药材甲几丁质脱乙酰酶家族基因的鉴定及其生理功能分析

项目名称： 药材甲几丁质脱乙酰酶家族基因的鉴定及其生理功能分析

项目编号： No.31501649

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 杨文佳

作者单位： 贵阳学院

项目金额： 22万元

中文摘要： 几丁质是昆虫表皮和中肠围食膜的主要成分，昆虫的生长发育依赖于体内几丁质合成和降解的精确控制。几丁质脱乙酰酶（Chitin deacetylases，CDAs）是参与几丁质降解的关键酶，调节昆虫的行为、变态发育及生殖等。由于几丁质不存在于高等动物中，故昆虫CDAs作为靶标研发新型杀虫剂备受关注，具有重要的研究价值。因此，本研究以中药材储藏期主要害虫药材甲为研究对象，在转录组测序基础上克隆CDA家族基因，利用qPCR技术对CDAs基因的时空表达谱以及应对胁迫的差异表达调控效应进行分析，并利用原位杂交确定其组织的基因定位，采用RNAi技术明确其在昆虫生长发育中的作用，同时结合关键基因体外表达产物的生化特性来揭示CDAs在药材甲体内的重要生理功能。研究结果将丰富我们对昆虫CDAs功能的认识，促进理解药材甲及其它甲虫种类变态、发育、繁殖的调控机制，为新型环境友好杀虫剂的创制提供理论依据。

中文关键词： 药材甲；几丁质脱乙酰酶；RNA干扰；基因表达；功能分析

英文摘要： Chitin is the main component of the integument and peritrophic matrix of the midgut in insects. The growth and development of insect reliances on rigorously controlling the synthesis and degradation of chitin. Chitin deacetylases (CDAs) are essential enzymes involved in the processes of chitin degradation. It has been documented that CDAs are responsible for numerous important physiological processes in many insects, such as regulating the behavior, molting, metamorphosis and reproduction. Due to chitin is not present in vertebrates, the insect chitin deacetylase has been paid more attention because of its potential as specific target for developing novel insecticides in pest control. Therefore, this study will be foucsing on Stegobium paniceum, a major insect pest in stored Chinese medicine material, cloning several full-length cDNA sequences of CDA family genes based on the obtained transcriptome database; detecting the expression patterns of CDA family genes by qPCR in different developmental stages and tissues, as well as in response to different stress; determining subcellular localization of those genes by in situ hybridization; elucidating the specific function of each gene during insect growth and development by RNAi. Meanwhile, the biochemical characteristics of their expression products in vitro will also be taken into consideration. The results will expand our knowledge on the functions of insect CDAs, facilitate our understanding of the development, reproduction and their regulation mechanisms in beetles, and provide basic information for the development of novel eco-friendly insecticides.

英文关键词： Stegobium paniceum;Chitin deacetylase;RNA interference;Gene expression;Functional analysis