Method in increasing oleic acid content of soybean and peanut seed by applying gene silent technology

A technology of gene silencing and peanut, applied in botany equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems of reducing the synthesis of linoleic acid and reducing catalytic activity, so as to increase the content of oleic acid, shorten the cycle, The effect of improving efficiency

Inactive Publication Date: 2006-01-25
NORTHEAST NORMAL UNIVERSITY
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Problems solved by technology

Δ12 fatty acid dehydrogenase is a key enzyme in fatty acid metabolism that catalyzes the formation of linoleic acid from oleic acid. The genetic modification of Δ12 fatty acid dehydrogenase gene FAD2-1 in soybean and peanut seeds by using the principle of gene silencing triggered by RNA inverted repeats leads to The gene is silenced, so that the formation of Δ12 fatty acid dehydrogenase is blocked and the catalytic activity is reduced, thereby reducing the synthesis of linoleic acid and accumulating a large amount of oleic acid

Method used

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  • Method in increasing oleic acid content of soybean and peanut seed by applying gene silent technology
  • Method in increasing oleic acid content of soybean and peanut seed by applying gene silent technology
  • Method in increasing oleic acid content of soybean and peanut seed by applying gene silent technology

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specific Embodiment approach

[0010] Take the application of gene silencing technology to increase the content of oleic acid in peanut seeds as an example:

[0011] 1. Materials used

[0012] Peanut material: The peanut varieties are Fenghua No. 1, Hehua No. 1 and 8130.

[0013] Strains and plasmids: Escherichia coli E.coli-DH5α, Agrobacterium LBA4404; cloning vectors pGEM-Teasy, pBI121 and pGLe-10.

[0014] Enzymes and chemical reagents: plant genome extraction kit; plasmid extraction kit, DNA restriction enzyme, T4DNA ligase, T4DNA polymerase, CIAP calf intestinal alkaline phosphorylase, pfu polymerase and DNA ladder; antibiotics ( Kanamycin, etc.) and other biochemical reagents.

[0015] 2. Technical methods

[0016] Cloning of FAD2-1 Gene Fragment and Construction of Expression Vector

[0017] Total DNA was prepared from peanut material and used as template for polymerase chain reaction (PCR). Two sets of PCR primers were designed with reference to the peanut FAD2-1 gene (AF272950) in GeneBank, an...

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Abstract

The invention belongs to plant breeding field, especially the biology engineering breeding to improve the quality of soybean and earthnut. It uses PCR method to clone the specific gene section from the gene of soybean and earthnut. According to gene silence principle, the gene section would be rebuilt into reverse repeating sequence structure, which would be built into the carrier that of promoter and terminator of the specific expression. The gene would be transferred, cultivated, and filtered by farm bacilli induce transformation. The invention could improve the quality of the soybean and the earthnut.

Description

technical field [0001] The invention belongs to the field of plant breeding, in particular to the field of bioengineering breeding for soybean and peanut quality improvement. The Δ12 fatty acid dehydrogenase gene FAD2-1 of soybean and peanut seeds was rebuilt by double-stranded RNA gene silencing technology, and the fatty acid content of soybean and peanut seeds was genetically improved by transgenic technology, and then the seeds with high oleic acid content were bred. Soy and peanut material. Background technique [0002] Double-stranded RNA gene silencing refers to a genetic phenomenon in which double-stranded RNA participates in the guidance of organisms, and targets exogenous and endogenous mRNAs for degradation, resulting in the inability to express specific genes. Double-stranded RNA gene silencing technology is currently the most effective and advanced technology for controlling gene expression, and is widely used in the research field of plant genetic engineering. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N15/82C12Q1/68A01H1/02A01H4/00
Inventor 许守民柳青刘立侠李桂民宋凯蔡一荣冯凯李望丰
Owner NORTHEAST NORMAL UNIVERSITY
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