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High-oleic acid peanut mutant gene AhFAD2B-814 and application thereof

An ahfad2b-814, mutated gene technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problem of peanut high oleic acid mutation gene that has not been published or published, and achieves increased genetic diversity and oleic acid content. , the effect of broadening the genetic basis

Inactive Publication Date: 2018-11-06
河北省农林科学院粮油作物研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, no peanut high oleic acid mutant gene and its application have been disclosed or published in China.

Method used

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  • High-oleic acid peanut mutant gene AhFAD2B-814 and application thereof
  • High-oleic acid peanut mutant gene AhFAD2B-814 and application thereof
  • High-oleic acid peanut mutant gene AhFAD2B-814 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1: Screening and Purification of High Oleic Acid Peanut Mutant Individual Plants

[0061] The collected 220 peanut germplasm resource materials and the created 150 germplasm were planted in the field, planted in rows according to the materials, sowed in a single seed, harvested after selfing, and harvested seeds in a single plant; in the second year, 3 seeds were selected for each material. A single plant was planted in a row in the field, and the self-inbred single plant was harvested as a single plant again, and the seeds were naturally air-dried and peeled off. Perten (Perten) DA7200 diode array near-infrared analyzer was used to detect the concentration of each individual plant seed. For the content of fatty acid, 9 rows with oleic acid content of each plant above 70% were screened out.

[0062] Utilize American Agilent 6890A gas chromatographic analyzer to further accurately detect the fatty acid composition and content of these strains, the steps are as fo...

Embodiment 2

[0072] Example 2: Cloning and sequencing of mutant nucleic acid sequence of high oleic acid peanut oleic acid dehydrogenase gene

[0073] (1) Identify and obtain candidate genes for high oleic traits in peanuts.

[0074] The biological pathway of plant fatty acid synthesis has been relatively clear. Studies have confirmed that there are two pathways for the biosynthesis of oleic acid: one uses stearic acid as a substrate to generate oleic acid under the catalysis of stearyl-CoA desaturase (SAD), which is the main pathway for oleic acid synthesis. The second way is to use palmi-enoic acid as a substrate to further extend the carbon chain under the action of fatty acyl-CoA to generate oleic acid. Part of the synthesized oleic acid is bound to the carbon frame of triglyceride to become the final storage oil, part of it is dehydrogenated under the action of oleic acid dehydrogenase to further form linoleic acid with double unsaturated bonds, and a small part is in the Eicosenoic...

Embodiment 3

[0093] Example 3: Expression and functional analysis of a novel mutant gene of high oleic acid peanut AhFAD2B in Saccharomyces cerevisiae

[0094] The above-mentioned peanut AhFAD2 gene belongs to the endoplasmic reticulum oleate dehydrogenase, and the first choice for its functional analysis is yeast, because yeast contains the electron transfer system and suitable membrane environment required by this type of dehydrogenase, and Saccharomyces cerevisiae Due to the lack of Δ12 (ω-6) and ω-3 fatty acid dehydrogenase, endogenous polyunsaturated fatty acids cannot be produced, and the intracellular fatty acid composition is simple, so it is the most ideal expression system for exogenous fatty acid dehydrogenase genes. INVScI / pYES2.0 is a commonly used Saccharomyces cerevisiae expression system for expressing exogenous fatty acid dehydrogenase.

[0095] According to the multiple cloning site of Saccharomyces cerevisiae expression vector pYES2.0, the restriction sites of KpnI and Eco...

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Abstract

The invention relates to a high-oleic acid peanut mutant gene AhFAD2B-814 and application thereof and belongs to the field of plant molecular breeding. A nucleotide sequence of the mutant gene AhFAD2B-814 is shown as SEQ ID NO.: 1 and consists of 1140 bases. The high-oleic acid peanut mutant gene AhFAD2B-814 and the application thereof provided by the invention have the benefits that a mutant nucleotide sequence of a peanut high-oleic acid mutant oleic acid dehydrogenase gene can be introduced into other peanut varieties or strains with common oleic acid contents by applying conventional breeding methods such as hybridization, backcrossing and the like, the oleic acid content of a target variety or strain introduced with the mutant nucleotide sequence of the gene AhFAD2B-814 is improved, the genetic diversity of a high-oleic acid peanut variety (strain) is significantly increased, and the genetic basis of a high-oleic acid peanut is widened; during high-oleic acid peanut breeding, by selecting a peanut material carrying an AhFAD2A mutant genotype as a parent, the selection on high-oleic acid characters of a breeding offspring is more purposeful, and the occurrence frequency of target traits is improved.

Description

technical field [0001] The invention belongs to the field of plant molecular breeding, and specifically relates to a nucleic acid sequence of a mutant gene encoding oleic acid dehydrogenase in high-oleic peanuts and capable of increasing the oleic acid content of peanuts and its application. Background technique [0002] Peanut is an important oil and economic crop in China. Peanut oil is a traditional edible oil in China and the main source of domestic edible vegetable oil. Peanut oil mainly contains palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1), linoleic acid (C18:2), behenic acid (C22:0) and arachidic acid ( C20:0) and other fatty acids. From the perspective of the composition of various fatty acids, unsaturated fatty acids account for about 80% (of which oleic acid is 36%-67%, linoleic acid is 15%-43%), and saturated fatty acids account for 20% (of which palmitic acid is 6%-11%) , stearic acid 2%-6%, arachidic acid 5%-7%, behenic acid 2%-3%). The com...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/02C12N15/11C12Q1/6895
CPCC12N9/001C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 陈四龙李玉荣刘义杰王瑾程增书宋亚辉张朋娟
Owner 河北省农林科学院粮油作物研究所
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