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Cloning and functional expression method of gene AhAP2ER related to drought stress of peanuts

A technology of drought stress and cloning method, which is applied in the field of cloning and functional expression of AhAP2ER gene under drought stress in peanuts, and can solve the problems of few AP2 subfamilies, few cloning and function studies, etc.

Inactive Publication Date: 2016-04-06
SHANDONG PEANUT RES INST
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Problems solved by technology

A large number of studies have shown that during various abiotic stresses, the accumulation of AP2 / EREBP family proteins plays a potential protective role in plants under stress conditions, but there are few reports on the cloning and functional studies of this class of genes trapped in peanuts, especially is less of the AP2 subfamily

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  • Cloning and functional expression method of gene AhAP2ER related to drought stress of peanuts
  • Cloning and functional expression method of gene AhAP2ER related to drought stress of peanuts
  • Cloning and functional expression method of gene AhAP2ER related to drought stress of peanuts

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Embodiment Construction

[0027] The technical route of the present invention will be described in further detail below in conjunction with specific examples, but it is not intended to further limit the present invention.

[0028] 1.1 Implementation materials:

[0029] Material preparation and treatment: The peanut variety J11 was selected as the material, and the peanut seeds were cultured and germinated in Hoagland culture medium. The conditions for germination and seedling growth were 16h light / 8h dark at a temperature of 26°C, and they were grown for 14 days for subsequent drought stress treatment. Drought stress was treated with PEG6000. Peanut roots were soaked in 20% PEG6000 solution. Peanut roots were taken as materials at 0h, 6h, 12h, 18h, 24h, 48h and 72h, and all materials were stored in a -80°C ultra-low temperature refrigerator for later use.

[0030] 1.2 RNA extraction and cDNA synthesis

[0031]All utensils must be treated to remove RNase. The ware was soaked in 0.1% DEPC for 12 hours...

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Abstract

The invention relates to the biotechnology field and in particular relates to a cloning and functional expression method of a gene AhAP2ER related to drought stress of peanuts. The gene AhAP2ER in drought stress response is synthesized through preparation and treatment of materials, extraction of RNA (ribonucleic acid), synthesis of cDNA (complementary deoxyribonucleic acid) and RACE (rapid-amplification of cDNA ends) cloning technology. Expression of the gene AhAP2ER under the condition of drought stress is also analyzed by using fluorescent quantitative RT-PCR (reverse transcription-polymerase chain reaction), thus finally drawing a conclusion that the gene AhAP2ER plays an important role in the adaptability of the peanuts to drought stress. The transgenic plants have obvious drought resistance characteristic compared with a control group by transferring the gene to the peanuts by transgenic means, indicating that the gene AhAP2ER can conduce to obviously improving the drought resistance of the peanuts.

Description

【Technical field】 [0001] The invention relates to the field of biological genetic engineering, in particular to a method for cloning and functional expression of AhAP2ER gene under drought stress in peanuts. 【Background technique】 [0002] my country is the world's largest peanut producer, with a total output of more than 15 million tons, accounting for more than 40% of the world's total peanut output, ranking first in the world. However, further development of the peanut industry is threatened by drought. According to statistics, my country's annual peanut production reduction caused by drought reaches 30% to 50%. In addition to the decline in yield, the drought may also lead to a series of consequences such as aflatoxin pollution and quality decline of peanuts. However, due to the narrow genetic basis of peanut variety resources and the lack of corresponding gene sources, it is difficult to breed high-resistant varieties using traditional breeding methods. With the rapi...

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Application Information

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IPC IPC(8): C12N15/29C12N15/10C12Q1/68
CPCC07K14/415C12N15/10C12Q1/6851C12Q2531/113C12Q2563/107C12Q2545/101
Inventor 赵小波单世华闫彩霞张廷婷李春娟张浩
Owner SHANDONG PEANUT RES INST
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