CN112442115B

CN112442115B - Application of wheat TaPRR95-B protein or coding gene thereof in regulating and controlling plant height

Info

Publication number: CN112442115B
Application number: CN202011446006.3A
Authority: CN
Inventors: 李爱丽; 陶姝; 毛龙; 耿帅锋
Original assignee: Institute of Crop Sciences of Chinese Academy of Agricultural Sciences
Current assignee: Institute of Crop Sciences of Chinese Academy of Agricultural Sciences
Priority date: 2020-12-08
Filing date: 2020-12-08
Publication date: 2022-03-04
Anticipated expiration: 2040-12-08
Also published as: CN112442115A

Abstract

The invention relates to the technical field of genetic engineering, in particular to application of wheat TaPRR95-B protein or a coding gene thereof in regulating and controlling plant height. The research of the invention finds that the wheat TaPRR95-B protein has a key effect on regulating the plant height of the plant, and when the coding gene of the wheat TaPRR95-B protein in the plant is knocked out, the plant height of the plant is obviously reduced. The invention further compares the phenotypes of the TaPRR95-B gene wild type and a tetraploid mutant thereof, proves that the knockout of the gene can obviously reduce the plant height of wheat, and provides a foundation for researching the molecular mechanism and application of the gene influencing the plant height.

Description

Application of wheat TaPRR95-B protein or coding gene thereof in regulating and controlling plant height

Technical Field

The invention relates to the technical field of genetic engineering, in particular to application of wheat TaPRR95-B protein or a coding gene thereof in regulating and controlling plant height.

Background

Wheat is an important food crop in the world and also a staple food for many people all over the world, so that the yield of wheat is closely related to national food safety. With the increase of population, the reduction of arable land area and the increase of production cost, how to increase the wheat yield to ensure the grain safety becomes an urgent problem to be solved. In the 70 s of the 20 th century, the green revolution dominated by the utilization of the wheat semi-dwarf gene brings great increase to the global wheat yield, shows that the plant height of wheat has important influence on the formation of the yield, researches on the genetic law of plant height development are helpful for understanding the action mechanism of yield formation, and has important significance for breeding lodging-resistant and high-yield varieties by utilizing the plant height gene and further improving the yield of wheat.

PRRs (Pseudo-Response modulators) family genes, with conserved amino-terminal PRR/RLD (Receiverlike domain) domain and a carboxy-terminal CCT domain, these 2 domains separated by 1 less conserved "variable" domain, the N-terminal Response-modulating acceptor domain is highly similar to the acceptor domain involved in its ASP phosphorylation signal transduction pathway, and the C-terminal CCT domain is similar to the NF-YA2 DNA binding region of yeast heme activating protein (HAP 2). The PRRs family gene influences the growth and development of plants, including the regulation and control of flowering time, the response to photosynthesis, heat shock response, oxidative stress response, the regulation of stomatal conductance, the metabolism of mitochondria, the response to cold stress and the like.

The PRRs genes are conserved in angiosperms and retain at least 5 copies of the PRR gene in their genome, which is divided into 3 clades before monocotyledonous and dicotyledonous differentiation: PRR1/TOC1(TIMING OF CAB2EXPRESSION1), PRR3 and PRR7, PRR5 and PRR9, although the amino acid sequences OF these genes are highly conserved in the PRR domain and CCT motif, several amino acid changes are present in the PRR domain OF 3 clades, which can lead to their functional differences. Currently, 5 PRR genes have been identified in arabidopsis and papaya, PRR1(TOC1), PRR3, PRR5, PRR7 and PRR9, respectively, and 5 orthologous PRR genes in rice, OsPRR 1/oscout 1, OsPRR37, OsPRR59, OsPRR73 and OsPRR95, respectively. Among these genes, OsPRR37 is more important because it affects the flowering period, plant height and spikelet number per ear of rice. However, little is known about the analysis of the structure and function of the PRR gene family in wheat, and the biological function and molecular mechanism of the gene in wheat are not clear.

Disclosure of Invention

In order to solve the problems in the prior art, the invention provides application of wheat TaPRR95-B protein or a coding gene thereof in regulating and controlling plant height. The research of the invention finds that the correlation between the B homologous chromosome gene of the TaPRR95 gene and the plant height can obviously reduce the plant height by knocking out the gene.

In a first aspect, the invention provides an application of wheat TaPRR95-B protein or a coding gene thereof in regulating and controlling plant height.

The invention further provides application of the wheat TaPRR95-B protein or the coding gene thereof in cultivation and preparation of transgenic plants or improvement of plant germplasm.

Furthermore, the wheat TaPRR95-B protein is selected from an amino acid sequence shown in any one of SEQ ID NO 1-3.

