Method for analyzing genetic diversity of Portunus trituberculatus by using DArT (diversity arrays technology) labelling technique

A technology for portunus trituberculatus and genetic diversity, which is applied in the field of analyzing the genetic diversity of portunus trituberculatus using DArT marker technology, can solve the problem of undisclosed genetic diversity of portunus trituberculatus geographic populations, and achieve low-cost effects

Active Publication Date: 2014-04-09
NINGBO UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

However, there is no relevant research report on the method of analyzing the genetic diversity of Portunus trituberculatus geographic populations using DArT marker technology at home and abroad.

Method used

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  • Method for analyzing genetic diversity of Portunus trituberculatus by using DArT (diversity arrays technology) labelling technique
  • Method for analyzing genetic diversity of Portunus trituberculatus by using DArT (diversity arrays technology) labelling technique
  • Method for analyzing genetic diversity of Portunus trituberculatus by using DArT (diversity arrays technology) labelling technique

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specific Embodiment 1

[0035] 1. Experimental Materials and Reagents

[0036] The present invention adopts the Portunus trituberculatus samples of 5 geographical populations, respectively taken from five different areas of my country from south to north: Dandong (DD), Yantai (YT), Qingdao (QD), Lianyungang (LYG), Ningbo (NB) ), representing the Yalu River Estuary population in the Yellow Sea; the Laizhou Bay population in the Bohai Sea; the wild population in the venue; the Haizhou Bay population; and the Zhoushan population in the East China Sea. The above five populations were all sampled in 2011. The average shell length of all samples was 11.51±1.22 cm, the average weight was 164.4±19.7 g, and the ratio of male to female was 1:1. The living body was transported back to the laboratory, and the giant claw was stored in a -80°C refrigerator for later use.

[0037] Escherichia coli TOP10F' was preserved by the laboratory, PstI, TaqI , T4 DNA ligase was purchased from Takara, ampicillin, X-gal (5-...

specific Embodiment 2

[0118] From the above 5 wild populations of Portunus trituberculatus Pst I / Taq I Representative fragments (shown in Example 4.2 above) were again hybridized to the polymorphism-enriched DArT chip and replicated once. According to the above data processing principles, a stable 0 / 1 matrix is ​​obtained. RESTDIST and NEIGHBOR programs with PHYLIP 3.695 software, according to 313 Pst I / Taq The Nei / Li method was used to calculate the genetic distance of I polymorphic markers and construct the unweighted group average method (UPGMA) phylogenetic diagram, and the SEQBOOT program was used for bootstrap analysis. The value at the bifurcation represents the confidence percentage obtained by 1,000 repeated sampling , only values ​​with a confidence level above 50% are displayed. Such as Figure 6 As shown in the phylogenetic tree, it shows the genetic relationship between the five geographical populations of Portunus trituberculatus, among which the populations of Qingdao ...

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Abstract

The invention discloses a method for analyzing genetic diversity of Portunus trituberculatus by using a DArT (diversity arrays technology) labelling technique. The method is characterized by comprising the steps of: constructing a representative DNA (deoxyribonucleic acid) fragment library of Portunus trituberculatus genome; pointing a representative genome DNA chip and setting a series of positive and negative contrasts on the chip; respectively hybridizing DNAs of Portunus trituberculatus of different geographical populations with the representative genome DNA chip and cleaning; finally, scanning through a chip scanner after the hybridizing step, extracting data through LuxScan3.0 software, and calculating to obtain polymorphic efficiency, polymorphic information content and genetic profiles. The method has the advantages of being capable of obtaining stable and clear genetic profiles, the method can be applied in fields of genetic diversity detection of Portunus trituberculatus, wild population and family identification, creation of genetic maps and fingerprints, and the like, and the method has the advantages of being low in cost, efficient, stable and accurate.

Description

technical field [0001] The invention relates to a method for analyzing the genetic diversity of Portunus trituberculatus, in particular to a method for analyzing the genetic diversity of Portunus trituberculatus by using DArT marker technology. Background technique [0002] Portunus trituberculatus ( Portunus trituberculatus ) belongs to Crustacea, Decapoda, Portunidae, Portunus, is an important large-scale marine economic crab, widely distributed in China, Japan, Korea and the Malaysian archipelago and other sea areas , it has the advantages of good meat quality, fast growth, high yield, high economic value, and rich farming profits. With the rapid development of mariculture, portunus trituberculatus farming has become the leading cultured species in coastal areas. [0003] At present, the crab seedlings of Portunus trituberculatus aquaculture mainly rely on wild resources. Due to environmental deterioration, disease and overfishing, wild resources have declined sharply, w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C40B50/06
CPCC12Q1/6827C12Q2563/107C12Q2545/101
Inventor 陈炯史雨红
Owner NINGBO UNIV
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