds, and plant cells, as well as their progeny. Plant cells include, but are not limited to, cells from seeds, suspension cultures, embryos, meristematic regions, callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen, and microspores. "Progeny" includes any subsequent generation of a plant.

[0025] In this application, the words "comprises," "comprising," or variations thereof are to be understood as including, in addition to the described elements, numbers, or steps, other elements, numbers, or steps. A "test plant" or "test plant cell" refers to a plant or plant cell in which a genetic modification has been effected, or a progeny of a plant or cell so modified that contains the modification. A "control," "control plant," or "control plant cell" provides a reference point for measuring phenotypic changes in the test plant or plant cell.

[0026] Negative or control plants can include, for example: (a) wild-type plants or cells, i.e., plants or cells having the same genotype as the genetically modified starting material that produced the test plant or cell; (b) plants or plant cells having the same genotype as the starting material but that have been transformed with an empty construct (i.e., with a construct that has no known effect on the trait of interest, such as a construct comprising a marker gene); (c) plants or plant cells that are non-transformed segregants of the test plant or plant cell; (d) plants or plant cells that are genetically identical to the test plant or plant cell but that have not been exposed to conditions or stimuli that would induce expression of the gene of interest; or (e) the test plant or plant cell itself, which is under conditions where the gene of interest is not expressed.

[0027] Those skilled in the art will readily recognize that advances in the field of molecular biology, such as site-specific and random mutagenesis, polymerase chain reaction methods, and protein engineering techniques, provide a wide range of appropriate tools and procedures for modifying or engineering the amino acid sequence and underlying gene sequence of proteins of agricultural interest.

[0028] In some embodiments, the nucleotide sequences of the present application can be altered to make conservative amino acid substitutions. The principles and examples of conservative amino acid substitutions are further described below. In certain embodiments, the nucleotide sequences of the present application can be substituted without changing the amino acid sequence according to the disclosed monocot codon preferences, for example, codons encoding the same amino acid sequence can be replaced with codons preferred by monocots without changing the amino acid sequence encoded by the nucleotide sequence. In some embodiments, part of the nucleotide sequence in the present application is replaced with different codons encoding the same amino acid sequence, thereby not changing the amino acid sequence encoded by the nucleotide sequence while changing the nucleotide sequence. Conservative variants include those sequences that encode the amino acid sequence of one of the proteins of the embodiments due to the degeneracy of the genetic code. In some embodiments, part of the nucleotide sequence in the present application is replaced according to the monocot codon preference. Those skilled in the art will recognize that amino acid additions and / or substitutions are generally based on the relative similarity of the amino acid side chain substituents, for example, the hydrophobicity, charge, size, etc. of the substituents. Exemplary amino acid substitution groups with various aforementioned properties are well known to those skilled in the art and include arginine and lysine; glutamic acid and aspartic acid; serine and threonine; glutamine and asparagine; and valine, leucine and isoleucine. Guidance on appropriate amino acid substitutions that do not affect the biological activity of the protein of interest can be found in the model of Dayhoff et al. (1978) Atlas of Protein Sequence and Structure (Natl. Biomed. Res. Found., Washington, DC) (incorporated herein by reference). Conservative substitutions such as replacing one amino acid with another amino acid having similar properties can be performed. Sequence identity identification includes hybridization techniques. For example, all or part of a known nucleotide sequence is used as a probe for selective hybridization with other corresponding nucleotide sequences present in cloned genomic DNA fragments or cDNA fragment groups (i.e., genomic libraries or cDNA libraries) from a selected organism. The hybridization probe can be a genomic DNA fragment, a cDNA fragment, an RNA fragment or other oligonucleotide, and can be marked with a detectable group such as 32P or other detectable markers. Thus, for example, a hybridization probe can be prepared by marking a synthetic oligonucleotide based on the embodiment sequence. The method for preparing hybridization probes and building cDNA and genomic libraries is generally known in the art. The hybridization of the sequence can be carried out under stringent conditions. As used herein, the term "stringent conditions" or "stringent hybridization conditions" represents following conditions, i.e., under these conditions, relative to hybridizing with other sequences, the probe will hybridize with its target sequence to a greater extent (e.g., at least 2 times, 5 times or 10 times of background) that can be detected.Stringent conditions are sequence-dependent and vary in different environments. By controlling hybridization stringency and / or controlling washing conditions, a target sequence that is 100% complementary to the probe can be identified (homologous probe method). Alternatively, stringent conditions can be adjusted to allow some sequence mismatches in order to detect lower similarities (heterologous probe method). Typically, the probe length is less than about 1000 or 500 nucleotides. Typically, stringent conditions are those in which the salt concentration is less than about 1.5 M Na ions, typically about 0.01 M to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3, and the temperature is: when used for short probes (e.g., 10 to 50 nucleotides), at least about 30°C; when used for long probes (e.g., greater than 50 nucleotides), at least about 60°C. Stringent conditions can also be achieved by adding destabilizing agents such as formamide. Exemplary low stringency conditions include hybridization at 37°C using 30% to 35% formamide buffer, 1M NaCl, 1% SDS (sodium dodecyl sulfate), and washing in 1× to 2× SSC (20× SSC = 3.0M NaCl / 0.3M trisodium citrate) at 50°C to 55°C. Exemplary moderate stringency conditions include hybridization at 37°C in 40% to 45% formamide, 1.0M NaCl, 1% SDS, and washing in 0.5× to 1× SSC at 55°C to 60°C. Exemplary high stringency conditions include hybridization at 37°C in 50% formamide, 1M NaCl, 1% SDS, and a final wash in 0.1× SSC at 60°C to 65°C for at least about 20 minutes. Optionally, the wash buffer may contain about 0.1% to about 1% SDS. Duration of hybridization is typically less than about 24 hours, typically about 4 hours to about 12 hours. Specificity generally depends on post-hybridization washes, with the key factors being the ionic strength and temperature of the final wash solution. The Tm (thermodynamic melting point) of a DNA-DNA hybrid can be approximated by the formula of Meinkoth and Wahl (1984) Anal. Biochem. 138:267-284: Tm = 81.5°C + 16.6 (log M) + 0.41 (% GC) - 0.61 (% formamide) - 500 / L, where M is the molar concentration of monovalent cations, % GC is the percentage of guanosine and cytosine nucleotides in the DNA, "% formamide" is the percentage of formamide in the hybridization solution, and L is the base pair length of the hybrid. The Tm is the temperature (under defined ionic strength and pH) at which 50% of the complementary target sequence hybridizes to a perfectly matched probe. Washes are typically performed at least until equilibrium is reached and low background levels of hybridization are achieved, such as for 2 hours, 1 hour, or 30 minutes. Each 1% mismatch should reduce the Tm by about 1°C; thus, the Tm, hybridization, and / or wash conditions can be adjusted to hybridize to sequences of the desired identity. For example, if sequences with ≥90% identity are desired, the Tm can be reduced by 10°C.Generally, stringent conditions are selected to be about 5°C lower than the Tm of the specific sequence and its complement at a defined ionic strength and pH. However, under very stringent conditions, hybridization and / or washing can be performed at 4°C below the Tm; under moderately stringent conditions, hybridization and / or washing can be performed at 6°C below the Tm; and under low stringency conditions, hybridization and / or washing can be performed at 11°C below the Tm.

