Peanut high-oleic-acid character gene selection method

A gene selection, high oleic acid technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as inability to analyze peanut germplasm

Inactive Publication Date: 2014-01-22
SHANDONG PEANUT RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to gain a deeper understanding of the genetic basis controlling peanut high oleic acid traits and improve the breeding efficiency of peanut varieties, breeders have developed some peanut varieties with high O / L ratio CAPS, Real-time PCR, AS-PCR (Allele-Specific PCR) and other marking methods, but these methods are limited to specific types of mutants and cannot analyze the rich and diverse peanut germplasm

Method used

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  • Peanut high-oleic-acid character gene selection method

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Effect test

Embodiment 1

[0018] For 3 peanut varieties (lines) Huayu 30, Luhua 12 and E11, take 1 piece of fresh peanut leaf (about 10 mg), put it in a 1.5mL EP tube, add liquid nitrogen, and quickly grind the leaf with a small plastic grinding rod into powder; add 400 μL extract (including Tris-HCl solution (PH8.0) with a concentration of 200mmol / L, sodium chloride solution with a concentration of 250mmol / L, EDTA solution with a concentration of 25mmol / L, and SDS solution with a concentration of 0.5%. Use after high-pressure steam sterilization. Before use, add 1% β-mercaptoethanol solution.), oscillate and mix, let stand for 10 minutes; add 200 μL 5mol / L potassium acetate solution (pH5.0 or not adjust pH), Shake and mix well, let stand for 15min; centrifuge at 12000r / min for 2min, take 400μL of the supernatant and transfer it to another 1.5mL EP tube, add an equal volume of isopropanol; mix gently and let stand at room temperature for 2 minutes, Centrifuge at 12000r / min for 5min, discard the superna...

Embodiment 2

[0020]For the three peanut varieties Huayu 19, Huayu 23 and Luhua 14, take 1 piece of fresh peanut leaf (about 10 mg), put it in a 1.5mL EP tube, add liquid nitrogen, and use a small plastic grinding rod to quickly grind the leaves into powder; add 400 μL extract solution (including Tris-HCl solution (PH8.0) with a concentration of 200mmol / L, sodium chloride solution with a concentration of 250mmol / L, EDTA solution with a concentration of 25mmol / L, and SDS solution with a concentration of 0.5%, and undergo high pressure Use after steam sterilization. Add 1% β-mercaptoethanol solution before use.), oscillate to mix, let stand for 10min; add 200μL 5mol / L potassium acetate solution (pH5.0 or not adjust pH), oscillate Mix well and let it stand for 15 minutes; centrifuge at 12000r / min for 2 minutes, take 400 μL of the supernatant and transfer it to another 1.5 mL EP tube, add an equal volume of isopropanol; mix gently and let stand at room temperature for 2 minutes. Centrifuge at 1...

Embodiment 3

[0022] For three peanut varieties (lines) E18, Huayu 32 and E16, take 1 piece of fresh peanut leaf (about 10 mg), put it in a 1.5mL EP tube, add liquid nitrogen, and use a small plastic grinding rod to quickly grind the leaves into powder; add 400 μL extract solution (including Tris-HCl solution (PH8.0) with a concentration of 200mmol / L, sodium chloride solution with a concentration of 250mmol / L, EDTA solution with a concentration of 25mmol / L, and SDS solution with a concentration of 0.5%, and undergo high pressure Use after steam sterilization. Add 1% β-mercaptoethanol solution before use.), oscillate to mix, let stand for 10min; add 200μL 5mol / L potassium acetate solution (pH5.0 or not adjust pH), oscillate Mix well and let it stand for 15 minutes; centrifuge at 12000r / min for 2 minutes, take 400 μL of the supernatant and transfer it to another 1.5 mL EP tube, add an equal volume of isopropanol; mix gently and let stand at room temperature for 2 minutes. Centrifuge at 12000r...

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Abstract

The invention discloses a peanut high-oleic-acid character gene selection method. The method comprises the steps of extracting leaf gene groups of different peanut breeds, and amplifying AhFAD2A and AhFAD2B genes from the gene groups by using a specificity primer; cutting and recycling specificity amplification stripes and sequencing to obtain the sequence of the gene; translating the obtained sequence, comparing and analyzing, and searching the point mutation or inserting site to judge the oleic acid content and specific value of oleic acid/linoleic acid (O/L) of the breed. The method is simple to operate, variety of peanuts germ plasm can be analyzed, the oleic acid content of single plant of peanut can be determined in the seedling stage, the breeding process can be shortened and the breeding effect of oleic acid can be improved by combining comprehensive agronomic character selection of the single plant.

Description

technical field [0001] The invention relates to a method for selecting genes for peanut high oleic acid traits, and belongs to the field of peanut biotechnology. Background technique [0002] my country is the world's largest peanut producer, consumer and exporter. Peanut has an important position and role in my country's national economy. Oleic acid is the main fatty acid in peanut oil, accounting for 39%-49% of the total peanut fatty acid. It can reduce harmful cholesterol in human lipid metabolism, but maintain the level of beneficial cholesterol, thereby slowing down atherosclerosis, effectively Prevent the occurrence of cardiovascular and cerebrovascular diseases such as coronary heart disease. In addition, the content of oleic acid is high, the anti-oxidation ability is strong, the product is not easy to oxidative rancidity, the shelf life is long, and the storage capacity is also strong. Therefore, the breeders of the world's major oil crops (soybean, rapeseed, pea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/13
Inventor 禹山林迟晓元杨庆利陈娜潘丽娟陈明娜王通王冕杨珍
Owner SHANDONG PEANUT RES INST
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