CN111808872B

CN111808872B - Gene DPY1 for regulating and controlling panicolaceae plant type and application and method thereof

Info

Publication number: CN111808872B
Application number: CN202010725748.3A
Authority: CN
Inventors: 刁现民; 赵美丞; 汤沙; 智慧; 何苗苗; 刘西岗; 吴传银
Original assignee: Institute of Genetics and Developmental Biology of CAS
Current assignee: Institute of Genetics and Developmental Biology of CAS
Priority date: 2020-07-24
Filing date: 2020-07-24
Publication date: 2022-06-24
Anticipated expiration: 2040-07-24
Also published as: CN111808872A

Abstract

The invention discloses a gene DPY1 for regulating and controlling the strain type of Panicoideae plants, and an application and a method thereof, wherein the gene DPY1 has a nucleotide sequence shown as SEQ ID NO1, or a nucleotide sequence which is at least 85% homologous with a coding sequence in SEQ ID No. 1. The gene for regulating the plant type has the length of 4622bp, comprises 11 exons and 10 introns, has an encoding sequence of 1902bp, has the protein size of 69.4kD, can improve the plant type and the single plant yield of millet, can be applied to Panicoideae plants including the millet, and can be implanted into the genome of a receptor cell of the Panicoideae plants to improve the plant type of the Panicoideae plants.

Description

Gene DPY1 for regulating and controlling panicolaceae plant type and application and method thereof

Technical Field

The invention relates to a gene for regulating millet plant type, in particular to a gene DPY1 for regulating panicolaceae plant type and application and a method thereof.

Background

Millet is an important regional food crop in northern China, and has the characteristics of drought resistance, water saving, barrenness resistance, C4 photosynthesis and the like (Diao et al, Frontiers of Agricultural Science and Engineering,2014,1(1), 16-20). The conventional breeding technology in the last 15 years enables the yield per unit of the millet to be continuously improved (Shanghai et al, China agricultural science, 2017,50(23),4469-4474), but compared with the corn, the plant type of the millet is not compact enough, particularly, after heading, the leaves are remarkably draped, and the close planting and the group yield of the millet are severely limited.

The reasonable close planting of cereal crops, including main grain crops such as wheat, rice, corn and minor grain crops such as sorghum and millet, can ensure the maximum yield. The close planting adaptability of plants is determined by the plant type to a great extent, and the shape construction of leaves of cereal crops is one of the important factors determining the plant type, which can include the length and width of the leaves, the included angle of stems and leaves and the drapability of the leaves. The compact and upright leaf type can obviously improve the plant type and the group yield of cereal crops, and is one of important agricultural traits of modern breeding.

At present, some leaf/plant type regulation key genes have been cloned in corn and rice, and leaf/plant type improvement and crop yield improvement in a close planting state are realized. For example, Up right Plant Architecture 2(UPA2) derived from teosinte was able to decrease the content of endogenous Brassinosteroids (BR) in maize and decrease the stem-leaf angle, thereby increasing the yield of maize under close planting (Tian et al, Science,2019,365(6454) 658-664); the regulation of rice BR synthetic gene DWARF11 and CATA transcription factor OsGATA7 can enhance the compactness of Plant type, thus showing the potential of increasing yield in a close planting state (Zhang et al, Plant Biotechnology Journal,2018,16, 1261-. Besides the included angle of the stem leaves, the drapability of the leaves is another important factor influencing the compactness of the plant. YABBY family transcription factors, such as corn dropping leaf1/2(drl1/2) (Strable et al, The Plant Cell,2017,29, 1622-.

Compared with corn and rice, the problem that the leaf draping degree is larger in the current main cultivars of millet is particularly obvious after heading. Therefore, the close planting and the later photosynthesis are not facilitated, and the further improvement of the millet yield is influenced. However, the key genes for controlling the leaf draping of the millet have not been cloned so far, which greatly influences the plant type improvement and yield breeding of the millet.

Disclosure of Invention

The invention aims to provide a gene DPY1 for regulating and controlling the plant type of Panicoideae plants and an application and a method thereof, which solve the problem that the leaf drapability of the existing millet plant type is high, can improve the plant type of millet, and obtain the plant type with relatively straight and upright leaves and thick stems and large ears in the mature period, thereby enhancing the close planting adaptability of crops.

In order to achieve the aim, the invention provides a gene DPY1 which regulates the Panicoideae plant type DPY1, wherein the gene DPY1 has a nucleotide sequence shown in SEQ ID NO1 or a nucleotide sequence which is at least 85 percent homologous with a coding sequence in SEQ ID No. 1.