[0029] In some embodiments, fragments of nucleotide sequences and the amino acid sequences they encode are also included. As used herein, the term "fragment" refers to a portion of the nucleotide sequence of a polynucleotide of an embodiment or a portion of the amino acid sequence of a polypeptide. Fragments of nucleotide sequences can encode protein fragments that retain the biological activity of a native or corresponding full-length protein and thus have protein activity. Mutant proteins include biologically active fragments of native proteins that contain contiguous amino acid residues that retain the biological activity of the native protein. Some embodiments also include transformed plant cells or transgenic plants that contain the nucleotide sequence of at least one embodiment. In some embodiments, plants are transformed using an expression vector that contains the nucleotide sequence of at least one embodiment and a promoter that drives expression in plant cells operably linked thereto. Transformed plant cells and transgenic plants refer to plant cells or plants that contain heterologous polynucleotides in their genomes. Generally speaking, the heterologous polynucleotides are stably integrated in the genome of the transformed plant cells or transgenic plants so that the polynucleotides are passed on to future generations. The heterologous polynucleotides can be integrated into the genome individually or as part of an expression vector. In some embodiments, the plants involved in the present application include plant cells, plant protoplasts, plant cell tissue cultures that can regenerate plants, plant calli, plant masses and plant cells, which are complete plants or parts of plants, such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruits, kernels, ears, cobs, shells, stalks, roots, root tips, anthers, etc. The present application also includes plant cells, protoplasts, tissues, calli, embryos, flowers, stems, fruits, leaves and roots derived from the transgenic plants of the present application or their progeny, and thus at least partially comprising the nucleotide sequence of the present application.

[0030] The term "amplification" in the context of nucleic acid amplification is any process in which additional copies of a selected nucleic acid (or its transcribed form) are produced. Common amplification methods include various polymerase-based replication methods, including polymerase chain reaction (PCR), ligase-mediated methods such as ligase chain reaction (LCR), and RNA polymerase-based amplification (e.g., by transcription) methods.

