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Rapid seed selection kit for high-oleic acid peanuts

A high oleic acid kit technology, applied in the field of high oleic peanut rapid selection kits, can solve the problems of large error in results and high cost, and achieve the effects of low cost, reduced detection cost, and reduced workload

Inactive Publication Date: 2019-02-01
SHANDONG PEANUT RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the above-mentioned shortcomings of the prior art, the purpose of the present invention is to provide a high-oleic acid peanut rapid selection kit, which is used to solve the problems of large result errors and high cost in the prior art

Method used

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  • Rapid seed selection kit for high-oleic acid peanuts
  • Rapid seed selection kit for high-oleic acid peanuts
  • Rapid seed selection kit for high-oleic acid peanuts

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1. Prepare the sample material of peanuts to be tested

[0044] 1.1 Design primers to detect different genotypes of FAD2A and FAD2B

[0045] FAD2A mutant genotype primer pair, including the forward primer FAD2A-A of the mutant FAD2A gene sequence, the 5' end of the primer is marked with FAM, the forward primer sequence is FAD2A-A: 5'-FAM-GTTTTGGGACAACACTTCTTT-3', reverse Primer FAD2-R, the 5' end of the primer is labeled with biotin (biotin), the sequence of the reverse primer is FAD2-R: 5'-biotin-CTCTGACTATGCATCAGAACTTGT-3', and the length of the product is 557bp;

[0046] FAD2B mutant genotype primer pair, including the forward primer FAD2B-A of the mutant FAD2B gene sequence, the 5' end of the primer is marked with VIC, the forward primer sequence is FAD2B-A: 5'-VIC-AACACTTGGYCGCGGTTT-3', reverse Primer FAD2-R, the 5' end of the primer is labeled with biotin (biotin), the sequence of the reverse primer is FAD2-R: 5'-biotin-CTCTGACTATGCATCAGAACTTGT-3', and the length...

Embodiment 2

[0066] 1. Prepare the sample material of peanuts to be tested

[0067] 1.1 Design primers to detect different genotypes of FAD2A and FAD2B

[0068] FAD2A mutant genotype primer pair, including the forward primer FAD2A-A of the mutant FAD2A gene sequence, the 5' end of the primer is marked with FAM, the forward primer sequence is FAD2A-A: 5'-FAM-GTTTTGGGACAACACTTCTTT-3', reverse Primer FAD2-R, the 5' end of the primer is labeled with biotin (biotin), the sequence of the reverse primer is FAD2-R: 5'-biotin-CTCTGACTATGCATCAGAACTTGT-3', and the length of the product is 557bp;

[0069] FAD2B mutant genotype primer pair, including the forward primer FAD2B-A of the mutant FAD2B gene sequence, the 5' end of the primer is marked with VIC, the forward primer sequence is FAD2B-A: 5'-VIC-AACACTTGGYCGCGGTTT-3', reverse Primer FAD2-R, the 5' end of the primer is labeled with biotin (biotin), the sequence of the reverse primer is FAD2-R: 5'-biotin-CTCTGACTATGCATCAGAACTTGT-3', and the length...

Embodiment 3

[0090] 1. Prepare the sample material of peanuts to be tested

[0091] 1.1 Design primers to detect different genotypes of FAD2A and FAD2B

[0092] FAD2A mutant genotype primer pair, including the forward primer FAD2A-A of the mutant FAD2A gene sequence, the 5' end of the primer is marked with FAM, the forward primer sequence is FAD2A-A: 5'-FAM-GTTTTGGGACAACACTTCTTT-3', reverse Primer FAD2-R, the 5' end of the primer is labeled with biotin (biotin), the sequence of the reverse primer is FAD2-R: 5'-biotin-CTCTGACTATGCATCAGAACTTGT-3', and the length of the product is 557bp;

[0093] FAD2B mutant genotype primer pair, including the forward primer FAD2B-A of the mutant FAD2B gene sequence, the 5' end of the primer is marked with VIC, the forward primer sequence is FAD2B-A: 5'-VIC-AACACTTGGYCGCGGTTT-3', reverse Primer FAD2-R, the 5' end of the primer is labeled with biotin (biotin), the sequence of the reverse primer is FAD2-R: 5'-biotin-CTCTGACTATGCATCAGAACTTGT-3', and the length o...

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Abstract

The invention provides a rapid seed selection kit for high-oleic acid peanuts. The research results show that fatty acid dehydrogenase gene (FAD2) is the key gene for controlling the oleic acid traitof peanuts, wherein FAD2 is co-encoded by two non-alleles FAD2A and FAD2B located on different genomes. According to the present invention, based on the genetic gene, multiplex amplification primers respectively for wild and mutant FAD2A gene and FAD2B gene markers are designed by using a colloidal gold test paper strip as a carrier according to the gene sequence conserved region in a peanut high-oleic-acid mutant gene so as to optimize the primer combinations and the reaction system; by using the principle of nucleic acid hybridization, the labeled amplification products are bound with the specific antibody on the colloidal gold test paper strip so as to establish the PCR amplification and colloidal gold lateral flow immunoassay hybridization kit for detecting FAD2A gene and FAD2B gene; and the kit has characteristics of simple operation, strong specificity and high detection sensitivity.

Description

technical field [0001] The invention relates to the field of peanut seed breeding screening, in particular to a high-oleic acid peanut rapid seed selection kit. Background technique [0002] my country is the world's largest peanut producer, consumer and exporter. Peanuts play an important role in my country's national economy. Oleic acid is the main fatty acid in peanut oil, accounting for 39% to 49% of the total peanut fatty acid. It can reduce harmful cholesterol in human body 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 resistance is also strong. Therefore, the breeders of the world's main oil crops (soybean, rapeseed, peanut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895
CPCC12Q1/6895C12Q2600/13C12Q2600/16C12Q2600/156
Inventor 迟晓元陈娜陈明娜潘丽娟王冕王通杨珍禹山林
Owner SHANDONG PEANUT RES INST
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