南瓜实蝇气味结合蛋白基因鉴定及组织分布  

Identification and Expression Distribution of Odorent Binding Proteins from Bactrocera tau

作　　者：孔琼[1] 王传铭[1] 汪斌伟 张晓媛 李珣[1] 谢昆[1] 舒茜 黄镜梅 袁盛勇[1] KONG Qiong;WANG Chuan-ming;WANG Bin-wei;ZHANG Xiao-yuan;LI Xun;XIE Kun;SHU Qian;HUANG Jing-mei;YUAN Sheng-yong(College of Biological and Agricultural Sciences,Honghe University/Key Laboratory for Research and Utilization of Characteristic Biological Resources in Southern Yunnan,Yunnan Mengzi 661199,China)

机构地区：[1]红河学院生物科学与农学学院/云南省高校滇南特色生物资源研究与利用重点实验室,云南蒙自661199

出　　处：《西南农业学报》2021年第12期2666-2672,共7页Southwest China Journal of Agricultural Sciences

基　　金：云南省自然科学基金“南瓜实蝇气味结合蛋白基因的克隆与功能研究”(2016FB069);云南省高校滇南特色水果病虫害绿色防控科技创新团队;红河学院大学生创新创业项目“南瓜实蝇气味结合蛋白基因克隆及表达差异分析”(DCXL181067、DCXL190031);红河学院生物学(农学)重点建设学科;国家级大创项目“南瓜实蝇化学感受蛋白基因的克隆鉴定及分子对接”(202010687023)。

摘　　要：【目的】克隆南瓜实蝇(Bactrocera tau)的气味结合蛋白(odorent binding proteins,OBPs)基因,并分析其相关生物学信息,为了解南瓜实蝇的嗅觉机制提供参考。【方法】基于南瓜实蝇转录组测序结果筛选获得OBPs基因并进行生物信息学分析,通过基因克隆测序验证鉴定OBPs基因,并采用sqRT-PCR检测OBPs在雌、雄虫不同组织中的分布情况。【结果】经PCR产物测序验证和BLAST比对后共鉴定得到6个南瓜实蝇OBPs基因,序列长度在498~955 bp,编码128~314 aa,预测分子量大小为14.67~36.24 kDa,等电点为4.69~8.16,且每个OBP序列的N端均具有16~26 aa组成的信号肽序列,属于1个Minus型和5个Classic型。不同类型的OBPs均具有各自典型保守的Cys位点。6个OBPs分化明显,序列相似度在30%~51%。系统进化树表明,6个OBPs与瓜实蝇(Zeugodacus cucurbitae)的OBPs亲缘关系较近,柑橘大实蝇(Bactrocera minax)次之。半定量RT-PCR检测显示,6个OBPs在雌、雄虫不同组织中均有表达,而OBP14在雄虫腹部不表达,OBP15在雌虫腹部不表达。【结论】上述研究结果为进一步研究南瓜实蝇嗅觉识别分子机制和引诱机制奠定了基础。【Objective】In order to provide reference for understanding olfactic mechanism of Bactrocera tau,the odorent binding protein(OBPs)of B.tau was cloned and its related bioinformatics was analyzed.【Method】Based on transcriptome sequencing results of B.cucurbita,the OBPs were screened,identified and analyzed by bioinformatic methods,and the expression distribution of these OBPs in different tissues of female and male was detected by semi-quantitative reverse transcription and polymerase chain reaction(sqRT-PCR).【Result】The 6 OBPs of B.tau were identified by sequencing and BLAST analysis,the full length of these genes were 498-955 bp,and could encode 128-314 aa,predicted molecular weights were 14.67-36.24 kDa,isoelectric point were 4.69-8.16,and the N-terminal of every OBP sequence containing signal peptide for 16-26 amino acid.The 6 OBPs belonged to two subfamilies with 1 Minus-C OBPs and 5 Classical-C OBPs,and these genes had their own typical conserved Cys sites.The six sequences of OBPs showed obvious differentiation,and their similarity ranged from 30%to 51%.Phylogenetic tree showed that these OBPs were closely related to that of Zeugodacus cucurbitae,followed by B.minax.These OBPs were expressed in different tissues of both males and females in Z.cucurbitae,while OBP14 was not expressed in abdomen of male,and OBP15 was not expressed in female abdomen.【Conclusion】These results would establish a foundation for further research on molecular and attracting of mechanisms in olfactory sensation from B.tau.

关 键 词：南瓜实蝇 气味结合蛋白 基因鉴定 表达分布 半定量PCR 

分 类 号：Q969[生物学—昆虫学]