Preferably, the gene DPY1 comprises 10 introns and 11 exons, wherein 11 exons have the nucleotide sequences shown in SEQ ID NO 2-12 respectively, and 10 introns have the nucleotide sequences shown in SEQ ID NO 13-22 respectively.

Another purpose of the invention is to provide a protein coded by the gene DPY1.

Preferably, the protein has an amino acid sequence as shown in seq.id NO 23.

The invention also aims to provide application of the gene DPY1 in improving the strain type of Panicoideae plants.

Preferably, the gene DPY1 is implanted into the zeaideae recipient cell genome.

Preferably, the Panicoideae plant comprises: any one of millet, corn and sorghum.

It is another object of the present invention to provide a method for improving the plant type of Panicoideae, which comprises: integrating the gene DPY1 into the genome of a recipient cell of a Panicoideae plant in an overexpression mode, and obtaining a single-copy homozygous progeny; wherein the overexpression mode is to use a promoter to drive the expression of the coding region of the gene DPY1.

Preferably, the promoter comprises: the Ubiquitin promoter or the CaMV 35S promoter.

The panicolaceae plant type regulating gene DPY1 and the application and the method thereof solve the problem of larger drapability of the conventional millet plant type leaves and have the following advantages:

the invention provides a novel gene for regulating plant type, which is 4622bp in length, comprises 11 exons and 10 introns, has an encoding sequence of 1902bp, has a protein size of 69.4kD, can improve the plant type (compact leaf type and thick stem) and the single plant yield of millet, and obtains the plant type with relatively upright leaves and large thick ears of the stem in the mature period, thereby having the potential of enhancing the close planting adaptability of crops.

Drawings

FIG. 1 shows the phenotypic results of dpy1 mutants according to Experimental example 1 of the present invention.

FIG. 2 is a schematic representation of the protein structure of DPY1 of the present invention.

FIG. 3 shows the result of the site cloning and functional verification of DPY1 in Experimental example 2.

FIG. 4 shows the effect of the overexpression of DPY1 gene on the plant type of millet in Experimental example 2 of the present invention.

FIG. 5 shows the effect of the overexpression of the DPY1 gene on the thickness of the millet stalks in the experimental example 2 of the present invention.

FIG. 6 is a graph showing the effect of the overexpression of DPY1 gene on the yield of individual millet plants in Experimental example 2 of the present invention.

FIG. 7 is a homology tree spectrum of DPY1 of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

A gene DPY1 for regulating the plant type of Panicoideae plant, wherein the gene DPY1 has a nucleotide sequence shown in SEQ ID NO1, or a nucleotide sequence which is at least 85% homologous with the coding sequence in SEQ ID No.1 (the similarity of the nucleotide sequence of DPY1 and the nucleotide sequence of an orthologous gene of corn and sorghum is more than 85%, Sobic.003G051100 is the DPY1 gene of sorghum, and the nucleotide sequence is shown in figure 7). DPY1 contains 10 introns and 11 exons, wherein 11 exons have the nucleotide sequences shown in SEQ ID NO 2-12 respectively, 10 introns have the nucleotide sequences shown in SEQ ID NO 13-22 respectively, the coding sequence (CDS) is 1902bp, the size of the coded protein is 69.4kD, and the amino acid sequence shown in SEQ ID NO23 is contained.

The gene sequence is implanted into the receptor cell genomes of millet and other Panicoideae plants, commercial and farmer varieties of millet (Setaria italica), corn (Zea mays) and Sorghum (Sorghum bicolor) can improve the plant type.

The following experimental examples are provided to describe the Panicoideae plant type DPY1 of the present invention in detail.

Experimental example 1 EMS mutagenesis to obtain mutants with severe leaf draping

In order to genetically improve the natural drapability of the leaves of the millet, the commercial millet variety Yugu No.1 is mutagenized by 1% EMS (ethyl methane sulfonate) for 8 hours, and the millet variety Yugu No.1 is sown in the field after being thoroughly cleaned by distilled water. At near 2000M₂Generations (mutagenesis second generation, i.e. mutagenesis current generation M)₁The mutant obtained by sowing the variant individuals obtained in (1) and growing) a mutant dropy leaf 1(dpy1) which shows a curled leaf-drape at the seedling stage and a severe leaf-drape at the mature stage was found in the family (see FIG. 1, A-B).

