Molecular markers and QTL sites of free proline content of strong winter Brassia campestris L.

A technology of Chinese cabbage-type winter rape and free proline, which is applied in the field of molecular biology and genetic breeding, can solve the problems that the research on molecular markers and QTL positioning has not been reported, so as to speed up the breeding process, improve the screening efficiency and save production cost effect

Active Publication Date: 2015-11-04
GANSU AGRI UNIV
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, there have been many studies on the content of free proline in Chinese cabbage-type winter rapeseed, but there have been no reports on its molecular markers and QTL mapping.

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
  • Molecular markers and QTL sites of free proline content of strong winter Brassia campestris L.
  • Molecular markers and QTL sites of free proline content of strong winter Brassia campestris L.
  • Molecular markers and QTL sites of free proline content of strong winter Brassia campestris L.

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Construction of isolated populations of strong winter Chinese cabbage-type winter rapeseed and determination of free proline content.

[0038] The specific construction of the isolated population used in this embodiment is as follows:

[0039] (1) In August 2010, multi-generation bagging and self-crossing of the parent rapeseed 'Longyou 7' with strong winter resistance and strong winter resistance and rapeseed 'Longyou 9' with strong winter resistance were used to configure the hybrid combination, harvested in May 2011 F1 generation seeds.

[0040] (2) F1 seeds were sown in August 2011, and bagged and self-crossed in April 2012 to obtain F2 generation seeds.

[0041] The F2 generation seeds were sown in August 2012. At the seedling stage, 103 plants were randomly selected to be listed and marked, and fresh young leaves were collected in early November, brought back to the laboratory in an ice box and stored in a -70°C ultra-low temperature refrigerator. Th...

Embodiment 2

[0042] Example 2: Extraction of total DNA from leaves of parents and F2 generation segregation populations.

[0043] Total leaf DNA was extracted by the CTAB method, and the specific steps were as follows:

[0044] (1) Pick healthy young leaves as materials for extracting rapeseed genomic DNA, wash them with distilled water, and dry them with paper.

[0045] (2) Take 0.5g of fresh leaves and place them in a mortar, add liquid nitrogen, grind them quickly until they are whitish powder, and put them into a 2ml centrifuge tube immediately.

[0046](3) Add 700ul of preheated 2×CTAB extraction buffer to infiltrate the powder and invert the centrifuge tube to fully disperse the powder. Place in a water bath at 65°C for 60 minutes, during which time gently mix 2-3 times.

[0047] (4) Take out the centrifuge tube, cool to room temperature, add 700ul of chloroform / isoamyl alcohol (24 / 1), gently invert to mix, and let stand for 10 min.

[0048] (5) Centrifuge at 13000rpm for 10min. ...

Embodiment 3

[0056] Example 3: Source, synthesis and polymorphism screening of SSR and InDel primers.

[0057] (1) 315 pairs were evenly selected from the rapeseed microsatellite primer sequences published on www.UK crop.net and all primers on the 10 chromosomes published on the Brassica Database website http: / / brassicadb.org / . Among them, there are 51 pairs of SSR primers and 264 pairs of InDel primers. Primers were synthesized by Shanghai Bioengineering Technology Co., Ltd.

[0058] (2) Randomly select 6 strains of DNA from each parent and mix them in equal amounts, and use them as templates for screening primers.

[0059] (3) PCR reaction system

[0060] The PCR reaction system is 10ul:

[0061] Mix Taq Enzyme 6ul

[0062] Upper primer 0.5ul

[0063] Lower primer 0.5ul

[0064] ddH2O 2ul

[0065] Template: 1ul

[0066] (4) PCR amplification program: pre-denaturation at 94°C for 4 minutes; denaturation at 94°C for 50s, annealing at 55-60°C for 50s (annealing temperature decreased...

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 discloses molecular markers and QTL sites of free proline content of strong winter Brassia campestris L. Screening comprises the following steps: (1) constructing an F2-generation segregation population through hybridization of parents obtained through multi-generation bagged self-crossing, i.e., strong winter superstrong-cold-resistance winter rape Longyou No. 7 and strong-cold-resistance winter rape Longyou No. 9; (2) extracting leaf DNAs of the two parents and the F2-generation segregation population by using a CTAB method; (3) carrying out polymorphism primer screening on the DNAs of the two parents by using SSR and InDel primer pairs; (4) constructing a genetic linkage map according to obtained genotype data of the F2-generation segregation population and carrying out QTL analysis; and (5) naming the obtained three QTL sites of the free proline content of strong winter Brassia campestris L. as Qfre.gsau-2A, Qfre.gsau-3A and Qfre.gsau-6A, respectively, and acquiring the six molecular marks linked with the free proline content of strong winter Brassia campestris L., i.e., BrID10709, BrID101165, BrID90131, BrID10157, BrID10397 and 8C0419-1.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and genetic breeding, and specifically relates to a QTL site of a free proline content gene of strong winter Chinese cabbage type winter rapeseed, and also relates to a molecular marker linked with the free proline content gene. Background technique [0002] Brassica napus has a long history of cultivation in my country and is rich in germplasm resources. It is one of the important oil crops in my country. But the Chinese cabbage-type rape is mainly spring-type, and there is a lack of germplasm of strong-winter-type Chinese cabbage-type rapeseed, especially winter rapeseed varieties suitable for planting in the northern arid and cold regions. The climate in northern my country is very cold, and winter rapeseed has excellent cold resistance to survive the winter safely. Therefore, cold-resistant breeding is the key to the development of winter rapeseed in the north. Breeding cold-resista...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 李学才孙万仓杨刚刘自刚曾秀存武军艳方彦孔德晶陈奇王志江
Owner GANSU AGRI UNIV
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