橘小实蝇神经肽sulfakinin的分子生物学特性及其表达模式  

Molecular characterization and expression profiling of sulfakinin in Bactrocera dorsalis

作　　者：栗洪飞 郑莉莎 石岩 王进军[1] 蒋红波[1] LI Hongfei;ZHENG Lisha;SHI Yan;WANG Jinjun;JIANG Hongbo(College of Plant Protection,Academy of Agricultural Science,Southwest University,Chongqing 400716,China)

机构地区：[1]西南大学植物保护学院西南大学农业科学研究院,重庆400716

出　　处：《生物安全学报》2018年第3期186-192,共7页Journal of biosafety

基　　金：国家自然科学基金面上项目(31572016)

摘　　要：【目的】分析橘小实蝇sulfakinin的时空表达模式及其对饥饿的响应,初步明确sulfakinin在调节橘小实蝇行为和生理方面的功能。【方法】利用RT-PCR技术克隆橘小实蝇神经肽sulfakinin的c DNA序列,采用实时定量PCR技术分析该基因在橘小实蝇不同发育阶段、成虫不同组织、幼虫不同组织及其在饥饿胁迫下的表达模式,并运用半定量RT-PCR技术进一步验证。【结果】实时定量PCR分析结果表明,sulfakinin在橘小实蝇的幼虫期和成虫期显著性高表达,卵期和蛹期几乎不表达。其中早期成虫表达量最高,约为晚期成虫的2倍,幼虫期表达量位于两者之间。在橘小实蝇幼虫、成虫不同组织中,sulfakinin在中枢神经系统的表达量显著高于其他组织,在触角的表达量仅次于中枢神经系统。此外,橘小实蝇在经饥饿处理24 h的过程中,sulfakinin的表达量均出现下调,其中从饥饿处理结果可知在6 h下降幅度最大,12 h和24 h表达量下降幅度减少。半定量RT-PCR技术进一步检测结果与实时定量PCR技术检测结果一致。【结论】sulfakinin在橘小实蝇中枢神经系统可能调节橘小实蝇成虫对饥饿胁迫的响应。其在触角中的高表达可能与橘小实蝇嗅觉的敏感性相关。本研究为进一步研究橘小实蝇sulfakinin的生理功能、评估其药靶潜力奠定了理论基础。[Aim] We investigated the temporal and spatial expression profiles of sulfakinin in Bactrocera dorsalis, as well as its response to starvation stress, to elucidate its vital functions in this fly＇s physiology and behavior.[Method] The cDNA of the B. dorsalis sulfakinin （Bacsulfakinin） was cloned by RT-PCR. QRT-PCR （real-time quantitative PCR）was employed to determine its expression patterns in different developmental stages, different tissues as well as the corresponding expression profiles under the stress of starvation. All the QRT-PCR results were further confirmed by semi-quantitative RT-PCR.[Result] QRT-PCR results indicated that sulfakinin was highly expressed in larvae, early adults and late-adults, while has little expression in the eggs and pupae. The expression of sulfakinin was highest in early adults, which was about twice as in late adults, and the expression level of sulfakinin in larvae was between the early adults and late-adults. The expression of sulfakinin was remarkably higher in the CNS （Central Nervous System） than in other tissues in both larvae and adults. Interestingly, the expression of sulfakinin was expressed the highest in the antennae. In addition, in the process of starvation for 24 h, the expression of sulfakinin decreased at all three time points tested which induced the fruit fly without food and water for 6, 12 and 24 hours. The expression level of sulfakinin decreased the most by 6 h from the starvation stress results.[Conclusion] Sulfakinin is possibly modulating the response of the oriental fruit fly upon starvation. The high expression level of sulfakinin in antenna may be related to olfactory sensitivity of the oriental fruit fly. This study laid a solid foundation for future studies of the physiological functions and assessment of target potential for sulfakinin.

关 键 词：橘小实蝇 神经肽 SULFAKININ 表达模式 分子克隆 

分 类 号：S433[农业科学—农业昆虫与害虫防治]