CN114395580A

CN114395580A - Gene for controlling plant height of corn

Info

Publication number: CN114395580A
Application number: CN202210202870.1A
Authority: CN
Inventors: 罗芸; 刘喻; 严建兵
Original assignee: Huazhong Agricultural University
Current assignee: Huazhong Agricultural University
Priority date: 2022-03-02
Filing date: 2022-03-02
Publication date: 2022-04-26
Anticipated expiration: 2042-03-02
Also published as: CN114395580B

Abstract

The invention relates to an application of ZmTPK2 gene in controlling the characteristics of plant height and/or ear height of corn, belonging to the field of molecular genetics. The invention discloses application of a ZmTPK2 gene in controlling the characteristics of plant height and/or ear height of corn. Further, the invention provides a method for reducing the plant height and/or ear height of corn.

Description

Gene for controlling plant height of corn

Technical Field

The invention relates to an application of ZmTPK2 gene in controlling the high character of corn plants, belonging to the field of molecular genetics.

Background

The dwarfing character brings breakthrough to crop yield, and the method for reducing the plant height of the crops has great application potential. In the main agronomic traits of corn, the plant height influences the lodging resistance, photosynthetic efficiency and harvest index of the corn and is closely related to the yield of the corn. Therefore, the plant height trait has important value in corn breeding practice and germplasm resource improvement work.

The plant height and ear position high traits are commonly controlled by the major genes and the micro-effect polygenes, and are expressed as typical quantitative trait inheritance. Although some genes for regulating the height of the corn plant are positioned and cloned (China southern agriculture university, corn ZmPIF3s mutant protein, a coding gene thereof and application thereof in breeding: CN201910273522.1[ P ] 2019-08-02.; Hangzhou Ruifeng Biotech limited company, CYP78A gene is applied to increasing the height of the corn plant and enhancing the growth vigor of the plant: CN201510230547.5[ P ] 2016-12-07.; China agricultural university, a gene related to the height of the corn plant, a coding protein thereof and application thereof: CN200410037404.4[ P ] 2005-01-26.; Chinese agricultural academy crop scientific research institute, a method for creating a corn dwarfing material by using a gene editing technology: CN201910371358.8[ P ] 2019-08-16.), more genes related to be further cloned.

Disclosure of Invention

In order to solve the problems, the invention provides a novel gene which can control the height of the maize plant and/or the height of the ear position, can reduce the height of the maize plant and/or the height of the ear position by inhibiting the expression or activity of a protein coded by the gene, and can be used for cultivating dwarf maize varieties.

The invention aims to provide a nucleic acid sequence of a gene ZmTPK2 influencing the plant height and/or ear height traits of corn and an amino acid sequence coded by the same.

The invention also aims to disclose a method for improving the plant height and/or ear height of the corn.

The third purpose of the invention is to disclose a mutant gene of dwarf maize.

In order to achieve the purpose, the invention adopts the following technical scheme:

the invention provides an application of a corn gene in regulating and controlling the plant height and/or ear height of corn, which is characterized in that: the gene sequence is shown in any one of SEQ ID NO.2-SEQ ID NO. 3. Wherein SEQ ID NO.2 is the genome sequence of the gene in corn, and SEQ ID NO.3 is the cDNA sequence of the gene. The identification result of the invention shows that after the gene is knocked out by using a gene editing means, the height of the corn plant and/or the height of the ear position are/is reduced, so that the technical effect of improving the height of the corn plant and/or the height of the ear position can be achieved by constructing the sequence shown by SEQ ID NO.2 or SEQ ID NO.3 on an overexpression vector, a corresponding gRNA recognition target or RNAi interference sequence can be designed according to the sequence shown by SEQ ID NO.2 or SEQ ID NO.3, and the normal expression of the gene is interfered in a gene editing or RNAi interference or T-DNA directional insertion mode, so that the technical effect of reducing the height of the corn plant and/or the height of the ear position is achieved.

