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Use of rice osmotic stress response protein OsU496A in improving osmotic stress resistance of rice

An osmotic stress, anti-osmotic technology, applied in the application, recombinant DNA technology, chemical instruments and methods, etc., can solve the problems of rice morphology, internal physiological environment and metabolism changes, plant death, plant growth retardation and other problems

Pending Publication Date: 2020-12-29
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Rice requires a huge amount of water throughout its growth period. At present, the world's agricultural water consumption accounts for more than 70% of the total fresh water. The osmotic stress caused by the shortage of water resources will not only hinder the growth and development of rice, but also lead to a decline in quality and yield.
In an osmotic stress environment, rice cannot obtain the water needed for normal growth, which will cause changes in the morphology, internal physiological environment and metabolism of rice, eventually leading to stunted growth and development of plants, and severe plant death

Method used

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  • Use of rice osmotic stress response protein OsU496A in improving osmotic stress resistance of rice
  • Use of rice osmotic stress response protein OsU496A in improving osmotic stress resistance of rice
  • Use of rice osmotic stress response protein OsU496A in improving osmotic stress resistance of rice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0083] Example 1: Construction of an interference expression vector and an overexpression vector of the OsU496A gene.

[0084] 1.1 Construct the interference expression vector of OsU496A gene;

[0085] The OsU496A interference expression vector is constructed by reversely connecting a segment of target sequence SEQ ID No: 4 at both ends of the intron of the vector. Specific method: Design specific amplification primers for the target sequence SEQIDNo: 4, add SacI and SpeI restriction sites at the end of the primers, and the primer sequences are F: 5'-GGTACCACTAGTAGCAGCCTCCACACCCTCG-3' and R: 5'-GGATCCGAGCTCTCGATCCACTTCCCCATA-3' . The total RNA of rice leaves was extracted using the Plant RNA Extraction Kit of Kangwei Century Company. The described rice leaf total RNA is according to the ReverAid of Quanshijin Company TM FirstStrandcDNASynthesisKit Product Description Perform reverse transcription to obtain the first-strand cDNA of rice leaves. Using cDNA as a template, the...

Embodiment example 2

[0096] Implementation case 2: Interfering expression of pTCK303-iU496A and overexpression of pTCK303-0sU496A recombinant vector transforming rice plants;

[0097] After verifying that the constructed interference expression pTCK303-iU496A and overexpression pTCK303-OsU496A recombinant vectors are correct, they were transferred into Agrobacterium EHA105. The specific steps are as follows:

[0098] (a) Add 2 μL of the recombinant plasmid to 100 μL of Agrobacterium competent, and mix slightly;

[0099] (b) Stand on ice for 5 minutes;

[0100] (c) Immerse in liquid nitrogen for 5 minutes;

[0101] (d) 37°C water bath for 5 minutes;

[0102] (e) Stand on ice for 5 minutes;

[0103] (f) Add 700 μL of liquid LB medium, shake at 28°C / 200rpm for 2-3 hours;

[0104] (g) Take 200 μL of bacterial liquid and spread it on the LB plate containing kanamycin and rifampicin;

[0105] (h) Inverted and placed in an incubator at 28°C for 2-3 days.

[0106] Then, the recombinant plasmids were...

Embodiment example 3

[0107] Implementation case 3: Phenotype analysis of OsU496A interference expression lines and overexpression lines under osmotic stress;

[0108] The RNAi-OsU496A, wild-type and OX-OsU496A seeds with consistent germination were cultured in IRRI nutrient solution for 10 days, and then treated with 50 mM mannitol solution. After treatment, the phenotypes of the strains were significantly different. The wilting degree of the RNAi-OsU496A strain was significantly lower than that of the wild type and the overexpression strain, and the wilting degree of the overexpression strain was the most serious. It can be seen that the interference expression strain is resistant to osmotic stress. ability is better than wild type, and wild type is better than overexpression ( figure 2 ).

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Abstract

The present invention discloses a use of rice osmotic stress response protein OsU496A in improving osmotic stress resistance of rice. Firstly, an interference expression vector of a specific fragmentof a coding gene of the rice osmotic stress response protein OsU496A containing positive and negative RNAi is introduced into rice cells, tissues or organs, expression amount of the coding gene of therice osmotic stress response protein OsU496A in the rice cells, tissues or organs is reduced, then the rice cells, tissues or organs are cultured into plants, so that the expression amount of the coding gene of the rice osmotic stress response protein OsU496A is reduced and the rice with good physiological indexes and improved osmotic stress resistance under osmotic stress is obtained. The use ofthe rice osmotic stress response protein OsU496A provides a new choice for genetic improvement and molecular breeding of the rice.

Description

technical field [0001] The invention relates to the application of a rice osmotic stress response protein OsU496A in improving the ability of rice to resist osmotic stress, and belongs to the field of plant genetic engineering. Background technique [0002] Rice is a crop with a wide range of planting. It is planted in 113 countries around the world. It is one of the staple foods for more than half of the world's population and the most important food crop in Asia. China is the world's largest rice producer and consumer, and has a long history of rice cultivation. In recent years, due to the popularization of fine varieties and the reform of farming system, the yield of rice has increased significantly, but abiotic stress such as osmotic stress still restricts the development of high yield, diversification and high quality of rice. Rice requires a huge amount of water throughout its growth period. At present, the world's agricultural water consumption accounts for more than...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8218C12N15/8271
Inventor 吕冰王家豪方是珍王艺霖陈洪晴陈开杨超陈云
Owner YANGZHOU UNIV
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