A primer for detecting the purity of Jinxia No. 3 Chinese cabbage seeds and its application

A technology of Chinese cabbage and seeds, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc. It can solve the problems of being easily affected by the environment, long cycle time, and labor intensive, etc., and achieve efficient and accurate quality control Effect

Active Publication Date: 2022-04-19
TIANJIN RES INST OF VEGETABLE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional method of identifying the purity of Chinese cabbage seeds generally adopts direct cultivation in the field. In a specific development period, the purity of the seeds is judged by the specificity and consistency of the traits.
Some Chinese cabbage varieties need to judge the purity of the seeds according to the characteristics of the balls, which often causes conflicts between the identification time of the seed purity and the peak season of seed sales, resulting in unnecessary losses

Method used

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  • A primer for detecting the purity of Jinxia No. 3 Chinese cabbage seeds and its application
  • A primer for detecting the purity of Jinxia No. 3 Chinese cabbage seeds and its application
  • A primer for detecting the purity of Jinxia No. 3 Chinese cabbage seeds and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Screening of InDel-labeled primers for purity identification

[0023] According to the resequencing data of the Chinese cabbage genome, multiple pairs of primers were designed to detect the polymorphism of Jinxia 3, the male parent and the female parent, and the primer pair A04-16 with co-dominant differences was screened out. The sequence is as follows:

[0024] F: 5'- CAG CTT TAC CTT GAA ACA TTT CCT CAA GTA G-3' (SEQ ID NO:1)

[0025] R: 5'-GAG ATC TAA CTT TGC ATT GTT GTG CTT TGG-3' (SEQ ID NO:2)

[0026] 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 155 bp can be produced in the parent material, a specific band of 188 bp can be produced in the maternal material, and a specific band of 188 bp and 155 bp c...

Embodiment 2

[0036] Comparative test of traditional field identification methods: the Jinxia No. 3 commercial variety to be tested produced by 5 contracted farmers in the seed production base was randomly selected from each household, and a single seed was sown in a plug for seedling cultivation. When it grew to 5 leaves and 1 heart, Customized to the field, the density is controlled at a row spacing of about 40 cm, and a plant spacing of about 35 cm. After about 35 days, when the ball formation rate reaches 90%, according to the performance traits of the sample to be tested, such as leaf color, leaf shape, plant height and ball wrapping, whether Comply with the uniformity and consistency of the Jinxia No. 3 commercial variety, judge whether it is the Jinxia No. 3 commercial variety, and count the purity of the seeds;

[0037]

[0038] Conclusion: By comparing the identification results of InDel molecular markers with the traditional field identification results, it is found that the ide...

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Abstract

The invention discloses an InDel marker-specific primer for detecting the purity of Jinxia No. 3 summer-sown Chinese cabbage hybrid, which is the nucleotide sequence shown in SEQ ID NO: 1‑2. The invention further discloses a method for identifying the purity of Jinxia 3 Chinese cabbage hybrid seed by using InDel marker primers. The specific primers and methods provided by the present invention can complete the identification of the seed purity of Jinxia No. 3 Chinese cabbage hybrid in 3-5 hours in the laboratory, and have the advantages of being fast, stable, and not affected by the environment, and can replace traditional field seed purity. The identification method is beneficial to the further popularization of summer-sown Chinese cabbage variety Jinxia 3 and the quality control of seeds.

Description

technical field [0001] The invention relates to the InDel molecular marker primer sequence and application for the purity identification of Chinese cabbage Jinxia No. 3 summer-sown hybrid seeds, and belongs to the field of biotechnology. Background technique [0002] Chinese cabbage ( Brassica rapa L. ssp. pekinensis ) belongs to Brassicaceae Brassica crops, and has been one of the vegetables with the largest planting area and output in my country for many years. With the improvement of people's living standards and changes in eating habits, the quality requirements for Chinese cabbage are getting higher and higher, and it is required that Chinese cabbage can be supplied throughout the year. Excellent varieties are the key to ensure the quality of Chinese cabbage, and ensuring the purity of seeds of excellent varieties is very important in the production of Chinese cabbage. [0003] The traditional method of identifying the purity of Chinese cabbage seeds generally adop...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 华德平王超楠黄志银张红范伟强李梅张斌刘晓晖
Owner TIANJIN RES INST OF VEGETABLE
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