In another aspect, the present invention provides a method for reducing plant height and/or ear height of corn, characterized in that: inhibiting the expression and/or activity of the protein shown by SEQ ID NO.1 in corn, and selecting the plants with reduced plant height and/or ear height of corn.

In some embodiments, the above-described method of inhibiting protein expression and/or activity comprises any one of gene editing, RNA interference, T-DNA insertion.

In some embodiments, the above gene editing employs the CRISPR/Cas9 method.

In some embodiments, the DNA sequence of the genomic target region in maize of the CRISPR/Cas9 method described above is shown as SEQ ID No.4 or SEQ ID No. 5.

In another aspect, the present invention also provides a kit for reducing plant height and/or ear height of corn, wherein the kit is characterized in that: including any of the following:

(1) an RNA molecule targeting the sequence shown in SEQ ID NO.4 or SEQ ID NO. 5; the RNA molecule can be an sgRNA molecule containing a gRNA, crRNAs and tracrRNA structure, or a complex formed by the gRNA, the crRNAs and the tracrRNA independently, or a complex comprising the gRNA and the crRNAs;

(2) a DNA molecule encoding the RNA of (1);

(3) a vector expressing the RNA of (1).

In some embodiments, the RNA molecule has a sequence as set forth in SEQ ID NO.6 or SEQ ID NO. 7.

In another aspect, the present invention also provides a maize mutant gene characterized by: the sequence of the mutant gene is shown as SEQ ID NO.8 or SEQ ID NO. 9.

In another aspect, the invention also provides application of the maize mutant gene in reducing the plant height and/or the ear height of maize. The mutant gene is introduced into other corn materials by a conventional hybridization transformation method, so that the technical effect of reducing the plant height and/or the ear position height of the receptor corn can be achieved.

Compared with the prior art, the ZmTPK2 gene and the protein coded by the ZmTPK2 gene have the function of regulating and controlling the plant height and/or ear height of corn, and the function is not reported in the prior published data and is not suggested by the technology. Knocking out the gene by using a CRISPR/Cas9 method can reduce the height of a corn plant and/or the height of a panicle position. The invention identifies the mutant gene sequence after gene knockout, and can further improve the plant height and/or ear height character of other corn varieties by using the mutant genes, thereby cultivating dwarf corn varieties and improving the lodging resistance of corn.

Drawings

FIG. 1 maize ear length QTL location results. 1-10 represent chromosome 1 through chromosome 10, respectively. The ordinate represents LOD value, and the abscissa represents genetic distance on chromosome

FIG. 2 Gene editing vector map. The main elements are labeled on the figures.

FIG. 3 display of target positions, target sequences (grey) and edited sequences for gene editing. "-" indicates a deletion.

Detailed Description

The following definitions and methods are provided to better define the present application and to guide those of ordinary skill in the art in the practice of the present application. Unless otherwise indicated, terms are to be understood in accordance with their ordinary usage by those of ordinary skill in the relevant art. All patent documents, academic papers, industry standards and other publications, etc., cited herein are incorporated by reference in their entirety.

As used herein, "maize" is any maize plant and includes all plant varieties that can be bred with maize, including whole plants, plant cells, plant organs, plant protoplasts, plant cell tissue cultures from which plants can be regenerated, plant calli, intact plant cells in plants or plant parts, such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruits, stems, roots, root tips, anthers, and the like. Unless otherwise indicated, nucleic acids are written from left to right in the 5 'to 3' direction; amino acid sequences are written from left to right in the amino to carboxy direction. Amino acids may be referred to herein by their commonly known three letter symbols or by the one letter symbols recommended by the IUPAC-IUB Biochemical nomenclature Commission. Similarly, nucleotides may be represented by commonly accepted single-letter codes. Numerical ranges include the numbers defining the range. As used herein, "nucleic acid" includes reference to deoxyribonucleotide or ribonucleotide polymers in either single-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 term "encode" or "encoded" when used in the context of a particular nucleic acid means that the nucleic acid contains the necessary information to direct translation of the nucleotide sequence into a particular protein. The information encoding the protein is represented using a codon. As used herein, "full-length sequence" in reference to a particular polynucleotide or protein encoded thereby refers to the entire nucleic acid sequence or the entire amino acid sequence having a native (non-synthetic) endogenous sequence. The full-length polynucleotide encodes the full-length, catalytically active form of the particular protein. The terms "polypeptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The term is used for amino acid polymers in which one or more amino acid residues are artificial chemical analogues of the corresponding naturally occurring amino acids. The term is also used for naturally occurring amino acid polymers. The terms "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 natural amino acids that can function in a similar manner as naturally occurring amino acids.