[0031] The term "inbred line" used in this article refers to a line with more uniform agronomic traits and a simpler genetic basis obtained by continuously eliminating poor ear rows and selecting individual plants with better agronomic traits for self-pollination over several generations under artificially controlled self-pollination.

[0032] "Geroplasm" refers to the genetic material of or derived from an individual (e.g., a plant), a group of individuals (e.g., a plant line, variety, or family), or a clone of a line, variety, species, or culture. Germplasm can be part of an organism or cell, or can be isolated from an organism or cell. Germplasm generally provides the genetic material and specific molecular makeup that provides the physical basis for some or all of the heritable traits of an organism or cell culture. As used herein, germplasm includes cells, seeds, or tissues from which new plants can be grown, or plant parts such as leaves, stems, pollen, or cells that can be cultured into whole plants.

[0033] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention. Without departing from the spirit and substance of the present invention, the modification or replacement of the inventive method, step or condition, all fall within the scope of the application. If not otherwise specified, the embodiments are according to conventional experimental conditions, such as the molecular cloning laboratory manual (Sambrook J & Russell DW, Molecular cloning: alaboratory manual, 2001) of Sambrook et al., or according to the conditions of manufacturer's instructions. If not otherwise specified, the chemical reagents used in the embodiments are conventional commercial reagents, and the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0034] Example 1 Study on the Function of T01 Transcript of GRMZM2G005732 Gene

[0035] In previous research, the inventors discovered that the GRMZM2G005732 gene (for details, see maizeGDB, http: / / www.maizegdb.org) affects the flowering period of maize. Overexpression of this gene (T01 transcript) delays maize flowering, while mutating this gene using gene editing technology advances maize flowering (see patent CN112646820 A for details).

[0036] The inventors further identified the yield traits of three maize lines overexpressing T01 transcripts. The results showed that, except for the change in flowering time, there were no significant differences in yield / biomass related traits such as plant height, ear height, aboveground biomass wet weight and aboveground biomass dry weight compared with the control (see Figure 1 ).

[0037] Example 2 Study on the Function of T03 Transcript of GRMZM2G005732 Gene

[0038] Further query the information of GRMZM2G005732 gene in maizeGDB database and find that there are 3 transcripts predicted by this gene. Since the public information does not clearly state whether these transcripts have different functions and what the specific functions are, the inventor obtains the nucleic acid molecules of other transcripts by artificial synthesis (direct cloning cannot be obtained) to further study its function. Since the amino acid sequence encoded by T02 transcript is very short, the present invention is referenced to the CDS sequence (as shown in SEQ ID NO.1) of T03 transcript, and the sequence GRMZM2G005732-T03 (as shown in SEQ ID NO.2) is artificially optimized and synthesized again according to the monocotyledon codon preference. The amino acid sequence encoded by the optimized sequence is consistent with the CDS sequence in the database as shown in SEQ ID NO.3.

[0039] According to the method in Example 3 of patent CN 112646820 A, the synthesized GRMZM2G005732-T03 CDS nucleic acid molecule after artificial codon optimization was constructed into an expression vector and transformed into corn.

[0040] Specifically: the vector uses pCAMBIA3300 as the backbone. The pCAMBIA3300 plasmid is double-digested with BamHI and SacI to recover the vector fragment; the artificially synthesized T03 is designed with BamHI and SacI restriction sites, and after restriction digestion, it is ligated between the BamHI and SacI sites of pCAMBIA3300 to construct a plant expression vector (see the vector map for details). Figure 2 The constructed vector contains the T03 expression cassette (containing the ZmUbi promoter and the nos terminator) and the bar gene expression cassette (containing the 35s promoter and the PolyA terminator). The vector plasmid was transformed into Agrobacterium tumefaciens EHA105.

