Corn casein kinase 2CK2[alpha]2 and application of coding gene thereof based on high-temperature stress response

A technology of casein kinase and coding gene, which is applied in the field of high temperature stress resistance of maize, and can solve the problems of lack of high temperature resistant gene resources and germplasm resources of maize

Active Publication Date: 2021-11-02
HENAN AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a corn casein kinase 2CK2α2 and the application of its coding gene based on high temperature stress response, so as to solve the current technical problem of lack of high temperature resistant gene resources and germplasm resources of corn

Method used

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  • Corn casein kinase 2CK2[alpha]2 and application of coding gene thereof based on high-temperature stress response
  • Corn casein kinase 2CK2[alpha]2 and application of coding gene thereof based on high-temperature stress response
  • Corn casein kinase 2CK2[alpha]2 and application of coding gene thereof based on high-temperature stress response

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Casein kinase 2 in response to high temperature in corn α Identification of subunits

[0024] 1. qRT-PCR identification of casein kinase 2 affected by high temperature in maize α Subunit

[0025] To determine the ZmCK2α1 、 a2 、 a3 、 Whether α4 is affected by high temperature, the three-leaf stage seedlings of the corn hybrid Zhengdan 958 were subjected to high temperature stress treatment for 0 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h and 10 h, and then the first The expression of the total RNA of the two leaves was analyzed by qRT-PCR; the results showed that ZmCK2α2 and ZmCK2α3 were significantly induced by high temperature stress, and the expression of ZmCK2α2 increased most significantly, and the expression level was the highest at 4 h of high temperature treatment, but high temperature stress had no effect on ZmCK2α1 and ZmCK2α4 expression basically had no effect (see figure 1 ).

[0026] 2. wxya 2 α 2 Analysis of cis-acting elements in gene promoter...

Embodiment 2

[0029] Embodiment two: wxya 2 α 2 High temperature resistance function test of genes

[0030] Malondialdehyde (MDA) content is the embodiment of the degree of peroxidation of the plant cell membrane. A high content of MDA indicates a high degree of peroxidation of the plant cell membrane and serious damage to the cell membrane. while H 2 o 2 The accumulation of content can directly or indirectly oxidize biological macromolecules such as nucleic acid and protein in the cell, and damage the cell membrane, thereby accelerating the aging and disintegration of cells. Catalase (CAT) can scavenge H 2 o 2 , is one of the important enzymatic defense systems in plants. to be sure ZmCK2α2 Whether it is involved in the resistance of maize to high temperature, using the transient transformation system of maize protoplasts, the overexpression vector OE -ZmCK 2 α 2 and wxya 2 α 2 dsRNA and distilled water were respectively transformed into maize leaf protoplasts, incubated at r...

Embodiment 3

[0032] Embodiment three: based on wxya 2 α Identification of high temperature resistance function of transgenic plants of 2

[0033] for further exploration wxya 2 α 2 The function under high temperature stress was obtained by using maize genetic transformation technology wxya 2 α 2 Overexpression homozygous lines. First, by homologous recombination wxya 2 α 2 Recombined into the binary expression vector pFGC5941 containing the 35S promoter, then transformed the plasmid with correct sequencing results into Agrobacterium GV3101, and finally transformed pFGC5941- ZmCK2α2 Maize was transformed with the vector, and T was obtained after herbicide selection 0 Positive plants were self-crossed to T 2 generation and obtained ears of transgenic plants.

[0034] 1. Detection of transgenic lines by qRT-PCR wxya 2 α 2 expression level

[0035] In order to determine the overexpression transgene wxya 2 α Whether the expression level of 2 is improved, the obtained T ...

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Abstract

The invention discloses corn casein kinase 2CK2[alpha]2 and application of a coding gene of the corn casein kinase 2CK2[alpha]2 based on high-temperature stress response, and aims to solve the technical problem that high-temperature-resistant gene resources and germplasm resources of corn are deficient at present. Based on the discovery that a corn casein kinase 2[alpha] subunit can respond to high temperature stress, overexpression vectors OE-ZmCK2[alpha]2 and pFGC5941-ZmCK2[alpha]2 overexpression vectors are constructed, ZmCK2[alpha]2 overexpression homozygous strains are obtained by using a corn genetic transformation technology, and strains with high expression quantity are screened out by using qRT-PCR (quantitative reverse transcription-polymerase chain reaction) detection and are used as breeding materials for breeding high-temperature-resistant corn varieties / lines. The invention determines that the corn casein kinase 2[alpha] subunit can respond to high-temperature stress, and preliminarily analyzes the action mechanism of the corn casein kinase 2[alpha] subunit under high-temperature stress, thereby being beneficial to understanding the heat-resistant mechanism of corn, and providing important gene resources and related germplasm resources for cultivating high-temperature-resistant corn varieties.

Description

technical field [0001] The invention relates to the technical field of corn high temperature stress resistance, in particular to the application of corn casein kinase 2CK2α2 and its coding gene based on high temperature stress response. Background technique [0002] Corn is the largest food crop in my country, and its output and area both rank first in food crops, and it plays an important role in national food security. However, with the warming of the global climate, high temperature and heat damage have occurred frequently in recent years, which has caused serious losses to corn production. Relevant studies have shown that the optimum temperature for maize reproductive growth is 25-28°C. Higher temperatures can accelerate its growth and development and shorten its growth period, but extreme high temperatures hinder its growth and development. For example, high temperatures above 32°C are not conducive to Its dry matter accumulates, and the high temperature above 35°C for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/12C12N15/54C12N15/82A01H5/00A01H6/46C12Q1/6895
CPCC12N9/12C12Y207/11001C12N15/8271C12Q1/6895C12Q2600/13C12Q2600/158
Inventor 胡秀丽刘艳培赵玉龙陈闪闪
Owner HENAN AGRICULTURAL UNIVERSITY
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