As used herein, the terms "isolated" and "purified" may be used interchangeably to refer to a nucleic acid or polypeptide, or biologically active portion thereof, that is substantially or essentially free of components that normally accompany or react with the nucleic acid or polypeptide as found in its naturally occurring environment. Thus, an isolated or purified nucleic acid or polypeptide produced by recombinant techniques is substantially free of other cellular material or culture medium, or is substantially free of chemical precursors or other chemicals when chemically synthesized. An "isolated" nucleic acid is typically free of sequences (such as sequences encoding proteins) that naturally flank the nucleic acid (i.e., sequences located at the 5 'and 3' ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For example, in various embodiments, an isolated nucleic acid may comprise less than about 0.5kb of nucleotide sequences that naturally flank the nucleic acid in the genomic DNA of the cell from which the nucleic acid is derived.

In this application, the words "comprise", "comprising" or variations thereof are to be understood as embracing elements, numbers or steps in addition to those described. By "subject plant" or "subject plant cell" is meant a plant or plant cell in which the genetic modification has been effected, or a progeny cell of the plant or cell so modified, which progeny cell comprises the modification. The "control" or "control plant cell" provides a reference point for measuring the phenotypic change of the test plant or plant cell. The control plant or plant cell may include, for example: (a) a wild-type plant or cell, i.e., a plant or cell having the same genotype as the starting material for the genetic alteration that produced the test plant or cell; (b) plants or plant cells having the same genotype as the starting material but which have been transformed with an empty construct (i.e., a construct that has no known effect on the trait of interest, such as a construct comprising a target gene); (c) a plant or plant cell that is a non-transformed isolate of a subject plant or plant cell; (d) a plant or plant cell that is genetically identical to the subject plant or plant cell but that has not been exposed to conditions or stimuli that induce expression of the gene of interest; or (e) the subject plant or plant cell itself, under conditions in which the gene of interest is not expressed.

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 suitable tools and procedures for engineering or engineering amino acid sequences and potential gene sequences of proteins of agricultural interest.

In some embodiments, changes may be made to the nucleotide sequences of the present application to make conservative amino acid substitutions. The principles and examples of conservative amino acid substitutions are further described below. In certain embodiments, substitutions that do not alter the amino acid sequence of the nucleotide sequences of the present application can be made in accordance with the codon preferences disclosed for monocots, e.g., codons encoding the same amino acid sequence can be substituted with monocot preferred codons without altering the amino acid sequence encoded by the nucleotide sequence. In some embodiments, a portion of the nucleotide sequence in this application is replaced with a different codon that encodes the same amino acid sequence, such that the nucleotide sequence is not altered while the amino acid sequence encoded thereby is not altered. Conservative variants include those sequences that, due to the degeneracy of the genetic code, encode the amino acid sequence of one of the proteins of the embodiments. In some embodiments, a partial nucleotide sequence herein is replaced according to monocot preferred codons. One 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, e.g., hydrophobicity, charge, size, etc., of the substituents. Exemplary amino acid substituent groups having various of the foregoing properties are 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 regarding suitable amino acid substitutions that do not affect the biological activity of the Protein of interest can be found in the model of the Atlas of Protein sequences and structures (Protein Sequence and Structure Atlas) (Natl. biomed. Res. Foundation, Washington, D.C.) (incorporated herein by reference). Conservative substitutions such as exchanging one amino acid for another with similar properties may be made. Identification of sequence identity includes hybridization techniques. For example, all or part of a known nucleotide sequence is used as a probe for selective hybridization to other corresponding nucleotide sequences present in a population of cloned genomic DNA fragments or cDNA fragments (i.e., a genomic library or cDNA library) from a selected organism.

