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Tomato glutaredoxin gene SlRX1 as well as cloning method and application thereof

A protein and gene technology, applied in the field of molecular biology and genetic engineering, can solve the problems of lack of genetic resources of stress-resistant plants, achieve the effect of strong theoretical research and applied research value, increase yield and quality, and enhance resistance

Inactive Publication Date: 2010-09-29
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a tomato glutaredoxin gene SlGRX1 and its cloning method and application in view of the current problems such as the molecular mechanism of plant stress resistance and the lack of stress-resistant plant gene resources

Method used

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  • Tomato glutaredoxin gene SlRX1 as well as cloning method and application thereof
  • Tomato glutaredoxin gene SlRX1 as well as cloning method and application thereof
  • Tomato glutaredoxin gene SlRX1 as well as cloning method and application thereof

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Embodiment 1

[0021] Cloning of embodiment 1 tomato glutaredoxin gene

[0022] According to the CGFS glutaredoxin-like genes of Arabidopsis and rice, a pair of degenerate primers were designed according to the conserved homologous sequence. The upstream primer T1 is: 5'-SAAAGTGGTTCTSTWYATGAARGG-3', and the downstream primer T2 is: 5'- ATATCWSAACCACCGAMMARYTC-3', where W stands for T or A, S stands for G or C, M stands for A or C, R stands for A or G, Y stands for C or T, N stands for A or C or G or T, M stands for A Or C, R stands for A or G, Y stands for C or T, D stands for A or G or T. After RT-PCR amplification, sequencing, and homology comparison with online databases such as NCBI, it was proved that the obtained fragment was the glutaredoxin gene of tomato. Using this fragment as the seed sequence, use the electronic cloning method to carry out extension splicing in the tomato EST database, and design primers according to the extended sequence. The upstream primer YGS2 is: 5'-CATGGCG...

Embodiment 2

[0023] The characteristics and function analysis and application of embodiment 2 SlGRX1 gene:

[0024] 1. Analysis of tissue expression characteristics of SlGRX1 gene

[0025] In order to analyze the expression of the gene in tomato, the total RNA in the leaves, roots, stems and petals of tomato was extracted with TRIzol reagent, reverse transcribed into cDNA, and quantitative real time RT-PCR was used to analyze the expression of SlGRX1 in different tissues of tomato Primer 5'-ATGCGAGCATCGTTGTTGC-3' / 5'-TCTATCTGGCTCCTCAACCACAC-3' was used to amplify the target SlGRX1 gene; primer 5'-CACGAGGTATAATCTAGGGTTTG-3' / 5'-ATTTTGTCAGGGTTGTATCCTAC-3' was used to amplify Internal reference gene EF-1-α. The analysis results show that in the detected tissues, the expression of this gene has been detected, and the expression level in the leaves is the highest, and the expression level in the roots is the lowest (see figure 1 ).

[0026] 2. Subcellular localization of SlGRX1 protein

[002...

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Abstract

The invention discloses a tomato glutaredoxin gene SlRX1 as well as a cloning method and application thereof. The tomato glutaredoxin gene SlRX1 is a first glutaredoxin gene separated from tomatoes, consists of 1,003 basic groups and is used for coding CGFS glutaredoxin forced and induced in environments of drought, high salt, and the like. The gene can be expressed in roots, stems, leaves and petals of the tomatoes and participates in an anti-nonbiostress reaction of plants. If the gene is silent in tomatoes, the tomatoes are more sensitive to the environments draught, high salt, oxidativestress, and the like, and if the gene is excessively expressed in arabidopsis thaliana, the resistance to the environments of draught, high salt and oxidativestress of transgenic arabidopsis thaliana. The gene can improve the resistance of plants to the environments of draught, high salt and oxidativestress and has strong theoretical research and application values.

Description

technical field [0001] The invention relates to the fields of molecular biology and genetic engineering, in particular to a tomato glutaredoxin gene SlGRX1 and its cloning method and application. The present invention relates to the cDNA sequence of tomato SlGRX1, and the protein coded by the cDNA belongs to CFGS glutaredoxin. Background technique [0002] Active oxygen is an important signal transduction factor in the process of plant growth and development, and plays an important role in promoting plant growth and metabolism, as well as high yield and quality of crops. However, excessive accumulation of reactive oxygen species in plants will destroy the biomacromolecules in the body, interfere with the normal signal transduction of plants and cause serious damage to plants, which may easily cause wilting or even death of plants. Various adverse environments for plant growth, such as drought, salt damage, chilling damage, heavy metals, ultraviolet radiation, ozone, mechani...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/10C07K14/415A01H5/00
Inventor 周雪平郭玉双吴建祥
Owner ZHEJIANG UNIV
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