mSNP technology based mixed sample detection method for detecting purity of zucchini seeds

Abstract

The invention relates to an mSNP technology based mixed sample detection method for detecting the purity of zucchini seeds. The method is carried out by utilizing a primer pair 1F / R-20F / R, and the gene sequence of the primer pair 1F / R-20F / R is as shown in SEQ ID No.1-40. According to the invention, an mSNP technology is adopted, so that more SNP variations can be detected under the condition that amplicons are not changed; and a mixed sample method is adopted for detection, the sequencing cost is reduced while the amplification workload is reduced, the detection speed is also increased, and detection of 1440 seeds can be completed within one day at most. According to the method, the single nucleotide polymorphism is directly read by using a sequencing technology, and a purity result is directly interpreted through a program, so that the result is visual and reliable, and the influence of subjective factors during the seed development period and the result interpretation is avoided.

Description

technical field [0001] The invention belongs to the field of seed purity detection, and in particular relates to a mixed-sample detection method for detecting the purity of zucchini seeds based on mSNP technology. Background technique [0002] Zucchini (Cucubita pepo L.) is an annual plant of Cucurbitaceae squash, native to southern North America, so it is also called American squash. Zucchini has both nutritional value and health care function, and is loved by the public. It is planted in large areas all over our country and is one of the main varieties of vegetables in our country. Except for a small amount of imported varieties such as French winter jade and American jasper, other seeds on the market mainly rely on domestic cultivation, and the seed production areas are mainly concentrated in northwest my country and northern Shanxi Province. The two most important indicators of seed quality in seed production and trade are seed purity and authenticity. Using hybrid vig...

AI Technical Summary

Problems solved by technology

RFLP is a molecular marker technology invented in 1974. It has the advantages of good repeatability, co-dominance, and not affected by growth and development stages in seed identification; the disadvantage is that it does not provide sufficient information for pedigree analysis, and it is difficult to identify. It is difficult to operate in general detection rooms using isotopes, etc.

Compared with RFLP, RAPD has the advantages of requiring less DNA, simpler operation, and no need to use isotopes; the disadvantage is that dominant markers cannot distinguish heterozygous and homozygous types, and poor stability.

AFLP is a technology invented by Zabeau in the Netherlands in 1992. This technology can analyze crops with larger genomes, and its polymorphism is far more than RFLP and RAPD. The disadvantage is that it needs isotopes, etc.

SSR markers are co-dominant markers, rich in polymorphisms, and good in stability; however, primers must be designed for single-copy sequences at both ends of the microsatellites at each chromosomal locus, and the development of primers is difficult

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