Microsatellite marker primer related to low temperature resistance of fugu rubripes and application thereof

A technology of redfin puffer and low temperature resistance, which is applied in the field of fish DNA marking technology and application, to shorten the breeding period and improve the breeding efficiency

Active Publication Date: 2021-02-05
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of molecular markers to assist in the cultivation of redfin puffer with low temperature tolerance is an important way to reduce costs and improve economic benefits. At present, the research on microsatellite molecular markers related to low temperature tolerance of redfin puffer has not been reported in China.

Method used

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  • Microsatellite marker primer related to low temperature resistance of fugu rubripes and application thereof
  • Microsatellite marker primer related to low temperature resistance of fugu rubripes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 1. Acquisition of low-temperature-resistant materials for pufferfish

[0018] The red-finned puffer samples used in the experiment came from Dalian Tianzheng Industrial Co., Ltd., and 68 juvenile fishes with strong physique, no damage and good vitality were selected, with a body length of 15.1cm±0.5cm and a weight of 150g. They were temporarily raised in the experimental tank for 2 weeks. 23 ± 0.5 ° C, using a heating rod to maintain temperature. After the temporary breeding is over, the temperature will be lowered by 5°C every day through natural sea water (water temperature 5°C), and 3°C on the last day, and finally reach 5°C and then maintain (23°C-18°C-13°C-8°C-5°C ). During the cooling period, the state of the experimental fish was observed every 20 minutes. When it was found that the dying fish were taken out in time, the caudal fins were cut and placed in absolute ethanol, and then frozen in a -40°C refrigerator for later use. The experiment was stopped when th...

Embodiment 2

[0037] Example 2: Microsatellite markers resistant to low temperature are applied to the breeding of low temperature resistant strains of puffer puffer

[0038]Before strengthening the broodstock, part of the caudal fin of the candidate parent was cut, fixed and stored in 95% absolute ethanol at a low temperature for extraction of genomic DNA. Using TIANGEN's Marine Animal DNA Extraction Kit, the tail fin DNA of the redfin puffer was extracted. The quality and integrity of the extracted DNA were determined by 1% agarose gel electrophoresis, and the concentration was determined by an ultraviolet spectrophotometer. Finally, it was diluted with double distilled water to about 50 ng / μL, and stored in a -40°C refrigerator.

[0039] Use the screened microsatellite marker fms100 primers for PCR amplification to find differential bands. The PCR reaction system is 20 μL: 1 μL DNA solution, 1 μL upstream primer, 1 μL downstream primer, 10 μL 2xTaq PCR Mix, 7 μL sterilized double distil...

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Abstract

The invention relates to a microsatellite marker related to low temperature resistance of fugu rubripes and application thereof, and belongs to the field of fish DNA marking technologies and application, particularly screening of low temperature resistance molecular markers of fugu rubripes and molecular marker-assisted breeding by utilizing the markers. A microsatellite marker (fms108) related tolow temperature resistance of the fugu rubripes is screened by using a BSA-SSR technology, and the primer sequences of the microsatellite marker (fms108) are shown by SEQUENCE LISTING 1 and SEQUENCELISTING 2. Breeding parents can be selected according to the marker, so that the low-temperature resistance of offspring is improved.

Description

technical field [0001] The invention belongs to the field of fish DNA marker technology and application, and specifically relates to a microsatellite marker primer related to the low temperature resistance of puffer puffer and its application. Background technique [0002] The red-fin puffer is an important fish for marine culture and export in northern my country. Its suitable growth temperature is 16°C-25°C. If the temperature is too low, the growth performance will decline or even die. For pufferfish, the land-sea relay and overwintering process will inevitably encounter temperature drop. In addition, the method of boiling water to increase the water temperature will also encounter problems such as increased cost and environmental protection pressure. Therefore, through the selection of strains with cold-resistant traits It can reduce the economic loss caused by temperature drop. [0003] Some progress has been made in the breeding of heat-resistant varieties of aquatic an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2600/124C12Q2600/172C12Q2531/113C12Q2565/125
Inventor 刘志峰马爱军袁晨浩王新安孙志宾
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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