[0041] Agrobacterium EHA105 containing the expression vector was spread on YEP solid culture medium and cultured in the dark at 28°C for 1-3 days. The cultured Agrobacterium was scraped off the plate and resuspended, and the OD550 was adjusted to 0.3 to prepare the infection solution for later use. Take the young ears of corn KN5585 (from Weimi Biotechnology (Jiangsu) Co., Ltd., variety right application number 20191002444) 9-12 days after pollination, remove the bracts, disinfect with 75% alcohol for 10 minutes, and remove the immature embryos into a centrifuge tube containing 2mL of resuspension solution, 100 immature embryos per tube, for later use. During infection, discard the resuspension, add 2mL of infection solution, gently invert the centrifuge tube several times to mix, and let it stand in the dark at room temperature for 5 minutes. After the infection is completed, the immature embryos are inoculated into the co-culture medium with the scutellum facing up and cultured in the dark at 20°C for 3 days. Transfer to the resting culture medium and culture in the dark at 28°C for 7 days. Then transfer it to the selection medium S1 containing 1.5 mg / L bialaphos and culture it in the dark at 28°C for 2 weeks. If the initial callus tissue has been obtained, transfer it to the screening medium S2 containing 3 mg / L bialaphos, and replace the S2 medium every two weeks. When the resistant callus tissue obtained by screening proliferates to a diameter of about 2 cm, transfer it to a dark differentiation medium and culture it in the dark at 25°C for 2-3 weeks. The differentiated coleoptile is transferred to a light differentiation medium and cultured under light at 25°C for 2 weeks. When the coleoptile forms complete seedlings and roots, the seedlings are transferred to a culture bottle to promote root growth and seedling growth. After 10 days, the seedlings are transplanted into nutrient pots and cultured in an indoor greenhouse. After the seedlings grow 1-2 new leaves, they are moved into large flower pots. The transgenic corn and the control were planted at the Hainan South Transgenic Experiment Base of Jilin Academy of Agricultural Sciences, and all materials were PCR tested to determine whether they were positive transgenic strains.

[0042] The phenotypic comparison of the materials with positive PCR test results and the negative control materials of KN5585 was carried out. It was found that the corn transformed with T03 had significantly increased plant height, ear height, number of stem nodes, thicker stems, and more leaves than the control KN5585 (see Figure 3 ).

[0043] Four strains were selected to investigate the traits of plant height, ear height, and aboveground biomass (wet weight and dry weight). Five strains with the same growth were selected for investigation from different strains and controls. For the aboveground biomass (wet weight), the plants were weighed immediately after being retrieved from the experimental field, and all tissues of the whole plant except the ear were weighed. The dry weight was measured again after the aboveground part was naturally air-dried for 30 days. The results showed that the plant height of the T03-transformed strains increased by 24% to 53% compared with the control, the ear height increased by 95% to 154%, the aboveground biomass wet weight increased by 81% to 134%, and the aboveground biomass dry weight increased by 60% to 98% ( Figure 4 ).

[0044] Combining phenotypic and agronomic trait data analysis, it was found that corn expressing T03 showed significant increases in plant height, ear height, and aboveground biomass compared to the negative control.

[0045] These results indicate that another transcript, T03, of the GRMZM2G005732 gene is associated with both yield and biomass traits in maize. Expressing the T03 transcript in maize can increase yield and / or biomass, thereby enabling the development of new high-yield / high-biomass maize germplasm. This provides a new approach for developing new high-yield / high-biomass maize germplasm.

[0046] Although the present invention has been described in detail above using general descriptions and specific embodiments, it will be apparent to those skilled in the art that modifications and improvements may be made based on the present invention. Therefore, such modifications and improvements, which do not depart from the spirit of the present invention, are intended to be within the scope of protection claimed herein.

Claims

1. An application of a nucleic acid molecule in increasing corn biomass, characterized in that: The nucleotide sequence of the nucleic acid molecule is shown in SEQ ID NO. 2; The increase in corn biomass is achieved by any one of the following: increasing plant height, and / or increasing ear height, and / or increasing the number of stem nodes, and / or thickening the stem, and / or increasing the number of leaves, and / or increasing the aboveground biomass.

2. An application of an expression cassette in increasing corn biomass, characterized in that: The expression cassette contains the nucleic acid molecule according to claim 1; The increase in corn biomass is achieved by any one of the following: increasing plant height, and / or increasing ear height, and / or increasing the number of stem nodes, and / or thickening the stem, and / or increasing the number of leaves, and / or increasing the aboveground biomass.

3. An application of an expression vector in increasing corn biomass, characterized in that: The expression vector contains the expression cassette according to claim 2; The increase in corn biomass is achieved by any one of the following: increasing plant height, and / or increasing ear height, and / or increasing the number of stem nodes, and / or thickening the stem, and / or increasing the number of leaves, and / or increasing the aboveground biomass.

4. A use of a host cell in increasing corn biomass, characterized in that: The host cell contains the expression vector according to claim 3; The increase in corn biomass is achieved by any one of the following: increasing plant height, and / or increasing ear height, and / or increasing the number of stem nodes, and / or thickening the stem, and / or increasing the number of leaves, and / or increasing the aboveground biomass; The host cell is an Agrobacterium cell.

5. A method for increasing corn biomass, characterized by: Expressing the nucleic acid molecule of claim 1 in corn and selecting corn plants with increased biomass; The increase in corn biomass is achieved by any one of the following: increasing plant height, and / or increasing ear height, and / or increasing the number of stem nodes, and / or thickening the stem, and / or increasing the number of leaves, and / or increasing the aboveground biomass.

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