In some embodiments, fragments of the nucleotide sequences and the amino acid sequences encoded thereby are also included. As used herein, the term "fragment" refers to a portion of the nucleotide sequence of a polynucleotide or a portion of the amino acid sequence of a polypeptide of an embodiment. Fragments of the nucleotide sequences may encode protein fragments that retain the biological activity of the native or corresponding full-length protein, and thus have protein activity. Mutant proteins include biologically active fragments of the native protein that comprise contiguous amino acid residues that retain the biological activity of the native protein. Some embodiments also include a transformed plant cell or transgenic plant comprising the nucleotide sequence of at least one embodiment. In some embodiments, plants are transformed with an expression vector comprising at least one embodiment of the nucleotide sequence and operably linked thereto a promoter that drives expression in plant cells. Transformed plant cells and transgenic plants refer to plant cells or plants that comprise a heterologous polynucleotide within their genome. Generally, the heterologous polynucleotide is stably integrated within the genome of the transformed plant cell or transgenic plant such that the polynucleotide is transmitted to progeny. The heterologous polynucleotide may be integrated into the genome alone or as part of an expression vector. In some embodiments, the plants to which the present application relates include plant cells, plant protoplasts, plant cell tissue cultures from which plants can be regenerated, plant calli, plant clumps, and plant cells, which are whole plants or parts of plants, such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruits, nuts, ears, cobs, husks, stalks, roots, root tips, anthers, and the like. The present application also includes plant cells, protoplasts, tissues, calli, embryos, and flowers, stems, fruits, leaves, and roots derived from the transgenic plants of the present application or progeny thereof, and thus comprising at least in part the nucleotide sequences of the present application.

The invention locates QTL influencing corn ear length character from RIL colony constructed by KUI3 and B77, and a major effective site is found in 58kb interval of No.6 chromosome. According to the gene function annotation information, the Zm00001d037916 is presumed to be a gene for controlling the corn ear length trait in the QTL interval and is named as ZmTPK 2.

The invention further verifies the function of the gene through a gene editing knockout experiment, and unexpectedly discovers that the plant height and the ear height of the corn are both reduced after the ZmTPK2 gene is edited.

The corn with reduced plant height and/or ear height can be used for cultivating dwarf corn varieties and increasing the lodging resistance of the corn. The invention further determines the mutant gene sequence after gene knockout, and the mutant genes can be transferred to other corn varieties to improve the plant height and/or ear height character of the receptor, thereby cultivating dwarf varieties and increasing the lodging resistance of corn.

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. Modifications or substitutions to methods, steps or conditions of the present invention may be made without departing from the spirit and substance of the invention and are intended to be included within the scope of the present application. Unless otherwise indicated, the examples follow conventional experimental conditions, such as the Molecular cloning laboratory Manual of Sambrook et al (Sambrook J & Russell D W, Molecular cloning: a laboratory Manual,2001), or the conditions as recommended by the manufacturer's instructions. Unless otherwise specified, the chemical reagents used in the examples are all conventional commercially available reagents, and the technical means used in the examples are conventional means well known to those skilled in the art.

Example 1 location of corn ear Length QTL

The invention constructs RIL group from short ear corn inbred line material KUI3 and long ear corn inbred line material B77 as parents, and by investigating corn ear length phenotype values at different planting time and places and utilizing genotype data and RNA-seq data of parent Illumina Maize SNP50 BeadChip, a QTL qEL6.2 for controlling ear length and grain number is identified in the interval range of 134.1-139.3Mb (V2 version) of corn sixth chromosome, and 18.92% of phenotypic variation can be explained. According to the positioning result, when the QTL is detected at 8 environmental points, the highest LOD value, the effect value and the explained phenotypic variation rate of the QTL are different, but the QTL can be detected at all 8 environmental points, which shows that the QTL is very stable and can act at a plurality of environmental points (figure 1).

