用PCR-RFLP方法鉴别赤拟谷盗和杂拟谷盗的研究  

Identification of Tribolium castaneum and Tribolium confusum based on PCR-RFLP

作　　者：贺艳萍 李嘉旭[1] 潘登 毕洁 汤秋玲 HE Yanping;LI Jiaxu;PAN Deng;BI Jie;TANG Qiuling(School of Food Science and Engineering,Wuhan Polytechnic University,Wuhan 430023,China;Key Laboratory for Deep Processing of Major Grain and Oil,Ministry of Education,Wuhan Polytechnic University,Wuhan 430023,China)

机构地区：[1]武汉轻工大学食品科学与工程学院,湖北武汉430023 [2]武汉轻工大学大宗粮油精深加工教育部重点实验室,湖北武汉430023

出　　处：《河南工业大学学报(自然科学版)》2025年第1期90-95,共6页Journal of Henan University of Technology:Natural Science Edition

基　　金：大宗粮油精深加工省部共建教育部重点实验室开放课题(DZLY2023006)。

摘　　要：为快速准确鉴定外部形态相似的赤拟谷盗(Tribolium castaneum(Herbst))和杂拟谷盗(Tribolium confusum Jacquelin du Val),分别对赤拟谷盗和杂拟谷盗的成虫和幼虫Cytb基因片段进行PCR扩增、测序,并分析两物种该基因片段的限制性内切酶位点差异,根据PCR产物酶切图谱的差异进行物种鉴定。结果表明:两种害虫成虫和幼虫所选Cytb基因扩增片段长度均为640 bp,两物种该基因片段序列同源性为77.19%,核苷酸转换发生83次,颠换发生63次,转换与颠换次数比为1.3;两物种该片段内Bst X I、Sal I、Xba I、Vsp I和Age I等5种限制性内切酶酶切具有显著差异,对Bst X I、Sal I、Xba I和Vsp I 4种酶切图谱进行了验证,其酶切图谱与软件分析结果一致,可以通过这4种限制酶的酶切图谱对赤拟谷盗和杂拟谷盗的成虫及幼虫进行快速准确鉴定。用PCR-RFLP方法可以准确鉴定赤拟谷盗和杂拟谷盗的幼虫和成虫,为精准防控提供科学支持。Utilizing the sequence differences of the cytochrome b(Cytb)gene in mitochondrial DNA allows for the rapid and accurate identification of the externally similar Tribolium castaneum(Herbst)and Tribolium confusum Jacquelin du Val,providing an essential foundation for improving precision control of stored grain pests.This study amplified,sequenced,and analyzed the Cytb gene fragments of adult and larval stages of T.castaneum and.T.confusum,respectively.The species were identified based on differences in restriction enzyme sites of these gene fragments.The amplified gene fragments of both pest species were 640 bp in length.The sequence homology of this gene fragment between the two species was 77.19%,with 83 nucleotide transitions and 63 transversions,resulting in a transition/transversion ratio of 1.3.Five restriction enzymes(Bst X I,Sal I,Xba I,Vsp I,and Age I)showed differences in the gene fragment between the two species.The digestion profiles of Bst X I,Sal I,Xba I,and Vsp I were validated,and their patterns matched the analysis results.These restriction enzyme digestion profiles enable rapid and accurate identification of T.castaneum and T.confusum at both adult and larval stages.The PCR-RFLP method can accurately identify the larvae and adults of T.castaneum and T.confusum,providing scientific support for precise pest control.

关 键 词：赤拟谷盗 杂拟谷盗 CYTB基因 酶切 鉴定 

分 类 号：TS210[轻工技术与工程—粮食、油脂及植物蛋白工程]