豌豆蚜触角转录组化学感受蛋白的鉴定、表达和结合特性分析  

Identification,expression and binding specificity of chemosensory proteins in the antennal transcriptome of the pea aphid,Acyrthosiphon pisum(Hemiptera:Aphididae)

作　　者：刘靖涛 王倩 赵瑞[2] 王欢欢 高洁 董辉 张永军[3] 丛斌[1] 谷少华[2] LIU Jing-Tao;WANG Qian;ZHAO Rui;WANG Huan-Huan;GAO Jie;DONG Hui;ZHANG Yong-Jun;CONG Bin;GU Shao-Hua(College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China;Department of Entomology, China Agricultural University, Beijing 100193, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China)

机构地区：[1]沈阳农业大学植物保护学院,沈阳110866 [2]中国农业大学昆虫学系,北京100193 [3]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193

出　　处：《昆虫学报》2022年第3期261-270,共10页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(31772164);植物病虫害生物学国家重点实验室开放基金(SKLOF202106);北京市青年拔尖人才项目(2015000021223ZK29)。

摘　　要：【目的】本研究旨在鉴定豌豆蚜Acyrthosiphon pisum触角转录组中化学感受蛋白(chemosensory protein,CSP)基因,明确触角中高表达的豌豆蚜CSP蛋白与蚜虫报警信息素、性信息素以及植物挥发物的分子结合特性。【方法】通过对豌豆蚜成蚜触角进行转录组测序,鉴定触角中候选CSP基因;采用RPKM值和半定量RT-PCR分别明确这些CSP基因在豌豆蚜成蚜触角以及不同组织(触角、口针、去除触角和口针的头、胸、腹、足和翅)中的表达谱;通过荧光竞争结合实验测定这些触角CSPs与蚜虫报警信息素和性信息素以及植物挥发物的结合特性。【结果】在豌豆蚜成蚜触角转录组中鉴定得到10个CSP基因。RPKM值和半定量RT-PCR分析结果表明ApisCSP2,ApisCSP4和ApisCSP5在豌豆蚜成蚜触角中表达量最高。荧光竞争结合实验表明ApisCSP4与报警信息素主要组分(E)-β-法尼烯[(E)-β-farnesene,EβF]和微量组分(-)-α-蒎烯结合力强,K_(i)值分别为2.2±0.8和4.9±0.7μmol/L;ApisCSP5与两种性信息素组分(+)-(4aS,7S,7aR)-荆芥内酯和(-)-(1R,4aS,7S,7aR)-荆芥醇结合力强,K_(i)值分别为4.8±0.9和2.6±0.6μmol/L。【结论】豌豆蚜成蚜触角转录组中鉴定到10个CSP基因,其中ApisCSP4和ApisCSP5可能分别参与了蚜虫报警信息素和性信息素的转运。【Aim】This study aims to identify the chemosensory protein(CSP)genes in the antennal transcriptome of the pea aphid,Acyrthosiphon pisum,and to investigate the binding characteristics of antennae-enriched CSPs of A.pisum with aphid alarm pheromones,sex pheromones and plant volatiles.【Methods】Based on the antennal transcriptome data of A.pisum,the CSP genes in antennae were identified.The expression profiles of these CSP genes in antennae and different tissues(antennae,stylet,head without antenna and stylet,thorax,abdomen,leg and wing)of A.pisum adults were assayed based on RPKM value and by semiquantitative RT-PCR,respectively.The binding characteristics of these CSPs in antennae with aphid alarm pheromones and sex pheromones,and plant volatiles were assayed by fluorescence competitive binding assay.【Results】A total of ten CSP genes were identified from the antennal transcriptome of A.pisum adults.The RPKM value and semiquantitative RT-PCR results revealed that ApisCSP2,ApisCSP4 and ApisCSP5 were highly expressed in the antennae of A.pisum adults.The fluorescence competitive binding assays indicated that ApisCSP4 showed strong binding affinities to the main component of aphid alarm pheromone(E)-β-farnesene(EβF),and the minor component(-)-α-pinene,with the K_(i) values of 2.2±0.8 and 4.9±0.7μmol/L,respectively.ApisCSP5 exhibited strong binding affinities to the two components of sex pheromones(+)-(4aS,7S,7aR)-nepetalactone and(-)-(1R,4aS,7S,7aR)-nepetalactol,with the K_(i) values of 4.8±0.9 and 2.6±0.6μmol/L,respectively.【Conclusion】There are ten CSP genes identified in the antennal transcriptome of A.pisum adults,among which ApisCSP4 and ApisCSP5 may be involved in the transport of aphid alarm pheromones and sex pheromones,respectively.

关 键 词：豌豆蚜 化学感受蛋白 组织表达 荧光竞争结合实验 报警信息素 性信息素 

分 类 号：Q966[生物学—昆虫学]