To pinpoint QTL qel6.2, the remaining hybrid material F6 (HIF) in the RIL population was selected: HZAU-551-1 is used as an initial positioning material, molecular markers IDP8645, IDP785, B4, B14 and the like are designed to screen background heterozygous segments, and finally materials heterozygous in the positioning segments and homozygous in the background are selected to verify the QTL and carry out next fine positioning. In the fine positioning process, a single-factor variance analysis is carried out on the near-isogenic line ear length phenotype after continuously screening new markers and new recombinant individual strains for propagation, so that the positioning section is narrowed. According to the results of the ear length phenotype analysis of different near isogenic lines, the QTL qEL6.2 is divided into two smaller QTLs which are respectively named as qEL6.2-1 and qEL6.2-2.

In order to clone the panicle length gene in qEL6.2-1, progeny of multiple different recombination types of maize within the mapped region were planted in Wuhan in spring 2018 and genotyped. From which NILs homozygous for different families in the same chromosome segment were obtained using multiple markers within qEL6.2-2 interval^KUI3Genotype and homozygous NIL^B77The genotype material and the new chromosome recombination individual plant are screened out for selfing. And (3) carrying out baoding planting on selfing progenies of different recombination types and new filial generations of the recombinant single plants in the spring of 2019 in Hebei, screening a new genotype homozygous material, and positioning qEL6.2-1 locus in the interval of the markers S155-S157 by utilizing further genotyping and fine positioning. Based on maize B73 reference genome (RefGenV4) annotation information, this interval is approximately 58kb, and contains an entire function annotation gene Zm00001d037915, and the promoter region of Zm00001d 037916.

Example 2 functional verification of Gene

The Zm00001d037916 gene encodes thiamine pyrophosphate kinase 2, which is named ZmTPK2 in the invention. According to annotation information on a maize reference genome, the amino acid sequence of the ZmTPK2 encoding protein is shown as SEQ ID NO.1, and the genome sequence is shown as SEQ ID NO. 2. In order to verify the function of the gene, the gene is knocked out by using a CRISPR/Cas9 gene editing technology. The method comprises the following specific steps:

the gRNA was designed by using the online software CRISPR-P2.0 (http:// CRISPR. hzau. edu. cn/CRISPR2/) to select two target editing sites in each CDS region of the gene (see Table 1 for sequence). About 2000bp of a fusion unit sequence of 'U6-promoter 1-gRNA 1-sgRNA-U6-promoter 2-gRNA 2-sgRNA' is directly synthesized, the fusion unit sequence is cut from an intermediate vector PUC57 by a double enzyme digestion system, and the cut fragment is connected to a framework vector CPB-ZmUbi-hspCs 9 by a homologous recombination method (figure 2). And transforming the recombinant vector into an escherichia coli competent cell DH5 alpha, detecting a target sequence, and transforming a maize inbred line KN5585 after the sequence detection is correct. The obtained T0 generation transformed plants were tested for two target sites of each gene. And (3) sequencing after PCR amplification, and analyzing the editing type according to the sequencing sequence.

TABLE 1 Gene edited target sequences and detection primer information

A total of 2 successfully edited lines were obtained, designated KO1 and KO2, respectively, and the target sequences were edited as shown in fig. 3. The sequences of the ZmTPK2 gene in KO1 and KO2 after mutation are shown in SEQ ID NO.8 and SEQ ID NO. 9.

Further analyzing the agronomic traits of the knockout strain, the invention unexpectedly discovers that when the gene is knocked out, the corn plant type is obviously reduced, the plant height and the ear height are obviously reduced, and the flowering period is not influenced (Table 2).

TABLE 2 influence of ZmTPK2 on plant type and flowering time

OE 1: 1 over-expressed material; KO1, KO 2: 2 knock-out materials; NT: a receptor control; n: number of materials. The phenotypic values are expressed as mean ± standard deviation, and significance was calculated using one-way anova.