The midveins in the leaves of dpy1 and wild type Yugu No.1 (WT) were observed: the median pulse width is significantly narrowed at dpy1 (WT: 0.5. + -. 0.12 cm; dpy1: 0.28. + -. 0.18cm) (see FIG. 1, C).

The leaves of dpy1 and wild type Yugu No.1 (WT) were cross-sectioned and stained with phloroglucinol (capable of specifically staining lignin) to find: the number of vascellum abaxial pachydic cells in the midvein was significantly reduced in DPY1 (see fig. 1, D-E), and the amount of lignin deposition was reduced (see fig. 1, E), indicating that the DPY1 gene affected the drapability of the leaf by affecting the division of vascellum pachydic cells and lignin deposition.

Experimental example 2 cloning and functional verification of DPY1 Gene

1. Cloning of the DPY1 Gene

For cloning DPY1, the mutant DPY1 is used as a female parent, the millet variety SSR41 is used as a male parent for hybridization, and the progeny leaf type is normally heterozygote F₁Generation, F₁F for selfing to form dpy1 × SSR41₂Generation genetic linkage population. At F₂The segregation ratio of wild type phenotype and mutant phenotype in segregating population was 3:1(2452:789, χ)²＝0.71,P>0.05), indicating that the mutant has a drape leaf pattern controlled by a recessive gene.

Next, a candidate gene Seita.5G121100 for DPY1 was obtained by map-based cloning with Mutmap sequencing analysis using 116 recessive individuals (see FIG. 3, A-B). The analysis of Seita.5G121100 encoded protein by using protein domain online analysis software SMART shows that it contains extracellular signal peptide, 5 LRRs (Leucine-Rich Repeat), transmembrane domain and kinase domain (the protein structure of DPY1 is shown in FIG. 2).

The DPY1 genome sequence is 4622bp (SEQ. ID NO 1), comprises 11 exons (SEQ. ID NO 2-12) and 10 introns (SEQ. ID NO 13-22), the coding sequence (CDS) is 1902bp, and the protein size is 69.4kD (SEQ. ID NO 23).

2. Functional verification of DPY1 gene

(1) Mutation of DPY1 Gene

Further sequencing finds that A-T mutation of the +882 deoxynucleotide of dpy1 causes disturbance of transcript shearing to cause early termination mutation, and further protein with an amino acid sequence of SEQ ID NO23 cannot be synthesized.

(2) CRISPR/CAS9 knock-out vector construction of DPY1 and plant genetic transformation

To further validate the function of seita.5g121100(DPY1), a knock-out target (CCCAGTTGTCAAGCACGTTA) was first designed at the second exon of DPY1 and ligated downstream of the OsU6 promoter, thus constructing the CRISPR/CAS9 knock-out vector for DPY1. Transformation of the vector into Yugu 1 at T₁Both individual editors showed a phenotype similar to DPY1 (see FIG. 3, C), and CR-1/-4 was a knock-out strain of the DPY1 gene with deletions of 1 and 26 nucleotides, respectively, in the second exon, thus creating a frameshift mutation.

(3) Construction of anaplerotic vector and anaplerotic verification of DPY1 gene

Carrying out PCR amplification by taking the genome DNA of Yugu 1 as a template, wherein the adopted synthetic primers are as follows:

DPY1-g-F (upstream primer, SEQ. ID NO 24):

TTACTTCTGCACTAGGTACCATGCAGCGGCGGCGG；

DPY1-g-R (downstream primer, SEQ. ID NO 25):

CGGACTTAAGACTAGTAACATCATATACAGGCATGCGC。

the PCR amplification product was a 7.0kb genomic fragment containing the entire genome of Seita.5G121100 and the upstream 2.5kb promoter region (see SEQ. ID NO. 26).

The 7.0kb genomic fragment was ligated to pCAMBIA1305-EGFP vector (DPY1:: DPY1-GFP) by double-restriction enzyme ligation (KpnI/SpeI), and the mutant phenotype of DPY1 could be fully restored by Agrobacterium transformation of DPY1 (see FIG. 3, C), and Re-1/-2 was a complementing transgenic plant which turned the 7kb genomic fragment of DPY 1(DPY 1:: DPY1-GFP) into the DPY1 mutant, indicating that Seita.5G121100 is the DPY1 gene.

(4) Homologous gene of DPY1 in maize genome

Alignment of the nucleotide sequence of DPY1 in the genome of maize revealed the presence of two copies of DPY1 of GRMZM2G010693(ZmDPY1.1) and GRMZM2G067675(ZmDPY1.2) in the maize genome, as shown in FIG. 7.

