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Development of Single Nucleotide Polymorphism (SNP) Markers to Establish Diagnostic System to Detect Disease-Resistant Strain of Apis mellifera (Hymenoptera: Apidae)
We developed single nucleotide polymorphism (SNP) markers and are establishing diagnostic systems to distinguish disease resistance- and susceptible-strains of honey bees using the SNPs. For development of SNP markers, whole genome was sequenced each from 20 individuals of “disease resistance-strain” and “susceptible-strain” of Apis mellifera ligustica using the Illumina HiSeq 2000 sequencer. Approximately, 344 and 294 million sequence reads were mapped to the honeybee reference assembly (Amel_4.5) for each strain, respectively. Among the total 2,246,428 SNPs yielded, 33 were found to be fixed between the two strains with all homozygosity. Sixteen of them were casually amplified and sequenced from randomly selected each 10 individual of honey bees from each strain and presented strain specific SNPs. These ten SNPs were used to diagnose the two strains either by original size difference, caused by indel-accompanying SNP, typical PCR-RFLP, or AS PCR.