The above results show that the ZmTPK2 mutant gene in KO1 and KO2 can reduce the plant height and/or ear height of corn. Any one of the two mutant sequences can be introduced into the conventional corn material by hybridization to reduce the height of the corn plant and/or the height of the ear position, so that dwarf corn varieties can be cultivated.

Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Sequence listing

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<211> 19

<212> DNA

<213> Zea mays

<400> 4

cccttcaacc gcaccgtca 19

<210> 5

<211> 20

<212> DNA

<213> Zea mays

<400> 5

ggatggatca gcggcggtgc 20

<210> 6

<211> 102

<212> DNA/RNA

<213> unknown (Artificial Synthesis)

<400> 6

ugacggugcg guugaagggg uuuuagagcu agaaauagca aguuaaaaua aggcuagucc 60

guuaucaacu ugaaaaagug gcaccgaguc ggugcuuuuu uu 102

<210> 7

<211> 103

<212> DNA/RNA

<213> unknown (Artificial Synthesis)

<400> 7

ggauggauca gcggcggugc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc 60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu uuu 103

<210> 8

<211> 2281

<212> DNA

<213> Zea mays

<400> 8

gcgtgattgc gtgaaagtct cttctggatc caaactaata aacaccacgc gagtacgccg 60

cgtgcgacct cgaggctcga gcttgacgcc cgcgtgccat ggctcccacg gcgccgacgc 120

aacggccgac gatgcgccac tcctccgcgt tcctcctccc tggatcagcg gcggtgcggg 180

tgtgcgcgga cggtggcgcc aaccgcgtct tcgacggcat gccggccctg ctccccggcc 240

aggaccccga cgaggttcgc gcgaggtacc attcttcgtt gctagcgtct ctgtcggttg 300

catcacattg cgtctcgttg tctcaagagt acaactgtat aagatcgtac cgggtgtgct 360

tgcctcctag tatttatttg gtcatatatg attttcccag tacaatctct gctcgagtct 420

gcccgcgact gcgtgcccag catatacttg atgtaatgca ctaatgcttg ggccttgggg 480

ggagtggtgg gctttgccaa cgagttgcct gctatgactc tactgttgtt gctctgcgcg 540

catccgattc tagagaagcg taaagggttc tcgtatcttc ccagttcaaa tgacttacta 600

gtaataatgt aatatgctgg tgctgacgtt gggcgccact tctcggtgaa acgtcagctt 660

actggcgaat taagggctgc gttggtgtta gtagtggttt ctgcattcac ttgggagcag 720

ttatctttta tatcatcaac gaatacgctc aaaccacaga gaaacttttt cttgtaaaga 780

tggaaggcta aaagttgctt caaatcgcac ttcgccgatg tgcattataa acagctagtt 840

aatggaattt ccgtttcact taccaagttt cgttatttct ttatgcagtt acaagccaga 900

tgtaatcaaa ggggatatgg actcagtaag accagaagtg aaggaacatt attccaactt 960

ggtaaagctg ctttaattta cattcattat tggtatgaat ctactaaaag ggtaaaacga 1020

attttaattt gggacggaag gagtactagc tagtttaact tctctagtgc ctcttttagt 1080

tctggtgatc aaattggcat ttcccttgta ccaatccttg ttgagagaaa aaaaagcaat 1140

ggtcttagag ctgtatatat cagatgaggg ttcatccgaa aggggataaa taacagtatc 1200

cttttttgtt ccgacttagc atagtatagc tggtggcatc tagtaagatg cagctagctg 1260

attgtgatta taatatctgc tttataatat aatggttgat gattcttgtt ccgacttatg 1320

cttgacatgt tttgtttctt ctgccctcca aaaggtgcat ctatatcatc gtatcatgga 1380

ggttcttgta ttgatagatc agtcttattg cataagatat tgaagctgta cattagggtt 1440

ttcattttcg acttcaggca tggtctttct gaaaacgaga tcctttctta tggtaattct 1500