Performing PCR amplification by using cDNA of a corn variety B73 as a template, wherein the adopted synthetic primers are as follows:

ZmDPY1.1-OE-F (upstream primer, SEQ. ID NO 27):

CGACTCTAGAGGATCCAATTGAATCTGCGGGGGTTGC；

ZmDPY1.1-OE-R (upstream primer, SEQ. ID NO 28):

GCTTGGCGCGACTAGTCCTCGGGCCGGAGAGCTCCAT。

PCR amplification using the cDNA of maize variety B73 as a template yielded ZmDPY1.1 of approximately 2.0kb, which was subsequently ligated downstream of the Ubiquitin promoter of the overexpression vector PTCK303 by the enzyme ligation method (BamHI/SpeI double digestion) (Ubi:: ZmDPY1-3FLAG), and the transformed millet (DPY1) by the Agrobacterium-mediated method was used to transform ZmDPY1-3FLAG, which was found to completely restore the mutant phenotype of DPY1 (see FIG. 3, C), and OX-2/-3 was a transgenic plant overexpressing ZmDPY1.1 in DPY 1(Ubi:: ZmDPY1-FLAG), indicating that DPY1 has a conserved plant-type regulatory function in Panicoideae crops.

Performing PCR amplification by using cDNA of millet (Yugu 1) as a template, wherein the adopted synthetic primers are as follows:

DPY1-OE-F (upstream primer, SEQ. ID NO 29):

CGACTCTAGAGGATCCTTGCCGAATAGAATCTGCGG；

DPY1-OE-R (upstream primer, SEQ. ID NO. 30):

GCTTGGCGCGACTAGTCTGTCtCCTCGGTCCCGAGA。

the cDNA of millet is taken as a template for PCR amplification to obtain DPY1 with about 2.0kb, the same enzyme digestion connection method is utilized to connect DPY1 to the downstream of a Ubiquitin promoter (Ubi:: DPY1-3FLAG) of an overexpression vector PTCK303, the Ubi:: DPY1-3FLAG is transformed into millet variety Yugu 1, OX-1/-2 is two transgenic plants overexpressing millet DPY1 under WT background (Yugu 1), and the overexpression DPY1 is found to improve the draping plant type of the millet, so that the leaves are relatively stiff and upright (see figure 4), and meanwhile, the stem thickness and the single plant yield can be increased (see figure 5-6).

While the present invention has been described in detail with reference to the preferred embodiments, it should be understood that the above description should not be taken as limiting the invention. Various modifications and alterations to this invention will become apparent to those skilled in the art upon reading the foregoing description. Accordingly, the scope of the invention should be determined from the following claims.

Sequence listing

<110> research center of agricultural resources of institute of genetics and developmental biology of Chinese academy of sciences

<120> Gene DPY1 regulating Panicoideae plant type, and application and method thereof

