Gene relevant to low-temperature, drought, ABA and high salt resistance of wheat and application thereof

A low-temperature resistance and gene technology, applied to the biological field, can solve the problem of unclear whether the stress resistance is related, and achieve the effect of enhancing the activity

Inactive Publication Date: 2019-03-08
CROP INST ANHUI PROV ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the deficiencies in the prior art, provide a kind of gene and its application related to wheat resi

Method used

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  • Gene relevant to low-temperature, drought, ABA and high salt resistance of wheat and application thereof
  • Gene relevant to low-temperature, drought, ABA and high salt resistance of wheat and application thereof
  • Gene relevant to low-temperature, drought, ABA and high salt resistance of wheat and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1. Analysis of expression pattern of wheat TaCOBL gene

[0052] Wheat seeds with full grains were selected, and 1 / 2 Hoagland medium was used for hydroponics to two leaves and one heart stage, and the following treatments were carried out: (1) Control (CK) did not carry out any stress treatment; (2) Stress treatment: a part of the seedlings were placed under normal temperature were used containing ABA (100 μmol·L -1 ), 20% PEG6000 (20%), NaCl (100 mmol·L -1 ) solution of Hoagland broth treatment. Another part of the seedlings was directly placed in a light incubator at a temperature of 4 °C. The leaves and roots of 1, 2, 6, 12 and 24 h of stress treatment were clipped and placed in liquid nitrogen for analysis of gene expression characteristics during the stress process. Total RNA was extracted according to the instructions of the RNA extraction kit. After concentration determination, the concentration of RNA extracted from different tissues was uniformly adju...

Embodiment 2

[0053] Embodiment 2, a kind of method of wheat TaCOBL gene promoter activity identification

[0054] Sequence analysis was performed on 20 wheat materials, and it was found that the gene located on chromosome 5B has two allelic variations among different materials, namely, there is an indel of four bases of TATA at the upstream regulatory region-550bp ( figure 2 ). The two allelic types were named Hap5B-a (with TATA) and Hap5B-b (with TATA missing), respectively. According to the full-length sequence of the gene on chromosome 5B, a pair of primers were designed to clone two alleles from Wanmai 38 (containing Hap5B-a allele) and Yangmai 158 (containing Hap5B-b allele). promoter sequence. The upstream primer is located at -1670 bp before ATG, and the downstream primer is the antisense strand sequence of 20 bp before ATG. Add a Hindll1 restriction site (AAGCTT) and a protective base ACG to the 5' end of the upstream primer, and add a BamHI restriction site (GGATCC) and a prot...

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Abstract

The invention discloses a gene relevant to low-temperature, drought, ABA and high salt resistance of wheat and application thereof, and relates to the technical field of biology. The invention provides expression mode analysis of the wheat TaCOBL gene. The result shows that the gene realizes the up-regulated expression through low-temperature induction and realizes down-regulated expression through drought, ABA and high salt induction. A promoter-GUS fusion expression double-ingredient vector is respectively built; according to the GUS active determination and tissue chemical dyeing result, under the low-temperature induction effect, the promoter of the gene can greatly improve the GUS gene expression level; the result shows that the promoter of the gene belongs to the induction type promoter; under the low-temperature induction, the activity of the Hap5B-a type promoter is higher than that of an Hap5B-b type promoter. The result proves that the activity of the promoter can be enhancedthrough the insertion of promoter region TATA four basic groups. The gene type Hap5B-a is an excellent allelic variation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a gene related to wheat resistance to cold, drought and the like. Background technique [0002] Wheat (Triticum aestivum L.) is an important food crop in my country, and its sown area and total output both account for about 22% of the national food crop, and it is also widely grown worldwide. Abiotic stresses such as low temperature, drought, and salinity severely restrict the production of wheat. Discovering the stress resistance gene resources of wheat and improving the stress resistance of wheat varieties is an important way to deal with adverse environmental stress. Mining and utilizing stress resistance genes and cultivating breakthrough varieties is an effective way to increase wheat yield. [0003] The COBRA gene affects the correct positioning of cellulose microfibrils in plant cell walls and the directional elongation of cells by encoding an important extracellular glycosy...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12Q1/6851
CPCC07K14/415C12Q1/6851C12Q2561/113C12Q2531/113
Inventor 刘方方万映秀张琪琪曹文昕李炎李耀
Owner CROP INST ANHUI PROV ACAD OF AGRI SCI
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