Novel method for authenticating Pacific oyster population

Abstract

The invention relates to a method for authenticating oyster population, in particular to a novel method for authenticating Pacific oyster population, comprising the steps of sample collection, genome extraction, primer design, PCR amplification, and EST-SSRs specific site authentication. In the step of sample collection, sample individuals from different populations of ostrea cucullata, Pacific oyster and crassostrea rivularis are collected; in the step of genome extraction, genomes of a plurality of samples of each oyster population are extracted; in the step of primer design, EST-SSRs primers are designed according to an expressed sequence tag (EST) sequence containing microsatellites; in the step of PCR amplification, the EST-SSRs primers are used to carry out the PCR amplification to obtain the PCR product with a target segment; and in the step of EST-SSRs specific site authentication, agarose gel electrophoresis is firstly carried out on the PCR product to obtain the PCR product with clear and stable bands, and then polyacrylamide gelelectrophoresis is carried out to detect polymorphic sites so as to authenticate the allele Pacific oyster population having the allele sites with specific EST microsatellite sequence. In the invention, an EST-SSRs molecule marker technology is utilized, the primers are reasonably designed, and two-step electrophoresis detections are adopted so as to achieve the purpose of authenticating the Pacific oyster population. The invention has the advantages of simple method, time saving and accurate analysis.

Description

Technical field: [0001] The invention relates to a new method for quickly and accurately identifying oyster populations. Background technique: [0002] Oyster shells have strong plasticity, and the external shape of shells varies greatly. Researchers are actively seeking other research methods in order to solve difficult problems in oyster classification. Most scholars believe that the structural differences of oyster molluscs are very small, and the taxonomic evidences that can be provided are few. The vast majority of oysters have a constant number of chromosomes (2n=20), and the difference in karyotype is minimal, which cannot provide evidence for distinguishing oyster species (see Ahmed M. Cytogenetics of oysters. Cytologia, 1973, 28: 337-346. ). In terms of biochemical means, protein and isoenzyme electrophoresis techniques are used to study genetic diversity analysis among genera and species identification among populations with overlapping distribution areas, but th...

AI Technical Summary

Problems solved by technology

Therefore, due to the lack of effective methods for identifying populations, the species names of oysters have been confused for a long period of time, and the phenomena of homonyms and heteronyms are very serious.

[0003] In addition, the frequent introduction and breeding of off-site species broke the geographical isolation between oyster species, which had a serious impact on the germplasm resources of oysters in my country, and further aggravated the difficulty of the existing oyster morphological classification.

The confusion in classification has seriously affected the cultivation and breeding of oysters, and brought a lot of inconvenience to the development of cross breeding