catgtattat caattgcagg gcaccagaat tgttgacgag tcacatgacc aggacacgac 1560

cgatttacac aaatgcatag cttttatagc tgagaattat tctgccacta acaaatctaa 1620

cgtaagataa tccaacccaa caatattcta atcaaattag tatggaagga caataatgta 1680

tctacatttt tttaaagcag ctcagcatct tggtccttgg agcacttggg ggaaggtttg 1740

atcacgagat gggaaacatc aatgtgcttc atctcttccc aagcatcaat atcgtcctcc 1800

tatcagatga ttgtctgatc tttctgctct cgaaaatgca tacccacgag attcacgttg 1860

aaaagtcgat cgagggaccc cactgtggat tgatcccgat tggcatgccg tcgactagca 1920

ccacgaccac cgggcttcga tggaatctag gttatcctcg tccttatcct tcctaatgca 1980

ggcttagcaa cagaagttga tgtttacaac gtgattacct ctgtacagat aataccagta 2040

tgagcttcgg aggattgatc agcacgtcaa atatcgtgga agaggacaag gtaacggtca 2100

cctcggattc agaccttatg tggacaacat cgctccggaa gtgaatctac actcgagtat 2160

agagtatcgt tatactgttg taatattaga gaaataagtg acatgctttt aatatatata 2220

tatatatata tatatatata tatatatata tatatatata tatatatata tatattaaat 2280

a 2281

<210> 9

<211> 2496

<212> DNA

<213> Zea mays

<400> 9

gcgtgattgc gtgaaagtct cttctggatc caaactaata aacaccacgc gagtacgccg 60

cgtgcgacct cgaggctcga gcttgacgcc cgcgtgccat ggctcccacg gcgccgacgc 120

aacggccgac gatgcgccac tcctccgcgt tcctcctccc tcaaccgcac cgtcatcgcc 180

cgcgcctggt cccgatgacg ccgccgtcgc ggtcgtcgtc ctcaaccagc cgctgccccg 240

cttcgcgccc ctcctctggt ccagaggtac gccgacggtg ctcctcccct cctccttgcc 300

tgaactcccc ccgtgtctca tctactggcg gcactgccgt ccctgaccct gactgatgcg 360

ctgtttccgt gcctggagat cagcggcggt gcgggtgtgc gcggacggtg gcgccaaccg 420

cgtcttcgac ggcatgccgg ccctgctccc cggccaggac cccgacgagg ttcgcgcgag 480

gtaccattct tcgttgctag cgtctctgtc ggttgcatca cattgcgtct cgttgtctca 540

agagtacaac tgtataagat cgtaccgggt gtgcttgcct cctagtattt atttggtcat 600

atatgatttt cccagtacaa tctctgctcg agtctgcccg cgactgcgtg cccagcatat 660

acttgatgta atgcactaat gcttgggcct tggggggagt ggtgggcttt gccaacgagt 720

tgcctgctat gactctactg ttgttgctct gcgcgcatcc gattctagag aagcgtaaag 780

ggttctcgta tcttcccagt tcaaatgact tactagtaat aatgtaatat gctggtgctg 840

acgttgggcg ccacttctcg gtgaaacgtc agcttactgg cgaattaagg gctgcgttgg 900

tgttagtagt ggtttctgca ttcacttggg agcagttatc ttttatatca tcaacgaata 960

cgctcaaacc acagagaaac tttttcttgt aaagatggaa ggctaaaagt tgcttcaaat 1020

cgcacttcgc cgatgtgcat tataaacagc tagttaatgg aatttccgtt tcacttacca 1080

agtttcgtta tttctttatg cagttacaag ccagatgtaa tcaaagggga tatggactca 1140

gtaagaccag aagtgaagga acattattcc aacttggtaa agctgcttta atttacattc 1200

attattggta tgaatctact aaaagggtaa aacgaatttt aatttgggac ggaaggagta 1260

ctagctagtt taacttctct agtgcctctt ttagttctgg tgatcaaatt ggcatttccc 1320

ttgtaccaat ccttgttgag agaaaaaaaa gcaatggtct tagagctgta tatatcagat 1380

gagggttcat ccgaaagggg ataaataaca gtatcctttt ttgttccgac ttagcatagt 1440

atagctggtg gcatctagta agatgcagct agctgattgt gattataata tctgctttat 1500

aatataatgg ttgatgattc ttgttccgac ttatgcttga catgttttgt ttcttctgcc 1560

ctccaaaagg tgcatctata tcatcgtatc atggaggttc ttgtattgat agatcagtct 1620

