Method for verifying anti-chalk disease traits of swarm by SNP marker
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A chalk disease and bee colony technology, applied in the field of using SNP markers to identify bee colony resistance to chalk disease, can solve the problems of less research and utilization, etc.
Inactive Publication Date: 2015-04-08
FUJIAN AGRI & FORESTRY UNIV
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[0009] As a new generation of biotechnology, although SNP technology has extremely high practical value, its application in the field of human diseases has been relatively mature, and some progress has been made, but the technology There are still few researches and utilizations on molecular markers of bee chalk resistance
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Embodiment 1
[0035] A method utilizing SNP markers to identify anti-chalk disease traits of Apis mellifera colony, comprising the following steps:
[0036] (1) Sampling of bee larvae: 3-day-old honeybee larvae were randomly collected from the Italian bee colonies without other obvious diseases to be investigated, 20 bee larvae per group, each individual was placed in a centrifuge tube, and then Freeze at -20°C for later use;
[0037] (2) Extraction of DNA from larval samples (using Sangon’s Animal Genome Rapid Extraction Kit, SK8222)
[0038] Place the larvae from step (1) in a centrifuge tube for tissue disruption, add 350 ul Buffer Digestion, shake evenly, and place in a water bath at 65 °C for 1 h until the cells are completely lysed.
[0039] Add 150 ul Buffer PA, mix thoroughly by inverting, and place in a -20°C refrigerator for 5 min.
[0040] Centrifuge at 10,000 rpm at room temperature for 5 min, transfer 400 ul of the supernatant to a new 1.5 ml centrifuge tube, if the supe...
Embodiment 2
[0076] The specific data are as follows:
[0077] Table 1A The effect of different genotypes on the disease resistance of bee colonies and the distribution of gene frequency
[0078]
[0079] Note: Chi-square test of the effect of different genotypes on the disease resistance of bee colonies P < 0.05.
[0080] Table 1B The effect of different genotypes on the disease resistance of bee colonies and the distribution of gene frequency
[0081]
[0082] Note: Chi-square test of the effect of different genotypes on the disease resistance of bee colonies P < 0.05.
[0083] Table 1C The effect of different genotypes on the disease resistance of bee colonies and the distribution of gene frequency
[0084]
[0085] Note: Chi-square test of the effect of different genotypes on the disease resistance of bee colonies P < 0.05.
[0086] (1) The statistical results of Table 1A, Table 1B, and Table 1C through Fisher's Exact Test (Fisher's Exact Test) show that there are s...
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Abstract
The invention relates to a method for verifying anti-chalk disease traits of a swarm by an SNP marker. The method comprises the following steps: collecting larva samples from Italian swarms to be observed, extracting larva DNA, synthesizing a target primer, carrying out PCR with each extracted individual DAN as a template, carrying out electrophoresis detection on a PCR product, sequencing, finally comparing and analyzing the obtained sequencing result, and verifying the anti-chalk disease traits by judging whether the occurrence frequency PC of a C allele of randomly captured larva individuals from the swarm has difference with the occurrence frequency PT of a T allele in the SNP marker C2587245T. According to the invention, aiming at the problem that the Italian swarms are severely affected by the chalk disease, the SNP (C2587245T) marker related to anti-chalk disease of Italian bee larva provided from the molecular biology level for scientifically, accurately and quickly verifying the strength of the anti-chalk disease performance of the Italian swarms, the breeding cycle of the anti-chalk disease breeding of the Italian bees can be greatly shortened, the bee breeding speed is accelerated and the loss in the beekeeping industry caused by the chalk disease is reduced.
Description
technical field [0001] The invention relates to a method for identifying chalk resistance traits of bee colonies, in particular to a method for identifying chalk resistance traits of bee colonies by using SNP markers. Background technique [0002] my country is a big beekeeping country. Chalk disease is a fungal disease of bee larvae, which is prevalent and occurs worldwide. The occurrence of chalk disease can reduce the number of newly emerged bees, reduce the productivity of the bee colony, and sometimes cause serious colony loss. At present, many scholars at home and abroad are involved in the research of chalk disease. For a long time, my country's bee industry scientific and technological workers have devoted themselves to the research of bee pathology, disease resistance breeding, etc., in order to discover the pathogenesis of chalk disease and the inheritance of chalk resistance. Marking, etc., serve for bee disease resistance breeding. The research on the behavioral...
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