飞蝗dpp基因在翅发育中的功能分析  

Functional Analysis of dpp Gene in Locust Wing Development

作　　者：武兆辰 张福斌 徐子龙 张晶 张建珍[2] 赵小明[2] WU Zhaochen;ZHANG Fubin;XU Zilong;ZHANG Jing;ZHANG Jianzhen;ZHAO Xiaoming(School of Life Sciences,Shanxi University,Taiyuan 030006,China;Institute of Applied Biology,Shanxi University,Taiyuan 030006,China)

机构地区：[1]山西大学生命科学学院,山西太原030006 [2]山西大学应用生物学研究所,山西太原030006

出　　处：《山西大学学报（自然科学版）》2022年第6期1632-1639,共8页Journal of Shanxi University(Natural Science Edition)

基　　金：山西大学第十八期本科生科研训练项目资助(2020018381);国家自然科学基金(31970469,31702067)。

摘　　要：Dpp(Decapentaplegic)是脊椎动物和无脊椎动物中非常重要且高度保守的一个信号通路,在细胞的生长和分化等方面发挥调控作用。在完全变态昆虫中,Dpp调控翅的生长发育,但对渐变态昆虫翅发育的作用机制尚不清楚。文章以飞蝗(Locusta migratoria)为研究对象,根据同源性搜索识别飞蝗dpp基因(Lmdpp),并对其编码的氨基酸序列与不同昆虫Dpp进行多序列比对。然后,利用RT-q PCR方法对Lmdpp在五龄飞蝗不同组织及不同发育天数的翅芽进行表达特性分析,结果显示Lmdpp在五龄每天的翅芽组织中均有表达,且在多个组织中高表达。利用RNA干扰技术探讨Lmdpp在飞蝗翅发育中的作用,结果显示,与对照组相比,注射Lmdpp基因的dsRNA(doublestranded RNA,dsRNA)的处理组约有31%的飞蝗在蜕皮后出现翅紊乱表型。苏木精-伊红(Hematoxylin-eosin,H&E)染色结果表明,沉默Lmdpp后的翅新表皮形成异常;RT-q PCR结果显示,沉默Lmdpp后导致飞蝗翅特异表皮蛋白基因LmAcp7、LmAcp8和LmAcp19(Adult cuticle protein,Acp)的表达量显著降低。结果表明,Lmdpp可能通过调控飞蝗翅特异表皮蛋白基因的表达参与翅的发育与形成,研究结果可为dpp在渐变态昆虫翅发育中的作用机制研究提供基础。Dpp(Decapentaplegic)pathway is a very important and highly conserved signaling pathway in both vertebrates and invertebrates,which plays a regulatory role in cell growth and differentiation.Dpp regulates wing development in complete metamorphosis insects,but its effect on wing development of incomplete metamorphosis insects remains unclear.In this study,taking Locusta migratoria as the research object,Lmdpp gene was identified based on homology search,and the amino acid sequence encoded by Lmdpp was aligned with that of different insects.Then,RT-qPCR method was used to analyze the expression of Lmdpp in different tissues and different development days of the fifth instar nymphs,and the results showed that Lmdpp was expressed in wing pads on each day of the fifth instar,and was highly expressed in several tissues tested.RNA interference technique was used to investigate the role of Lmdpp in the wing development of L.migratoria.The results showed that compared with the control group,about 31%locusts in the treatment group injected with dsRNA(double-stranded RNA,dsRNA)of Lmdpp showed abnormal wing phenotype after molting.The Hematoxylin-eosin(H&E)staining showed that the new epidermis of wings formed abnormally after silencing Lmdpp.The RT-qPCR results showed that the expression of wing specific cuticle protein genes LmAcp7,LmAcp8 and LmAcp19(Adult cuticle protein,Acp)were significantly decreased after silencing Lmdpp.These results suggest that Lmdpp may be involved in wing development and formation by regulating the expression of wing specific cuticle protein genes of locusts,providing a basis for the study of the mechanism of Dpp in wing development of incomplete metamorphosis insects.

关 键 词：飞蝗 翅发育 DECAPENTAPLEGIC RNA干扰 

分 类 号：O436[机械工程—光学工程]