中文摘要：

【目的】海藻糖酶（trehalase,Tre）是昆虫体内海藻糖代谢的一个关键酶,包括可溶型（Tre1）和膜结合型（Tre2）两种类型,在昆虫发育和能量调节中具有重要作用。本研究旨在克隆稻纵卷叶螟Cnaphalocrocis medinalis海藻糖酶基因（Cm Tre）,解析其在稻纵卷叶螟不同发育阶段和不同组织中的表达模式,分析该基因及酶蛋白的分子特征。【方法】通过稻纵卷叶螟转录组数据结合RACE技术,克隆稻纵卷叶螟Cm Tre的全长c DNA序列,并对该基因进行生物信息学分析;采用实时荧光定量PCR（RT-q PCR）解析Cm Tre在稻纵卷叶螟不同发育阶段及成虫不同组织部位的m RNA表达模式。【结果】获得两种类型的稻纵卷叶螟Cm Tre基因,即可溶型海藻糖酶基因Cm Tre1和膜结合型海藻糖酶基因Cm Tre2。Cm Tre1的全长c DNA长度为2 364 bp,开放阅读框长1 704 bp,编码567个氨基酸;Cm Tre2的全长c DNA长度为2 079 bp,开放阅读框长1 923 bp,编码640个氨基酸。生物信息学分析表明,Cm Tre前端有一个信号肽,Cm Tre1无跨膜结构,Cm Tre2有一个跨膜结构。同源性和聚类分析表明,Cm Tre1和Cm Tre2的氨基酸序列与竹蠹螟Omphisa fuscidentalis海藻糖酶Tre1和Tre2氨基酸序列的一致性最高,分别为74%和79%。同源建模预测结果显示,Cm Tre1的三维分子结构包含19个α螺旋和2个β折叠片;Cm Tre2的三维分子结构含有23个α螺旋,没有β折叠片。RT-q PCR结果显示,Cm Tre在稻纵卷叶螟整个发育历期都有表达,在成虫期表达水平最高,在整个幼虫期都有相对稳定的表达;Cm Tre1在蛹期表达水平最低,Cm Tre2在5龄幼虫时期表达水平最低。Cm Tre在所检测的成虫组织（中肠、体壁、马氏管、头、卵巢、脂肪体、肌肉和精巢）中均有表达;Cm Tre1在中肠和体壁中的表达水平较高,Cm Tre2在肌肉和中肠中的表达水平较高。【结论】本研究克隆了稻纵卷叶螟两种类型的海藻糖酶基因,分析了其

英文摘要：

【Aim 】 Trehalase（ Tre） is a key enzyme in trehalose metabolism of insects and plays important roles in the development and regulation of energy. Insects possess two types of trehalases,i. e.,soluble trehalase（ Tre1） and membrane-bound trehalase（ Tre2）. This study aims to clone trehalase gene（ Cm Tre） from the rice leaf folder,Cnaphalocrocis medinalis,to clarify its expression patterns in different tissues and developmental stages,and to analyze the molecular characteristics of the two types of the gene and their products. 【Methods】Based on the transcriptome data of C. medinalis,the full-length c DNA of Cm Tre was cloned using the rapid amplification of c DNA ends（ RACE）-PCR and analyzed by bioinformatics. Cm Tre m RNA expression levels in different tissues of adults and developmental stages of C. medinalis were detected by using real-time quantitative PCR（ RT-q PCR）. 【Results】We cloned two types of Cm Tre,i. e.,soluble trehalase gene Cm Tre1 and membrane-bound trehalase gene Cm Tre2. The full-length c DNA of Cm Tre1 is 2 364 bp containing a 1 704 bp open reading frame（ ORF） that encodes567 amino acids,while that of Cm Tre2 is 2 079 bp containing a 1 923 bp ORF that encodes 640 amino acids. Bioinformatics analysis indicated that Cm Tre includes a signal peptide and Cm Tre1 has no transmembrane domain,whereas Cm Tre2 contains a transmembrane domain. Homology and phylogenetic analyses showed that the amino acid sequences of Cm Tre1 and Cm Tre2 have the highest identities with those of Tre1 and Tre2 from Omphisa fuscidentalis,which are 74% and 79%,respectively. Homology modeling analysis demonstrated that the tertiary structure of Cm Tre1 is composed of 19 α-helices and 2 β-sheets while that of Cm Tre2 is composed of 23 α-helices and without β-sheet. RT-q PCR revealed that Cm Tre was expressed throughout all developmental stages of C. medinalis,with the highest expression level in adult and a relatively stable expression level in larval stage. Cm Tre1 was expressed at the l