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Method for screening fertility-related molecular markers of recessive nucleic sterile line of non-heading Chinese cabbages through SNP technology and SSR technology together and application

A related molecular marker, recessive nuclear sterility technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., to achieve the effect of saving investment and shortening breeding years

Active Publication Date: 2016-03-30
WUHAN VEGETABLE RES INST
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing research results are mainly used in Chinese cabbage breeding, and there are few reports on the development of molecular markers for recessive sterility materials in Chinese cabbage.

Method used

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  • Method for screening fertility-related molecular markers of recessive nucleic sterile line of non-heading Chinese cabbages through SNP technology and SSR technology together and application
  • Method for screening fertility-related molecular markers of recessive nucleic sterile line of non-heading Chinese cabbages through SNP technology and SSR technology together and application
  • Method for screening fertility-related molecular markers of recessive nucleic sterile line of non-heading Chinese cabbages through SNP technology and SSR technology together and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Molecular marker screening of non-heading Chinese cabbage recessive male sterile line fertility gene linkage

[0056] 1. DNA extraction

[0057] The numbers of each individual plant in the F2 generation segregation population were listed, and 0.5g of fresh leaves were selected at the seedling stage, packed in a 2.0ml centrifuge tube, and stored at -20°C or -70°C for later use. Genomic DNA was extracted using the CTAB method (Luetal, 2004). The specific steps are: pour the leaves into a mortar, add 700-800 μl of 2% CTAB buffer solution (2% CTAB; EDTA: 20mmol / L; pH=8.0Tris-HCl: 100mmol / L; NaCl: 1.4mol / L; PVP1%), ground into a homogenous slurry. Water bath at 65°C for 60 minutes, with gentle shaking every 15 minutes. Leave it at room temperature for 10 min, add an equal volume of chloroform:isoamyl alcohol (24:1), shake gently, and mix well. Centrifuge at 12,000r / min for 15min, take it out gently, pipette the supernatant into a new 2ml centrifuge tube, add 1...

Embodiment 2

[0099] Example 2: Using molecular markers linked to the fertility gene of Banana A of non-heading Chinese cabbage to assist in the selection of excellent sterile lines and transfection, the steps are as follows:

[0100] a) The non-heading Chinese cabbage recessive male sterile line Bianjiaohuang A was used as the female parent, and the fertile inbred line of non-heading Chinese cabbage with excellent agronomic traits was selected as the male parent, and the F1 generation was produced by crossing.

[0101] b) The F1 generation is used as the female parent, and the above-mentioned excellent fertile inbred line is used as the male parent to backcross to produce the BC1F1 generation.

[0102] c) Plant BC1F1 generation plants, take samples from individual plants, extract DNA samples from individual plants, use labeled SSR60-1, SSR60-2 and SSR8-2 primers to perform single-plant PCR amplification, 6% polyacrylamide gel electrophoresis, and silver staining After developing, the ampli...

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Abstract

The invention relates to a method for screening fertility-related molecular markers of a recessive nucleic sterile line of non-heading Chinese cabbages through the SNP technology and the SSR technology together and application. An F2 generation segregation population is obtained through dwarf hybridization of a yellow dwarf Chinese cabbage A sterile line and He B and serves as a researching object, series-and-fertility-linked SNP markers capable of being subjected to electrophoresis detection are obtained through conversion via the SNP chip technology, and SSR molecular markers linked with fertility genes are developed through chip SNP marker information. A single plant genetype in a sterile line transferred descendant group can be rapidly identified through the obtained SSR molecular markers, convenience is provided for rapid breeding of the sterile line and hybrid seed production in a later stage, and meanwhile a foundation is laid for fine localization and clone of genes related to recessive cell nucleus male sterility of the non-heading Chinese cabbages.

Description

technical field [0001] The invention belongs to the field of non-heading Chinese cabbage breeding and biotechnology, and specifically relates to a method and application of a method for jointly screening non-heading Chinese cabbage recessive male sterile line fertility-related molecular markers using SNP chip technology and SSR marker technology. Background technique [0002] Non-heading cabbage (Brassicacampestrisssp. Chinensis Makino), also known as Chinese cabbage, green vegetables, and feather vegetables, is widely cultivated in East Asia and is an important leafy vegetable. Non-heading Chinese cabbage belongs to Brassicaceae Brassica subspecies, and is a typical cross-pollination crop with obvious heterosis (Deng Xiaohui, 2007). At present, the use of male sterile lines for seed production is an effective way to utilize plant heterosis. Therefore, the research on the selection and application of male sterile lines in Chinese cabbage has received great attention from bre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6827C12Q1/6895C12Q2600/156
Inventor 李霞周国林汪爱华胡侦华黄兴学王斌才邓耀华张润花
Owner WUHAN VEGETABLE RES INST
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