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Discriminating method of inherited characteristics of Kyrgyzstan cichlidae and application thereof

A kind of technology of australis and fish, applied in the field of biometric identification, can solve the problem of difficulty in distinguishing between scientific and technological personnel and consumers

Inactive Publication Date: 2011-06-15
李思发
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008]However, although the shape of argyle tilapia is between the parents, experienced professionals can identify it, but it is difficult for ordinary technicians and consumers to distinguish

Method used

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  • Discriminating method of inherited characteristics of Kyrgyzstan cichlidae and application thereof
  • Discriminating method of inherited characteristics of Kyrgyzstan cichlidae and application thereof
  • Discriminating method of inherited characteristics of Kyrgyzstan cichlidae and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Karyotype analysis

[0063] 1.1 method

[0064] Phytohemagglutinin (PHA) was injected in vivo to prepare chromosome slide specimens. Take 10 healthy fish, weighing about 100g. The experimental fish were temporarily kept in an indoor temperature-controlled, inflatable aquarium one week before the experiment, and the water temperature was about 30°C. PHA was injected at the pectoral fin of 10μg / g fish body weight. After 16 to 18 hours, colchicine solution was injected at 2μg / g fish body weight. After 3h, the gills were cut and bloodletted. The head and kidney were placed in a dish filled with physiological saline and ground to make the The cells freed up. Aspirate the cell suspension in 0.075mol / L KCL solution at 37°C for 30 min. Subsequently, it was centrifuged at 1000r / min for 8min, the supernatant was discarded, and the newly prepared Carnoy's solution (methanol: glacial acetic acid=3:1) was added to fix for 15min and then centrifuged for 8min, then the fixati...

Embodiment 2

[0083] Example 2 Isozyme

[0084] 2.1 Method

[0085] Choose 30 fish of good size, healthy and weighing about 100g. According to the National Standard of the People's Republic of China (GB / T18654.13-2008): "Cultured Fish Germplasm Inspection Part 13: Isozyme Electrophoresis Analysis".

[0086] In short, this includes: obtaining fish tissues (respectively muscle, liver, kidney, heart, and eye tissues), after homogenizing, centrifuging the homogenized sample at 15000 rpm until the supernatant is clear, and taking the form after centrifugation The supernatant was detected by isozyme electrophoresis.

[0087] 2.2 Results

[0088] The ADH, LDH, IDH and equivalent enzymes in the muscle, liver, kidney, heart, and eye tissues of Tilapia and its parents were detected by polyacrylamide gel electrophoresis.

[0089] The results showed that for ADH and IDH, Tilapia and its two parents did not show obvious polymorphism, but for LDH, it showed obvious polymorphism.

[0090] As shown in Figure 3, the ...

Embodiment 3

[0091] Example 3 Microsatellite Marker

[0092] 3.1 Method

[0093] Thirty fish were randomly sampled, caudal fins were cut, numbered and stored in 95% ethanol for later use. Genomic DNA extraction is carried out by the conventional "phenol-chloroform" method, the specific steps are: take about 0.1g of the tail fin and add 400μL STE (30mmol / L Tris-HCl, pH8.0, 200mmol / L EDTA, 50mmol / L NaCl) buffer , After mixing, add proteinase K with a final concentration of 1% SDS 200μg / mL, and let it act overnight at about 55°C. Add an equal volume of saturated phenol and mix well for 1h, centrifuge at 10,000r for 10min, absorb the supernatant and add an equal volume of the mixed solution (phenol:chloroform:isoamyl alcohol=25:24:1), mix well for 0.5h, absorb the supernatant Add an equal volume of the mixed solution (chloroform: isoamyl alcohol = 24:1), mix well for 0.5h, discard the lower layer; then add 2 times the volume of absolute ethanol (preferably frozen) to precipitate the DNA, and then...

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Abstract

The invention provides a discriminating method of the inherited characteristics of Kyrgyzstan cichlidae and application thereof, in particular to a discriminating method of the inherited characteristics of a new specie-Kyrgyzstan cichlidae tilapia (an intergeneric distant hybridization and selective breeding offspring of tilapia nilotica and sarotherodon melanotheron) suitable for mariculture andapplication thereof. The discriminating method comprises the following steps of: detecting isoenzymes, karyotypes and / or microsatellite markers of fishes to be detected; and comparing a detection result with a positive control or a standard substance so as to determine or judge whether the detected fishes are Kyrgyzstan cichlidae tilapias or not, wherein the isoenzymes are lactate dehydrogenases.The discriminating method can reliably and accurately discriminate the Kyrgyzstan cichlidae tilapias.

Description

Technical field [0001] The invention relates to the field of biological identification. More specifically, the present invention relates to the genetic characteristics of a new species suitable for marine aquaculture-tilapia (intergeneric cross between Nile tilapia and Salola tilapia and breeding offspring). Method and its application. Background technique [0002] At present, there are some urgent problems in aquaculture production in the salty and sea waters of our country. For example, sea cage aquaculture lacks suitable breeding objects and breeding models suitable for China's national conditions. The existing breeding objects are almost all carnivorous fish. , The source of seedlings is difficult, and the bait is not easy to solve. Not only the breeding cost is high, the breeding risk is high, but also because of the large and long-term feeding of chilled trash fish, it is also prone to ecological and environmental problems; the main problem of saltwater breeding in the cen...

Claims

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Application Information

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IPC IPC(8): C12Q1/32C12Q1/68
Inventor 李思发蔡完其许映芳
Owner 李思发
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