Furthermore, the encoding gene of the wheat TaPRR95-B protein is selected from any one of the nucleotide sequences shown in SEQ ID NO. 4-6.

Further, the plant is a monocotyledon or dicotyledon, preferably wheat.

In a second aspect, the invention provides a SNP locus linked to a coding gene of a wheat TaPRR95-B protein, wherein the SNP locus is located at the 505806276 th position of a 5 th chromosome.

The invention further provides application of the SNP locus in identifying a coding gene of wheat TaPRR95-B protein, wherein the coding gene of the wheat TaPRR95-B protein is selected from any nucleotide sequence shown in SEQ ID NO. 4-6.

In a third aspect, the present invention provides a method for regulating plant height of a plant, comprising:

regulating and controlling the expression level of a coding gene of wheat TaPRR95-B protein in a plant, wherein the wheat TaPRR95-B protein has an amino acid sequence shown in any one of SEQ ID NO. 1-3.

Further, reducing the plant height of said plant by reducing the expression level of a gene encoding said wheat TaPRR95-B protein in said plant; or, breeding shorter plant lines by crossing the gene-knocked-out line encoding the TaPRR95-B protein of wheat with other lines.

Further, the reducing of the expression level of the coding gene of the wheat TaPRR95-B protein in the plant is specifically:

knocking out a gene encoding said wheat TaPRR95-B protein in said plant by CRISP/CAS9 technology.

The invention has the following beneficial effects:

the research of the invention finds that the B homologous chromosome gene of the TaPRR95 gene is related to the plant height of the plant, and further proves that the knockout of the gene can obviously reduce the plant height of wheat by comparing the tetraploid mutant with the wild type phenotype, thereby providing a foundation for researching the molecular mechanism and application of the gene influencing the plant height.

Drawings

FIG. 1 shows the expression of TaPRR95-B gene in stem, ear and anther;

FIG. 2 shows the plant height comparison between the PRR95-B gene mutant and the wild type, wherein WT is the wild type, and kronos1181 and kronos2031 are mutants.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

Example 1 genotyping and plant height Association analysis of TaPRR95-B Gene in core germplasm materials

1. Wheat micro-core seed materials (105 parts of bred varieties and 157 parts of local varieties) distributed in different wheat areas in China are selected as the discovery materials of polymorphic SNP sites.

2. SNP for obtaining micro-core seed material by capturing exons

DNA of 262 parts of material was subjected to library construction and sequencing by using a Roche exon Capture Kit (SeqCap Pure Capture Bead Kit,06977952001, Roche), and the specific method is described in the Kit specification.

3. The genotype of the gene in 262 material was obtained by whole genome sequence alignment with BWA software, mutation detection and genotyping with GATK software.

4. The association analysis is carried out by using plant height phenotype data of 262 parts of micro-core seed materials of three-year three-point (Luo Yang Henan 2002, Luo Yang Henan 2005, Beijing 2006) in the plant height. The result of the association between the TaPRR95-B gene and the plant height is shown in Table 1, and it can be seen that the SNP site of the TaPRR95-B gene is significantly related to the plant height trait of wheat material.

TABLE 1 SNP sites with significant association of plant heights

5. According to the TravesCS 5B01G320500 gene number, the homologous gene numbers TravesCS 5A01G320300 and TravesCS 5D01G326200 are obtained by comparison on a wheat genome database website, and the gene sequences of the TaPRR95 gene on A, B, D homologous chromosomes are obtained and are respectively shown as SEQ ID NO.4-6, and the gene is positioned on a fifth chromosome homologous group.

According to the gene number, the expression of the gene is inquired in a wheat transcriptome database, and the expression of the gene in stems, ears and anthers is found as shown in figure 1.

Example 2 phenotypic statistics of PRR95-B Gene mutants

In order to verify that the function of the TaPRR95 gene is related to plant height, in the example, an EMS induction library of tetraploid wheat is used to obtain a mutant of the gene B copy, and the specific information is shown in the following table:

TABLE 2 mutation information of PRR95-B Gene

This example compares the plant heights of wild type and mutant and gives the results shown in FIG. 2: compared with the height of a wild Kronos plant, the plant heights of mutant strains Kronos1181 and Kronos2031 are obviously reduced, which shows that the PRR95-B gene can obviously reduce the plant height of wheat.