tattgcataa gatattgaag ctgtacatta gggttttcat tttcgacttc aggcatggtc 1680

tttctgaaaa cgagatcctt tcttatggta attctcatgt attatcaatt gcagggcacc 1740

agaattgttg acgagtcaca tgaccaggac acgaccgatt tacacaaatg catagctttt 1800

atagctgaga attattctgc cactaacaaa tctaacgtaa gataatccaa cccaacaata 1860

ttctaatcaa attagtatgg aaggacaata atgtatctac atttttttaa agcagctcag 1920

catcttggtc cttggagcac ttgggggaag gtttgatcac gagatgggaa acatcaatgt 1980

gcttcatctc ttcccaagca tcaatatcgt cctcctatca gatgattgtc tgatctttct 2040

gctctcgaaa atgcataccc acgagattca cgttgaaaag tcgatcgagg gaccccactg 2100

tggattgatc ccgattggca tgccgtcgac tagcaccacg accaccgggc ttcgatggaa 2160

tctaggttat cctcgtcctt atccttccta atgcaggctt agcaacagaa gttgatgttt 2220

acaacgtgat tacctctgta cagataatac cagtatgagc ttcggaggat tgatcagcac 2280

gtcaaatatc gtggaagagg acaaggtaac ggtcacctcg gattcagacc ttatgtggac 2340

aacatcgctc cggaagtgaa tctacactcg agtatagagt atcgttatac tgttgtaata 2400

ttagagaaat aagtgacatg cttttaatat atatatatat atatatatat atatatatat 2460

atatatatat atatatatat atatatatat taaata 2496

Claims

1. The application of the corn gene in regulating and controlling the plant height and/or ear height of corn is characterized in that: the gene sequence is shown in any one of SEQ ID NO.2-SEQ ID NO. 3.

2. A method for reducing the plant height and/or the ear height of corn is characterized in that: inhibiting the expression and/or activity of the protein shown by SEQ ID NO.1 in corn, and selecting the plants with reduced plant height and/or ear height of corn.

3. The method for reducing the plant height and/or ear height of corn according to claim 2, wherein the plant height and/or ear height of corn is as follows: the method for inhibiting the expression and/or activity of the protein comprises any one of gene editing, RNA interference and T-DNA insertion.

4. The method for reducing the plant height and/or ear height of corn according to claim 3, wherein the plant height and/or ear height of corn is as follows: the gene editing adopts a CRISPR/Cas9 method.

5. The method for reducing the plant height and/or ear height of corn according to claim 4, wherein the plant height and/or ear height of corn is as follows: the DNA sequence of the genome target region of the CRISPR/Cas9 method in maize is shown as SEQ ID NO.4 or SEQ ID NO. 5.

6. A kit for reducing the plant height and/or the ear height of corn is characterized in that: including any of the following:

(1) an RNA molecule targeting the sequence shown in SEQ ID NO.4 or SEQ ID NO. 5;

(2) a DNA molecule encoding the RNA of (1);

(3) a vector expressing the RNA of (1).

7. The kit of claim 6, wherein: the sequence of the RNA molecule is shown as SEQ ID NO.6 or SEQ ID NO. 7.

8. A maize mutant gene characterized by: the sequence of the mutant gene is shown as SEQ ID NO.8 or SEQ ID NO. 9.

9. The use of the maize mutant gene of claim 8 for reducing plant height and/or ear height of maize.