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Application of rice seryl tRNA synthetase gene STS11 and protein encoded by gene

A serine and synthase technology, which is used in applications, genetic engineering, plant genetic improvement, etc.

Active Publication Date: 2020-03-13
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These results indicate that seryl tRNA synthetase plays a key role in the development of tobacco chloroplasts, but it has not been reported that this gene affects chloroplast development in rice

Method used

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  • Application of rice seryl tRNA synthetase gene STS11 and protein encoded by gene
  • Application of rice seryl tRNA synthetase gene STS11 and protein encoded by gene
  • Application of rice seryl tRNA synthetase gene STS11 and protein encoded by gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] 1. Rice material

[0040] The japonica rice variety is (Oryza sativa L.japonica) "Wuyujing 7", the indica rice variety is (Oryzasativa L.indica) "9311", the temperature-sensitive chlorophyll-deficient mutant derived from Wuyujing 7, and the STS11 gene transgenic line .

[0041] 2. SSR and STS markers localize the STS11 gene

[0042] Genomic DNA for gene localization was extracted from rice leaves by a rapid extraction method of rice trace DNA. Take 0.2g of rice leaves, freeze them with liquid nitrogen, grind them into powder in a small mortar with a diameter of 5cm, transfer them to a 1.5ml centrifuge tube to extract DNA, and dissolve the obtained DNA precipitate in 150μl ultrapure water. 2 μl of DNA sample was used for each PCR reaction.

[0043] Preliminary mapping of the STS11 gene: crossing the temperature-sensitive chlorophyll-deficient mutant tcm11 with the indica rice 9311, F 1 The leaves of all plants in the generation were normal phenotypes, and there was n...

Embodiment 2

[0050] Plant transformation and functional analysis:

[0051] In order to verify whether ORF13 is a candidate gene, we used PCR to amplify the full-length sequence of ORF13 genome including 2056 bp before ATG, full-length ORF13 and 860 bp downstream of the stop codon in Wuyujing 7 genome, and then ligated this fragment into a binary expression vector In pCAMBIA1300 ( Figure 5 ), the gene was transformed into tcm11 mutant by Agrobacterium-mediated plant transgenic method, and a total of 26 transgene complementary plants were obtained. Under the high temperature treatment of these transgenic lines, the phenotype and chlorophyll content of all positive plants were consistent with the wild type Wuyujing 7 ( Figure 6 , Figure 7 ). This shows that ORF13 is the candidate gene STS11 controlling the traits of the mutant.

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Abstract

The invention discloses application of a rice seryl tRNA synthetase gene STS11 (Seq ID No: 1, 2) and a protein (Seq ID No: 3) encoded by the gene. According to the invention, a seryl tRNA synthetase gene STS11 is cloned in rice for the first time by adopting a map-based cloning technology, and the gene function is identified through a transgenic complementation experiment. Through cloning and application of the STS11 gene, the development of rice chloroplasts can be effectively regulated and the high temperature tolerance breeding can be carried out by regulating the development of rice chloroplasts.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to the application of a rice seryl tRNA synthetase gene STS11 and its encoded protein. Background technique [0002] Rice is one of the three major food crops in my country, and rice is a favorite food in the world, especially in Asian countries. my country's rice planting area reaches 450 million mu, ranking second in the world. Among my country's 1.4 billion population, more than 60% take rice as the main grain. Chloroplast is the place where plants carry out photosynthesis, which converts light energy into chemical energy through photosynthesis to provide energy for plant growth. In recent years, with global warming, high temperature and heat damage have occurred frequently, which often damages chloroplasts and prevents normal photosynthesis, resulting in a significant reduction in rice production. Therefore, under high temperature environment, the normal development o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/82C12N5/10A01H5/12A01H6/46
CPCC12N9/1085C12N15/825C12N15/8271C12Y205/01073
Inventor 高振宇钱前曾大力房国楠郭龙彪张光恒朱丽任德勇张强
Owner CHINA NAT RICE RES INST
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