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<211> 392

<212> DNA

<213> DPY1-exon10

<400> 11

aacatgtaaa tggcaagcca gctctagatt ggtcgaggag aaagagaata gcactaggca 60

cagcacgagg tctgctgtac ctgcatgagc agtgcgatcc aaaaataatc catcgtgacg 120

taaaagcctc caatgtgctt cttgatgaat atttcgaagc aattgtggga gattttggat 180

tggcaaaact tttggatcac caggaatctc atgttaccac tgcagtgcgt gggactgttg 240

gacatatagc accggagtat ctgtcaactg gccagtcatc agagaagact gatgtgtttg 300

gatttggagt tctcttagtt gagttgatca ctggtcagaa ggcattggat tttggaagag 360

tagcaaatca gaagggtggc gtgctggatt gg 392

<210> 12

<211> 321

<212> DNA

<213> DPY1-exon11

<400> 12

gttaagaaac ttcatcaaga aaagcagcta agcatgatgg tggacaaaga cctgggtagc 60

aactacgaca gggttgagtt ggaggaaatg gtccaggtgg ctttgttatg cacacaatac 120

tacccatctc accgccctcg aatgtctgag gtaatcagga tgctggaagg tgacgggctt 180

gcagagaaat gggaagcatc acagaatgtg gacacgccaa agtccgtctc atcggagctc 240

ctgcctccaa agtacatgga tttcgccgcg gacgagtcct cgctcggcct agaagccatg 300

gagctctcgg gaccgaggtg a 321

<210> 13

<211> 346

<212> DNA

<213> DPY1-Introns1

<400> 13

gtacctaaca ttctctcctc ccatgtcttc tcgaaaatct taacaaaagg ccatggccga 60

ccttttccta ttttggaaaa ataaaaggac gaaaggccaa ctttttccat tcagccaaca 120

gaggattttc aggaatttgt tgaaaacatt cccctcctgc aatcaatcgc tggtttgaag 180

ttggggtaaa agatctcatt ttggtgaagg acactaaagg aaatgcagct gctgtaccta 240

tacttcctca tcagtcctct actcatcttg cttgtcgagt tctagctgtt tcttggcatt 300

acgaatttgg cgaagttgaa cctgcctgcc tgtggctgta ctgcag 346

<210> 14

<211> 93

<212> DNA

<213> DPY1-Introns2

<400> 14

gtacgttatg ctaccttgtt catgcccatt ccccggagca ccaacgggct ctgctttggg 60

ctgatcttct tacatgttct tcttcacttg cag 93

<210> 15

<211> 121

<212> DNA

<213> DPY1-Introns3

<400> 15

gtaagtttac tgatcccctg ttgtttacct ctggcacttc aaaattcaaa tttgaggttt 60

cttgagtgga ccaatctggt tgaagaaatg actgaaagta ctcggctttg cggatcaaca 120

g 121

<210> 16

<211> 99

<212> DNA

<213> DPY1-Introns4

<400> 16

gtaagatttt tccttcccca ttcttgattt tcaacagcgt tgtcattgta cacggtgatt 60

tcatttttgc tcattgtcta gttctcttgt tgccgccag 99

<210> 17

<211> 82

<212> DNA

<213> DPY1-Introns5

<400> 17

gtatgctctg caatcccttt ttgttgttcg tttttctctt cccctacttt tgtactaatt 60

cccgaagtgt tttgttttat ag 82

<210> 18

<211> 136

<212> DNA

<213> DPY1-Introns6

<400> 18

gtaagtaatt tcaacttatt ccttttcatg ggtattttta tttcggcccg aacactacca 60

ctactgtaac tacatgttat gccatggact aatgttcact taatcaagct ttcattttta 120

ctgtttgttc atacag 136

<210> 19

<211> 108

<212> DNA

<213> DPY1-Introns7

<400> 19

gtgagttgtt tttctcatca ttcattttcg ggaaatatgc atttattaca gaggaattgg 60

cttgcctttt cttctgcgtg acactcacat atgtctcctt ttcctcag 108

<210> 20

<211> 645

<212> DNA

<213> DPY1-Introns8

<400> 20

gtaatattct attcaaaccc gttcttatgt gatcatgata ttaaccacct agatatgaat 60

tgtattagat caatgtagta attgtagcaa tggctatggc tagcagttta taatttatga 120

tcttggagct ttttccttct gatcttccac agtaaattta catggatgta aagatatgca 180

tactgtctaa gtgttctatg tcagtttgag ctactgaggt ttcaataagc aggctaatta 240

ggctttgaca tgtttcctca atagcaatta agcattgctc taatattctg ccatatgatc 300

tccgttagga gttttgaaac cactgatgaa tttgaagata caatagacgc atctctcaac 360

attagttcta tgtggttacc atcaataatg cgagtataca acaaaaaaac tttttgtttc 420

