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Primer for identifying purity of brassica rapa L. ssp. chinensis makino sujun 316 seeds and application

A technology for headed cabbage and seeds, which is applied to primer sequences and application fields for identifying the purity of non-headed cabbage Sujun 316 hybrid seeds, can solve problems such as the conflict between the identification time of seed purity and the sales cycle, and achieves an environment that is not affected by the environment. Effect

Active Publication Date: 2019-05-31
TIANJIN RES INST OF VEGETABLE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This variety has been using traditional field methods to identify seed purity, often causing conflicts between the timing of seed purity identification and the sales cycle

Method used

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  • Primer for identifying purity of brassica rapa L. ssp. chinensis makino sujun 316 seeds and application
  • Primer for identifying purity of brassica rapa L. ssp. chinensis makino sujun 316 seeds and application
  • Primer for identifying purity of brassica rapa L. ssp. chinensis makino sujun 316 seeds and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1. Screening of InDel primers for purity identification

[0027] According to the resequencing data of the Chinese cabbage genome, multiple pairs of primers were designed, polymorphism detection was performed on three individual plants of Sujun 316, the male parent and the female parent, and the primer pair A10-2 with co-dominant differences was screened out. The sequence is as follows :

[0028] F: 5'-ATG CTT GGA GAT GAG ACA TGG ATC AAG-3' (SEQ ID NO: 1)

[0029] R: 5'- TTG CTC ATA TCA CCA TCA TAC AGA TAG GG-3' (SEQ ID NO: 2)

[0030] The marker has good reproducibility and clear bands. Using this primer as a specific primer, the genomic DNA of the sample to be tested is used as a template for PCR amplification, and the amplified products are separated by electrophoresis and stained for detection. The results show that: A specific band of 133 bp can be amplified in the parent material, a specific band of 104 bp can be amplified in the maternal material, and a specifi...

Embodiment 2

[0042] Comparative test of traditional identification methods: the Sujun 316 commercial variety to be tested produced by 5 contracted farmers in the seed production base was randomly selected about 100 seeds from each household, and the open field was ditched, and the seeds were sown at a density of about 200 plants per square meter. About 3 days after sowing, all the seedlings will emerge, and when they grow to the stage of 7 leaves and 1 heart, according to the performance characteristics of the samples to be tested, such as plant height, leaf color, leaf shape, petiole color, length, and width, etc., whether they conform to the uniformity of Sujun 316 commercial varieties and consistency, judge whether the sample to be tested is Jinyan Express No. 1 commercial variety, and count the seed purity of the sample to be tested.

[0043]

[0044] Conclusion: the result of using the present invention to identify the seed purity of Sujun 316 non-heading Chinese cabbage hybrid is c...

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Abstract

The invention discloses an InDel molecular marker specific primer for detecting the purity of sujun 316 brassica rapa L. ssp. chinensis makino hybrid seeds. The primer is a nucleotide sequence shown in SEQ ID NO:1-2. The invention further discloses a method for identifying the purity of the sujun 316 brassica rapa L. ssp. chinensis makino hybrid seeds through the specific primer. According to thespecific primer and the method, the purity of the brassica rapa L. ssp. chinensis makino sujun 316 hybrid seeds can be identified in a laboratory within 3-5 hours, and the method has the advantages that the method is quick to use and stable and is not influenced by the environment. The method can replace a traditional field seed purity identification method and is beneficial to efficient and accurate qualify control over the sujun 316 brassica rapa L. ssp. chinensis makino hybrid seeds.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a primer sequence and application for identifying the seed purity of non-heading Chinese cabbage Sujun 316 hybrid. Background technique [0002] Non-heading cabbage ( Brassica rapa L. ssp. chinensis Makino) belongs to Brassicaceae Brassica crops, also known as Chinese cabbage, green vegetables, and rapeseed (northern China). An important national leafy green vegetable, and its planting area in Japan, Korea and other foreign countries is also increasing. [0003] The traditional method of identifying the purity of non-heading cabbage seeds is generally planted in the field, and after a certain period of growth, the growth characteristics are observed to identify the purity of the seeds, which has the disadvantages of long cycle time, large amount of labor, and susceptibility to environmental influences. With the development of molecular biology, molecular marker-assisted breeding ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
Inventor 黄志银王超楠李梅华德平张红范伟强张斌刘晓晖
Owner TIANJIN RES INST OF VEGETABLE
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