Molecular marker for identifying hereditary sex of haliotis diversicolor and application of molecular marker

A technology of molecular markers and variegated abalone, which is applied in the field of molecular markers to identify the genetic sex of variegated abalone, can solve the problems that the gender molecular markers of variegated abalone have not been reported, and achieve a wide range of applicable materials, improve breeding benefits, and detect short cycle effect

CN114231608APending Publication Date: 2022-03-25JIMEI UNIV
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Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2022-03-25

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Abstract

The invention discloses a molecular marker for identifying hereditary sex of haliotis diversicolor and application of the molecular marker. The molecular marker shows insertion / deletion length polymorphism of a nucleotide sequence as shown in SEQ ID NO: 1. One of a pair of homologous chromosomes of a haliotis diversicolor individual is lack of the SEQ ID NO: 1 nucleotide sequence, and the genetic sex is male; a pair of homologous chromosomes of a haliotis diversicolor individual do not lack the SEQ ID NO: 1 nucleotide sequence, and the genetic sex is female. During detection, PCR amplification is performed by adopting SEQ ID NO: 2 and SEQ ID NO: 3 as primers, and when an amplification product is two fragments, the lengths of the two fragments are 266bp and 243bp respectively, the genetic sex of the abalone to be detected is male; when the amplification product only has one fragment and the length is 266bp, the genetic sex of the abalone to be detected is female. The method is suitable for both before and after gonad differentiation of haliotis diversicolor.
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Description

technical field

[0001] The invention relates to shellfish genetic sex identification and sex control technology in the technical field of aquatic organisms, in particular to a molecular marker for identifying the genetic sex of variegated abalone and its application. Background technique

[0002] The sex of aquatic organisms can be determined by genetic or environmental factors. Even species with genetically determined sex are prone to sexual reversal. Therefore, the sex of genetically determined aquatic organisms includes two levels of genetic sex and biological sex. Genetic sex is the genetic composition that determines the direction of sex, and biological sex is the sex that is actually expressed in gonads. In aquaculture and sex-controlled breeding, there are often cases where the genetic sex of the research object is inconsistent with the biological sex, and it is necessary to identify the genetic sex of the research object.

[0003] Economic traits in aquatic organi...

Examples

Embodiment 1

[0035] The applicant of the present invention took the variegated abalone collected by Jinjiang Fuda Abalone Aquatic Products Co., Ltd. in Jinjiang, Fujian, China as samples, and constructed the genome resequencing data of variegated abalone, with 11 groups of male and female, analyzed the resequencing data, and screened out the variegated abalone. The sexually dimorphic insertion-deletion site (Indel) of Sebao (that is, the site where all females are homozygous and all males are heterozygous). The results showed that the most sexually dimorphic loci were found on chromosome 3 and chromosome 9 (see Table 1).

[0036] Table 1 Chromosome distribution results of sexually dimorphic InDel in variegated abalone

[0037] chromosome number Number (percentage) 2 1(4.8％) 3 8(38.1％) 4 2(9.5％) 9 6(28.6％) 12 3(14.3％) 13 1(4.8％) Total 21

[0038]Further detection of sexually dimorphic InDel on chromosomes 3 and 9 revealed a 23bp-specific ...

Embodiment 2

[0040] Using the primer pair SC9 (SEQ ID NO: 2 and SEQ ID NO: 3) described in the present invention on the genomic DNA of the variegated abalone (from Jinjiang Fuda Abalone Aquatic Products Co., Ltd. in Jinjiang, Fujian, China) whose sex has been determined through the detection of gonads PCR amplification is carried out, and the accuracy and specificity of the method of the present invention are verified by comparing the agarose electrophoresis patterns. The specific method steps are as follows:

[0041] Take 24 variegated abalone females and 24 males to check the gonads to determine the gender, cut out the muscle tissue, put it in 100% ethanol solution and fix it, store it at -20°C, and use it for genome extraction; use phenol-chloroform method to extract the genome DNA, the resulting genomic DNA was uniformly diluted to 30ng / μL as a PCR template.

[0042] Primer F: 5'-TACAAGTGTTCCAGTCGTGA-3', SEQ ID NO: 2

[0043] Primer R: 5'-CACCTGTAGTTCAACACTCAG-3', SEQ ID NO: 3

[00...