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Arabidopsis tir2 Application of Genes in Improving Plant Salt Stress Resistance

A technology of Arabidopsis thaliana and plant salt, applied in the field of plant genetic engineering, can solve the problems of undiscovered functions and effects, and achieve the effects of improving plant salt stress resistance, increasing growth length, and increasing germination quantity and speed

Active Publication Date: 2022-04-01
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there are no relevant reports on the function and role of genes encoding key enzymes in the auxin synthesis pathway in plants under salt stress.

Method used

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  • Arabidopsis <i>tir2</i> Application of Genes in Improving Plant Salt Stress Resistance
  • Arabidopsis <i>tir2</i> Application of Genes in Improving Plant Salt Stress Resistance
  • Arabidopsis <i>tir2</i> Application of Genes in Improving Plant Salt Stress Resistance

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Arabidopsis TIR2 Obtaining of Gene Transgenic Line OE141

[0027] 1. Arabidopsis TIR2 Acquisition of genetic mutants

[0028] Acquired in the Arabidopsis database TIR2 The CDS sequence of the gene is 1173 nt in length and encodes a protein containing 391 amino acids. The nucleotide sequence of the gene is shown in SEQ ID NO:1, and the amino acid sequence is shown in SEQ ID NO:2. Arabidopsis TIR2 Gene T-DNA insertion mutant ( tir2-3 ) was obtained from the Arabidopsis Resource Center, and then subjected to homozygous screening, expression, analysis and final identification.

[0029] 2. Arabidopsis TIR2 Obtaining of Gene Transgenic Line OE141

[0030] 1. Arabidopsis TIR2 Obtaining of Gene Overexpression Lines

[0031] (1) Construction of a constitutive expression promoter driven by 35S TIR2 The overexpression vector (35S:: TIR2 ): According to Arabidopsis TIR2 Gene CDS sequence, design gene-specific primers, the sequence is as follows (5'-3'): ...

Embodiment 2

[0047] Arabidopsis thaliana under salt stress in embodiment 2 TIR2 Effects of genes on seed germination and cotyledon unfolding

[0048] 1. Preparation of culture medium

[0049] (1) Configuration of 1 / 2MS(-) medium for plant tissue culture: weigh 2.37 g / L drug MS, 15 g / L sucrose, dissolve in ultrapure water and dilute to 1000 ml, add 1M KOH solution and dilute hydrochloric acid to adjust the pH to 5.80;

[0050] Configuration of high-salt medium: Add 58.44g / L sodium chloride on the basis of 1 / 2MS(-) medium formula, and then it can be configured as a high-salt medium containing 150 mM NaCl.

[0051] (2) Subpackage: Dispense each 200 ml of liquid medium into 250 ml Erlenmeyer flasks, add 1.6 g of agarose, and sterilize at 121°C for 20 minutes under high temperature and high pressure.

[0052] (3) After the sterilization, when the culture medium is cooled to 50-60°C, pour it into a glass plate, and divide into 10 glass plates per 200 ml. After the culture medium is solidifie...

Embodiment 3

[0056] Example 3: Arabidopsis under salt stress TIR2 Effect of genes on seedling growth phenotype

[0057] 1, the preparation of culture medium: with embodiment 2.

[0058] 2. Seedling growth phenotype detection

[0059] The medium was divided into two groups, 1 / 2 MS(-) medium and high-salt medium were added as the control group CK and the salt stress group respectively, and 4 replicates were set for each group.

[0060] Take wild-type strain WT, T-DNA mutant strain ( tir2-3 ) and the overexpressed homozygous strain (OE141) were used as treatment materials. Arabidopsis thaliana seeds with the same size and plump grains were selected at the same period, sterilized with chlorine gas for 5 hours, and then spread evenly and dispersedly on 1 / 2MS (-) medium. Seal the medium, stratify in the dark at 4°C for three days, then grow vertically at 22°C for 3 days, then select seedlings with consistent root length and transfer them to 1 / 2MS(-) medium and high-salt medium respectively, ...

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Abstract

The invention provides Arabidopsis TIR2 Application of genes in improving plant resistance to salt stress. said TIR2 The interacting proteins of the genes include IAA4 and IAA8, and the present invention obtains Arabidopsis thaliana through genetic engineering technology TIR2 The overexpression vector of the gene, and thus obtained the overexpression strain of Arabidopsis thaliana, and finally obtained the Arabidopsis thaliana after screening TIR2 Gene overexpression homozygous line; the Arabidopsis TIR2 Gene overexpression homozygous lines show salt tolerance, have good plant salt stress resistance, can not only increase the rate of seed germination and cotyledon expansion rate, shorten the time of seed germination and cotyledon expansion, but also increase the length of root system of adult plant seedlings . The present invention confirms Arabidopsis thaliana through experiments TIR2 The gene has potential application value in improving plant salt stress resistance, and provides a basis for the utilization of Arabidopsis thaliana TIR2 Genetic breeding of salt-tolerant and high-yielding crop varieties has laid a good theoretical and application foundation.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to Arabidopsis TIR2 Application of genes in improving plant resistance to salt stress. Background technique [0002] Salt stress is one of the abiotic stress factors affecting crop yield. Large areas of saline-alkali land and salinized soil make it difficult for most crop varieties to be affected by varying degrees of salt damage during the growth process, making it difficult to develop their yield potential. Salt resistance of plants is a complex quantitative trait determined by multiple genes. Different plants have different salt tolerance methods and mechanisms, and their tissues or cells have different salt tolerance responses. With the continuous reduction of cultivated land and the continuous increase of population, it is of great practical significance to study the salt-tolerant mechanism of crops and cultivate salt-tolerant varieties for the development and effecti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00A01H6/20
CPCC12N15/8273C07K14/415
Inventor 黄金光蔡慧娴郑成超
Owner SHANDONG AGRICULTURAL UNIVERSITY
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