Two microsatellite markers related to rapid growth of pseudosciaena crocea, and preparation methods thereof

A technology of microsatellite markers and large yellow croaker, applied in biochemical equipment and methods, DNA preparation, microbial measurement/inspection, etc., can solve the problems of lack of systematic research, shorten the breeding process and improve the efficiency of breeding

Inactive Publication Date: 2013-07-31
JIMEI UNIV
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

The above studies are all based on a group of studies, and there is no systematic research on its generality in other groups

Method used

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  • Two microsatellite markers related to rapid growth of pseudosciaena crocea, and preparation methods thereof
  • Two microsatellite markers related to rapid growth of pseudosciaena crocea, and preparation methods thereof
  • Two microsatellite markers related to rapid growth of pseudosciaena crocea, and preparation methods thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: Screening method for fast and long microsatellite markers

[0016] The large yellow croaker material used is a full-sibling family (referred to as FS) bred by artificial insemination in an experimental base in Fujian Province. 208 large yellow croakers were collected from FS, and the heaviest and lightest large yellow croakers were selected based on body weight. There were 30 large yellow croakers, and the body length, height and weight of each large yellow croaker were measured. Part of the caudal fin was cut and fixed in 95% ethanol for DNA extraction.

[0017] DNA extraction and detection Use conventional phenol / chloroform / isoamyl alcohol method to extract DNA, UV spectrophotometer to measure OD 260 with OD 280 , determine the quality of the DNA and adjust the DNA concentration to 50ng / μL, and store it at -20°C for later use.

[0018] PCR and detection PCR reaction system: the total reaction volume is 20μL, including 1.0μL (50ng) template DNA, 2.0μL 10×...

Embodiment 2

[0030] Example 2: Preliminary verification of microsatellite markers

[0031] In order to verify the effectiveness of the screening markers, another 60 families were selected from the above-mentioned families for fast and long marker screening for verification analysis, and the experimental procedure was as described in Example 1. This time, only the relevant sites in the preliminary screening were analyzed, namely LYC0088, LYC0143 and LYC0200, and the primer sequences used were SEQUENCE LISTING1-SEQUENCELISTING6 in the sequence list. It has been verified that there are two loci related to growth traits, namely loci LYC0088 and LYC0143. These two loci are significantly correlated with the length, height and weight of large yellow croaker (P<0.01). The fast-growing markers of large yellow croaker are preliminary The verification results are shown in Table 2.

[0032] Table 2

[0033]

Embodiment 3

[0034] Example 3: Revalidation of microsatellite markers

[0035] The 2 microsatellite markers screened were used to genotype the largest 30 fish and the smallest 30 fish in another large yellow croaker population (the progeny of mixed random mating of 4 females and 4 males), and a total of 60 individuals were combined. The phenotypic data of the three traits of body length, body height, and body weight of each individual were tested for significance, and the results of the re-verification of the fast-length marker of large yellow croaker can be seen in Table 3.

[0036] table 3

[0037]

[0038] It can be seen from the test results that LYC0088 and LYC0143 are still closely related to growth traits, among which LYC0088 has a very significant correlation with body height and weight (P<0.01), and a significant correlation with body length (P<0.05), and LYC0143 has a significant correlation with body length and body weight. The correlation between body weight was significant...

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Abstract

The invention provides two microsatellite markers related to rapid growth of pseudosciaena crocea, and preparation methods thereof, and relates to molecular breeding of the pseudosciaena crocea. The two microsatellite markers related to the rapid growth of the pseudosciaena crocea are LYC0088 and LYC0143. Primer sequences of the LYC0088 are shown as SEQUENCE LISTING 1 and SEQUENCE LISTING 2; and primer sequences of the LYC0143 are shown as SEQUENCE LISTING 3 and SEQUENCE LISTING 4. The preparation methods comprise the steps of 1) screening microsatellite markers related to the rapid growth; 2) preliminary verifying the microsatellite markers related to the rapid growth; and 3) verifying again. The two microsatellite markers related to the rapid growth of the pseudosciaena crocea can be applied in selective breeding of body length, body height and body mass of the pseudosciaena crocea and in screening of the microsatellite markers related to the rapid growth of the pseudosciaena crocea.

Description

technical field [0001] The invention relates to molecular breeding of large yellow croaker, in particular to two microsatellite markers related to the rapid growth of large yellow croaker and a preparation method thereof. Background technique [0002] Large yellow croaker is an important marine cultured fish in my country. Since the breakthrough of artificial breeding in 1987, the scale and scope of large yellow croaker farming has continued to expand. By 2010, it has formed an industry with an annual output of 86,000 tons and an output value of about 6.5 billion yuan. However, the following problems such as germplasm degradation, slow growth, and frequent disease are becoming more and more serious. Therefore, genetic improvement of large yellow croaker is urgently needed. With the support of various projects, Jimei University, Ningde Fishery Technology Promotion Station and other units have carried out the artificial breeding technology of large yellow croaker from the "Ten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 刘贤德王志勇隋班良蔡明夷
Owner JIMEI UNIV
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