High-oleic-acid-content peanut molecular marker, assistant selection back cross breeding method and application of back cross breeding method

A molecular marker and backcross breeding technology, applied in the field of agricultural bioengineering, can solve the problems of high cost of fluorescent labeling, unstable results, cumbersome methods, etc., and achieve the effect of high accuracy of genotype selection, stable results, and simple operation

Active Publication Date: 2013-12-25
HENAN ACAD OF AGRI SCI
View PDF2 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to use high oleic acid mutants as hybrid parents for molecular marker-assisted selection (MAS), Barkley et al. (2010) developed real-time quantitative PCR at the F435 mutation site method for high oleic acid genotyping; Chen et

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • High-oleic-acid-content peanut molecular marker, assistant selection back cross breeding method and application of back cross breeding method
  • High-oleic-acid-content peanut molecular marker, assistant selection back cross breeding method and application of back cross breeding method
  • High-oleic-acid-content peanut molecular marker, assistant selection back cross breeding method and application of back cross breeding method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1. High oleic acid peanut molecular marker

[0044] (1) Molecular marker primer sequence and band pattern

[0045] 1. ahFAD2B Site variant specific primers (to detect MITE insertions)

[0046] Design the forward primer MITE-INS-F at 63 bp of the MITE insertion sequence of the peanut FAD2 mutant gene, and design the reverse primer MITE-INS-R at the downstream 472 bp of the MITE insertion to detect the MITE insertion; if there is a site mutation, the expected band of 613 bp will be amplified; if there is no site variation, no band will be amplified, and the primers obtained are as follows:

[0047] Forward primer MITE-INS-F: 5’……GGATGATGGATTGTATGG…….3’

[0048] Reverse primer MITE-INS-R: 5'.....CTCTGACTATGCATCAG...3'.

[0049] 2. ahFAD2B Site-free specific primers (detection of wild-type without MITE insertion)

[0050] Design the forward primer WILD-F at -80 bp of the peanut FAD2 wild-type gene, and design the reverse primer WILD-R at 654 bp. If there ...

Embodiment 2

[0058] Example 2. Genotyping and Verification of Oleic Acid Content Using Molecular Markers

[0059] 1. Genotyping and oleic acid content of Yuanza 9102×wt09-0023 offspring

[0060] The DNA of the F3:4 individual offspring of the Yuanza 9102×wt09-0023 hybrid combination was extracted, and the above-mentioned designed ahFAD2B The two pairs of molecular marker primers at the locus were amplified by PCR respectively, and the band patterns of the amplification results were as follows: figure 1 , figure 2 (The three-digit strain number in the figure is Yuanza 9102×wt09-0023, and the four-digit strain number is Fuhua 12×wt09-0023). Combined with the sequencing results of FAD2A, the band type of FAD2B was compared with the genotype and oleic acid content (Table 2).

[0061] The results showed that under the AA background, the genotypes of the B genome changed from BB, Bb to bb, and the oleic acid content increased from 31.9%, 38.1% to 63.3%; The acid content increased from 3...

Embodiment 3

[0069] Example 3: Backcross Breeding Method of High Oleic Acid Peanut Using Molecular Markers , see image 3 , the method includes the following steps:

[0070] (1) Use parents with ordinary oleic acid content (oleic acid content 70%) were crossed to get F 1 ;

[0071] (2) Select offspring F 1 As the male parent, backcross with the peanut parent of genotype AABB to obtain the offspring BC 1 f 1 ;

[0072] (3) Extract peanut BC 1 f 1 For the DNA of a single plant, first use a pair of primers in FAD2A to amplify the FAD2A gene fragment, wherein the forward primer is aF19: GATTACTGATTATTGACTT, and the reverse primer is R3: CCCTGGTGGATTGTTCA,

[0073] Sequencing after amplification to select a single plant containing the Aa genotype;

[0074] Then use the peanut ahFAD2B The mutation-specific primers and non-variation-specific primers at the locus were used to perform PCR detection on the individual plants containing the Aa genotype, and the two markers were dominant,...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a high-oleic-acid-content molecular marker, a back cross breeding method for carrying out assistant selection by utilizing the molecular marker, and an application of the back cross breeding method. Sequences of variant specific primers of the molecular marker are as follows: a forward primer MITE-INS-F, and a reverse primer MITE-INS-R; the sequences of non-variant specific primers are as follows: a forward primer WILD-F, and a reverse primer WILD-R. The back cross breeding method utilizing the molecular maker comprises the following steps: carrying out hybridization on a peanut parent of a gene type AABB and a parent of a gene type aabb, then carrying out cross breeding; amplifying an FAD2A gene segment by utilizing one pair of primers in a genome, and selecting a single plant containing a genotype Aa; carrying out PCR (polymerase chain reaction) detection by utilizing a primer, at a site ahFAD2B, of a peanut as a marker, and selecting a single plant containing a genotype Bb; carrying out back crossing on the single plant which is dominant in two markers, namely the single plant containing the genotype AaBb, and the parent AABB until a high-oleic-acid-content single plant which is of a genotype aabb and is consistent with a recurrent parent in other economical characters is selected from later generations, and then carrying out self-crossing and amplifying propagation. The high-oleic-acid-content molecular marker has the advantages of being simple in method, low in cost and stable in result; when the high-oleic-acid-content molecular marker is used for carrying out the back cross breeding, the accuracy of character selection can be obviously improved, the breeding cost is reduced, and the breeding efficiency is improved.

Description

technical field [0001] The invention belongs to the field of agricultural bioengineering, and in particular relates to a molecular marker for high oleic acid, a backcross breeding method for assisting selection by using the marker and its application. Background technique [0002] Cultivated peanuts ( Arachis hypogaea ), also known as groundnut, Papilionaceae, is an annual herb. Peanuts are originally produced in South America, and their planting areas are mainly distributed in Asia, Africa and America, of which Asia accounts for about 60%, Africa accounts for about 30%, and America accounts for about 5.5%. my country is the world's largest peanut producer, consumer and exporter, and peanuts are grown in all provinces, municipalities and autonomous regions. Peanut is an important oil crop, and its fatty acid composition has a great influence on the nutritional quality and storage quality of peanut. Oleic acid and linoleic acid in peanut oil account for about 80% of the to...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/11A01H1/02A01H1/04C12Q1/68
Inventor 张新友齐飞艳黄冰艳刘华韩锁义汤丰收董文召徐静
Owner HENAN ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap