Cloning and function-expressing methods of peanut adversity stress AhROLP1 gene
A technology of adversity stress and cloning method, which is applied in the field of cloning and functional expression of AhROLP1 gene under adversity stress in peanut, and can solve the problem that the research on abiotic stress function has not yet been reported.
Active Publication Date: 2014-02-05
SHANDONG PEANUT RES INST
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At present, there are few studies on the function of this gene, and its function in abiotic stress has not been reported
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[0030] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but it is not intended to further limit the present invention.
[0031] The concrete implementation process of the present invention is:
[0032] 1 Experimental materials and methods
[0033] 1.1 Experimental materials
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Abstract
The invention discloses cloning and function-expressing methods of a peanut adversity stress AhROLP1 gene. The adversity stress AhROLP1 gene is synthesized by preparing and processing materials, extracting RNA (Ribonucleic Acid), synthesizing cDNA, and cloning with RT-PCR (Reverse Transcription-Polymerase Chain Reaction). An open reading frame of the gene is 1620 bp, and 540 amino acids in total are coded. An amino acid sequence of the gene has homology of over 70% with reticuline oxidase of plants such as chickpeas, soybeans, wild strawberries, grapes and tomatoes. An expression mode of the AhROLP1 under low temperature, salt stress and drought stress is verified through fluorescent quantitative PCR (Polymerase Chain Reaction), and the result indicates that transcriptional level of the gene is obviously increased under adversity stress of three non-living things, and then is kept at a higher level all the time. The gene is lead into arabidopsis through transgenic means, and a transgenic plant has obvious cold resistance, salt resistance and drought resistance in comparison with a control group.
Description
technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for cloning and functional expression of AhROLP1 gene under adversity stress in peanuts. Background technique [0002] Peanut is a leguminous arachis crop, rich in oil and protein, and is an important oil crop and economic crop in my country. The yield and quality of peanuts are seriously affected by drought, saline-alkali and other stresses, and the annual reduction rate of peanuts caused by drought in the whole country reaches more than 20%. However, due to the narrow genetic basis of peanut variety resources and the lack of high drought-resistant, cold-resistant, and salt-tolerant gene sources, it is difficult to breed high-resistant varieties using conventional breeding methods. With the rapid development of molecular biology, the use of transgenic technology to improve plant stress tolerance has achieved remarkable results in recent years, and a deeper understanding of s...
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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N15/10C12Q1/68C12Q1/26
Inventor 陈娜禹山林杨庆利迟晓元潘丽娟陈明娜王通王冕杨珍
Owner SHANDONG PEANUT RES INST
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