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Sex determination method for apostichopus japonicus

An identification method and imitation technology of japonicus, applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve the problems of unfavorable promotion and high equipment requirements, and achieve simple equipment and technical requirements, easy operation, and intuitive results Effect

Active Publication Date: 2017-05-17
LIAONING OCEAN & FISHERIES SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, it has been reported that the sex of japonicus japonicus can be determined by scanning the gonads of japonicus japonicus with a medical ultrasonic scanner. Not conducive to issues such as promotion in production and research

Method used

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  • Sex determination method for apostichopus japonicus
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  • Sex determination method for apostichopus japonicus

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Embodiment 1

[0026] On June 19, 2015 and June 23, 2015, 40 imitation sea cucumbers were collected in Xingcheng and Jinzhou, Liaoning, respectively, weighing 215±28.8g and 264±22.5g, respectively, and kept until Available June 25, 2015.

[0027] (1) Preparation of coelomic fluid supernatant and coelomocyte disrupted fluid supernatant

[0028] Penetrate the abdomen of sea cucumber with a sterile syringe to extract 100 μL of body cavity fluid, centrifuge (4°C, 3500 rpm, 5 min), and take the supernatant as the body cavity fluid supernatant; take the cell pellet and resuspend it in 10 μL of distilled water, freeze and thaw at -20°C for 2 centrifuge (4° C., 12000 rpm, 5 min) to take the supernatant, which is used as the supernatant of coelomocyte disrupted liquid.

[0029] (2) SDS-PAGE and male identification were performed using body cavity fluid supernatant as a sample

[0030] ① Prepare Tris-glycine (containing SDS) buffer solution (3.03g Tris, 14.41g glycine, 1g SDS, 1L distilled water). ...

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Abstract

The invention discloses a sex determination method for apostichopus japonicas. The method comprises the following steps: (1) taking apostichopus japonicas coelomic fluid, and respectively preparing coelomic fluid supernatant and coelomocyte breaking fluid supernatant; (2) taking the coelomic fluid supernatant to carry out SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis), and taking the coelomocyte breaking fluid supernatant to carry out native-PAGE; (3) judging whether SDS-PAGE gel contains a 50kDa protein band or not to determine whether the apostichopus japonicas is male or not, and judging whether a phenoloxidase zymogram after the native-PAGE is carried out contains three bands or not to determine whether the apostichopus japonicas is female or not. The sex determination method has the following advantages: 1) the apostichopus japonicas is slightly harmed; 2) reagent, equipment and technical requirements are simple, and a result is visual; 3) determination speed is high, and handling capacity is high; 4) two technologies are used for respectively determining male and female apostichopus japonicas, and accuracy is 90% or above.

Description

technical field [0001] The invention belongs to the technical field of aquaculture, and in particular relates to a sex identification method for imitation sea cucumber. Background technique [0002] Apostichopus japonicus belongs to the phylum Echinodermata, an invertebrate, and is one of the most important economic species for breeding in my country's coastal waters. In the production process of sea cucumber seedlings, the proportion of male species participating in female species ginseng is often controlled within the range of 1 / 10 to 1 / 2. If the proportion of male species participating in female species ginseng is too high, it will affect the hatching of fertilized eggs. The selection of ginseng requires relatively less number of male ginseng. However, although japonicus japonicus is a dioecious organism, there is no obvious difference in the appearance of female and male individuals. It is impossible to determine the sex of japonicus japonicus from the appearance of the...

Claims

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Application Information

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IPC IPC(8): G01N27/447
CPCG01N27/447
Inventor 蒋经伟周遵春董颖高杉孙红娟陈仲苏鹤声
Owner LIAONING OCEAN & FISHERIES SCI RES INST
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