Molecular marker in close linkage with head cabbage waxless bright green gene cgl-4 and application of molecular marker

A technology of head cabbage and molecular markers, which is applied to the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of inability to apply assisted selection work, loose linkage relationship, low selection efficiency, etc., and achieve Improve breeding efficiency, reduce workload, and overcome the effect of long time period

Active Publication Date: 2016-12-07
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

Although Liu Dongming et al. (2014) have studied the molecular marker of the wax-free gene cgl-4 and obtained a molecular marker linked to the gene controlling the wax-free trait, but this marker is not related to the wax-free gene cgl- The genetic distance among the 4 is relatively far, 4.7cM, and the linkage relationship is not close. In the selection breeding work, it is prone to the problems of offspring selection error and low selection efficiency, which cannot be applied to the actual assisted selection work.

Method used

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  • Molecular marker in close linkage with head cabbage waxless bright green gene cgl-4 and application of molecular marker
  • Molecular marker in close linkage with head cabbage waxless bright green gene cgl-4 and application of molecular marker
  • Molecular marker in close linkage with head cabbage waxless bright green gene cgl-4 and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1, the acquisition of molecular marker BCYM000140 according to the present invention

[0042] (1) Construction of F2 generation segregation population

[0043] Using head cabbage non-wax powder mutant material LD10 and wax powder wild-type material 21-3 to prepare a hybrid combination, all individual plants of the F1 population showed the wax powder trait. The F1 generation was self-crossed to produce the F2 generation population, a total of 898 individuals, and the individual plant phenotype was identified, including 667 individual plants with wax powder and 231 individual plants without wax powder. The chi-square test verified that the segregation ratio of 3:1 .

[0044] (2) Extraction of head cabbage genomic DNA

[0045] Genomic DNA was extracted from leaves of parents, F1 and F2 populations by CTAB method. Take a size of about 1cm 2 Put the true leaves into a 2mL centrifuge tube, add 750μL CTAB lysis buffer, place the samples in a sampler for 5 minutes, ...

Embodiment 2

[0058] Embodiment 2, the assembly of kit of the present invention

[0059] Based on the molecular marker BCYM000140 obtained in Example 1, the kit described in the present invention was assembled. The kit includes upstream and downstream primers having sequences shown in Seq ID No.1 and Seq ID No.2 for amplifying the molecular marker BCYM000140.

[0060] The kit also includes conventional reagents for PCR reactions and / or electrophoresis. Specifically, the conventional reagents used in the PCR reaction include: 10×Buffer, dNTP, Taq enzyme, etc., and may also be common commercially available PCR kits, PCR reaction Mix, etc.

Embodiment 3

[0061] Example 3. Using the marker or kit of the present invention to carry out assisted selection breeding of bright green cabbage without wax powder

[0062] The wild-type 21-3 was crossed with the wax-free bright green genetic material LD10. Combined with molecular marker-assisted selection, the wax-free bright green gene cgl-4 of LD10 was introduced into 21-3 through multiple generations of backcrossing. The individual plants with LD10 band type in the progeny population were used for breeding improvement.

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Abstract

The invention provides a molecular marker in close linkage with head cabbage waxless bright green gene cgl-4 and application of the molecular marker. The molecular marker is characterized in that nucleotide sequences of a sense primer and a reverse primer of the molecular marker BCYM000140 are as shown in Seq ID No.1 and Seq ID No.2. The invention further provides a method and a kit for screening head cabbage genetic resources containing the head cabbage waxless bright green gene cgl-4 on the basis of the molecular marker. The marker or the kit has advantages of high specificity and accuracy and is capable of simply and quickly find out whether a tested material contains the waxless bright green gene cgl-4 or not in an early growth period of plants to make it convenient for targeted execution of subsequent breeding operations.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a molecular marker closely linked with cgl-4, a wax-free bright green gene of head cabbage, and an application thereof. Background technique [0002] Heading cabbage (Brassica oleracea var.capitata L.) belongs to Brassica genus Brassicaceae. It has the characteristics of rich nutrient content, wide adaptability, easy cultivation, and transportation resistance. It plays an important role in my country's annual vegetable supply and export earnings. Status, my country's annual cultivation area is about 900,000 hm 2 (Fang Zhiyuan, 2008; Yang Limei et al., 2011). Ball color green is a very important commercial trait of cabbage, and it is also one of the main breeding goals of breeders. Due to the white wax powder covering the surface of the cabbage plant, the leaves of the cabbage generally appear gray-green or dark green. The synthesis and transportation of wax powder on th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 杨丽梅刘东明
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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