SSR molecular marker primers interlinked with cabbage yellow testa gene Brsc-ye and application of primers

A molecular marker, cabbage technology, applied in the fields of vegetable breeding and molecular genetics, can solve the problems of time-consuming and laborious, delaying the breeding process, etc., and achieve the effects of improving the selection efficiency, speeding up the breeding process, and convenient detection.

Inactive Publication Date: 2015-05-27
NORTHWEST A & F UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

However, the color of the cabbage seed coat needs to be bloomed and hardened and the seeds mature, and it needs to be observed plant by plant in the field, which is time-consuming and laborious to operate, which greatly delays the breeding process

Method used

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  • SSR molecular marker primers interlinked with cabbage yellow testa gene Brsc-ye and application of primers
  • SSR molecular marker primers interlinked with cabbage yellow testa gene Brsc-ye and application of primers
  • SSR molecular marker primers interlinked with cabbage yellow testa gene Brsc-ye and application of primers

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Experimental program
Comparison scheme
Effect test

Embodiment

[0090] Embodiment: the application of SSR309 primer

[0091] (1) Using CTAB method to extract Chinese cabbage genomic DNA, the extraction steps are as follows:

[0092] A. Take 0.2g of fresh young leaves with the main vein removed, stuff them into a 2mL centrifuge tube with steel balls (the steel balls need to be cleaned by 75% alcohol by mass fraction), put them into liquid nitrogen for quick freezing, and use a tissue grinder ground into powder;

[0093] B. Add 700 μl of CTAB extract (CTAB: 2%, Tris-HCl (pH8.0): 100mmol / L, EDTA: 20mmol / L, NaCl: 1.4mol / L) preheated at 65°C to the centrifuge tube, Then add 8 μl of β-mercaptoethanol and mix quickly;

[0094] C. Then put the centrifuge tube into a water bath at 65°C, shake it every 5-10 minutes, and put it in the water bath for 45 minutes;

[0095] D. Take out the centrifuge tube, add an equal volume of a mixture of phenol, chloroform and isoamyl alcohol (phenol: chloroform: isoamyl alcohol = 25:24:1), shake well for 15 minut...

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Abstract

The invention discloses SSR molecular marker primers interlinked with a cabbage yellow testa gene Brsc-ye and an application of the primers. The marker primer is prepared by the following steps: with genome DNA of yellow-seed cabbages and ordinary brown-seed cabbage as well as a Fe segregation group thereof as a template, carrying out polymorphic primer screening on a yellow-seed cabbage parent and a brown-seed cabbage parent by using 520 SSR primer pairs, subsequently analyzing single plant of the F2 group, screening molecular markers interlinked with the cabbage yellow testa gene Brsc-ye so as to successfully obtain 7 pairs of SSR marker primers interlinked with Brsc-ye gene, and building a molecular genetic map of the cabbage yellow testa gene Brsc-ye. The interlink analysis discovers that the genetic distances of two SSR markers which are in close linkage with two sides of the Brsc-ye gene are 0.17cM and 0.29cM respectively; the molecular markers have the advantages of convenience in detection, stability in amplification and high in repeatability and accuracy; the reference is provided for molecular-assisted selective breeding of the yellow testa characteristics of cabbage and rape; the seed breeding process is accelerated.

Description

technical field [0001] The invention belongs to the fields of vegetable breeding and molecular genetics, and specifically relates to the development and application of SSR molecular marker primers linked with cabbage yellow seed coat gene Brsc-ye. Background technique [0002] Chinese cabbage (Brassica rapa L.) belongs to Brassicaceae Brassica Brassica subspecies. It is one of the main vegetable crops originated in my country and one of the three major rapeseeds in the world. It is closely related to people's daily life. , with the improvement of people's living standards, people's requirements for oil quality are increasing day by day. Therefore, the breeding of yellow seed coat Chinese cabbage has been paid more and more attention by domestic and foreign scholars and breeders. [0003] Studies have shown that rapeseeds with yellow seed coats have thin seed coats, so they have high oil content. Not only that, the oil produced by yellow seed coat rapeseeds has low crude fibe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 张鲁刚任延靖孙敬贤吴俊清赵静
Owner NORTHWEST A & F UNIV
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