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

<110> institute of crop science of Chinese academy of agricultural sciences

Application of <120> wheat TaPRR95-B protein or coding gene thereof in regulating and controlling plant height

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<213> wheat (Triticum aestivum)

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gtggaccagg acgggaggga ggaggaggtg cgccgggcgc tgccgatgat gcccgtcagg 120

gtgctgctcg ccgagggcga cgactccacg cgccacgtca tctccgcgct gctccgcaag 180

tgcggctacc acgtttctgc agcttctgat ggtgtcaagg cctgggaatt actaaaggaa 240

aaatcgttca aaatagatct cgttctcact gaagttgaac ttcctttgat gtctgggttc 300

ctcctactct ccacgatcat ggaacatgac gcgtgcaaga atatccctgt cataatgatg 360

tcttcgaatg atgcagttag catggttttc aaatgcatgc tgaagggcgc agccgatttc 420

cttgtcaagc cgatacgaaa gaatgagtta cggaacttgt ggcagcacgt ttggagaaaa 480

caacttgcaa atggtgagat cgatgtgcag cagatacaac aagaagaaaa tgtcgcagaa 540

cagcacggac aaaagactga aatgacaaaa gctgaacatt ccatacaaaa tgtcgtccgt 600

agaaatagag attgcagtga acaagaaagt gatgctcaaa gttcttgtac aagatcagaa 660

ccagaggctg agagcaagca caccaacagt ttcttggagt ttaagcagat aactgaaagg 720

caatcatcta ctgaccccaa aaacacactg gaaaatagag accctgaaaa ccctagtgat 780

aataagcgca agaaagcctc gacagatatc gaggtagtcc atataattga tgatgaacca 840

aagcctagca caccgatgga ggtggatatt gtgagaacaa accctcaagg aaacggcgat 900

aagtggttct ccatcccagc ccatcagttg gagctttctc tcagaagatc tgactatggc 960

agatcagagg accaagagaa aaatgatacg agaaccctga accactcaac ttcatctgcc 1020

ttttcattgt acaactgcag gcctatatcc tcttttggta acgctggtga tgctcagcca 1080

tgcagcacct cagcaacaca cgcagatctg caaaacaaaa atggagattc ggcagctccc 1140

tttcaagaca aggccgaccc aatttgccat cctatcagag ttgtagcact tcccgttcct 1200

gttggaggcc tcacgtttga cgggcaacca ttctggagtg gtgcacctgt ggcacccctt 1260

ctctatccac agtcaggtcc tcctatttgg aacggcagaa ctccagtgtc gcaagaggtg 1320

gacacgcaag caacttcatc gcagcagagt gatcccacag agatggactg tcagcagact 1380

gaaagcacac agcgacaaga agtgctacct tcaccgactg caaatgaaaa gcatctgcac 1440

gttgaaatcc cctcagacag caaccctcag caggtttcac ccatggccag cgaaagcgga 1500

accggaagta gtactgtgtt gaacaactct ggcaatgctc tcagtggaag tgcctgtgga 1560

agctcttcca accggatcgc cacccctacc gaacagtgta acgcatctga cggtgctact 1620

gaaaatccaa gcatggaggg ctcgcatcag ctgagccaac gagagattgc actgaacaag 1680

tttcggctca agaggaaaga aaggtgcttc gagaagaagg tgcggtacca gagcaggaaa 1740

ctacttgcag agcagcgtcc ccgggtgaag ggtcagtttg ttcggcaaga gcagaacatc 1800

caagcaagct ag 1812

Claims

1. The application of wheat TaPRR95-B protein or its coding gene in regulating plant height; the wheat TaPRR95-B protein is selected from an amino acid sequence shown in any one of SEQ ID NO 1-3; the plant is wheat.

2. The application of the wheat TaPRR95-B protein or the coding gene thereof in cultivating and preparing transgenic plants or improving plant germplasm; the wheat TaPRR95-B protein is selected from an amino acid sequence shown in any one of SEQ ID NO 1-3; the plant is wheat.

3. The use of claim 1 or 2, wherein the gene encoding the wheat TaPRR95-B protein is selected from the nucleotide sequence shown in any one of SEQ ID NO 4-6.

4. An application of SNP locus in identifying a coding gene of wheat TaPRR95-B protein is characterized in that the coding gene of the wheat TaPRR95-B protein is selected from a nucleotide sequence shown in any one of SEQ ID NO 4-6;

the SNP locus is linked with a coding gene of wheat TaPRR95-B protein, and the SNP locus is positioned at the 505806276 th site of a No. 5 chromosome.

5. A method for regulating plant height of a plant, which is characterized by comprising the following steps:

regulating and controlling the expression level of a coding gene of wheat TaPRR95-B protein in a plant, wherein the wheat TaPRR95-B protein is selected from an amino acid sequence shown as any one of SEQ ID NO 1-3;

the plant is wheat.

6. The method according to claim 5, wherein the plant height of said plant is reduced by reducing the expression level of a gene encoding said wheat TaPRR95-B protein in said plant; or, breeding shorter plant lines by crossing the gene-knocked-out line encoding the TaPRR95-B protein of wheat with other lines.

7. Method according to claim 6, wherein said reducing the expression level in said plant of a gene encoding said wheat TaPRR95-B protein is in particular: