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Small peptide for regulating and controlling sizes of plant seeds and organs, coding gene and application thereof

A technology for plant seeds and organs, which is applied in the field of small peptides and their coding genes for regulating the size of plant seeds and organs, and can solve the problems of difficult research on biological functions, restrictions on the development and application of small peptides, and the large number of small peptides in plants, etc. question

Active Publication Date: 2022-01-28
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still a large number of small peptides with unknown functions that have not been identified in plants, and the development and application of small peptides are greatly limited due to the large number of small peptides in plants and the difficulty in studying their biological functions.

Method used

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  • Small peptide for regulating and controlling sizes of plant seeds and organs, coding gene and application thereof
  • Small peptide for regulating and controlling sizes of plant seeds and organs, coding gene and application thereof
  • Small peptide for regulating and controlling sizes of plant seeds and organs, coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Example 1 Screening of atzsp1 mutants and identification of AtZSP1 gene

[0112] 1. Screening of atzsp1 mutants

[0113] The invention firstly screens and classifies unknown small peptides with amino acid length less than 150 in Arabidopsis thaliana, and there are 3698 unidentified genes in total. In order to obtain the efficiency of effective mutants, the scope was further narrowed, and a single-copy gene with an amino acid length of less than 100 was identified, and the gene inserted in the exon was ordered from the official website of TAIR (http: / / www.arabidopsi s.org / ). T-DNA insertion mutants. After the homozygous mutants were screened by PCR, a preliminary phenotype observation was carried out, and a gene involved in the regulation of Arabidopsis seed and organ size was identified, named AtZSP1 gene. The amino acid sequence encoded by the AtZSP1 gene is the polypeptide AtZSP1 shown in SEQ ID No. 1 in the sequence listing. The genomic gene of AtZSP1 is the doubl...

Embodiment 2A

[0132] The acquisition of embodiment 2 AtZSP1 gene overexpression plant

[0133] In order to further study the function of AtZSP1 gene, the overexpression plants of AtZSP1 gene were constructed.

[0134] The specific process is:

[0135] (1) Construction of overexpression vector

[0136] Using the cDNA of wild-type Arabidopsis as a template, PCR was performed with primers 5'-CGA CTGCAG ATGCAAAAATCGTTCTCGTTG-3' and 5'-GAG ACTAGT GGAATGGGGAGTGGAATCAG-3' (respectively containing two restriction enzyme sites, PstI and SpeI) Amplified, the PCR product was purified and recovered, digested with PstI and SpeI, and then ligated to the pSuper1300 vector ( Figure 8 ). The ligation product was transformed into Escherichia coli competent DH5α cells, cultured on LB solid medium containing 50 μg / mL kanamycin, and sequenced after colony PCR to obtain the correct clone. The recombinant overexpression vector was named p35S::AtZSP1. p35S::AtZSP1 replaces the fragment (small fragment) betwe...

Embodiment 3a

[0156] Phenotype observation of embodiment 3atzsp1 mutant and AtZSP1 gene overexpression plant

[0157] 1. Seed size of mutants and overexpressed plants

[0158] The experiment was repeated three times, and 30 seeds were taken from each line for each repetition.

[0159]By stereoscopic observation, the seed size of strain atzsp1-1 and strain atzsp1-2 mutants was significantly smaller than that of the wild type, on the contrary, the strain p35S::AtZSP1#1 and strain p35S::AtZSP1 Seeds of #2 were significantly larger than wild type. In order to quantify the degree of seed change, we counted the seed area of ​​each line, and the results showed that the seed area of ​​the mutant line atzsp1-1 and atzsp1-2 was smaller than that of the wild type, and similarly, the strains overexpressing p35S:AtZSP1 The seeds of both line p35S::AtZSP1#1 and line p35S::AtZSP1#2 were larger than wild type. The result is as Figure 10 shown. In addition, the 100-seed weight per plant of each strain...

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Abstract

The invention discloses a small peptide for regulating and controlling the sizes of plant seeds and organs, a coding gene and application thereof. The invention specifically discloses a polypeptide AtZSP1 (AtZSP1 small peptide) with an amino acid sequence of SEQ ID No.1 and an application of the coding gene of the polypeptide AtZSP1 in regulating and controlling the sizes of plant seeds and organs. According to the invention, a gene participating in regulation and control of the sizes of arabidopsis seeds and organs is successfully identified for the first time by inserting T-DNA into a mutant library and adopting a reverse genetics method according to bioinformatics analysis, the gene is named as the AtZSP1 gene, the sizes of the plant seeds and / or organs are increased through a genetic engineering method, an AtZSP1 overexpression plant is obtained, high-quality gene resources are enriched for the development and application fields of the small peptides, and the small peptides can be well applied to improvement of crop varieties and increase of crop yield, and have important practical value for plant breeding work.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to small peptides for regulating the size of plant seeds and organs, coding genes and applications thereof. Background technique [0002] Seed size is one of the main factors affecting crop yield, directly determines crop yield, and has long been an important goal of many crop breeding and improvement. In addition, the size of plant organs is also an important trait in sciences such as agronomy, horticulture, and herbalism. Therefore, the study of the regulatory mechanism of seed and organ size has become an important research field in plant science. The size of plant organs is determined by both cell proliferation and cell expansion, the former increases cell number through cell division, and the latter increases cell size through cell growth. Many genes involved in the regulation of cell proliferation and cell growth, thereby affecting plant organ size have been identifi...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/84C12N15/55A01H5/10A01H5/00A01H6/20
CPCC07K14/415C12N15/8205C12N15/8218C12N15/8261C12N9/22Y02A40/146
Inventor 张学琴曾月娟申思敏陈立群叶德
Owner CHINA AGRI UNIV
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