取食不同辣椒品种果实成分对棉铃虫幼虫唾腺蛋白及相应基因表达的影响  

Expression profiling of salivary gland proteins and the corresponding genes in Helicoverpa armigera( Lepidoptera: Noctuidae) larvae feeding on artificial diets containing fruits of different pepper varieties

作　　者：毛培[1] 杜孟芳[1] 姚双艳 蔡永萍[1] 郭线茹[1] 原国辉[1] 罗梅浩[1] 

机构地区：[1]河南农业大学植物保护学院,郑州450002

出　　处：《昆虫学报》2014年第10期1133-1145,共13页Acta Entomologica Sinica

基　　金：河南省杰出青年科学基金项目(074100510013)

摘　　要：【目的】棉铃虫Helicoverpa armigera(Hübner)在不同辣椒Capsicum annuum L.品种上的寄主适合度存在较大差异。本研究的目的是弄清不同辣椒品种对棉铃虫唾腺蛋白及相应基因表达的影响。【方法】通过双向电泳和实时荧光定量PCR(real-time quantitative PCR)的方法,探讨棉铃虫5龄幼虫取食添加辣椒碱含量不同的4个辣椒品种果实成分的人工饲料后,其唾腺中蛋白及相应基因表达的变化。【结果】双向电泳结果表明:取食4个处理的人工饲料后,棉铃虫5龄幼虫唾腺中蛋白质表达存在显著差异(P<0.05),定量分析发现28个蛋白点在不同处理间差异表达。通过ESI-Q-TOF MS分析鉴定出15种差异表达蛋白,其中β-呋喃果糖苷酶(β-fructofuranosidase)、热休克蛋白70(HSP70)和翻译起始因子5A(e IF5A)在不同处理间差异较大。进一步荧光定量PCR分析显示:人工饲料中添加辣椒碱含量较高的果实成分能够促进棉铃虫β-fructofuranosidase基因的表达,添加辣椒碱含量较低的果实成分则抑制了该基因的表达;4个处理对棉铃虫hsp70基因的表达均表现出持续抑制,对e IF5A基因则均表现出一定程度的抑制。【结论】该结果为丰富和拓宽β-fructofuranosidase,hsp70和e IF5A在昆虫中的功能研究、揭示棉铃虫对辣椒的寄主适应机制和生产上抗虫辣椒品种的选育提供了重要的基础材料。[ Aim ] The fitness of Helicoverpa armigera is dependent on different pepper （Capsicum annuum L. ） varieties. This study aims to explore the effect of different pepper varieties on the expression of salivary gland proteins and the corresponding genes in the 5th instar larvae of H. armigera. [ Methods] We used two-dimensional polyacrylamide gel electrophoresis （2-DE） and real-time quantitative PCR （qPCR） to examine the changes in the expression profiles of salivary gland proteins and the corresponding genes in the 5th instar larvae of H. armigera feeding on artificial diets containing fruits of commercial pepper varieties with different levels of capsaicinoids, respectively. [ Results ] The results indicated that the expression of proteins in the salivary glands of the 5th instar larvae of H. armigera was strongly influenced by the different pepper varieties, significant difference in the expression levels of 28 proteins was found among different treatments through quantitative analysis （P 〈 0. 05 ） and 15 differentially expressed proteins were successfully determined by electrospray ionization quadrupole time- of-flight mass spectrometry （ ESI-Q-TOF MS）. Real-time PCR analysis manifested that the higher level of capsaicin promoted the expression of β-fructofuranosidase, and vice versa. Continuous exposure to four pepper treatments inhibited the expression of hsp70 all the time and inhibited the expression of elF5A to a certain extent. [ Conclusion] The functional studies of β-fructofuranosidase, hsp70 and elF5A have been enriched and widened. The results revealed the adaptive mechanism of H. armigera in different pepper varieties, providing a sound basis for selection of pepper varieties resistant to insects.

关 键 词：棉铃虫 辣椒 辣椒碱 唾腺蛋白 Β-呋喃果糖苷酶 热休克蛋白70 翻译起始因子5A 

分 类 号：Q966[生物学—昆虫学]