ttttaagttt ttttactagt atgatcttcc aattactttt ggtatgtggg ttatttcaca 480

atcatgatgt ccccgttact ttctgtgctt ttcttagcct ttcaacaatc atgagctatt 540

ctttttaccc ttctaggact ttcttcgata tctttagcat gcttttttac tttctttctc 600

ttgcgtgcta tggtaattga acctgataca ttcttcaatg tgcag 645

<210> 21

<211> 889

<212> DNA

<213> DPY1-Introns9

<400> 21

gtttgtctct ctcttcttgc tcgtgacttc catgtgcata ttttgctgcc agttgaccca 60

atttctttga agcacacaag tcccgcttct tttttttttt aaaaaaagaa gcactttgtt 120

acaagaggga aaaaatgact atcaacgttg atcaaaattt tgtatgcgta aaatattaac 180

accttggcat ttttgctcaa tccaatatga aatatttacc ctatttttcg atgtatcata 240

gttgatgtga aatgtctgtg ttccaacatt tgattagtgt tgctgcacag agttactgaa 300

tgtagtttta tgttgtctga cataccttag gtttagtgca aacgggcaag ctagcatgtt 360

ttcttgtatt gtagctgatc ttgttgcact gcacagacag tcaccacacg ctcaatttac 420

tgttgaaatc ataataagta catgtggttc tgtgcgtggt ctaatgtgaa aaatcagatg 480

agtgatctgg atattgcagt ctaaaaatct attagcctta tttagttatt tatacactga 540

tttgaatacc cggctgcttt cagtcatatt atgccaggca aacagcttgg cttctccatt 600

ctggtgtcct tttattgaaa acataatggt ggtgttaact gttgagttat gttggtcaaa 660

ttaaatccta ctcccattct tgttgtggaa ttctgcagta atatggccat gctatatatt 720

gcattcaagc ctagttttgg aatataaact gaagaaattc aaacatcaca tatccagcta 780

tctttaactc ataacatggg ggtatcaata acagcaaaca ccatatgagc atcgtccaag 840

tatattatcc actatgcatg caaattaact ttctttcgct tttcttcag 889

<210> 22

<211> 201

<212> DNA

<213> DPY1-Introns10

<400> 22

gtaagtgtgc gtcttatggt ttactattct gatttgcata tttgattcaa agctttaaac 60

aaaaagaagt ctacctttgc ttttatacct tctttggctc tgcccttcag ttcagtcttg 120

cagagttttt tttcgttatt acaggctcat ttgtttgtat ttccatgaac atatttcctc 180

tttacttttg tctggttaca g 201

<210> 23

<211> 633

<212> PRT

<213> DPY1-protein

<400> 23

Met Trp Val Pro Val Ala Pro Met Trp Met Arg Trp Trp Val Val Val

1 5 10 15

Ala Gly Leu Leu Ala Val Ile Leu Pro Pro Ser Thr Ala Thr Leu Ser

20 25 30

Pro Ala Gly Ile Asn Tyr Glu Val Val Ala Leu Met Ala Ile Lys Thr

35 40 45

Glu Leu Glu Asp Pro His Asn Val Leu Asp Asn Trp Asp Ile Asn Ser

50 55 60

Val Asp Pro Cys Ser Trp Arg Met Val Thr Cys Ser Ser Asp Gly Tyr

65 70 75 80

Val Ser Ala Leu Gly Leu Pro Ser Gln Ser Leu Ser Gly Lys Leu Ser

85 90 95

Pro Gly Ile Gly Asn Leu Thr Arg Leu Gln Ser Val Leu Leu Gln Asn

100 105 110

Asn Ala Ile Ser Gly Ser Ile Pro Gly Thr Ile Gly Arg Leu Gly Met

115 120 125

Leu Lys Thr Leu Asp Met Ser Asp Asn Gln Leu Thr Gly Ser Ile Pro

130 135 140

Ser Ser Leu Gly Asn Leu Arg Asn Leu Asn Tyr Leu Lys Leu Asn Asn

145 150 155 160

Asn Ser Leu Ser Gly Val Leu Pro Asp Ser Leu Ala Thr Ile Asp Gly

165 170 175

Leu Ala Leu Val Asp Leu Ser Phe Asn Asn Leu Ser Gly Pro Leu Pro

180 185 190

Lys Ile Ser Ala Arg Thr Phe Ile Ile Ala Gly Asn Pro Met Ile Cys

195 200 205

Gly Ala Lys Ser Gly Asp Asn Cys Ser Ser Val Ser Leu Asp Pro Leu

210 215 220

Ser Tyr Pro Pro Asp Asp Leu Lys Thr Gln Pro Gln Gln Gly Ile Val

225 230 235 240

Lys Gly His Arg Ile Ala Thr Ile Cys Gly Ala Thr Val Gly Ser Val

245 250 255

Ala Phe Ala Thr Ile Val Val Gly Met Leu Leu Trp Trp Arg His Arg

260 265 270

Arg Asn Gln Gln Ile Phe Phe Asp Val Asn Asp Gln Tyr Asp Pro Glu

275 280 285

Val Cys Leu Gly His Leu Lys Arg Tyr Ala Phe Lys Glu Leu Arg Ala

