A novel diagnostic gene region for distinguishing between two pest fruit flies:Bactrocera tryoni(Froggatt)and Bactrocera neohumeralis(Hardy)(Diptera:Tephritidae)  

作　　者：Melissa L.Starkie Elizabeth V.Fowler Alexander M.Piper Xiaocheng Zhu Pauline Wyatt David Gopurenko Matt N.Krosch Francesca Strutt Karen F.Armstrong Hamish Patrick Mark K.Schutze Mark J.Blacket 

机构地区：[1]Department of Agriculture and Fisheries,Biosecurity,Brisbane,Queensland,Australia [2]Agriculture Victoria,AgriBio,Bundoora,Victoria,Australia [3]Department of Primary Industries,Wagga Wagga Agricultural Institute,Wagga Wagga,New South Wales,Australia [4]Forensic Services Group,Queensland Police Service,Brisbane,Queensland,Australia [5]Department of Pest-management and Conservation,Lincoln University,Lincoln,New Zealand [6]Better Border Biosecurity(B3,B3nz.org.nz),Lincoln,New Zealand

出　　处：《Insect Science》2024年第4期1285-1295,共11页昆虫科学（英文版）

基　　金：We thank the Tertiary Education Commission of New Zealand for funding to H.J.P.;Stuart Gilchrist for enabling GBS alignments to the prepublication B.tryoni reference genome.This study was conducted as part of the"Phenology,demography,and distribution of Australia's fruit flies"project,funded through the Strengthening Australia's Fruit Fly System Research Program;Funding for the program is provided by the Australian Government,with contributions matched from state and territory governments.

摘　　要：Bactrocera tryoni and Bactrocera neohumeralis are morphologically similar sibling pest fruit fly species that possess different biological attributes,geographic distributions,and host ranges.The need to differentiate between the two species is critical for accurate pest status assessment,management,biosecurity,and maintenance of reference colonies.While morphologically similar,adults may be separated based on subtle characters;however,some characters exhibit intraspecific variability,creating overlap between the two species.Additionally,there is currently no single molecular marker or rapid diagnostic assay that can reliably distinguish between B.neohumeralis and B.tryoni;therefore,ambiguous samples remain undiagnosed.Here we report the first molecular marker that can consistently distinguish between B.tryoni and B.neohumeralis.Our diagnostic region consists of two adjacent single nucleotide polymorphisms(SNPs)within the pangolin(pan)gene region.We confirmed the genotypes of each species are consistent across their distributional range,then developed a tetra-primer amplification refractory mutation system(ARMS)PCR assay for rapid diagnosis of the species.The assay utilizes four primers in multiplex,with two outer universal primers,and two internal primers:one designed to target two adjacent SNPs(AA)present in B.tryoni and the other targeting adjacent SNPs present in B.neohumeralis(GG).The assay accurately discriminates between the two species,but their SNP genotypes are shared with other nontarget tephritid fruit fly species.Therefore,this assay is most suited to adult diagnostics where species confirmation is necessary in determining ambiguous surveillance trap catches;maintaining pure colony lines;and in Sterile Insect Technique management responses.

关 键 词：ARMS-PCR COLONY molecular diagnostics PANGOLIN tetra-primer 

分 类 号：S41[农业科学—植物保护]