华北大黑鳃金龟生物钟基因HoPer的克隆和表达模式分析  

Cloning and expression profiling of circadian clock gene HoPer in Holotrichia oblita(Coleoptera:Melolonthidae)

作　　者：谢明惠 钟永志 林璐璐 张光玲 苏卫华 倪皖莉[2] 陈浩梁 XIE Ming-Hui;ZHONG Yong-Zhi;LIN Lu-Lu;ZHANG Guang-Ling;SU Wei-Hua;NI Wan-Li;CHEN Hao-Liang(Institute of Plant Protection and Agro-products Safety,Anhui Academy of Agricultural Sciences,Hefei 230001,China;Crop Research Institute,Anhui Academy of Agricultural Sciences,Hefei 230001,China)

机构地区：[1]安徽省农业科学院植物保护与农产品质量安全研究所,合肥230001 [2]安徽省农业科学院作物研究所,合肥230001

出　　处：《环境昆虫学报》2024年第2期471-479,共9页Journal of Environmental Entomology

基　　金：国家重点研发计划(2018YFD0201009);安徽省自然科学基金(1908085QC137);安徽省重点研发计划(202004a06020034)。

摘　　要：为明确华北大黑鳃金龟Holotrichia oblita周期蛋白基因(Period,Per)的表达模式,本研究采用RACE技术从2龄幼虫中克隆到生物钟基因HoPer,并进行生物信息学分析;再采用RT-qPCR技术检测HoPer在不同发育阶段、雌雄成虫不同组织以及不同光周期下的表达水平。结果表明:HoPer编码1 172个氨基酸,蛋白分子量129.67 kDa,等电点为5.62。HoPer推导出的氨基酸序列中具有2个PAS和1个Period C结构域,是昆虫PER蛋白的典型结构特征。表达模式分析得出:HoPer在各个发育阶段均有表达,但在卵中的表达量显著高于其他发育时期;HoPer在雌雄成虫的头和触角中表达水平最高,其次为翅和足,在胸和腹中表达水平最低;雌雄成虫中HoPer在试验设定的5个光周期中未发现明显的表达差异。本研究为进一步阐明华北大黑鳃金龟HoPer基因功能以及其在生物钟网络中的调控作用提供了一定的参考。In order to clarify the expression pattern of the period gene(Period,Per)of Holotrichia oblita,this study used RACE to clone the circadian clock gene HoPer from the second instar larvae,and conducted bioinformatics analysis.RT-qPCR was used to detect the expression levels of HoPer in different developmental stages,different tissues of male and female adults and different photoperiods.The results showed that HoPer encoded 1172 amino acids,with a molecular weight of 129.67 kDa and an isoelectric point of 5.62.The amino acid sequence deduced by HoPer had two PAS and one period C domain,which were typical structural characteristics of insect PER proteins.Expression pattern analysis showed that HoPer was expressed in all developmental stages,but the expression in eggs was significantly higher than that in other developmental stages.The expression level of HoPer in the head and antennae of male and female adults were in the top two,followed by wings and leg,and throax and abdomen were the last two.There was no significant difference in HoPer expression between male and female adults across the five photoperiods.This study provides some reference for further study on the function of HoPer gene and its regulatory role in the biological clock network.

关 键 词：华北大黑鳃金龟 生物钟基因 PER 昼夜节律 

分 类 号：Q963[生物学—昆虫学] S433[农业科学—农业昆虫与害虫防治]