290 295 300

Ala Thr Asn Asn Phe Asn Ser Lys Asn Ile Leu Gly Glu Gly Gly Tyr

305 310 315 320

Gly Ile Val Tyr Lys Gly Tyr Leu Arg Asp Gly Ser Val Val Ala Val

325 330 335

Lys Arg Leu Lys Asp Tyr Asn Ala Val Gly Gly Glu Val Gln Phe Gln

340 345 350

Thr Glu Val Glu Val Ile Ser Leu Ala Val His Arg Asn Leu Leu Arg

355 360 365

Leu Ile Gly Phe Cys Thr Thr Glu Cys Glu Arg Leu Leu Val Tyr Pro

370 375 380

Tyr Met Pro Asn Gly Ser Val Ala Ser Gln Leu Arg Glu His Val Asn

385 390 395 400

Gly Lys Pro Ala Leu Asp Trp Ser Arg Arg Lys Arg Ile Ala Leu Gly

405 410 415

Thr Ala Arg Gly Leu Leu Tyr Leu His Glu Gln Cys Asp Pro Lys Ile

420 425 430

Ile His Arg Asp Val Lys Ala Ser Asn Val Leu Leu Asp Glu Tyr Phe

435 440 445

Glu Ala Ile Val Gly Asp Phe Gly Leu Ala Lys Leu Leu Asp His Gln

450 455 460

Glu Ser His Val Thr Thr Ala Val Arg Gly Thr Val Gly His Ile Ala

465 470 475 480

Pro Glu Tyr Leu Ser Thr Gly Gln Ser Ser Glu Lys Thr Asp Val Phe

485 490 495

Gly Phe Gly Val Leu Leu Val Glu Leu Ile Thr Gly Gln Lys Ala Leu

500 505 510

Asp Phe Gly Arg Val Ala Asn Gln Lys Gly Gly Val Leu Asp Trp Val

515 520 525

Lys Lys Leu His Gln Glu Lys Gln Leu Ser Met Met Val Asp Lys Asp

530 535 540

Leu Gly Ser Asn Tyr Asp Arg Val Glu Leu Glu Glu Met Val Gln Val

545 550 555 560

Ala Leu Leu Cys Thr Gln Tyr Tyr Pro Ser His Arg Pro Arg Met Ser

565 570 575

Glu Val Ile Arg Met Leu Glu Gly Asp Gly Leu Ala Glu Lys Trp Glu

580 585 590

Ala Ser Gln Asn Val Asp Thr Pro Lys Ser Val Ser Ser Glu Leu Leu

595 600 605

Pro Pro Lys Tyr Met Asp Phe Ala Ala Asp Glu Ser Ser Leu Gly Leu

610 615 620

Glu Ala Met Glu Leu Ser Gly Pro Arg

625 630

<210> 24

<211> 35

<212> DNA

<213> Artificial Sequence

<400> 24

ttacttctgc actaggtacc atgcagcggc ggcgg 35

<210> 25

<211> 38

<212> DNA

<213> Artificial Sequence

<400> 25

cggacttaag actagtaaca tcatatacag gcatgcgc 38

<210> 26

<211> 2500

<212> DNA

<213> Artificial Sequence

<400> 26

tagtctacat ttaatacttc taattagtat ctaaacattc gatgtgacgg gtgcttaaat 60

ttaagttagt gaaccaaacc aggcctaagg tcgatctaaa aataaaagtt gagagactaa 120

gttaacaatc taatataaag tgatgaactt gattcgaggg taaagttcag aatcaaaacg 180

ctattttacc gattggtcat gactgaatgc catccttcag agttcagata caagcaatca 240

caaacacatc gataaaaaaa ctcatctatg aacgagctga atcttcgaac gtcatccatt 300

tccacacaga acacacagta gcaagcgagg aacaaccggc tgtccggctc agccgggggg 360

accggatgtc gctggataat ataaatataa taaatataaa aaaataaaca gactagattc 420

taccactacc accgtgacac tgatggacaa gagagagagg gagaggcagg ccgggacggt 480

gaactggtga cggtgacgag cagcgcgaca agaaaggacg ggacaaaatc aaaggcgggc 540

atgtgactcg ccagtgccca gcaaagcctt ttttaccttc atccccccct ctccgctcgc 600

ccttttctat attccccatc attctcttct ttatattacc tagatagccg tgcgcgcagt 660

acgcctcggc ttcgtagact agttcatgac aaacatgcgc cctgtcatta ccgctgtcgt 720

tgcgcagatg atgacaacca gagagaacac cggtgaaaga ttgatgaaat cctatgctaa 780

caggcaggca gatggggggg tttatctttg ctaaacgcgc acagctgccg gtaacatgca 840

ctaactccta gtaccggtat atagtacgag taaattttag catattgcta gcagcagcgg 900

tacattatga ttcccgatga gagtgcatta gcactactgc ttgttcgcct ctcgaattgc 960

aggcgttgga gcatctgccg tcttttcttg aacacaaggc cgtccatgcc cttggaaagc 1020

tgtgaaaaat accaaattca cggagttcaa gtgtccaacc aaacaaaaag atgaaaagag 1080

attaacaaag attctaggga gatggcaaat ccagcaagaa catttttttg gaaacttaag 1140

caagaacatt tccttttttc tgaaaacgta tgcacctttt tcggaaaatc taggggatgg 1200

aaaggaacac accattttgg tcccgcaact ttaacctgag ggttaagttc gtccctcaac 1260

attttagata ggccaatttg atccctcaac ttctgttttt agtcaaactt gtccttatgc 1320

tgacatggtg cactttgttg gtacgagact aatgtgaaaa ttccttttta cccttggctc 1380

atgtaggcta agcatgtatg tgttattatg ctcatattct cttgacatat acacgcaaaa 1440

atatattggt taactaacac ataatacggt ttaaaaaaaa tgtatgtacc aaaactcttt 1500

aaagtcatgc atgtgctcgt acgaattaaa tatgaaaaat ttatgtaccc aaacaaatat 1560

ttattcatac tcttttggta tacaactcaa tatatgtact catactttaa aatctaaaat 1620

caacatgtgc tcacatacgt atatgtattc atactttctc agtaaatatg tataaatatt 1680

tatgtgctcg tacttgattt ggatcatact gttattgtac ttaatgatgg aaattgagga 1740

taaatattta tgtgctcgta cttgatttgg atcatactgt tattgtactt aatgacggaa 1800

attgaggaga ggaagaaaga gctaagggta aaaatgacct tttgtattag ttttgtatta 1860

aaatgggact ctcaccagat gagttttaaa aaataaaaga tgagggatcg accgatttaa 1920

agttgaggga cgaatttgat ccttaaaaga aagttgaggg accaaaatgg acattttgcc 1980

ggaaaggaag gaggaggtca ggagcatgcg cgcccgtggc aggtcaggcc ggtttggggg 2040

aaaagcgctt gtcggtttgc agcggcgtgg agcgggtggg gagcatcagc ggcgtcagcg 2100

gacagcggcc gcggcagcca gccagcacca gcagcatggt ggtggattga taggtgggga 2160

aggggcctgc cgctgctgct gctgccgctg ccgctgctcg ttggcgtttc ccttcccctc 2220

ttgcccccac tacccacccc caccatcttc ctctcgggcg gtcgcgtgaa ctccccctct 2280

cccctcgctt tcgcgctgcg tgcgcggtgg catcattggc tatctggatt ggtccgggtt 2340

gggtagtaga gaggccggcc ttcttctgct tcttccctgc tgctggcggc ggcggagcgg 2400

aattgtggtg tggagctgtg gggggactgg cgtgcagcgt gctcctcctc ttcttgcaga 2460

tcatcgtttg ccgaattgaa tctgcggggc tgtgcgggcg 2500

<210> 27

<211> 37

<212> DNA

<213> Artificial Sequence

<400> 27

cgactctaga ggatccaatt gaatctgcgg gggttgc 37

<210> 28

<211> 37

<212> DNA

<213> Artificial Sequence

<400> 28

gcttggcgcg actagtcctc gggccggaga gctccat 37

<210> 29

<211> 36

<212> DNA

<213> Artificial Sequence

<400> 29

cgactctaga ggatccttgc cgaatagaat ctgcgg 36

<210> 30

<211> 36

<212> DNA

<213> Artificial Sequence

<400> 30

gcttggcgcg actagtctgt ctcctcggtc ccgaga 36

Claims

1. GeneDPY1The application of the gene in improving the plant type of the Panicoideae plant is characterized in that the geneDPY1The nucleotide sequence of (A) is shown in SEQ ID NO. 1.

2. The use of claim 1, wherein said gene is administeredDPY1Implanted into the genome of recipient cells of Panicoideae plants.

3. Use according to claim 1 or 2, characterized in that the Panicoideae plant comprises: any one of millet, corn and sorghum.

4. A method of modifying the plant type of a panicolaceae plant, the method comprising: the gene is transformed into a geneDPY1Using a super watchIntegrating the expression mode into the genome of the recipient cell of the panicolaceae plant and obtaining a single-copy homozygous progeny; wherein the overexpression mode is realized by using promoter to start a geneDPY1Expression of the coding region of (a); wherein the geneDPY1The nucleotide sequence of (A) is shown in SEQ ID NO. 1.

5. The method of claim 4, wherein the promoter comprises: the Ubiquitin promoter or the CaMV 35S promoter.