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SNP molecular marker for detecting lobed-leaf trait of brassica campestris vegetables and applications thereof

A molecular marker, vegetable leaf technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of developmental period limitation and low efficiency

Active Publication Date: 2019-04-12
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The leaf margin cracking traits of Brassica vegetables can only be fully manifested when the true leaves are fully developed. The traditional method is limited by the developmental period and the efficiency is low.

Method used

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  • SNP molecular marker for detecting lobed-leaf trait of brassica campestris vegetables and applications thereof
  • SNP molecular marker for detecting lobed-leaf trait of brassica campestris vegetables and applications thereof
  • SNP molecular marker for detecting lobed-leaf trait of brassica campestris vegetables and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0182] Acquisition of molecular markers and their primers

[0183] 1. Obtain molecular markers:

[0184] The implementation basis of the present invention is: the inventor's research group resequenced a large number of Brassica vegetable varieties with different leaf margin phenotypes through the Illumina HiSeq2000 platform to obtain SNP variation sites between a large number of varieties.

[0185] The implementation content is: after the candidate genes are obtained through QTL analysis and fine mapping of the leaf margin cleavage phenotype, the well-typed SNP loci located on the candidate genes are used to analyze the SNP loci of the leaf margin cleavage traits, so The molecular marker of the invention can accurately detect the leaf type of the variety.

[0186] Step 1: Obtain the main effect QTL: In order to better and faster screen and obtain Brassica vegetable resources with leaf margin cracks, and promote the practice of molecular marker-assisted selection breeding, thr...

Embodiment 2

[0215] This example is to use the supporting reagents of UK LGC Company to detect the application of the A1015797093A / G site in the identification of leaf margin cracks and whole margin materials, that is, use the primers obtained in Example 1 to detect 73 materials in Example 1 leaf shape to verify the detection effect. The detection method of the present embodiment comprises the following steps:

[0216] 1. DNA extraction

[0217] Genomic DNA of 73 DH populations were extracted by conventional CTAB method.

[0218] Specifically:

[0219] The leaves of the above 73 varieties at the seedling stage were picked, frozen at -40°C for more than 1 day, and then placed in a vacuum dryer (CoolSafe 55-4) for dehydration; then 20 μg of dry powdered leaves were taken, and DNA was extracted according to the CTAB method. as follows;

[0220] i. Add CTAB extract (2%CTAB, 1.4mM NaCl, 100mM Tris‐HCl pH8.0, 20mM EDTA pH8.0, 1%PVP‐40, 0.2%β‐mercaptoethanol) in the dry powdery leaves, Mix w...

Embodiment 3

[0240] This example is to use the supporting reagents of the British LGC company to detect the application of the A1015797093A / G site in the identification of leaf margin cracks and whole margin materials, that is, use the primers obtained in Example 1 to detect other varieties of Brassica vegetables Leaf shape to verify the detection effect.

[0241] The varieties in this example are: Turnip varieties with leaf edge cracks: Zaosheng Kanamachi, Baili, Xinxingyuan, Hongyuan, Tsuda, Baiyu, Changhuang, purple top white globe, ECD‐01, ECD‐02, ECD‐ 03, ECD-04, a total of 12 copies, whole-edge brassica vegetable varieties: Xiacui, Chunquan, Xingdu Youfeng, Xiabao, Fuqiang, 536, Jiaoerye, Youxiawang, Xiazi, Eryue Bai, Improved Qingza No. 3, Jinqing 60, a total of 12 copies.

[0242] The operation of the detection method of this embodiment includes the following steps:.

[0243] 1. DNA extraction: the same operation steps and parameters as in Example 2.

[0244] 2. Based on competi...

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Abstract

The invention belongs to the field of biotechnology, and particularly relates to a SNP molecular marker for detecting lobed-leaf trait of brassica campestris vegetables and applications thereof. The SNP molecular marker is selected from: (a) the 358th locus of a template strand of Bra009510 gene, wherein the rna polymorphism is G or A; and (b) a homologous gene homologous to the Bra009510 in a brassica campestris vegetable variety with an integrifolious trait maintained, wherein the rna polymorphism corresponding to the 358th locus of SEQ ID NO.1 is G, and / or a homologous gene homologous to the Bra009510 in a brassica campestris variety with a lobed-leaf trait, wherein the rna polymorphism corresponding to the 358th locus of the SEQ ID NO.1 is A. The brassica campestris vegetable variety with an integrifolious trait maintained and / or the brassica campestris variety with a lobed-leaf trait are selected from: natural varieties, hybrid varieties and artificial mutagen varieties. The sequence of the template strand of the Bra009510 gene is as shown in the SEQ ID NO.1. The molecular marker provided by the invention and corresponding primers have a selection accuracy of 100% whether in lobed-leaf or integrifolious materials, and can be used for molecular marker-assisted selection breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a SNP molecular marker for detecting leaf margin cracking traits of brassica vegetables and an application thereof. Background technique [0002] Brassica vegetables include Chinese cabbage, turnip and pakchoi, which have the characteristics of various types, rich genetic resources, wide cultivation area, and short growth cycle. They are an important part of vegetables in my country. As an important leafy vegetable, leaf edge shape directly affects the commerciality of Brassica vegetables, and as market demand changes, studying the molecular mechanism of leaf edge shape and creating new germplasm with specific leaf shape of Brassica vegetables is a solution to the market. An effective means of diversifying demand for Brassica vegetable varieties. [0003] Leaf margin cleavage is a manifestation of plant adaptability to the environment. First of all, the moderate crackin...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 于拴仓李盼苏同兵张凤兰李佩荣张彬辛晓云余阳俊张德双赵岫云汪维红
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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