中华稻蝗几丁质脱乙酰基酶1基因的分子特性及功能  

Molecular characterization and functional analysis of chitin deacetylase 1 gene in Oxya chinensis (Orthoptera: Acrididae)

作　　者：丁国伟[1,2] 于荣荣[1,2] 杨美玲[1,2] 马恩波[1,2] 杨静[2,3] 张建珍[1,2] 

机构地区：[1]山西大学应用生物学研究所,太原030006 [2]农业有害生物综合治理山西省重点实验室,太原030006 [3]山西省农业科学院植物保护研究所,太原030031

出　　处：《昆虫学报》2014年第11期1265-1271,共7页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(31272380);教育部博士点专项基金(博导类)(20121401110008)

摘　　要：【目的】通过研究中华稻蝗Oxya chinensis几丁质脱乙酰基酶1(chitin deacetylase 1,CDA1)基因的分子特性和生物学功能,为新型农药靶标筛选提供理论基础。【方法】在中华稻蝗转录组数据库搜索获得几丁质脱乙酰基酶基因片段,用生物信息学方法对其进行同源比对分析并作聚类关系图;利用c DNA末端快速扩增(rapidamplification of c DNA ends,RACE)获得该基因c DNA全长序列;通过Real-time quantitative PCR(q PCR)方法检测Oc CDA1在5龄若虫不同组织部位和不同发育时期的表达情况,并采用RNAi技术研究Oc CDA1在飞蝗生长发育中的功能。【结果】获得中华稻蝗几丁质脱乙酰基酶1基因全长c DNA,命名为Oc CDA1(Gen Bank登录号:KP271171)。Oc CDA1在前肠、体壁、后肠和脂肪体组织中高表达,在5龄第1天表达量显著高于以后几天。RNAi结果显示,注射该基因的dsRNA后,Oc CDA1基因的表达量降低了93.3%。Oc CDA1基因沉默后虫体出现进食减少、发育迟缓现象,总死亡率达到95.2%,其中38.1%的试虫在蜕皮前死亡,57.1%出现因蜕皮困难而死亡的表型。【结论】Oc CDA1在中华稻蝗的发育中起着重要作用,该基因沉默引起虫体进食减少和蜕皮异常从而导致死亡。Abstract： [ Aim ] Molecular characterization and functional analysis of chitin deacetylase 1 gene in Oxya chinensis were conducted so as to provide the insight into novel target identification for pest management. [ Methods ] CDA-like gene fragments were obtained from an O. chinensis transeriptome database and the sequences of these fragments were analyzed by bioinformatics and homology analysis methods. A full- length eDNA sequence of OcCDA1 was obtained by rapid-amplification of eDNA ends （RACE）. Real- time quantitative PCR （qPCR） was applied to detect the relative expression levels of OcCDA1 in different tissues of the 5th instar nymphs and at different developmental stages. RNA interference was performed to study the function of OcCDA1 during the growth and development of O. chinensis. [ Results ] The full- length eDNA sequence of chitin deacetylase 1 gene in O. chinensis was cloned and named as OcCDAl （GenBank accession number： KP271171） . Gene expression profiling in tissues showed that OcCDAlwas highly expressed in foregut, integument, hindgut and fat body, and the expression level of OcCDA1 was the highest in the integument of the 1st day of the 5th instar nymphs. After the 5th instar nymphs were injected with OcCDAl dsRNA, the relative expression/eve/of OcCDAl in the nymphs decreased by 93.3% as compared with that in the control （injected with dsGFP）. Significantly down-regulated expression of OcCDAl resulted in reduced feeding, slow development, and high mortality （95.2%） including 38.1% nymphs died before molting and 57.1% nymphs failed to detach from the old cuticle during molting process and died eventually. [ Conclusion] The results demonstrate that OcCDAl plays a key role in molting process of O. chinensis, and silencing OcCDAl may result in high mortality with feeding reduction or molting defect phenotype.

关 键 词：中华稻蝗 几丁质脱乙酰基酶1 基因表达 功能分析 RNA干扰 

分 类 号：Q966[生物学—昆虫学]