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

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...

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Bibliographic Details
Published in:Experimental & applied acarology 2016-11, Vol.70 (3), p.287-297
Main Authors: Gajic, Bojan, Stevanovic, Jevrosima, Radulovic, Eljko, Kulisic, Zoran, Vejnovic, Branislav, Glavinic, Uros, Stanimirovic, Zoran
Format: Article
Language:English
Subjects:
Animal Ecology
Animal Genetics and Genomics
Animal Systematics/Taxonomy/Biogeography
Animals
Biomedical and Life Sciences
Cytochrome
DNA, Mitochondrial - genetics
Entomology
Female
Haplotypes
Life Sciences
Mites
Mitochondrial DNA
Nucleic Acid Amplification Techniques - economics
Polymerase Chain Reaction - economics
Polymorphism, Restriction Fragment Length
Sequence Analysis, DNA
Serbia
Varroa destructor
Varroidae - genetics
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Summary: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.
ISSN:0168-8162
1572-9702
DOI:10.1007/s10493-016-0086-6