Haplotype identification and detection of mitochondrial DNA heteroplasmy in Varroa destructor mites using ARMS and PCR–RFLP methods

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• 作者：Bojan Gajić ; Jevrosima Stevanović ; Željko Radulović…
• 关键词：Varroa destructor ; Haplotype ; mtDNA heteroplasmy ; ARMS ; PCR–RFLP
• 刊名：Experimental and Applied Acarology
• 出版年：2016
• 出版时间：November 2016
• 年：2016
• 卷：70
• 期：3
• 页码：287-297
• 全文大小：605 KB
• 刊物主题：Entomology; Animal Systematics/Taxonomy/Biogeography; Animal Genetics and Genomics; Animal Ecology; Life Sciences, general;
• 出版者：Springer Netherlands
• ISSN：1572-9702
• 卷排序：70

文摘

In the present study, amplification refractory mutation system (ARMS) and polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) methods were used for identification of recently described Serbia 1 (S1) and Peshter 1 (P1) mitochondrial haplotypes of Varroa destructor. Based on single nucleotide polymorphisms (SNPs) within cytochrome oxidase 1 (cox1) and cytochrome b (cytb) gene sequences, a total of 64 adult V. destructor females were analyzed from locations where the S1 and P1 haplotypes had been detected previously. Results of haplotype identification obtained by ARMS and PCR–RFLP methods were completely consistent with the sequencing data. Furthermore, in some analyzed samples the occurrence of site heteroplasmy at haplotype-defining sites was detected, as it was confirmed by double peaks in the sequence chromatograms. Neither mites with simultaneous nucleotide variability, nor those with combined SNP and heteroplasmy in cox1 and cytb were found. Given that this is the first occurrence of site heteroplasmy in V. destructor, the origin of this phenomenon and possible specific traits of heteroplasmic mites have yet to be determined.