绿僵菌微循环产孢相关基因MaAGA、Pyk、PP5的克隆和功能分析-生物学专业毕业论文.docxVIP

重庆大学硕士学位论文中文摘要 重庆大学硕士学位论文 中文摘要 PAGE PAGE IV 种形态，而野生菌株只有椭圆状孢子产生。在菌落大小上，干扰菌株与野生菌株 并无显著性差异产生，但是干扰菌株的周边具有更多的白色菌丝。 （三）PP5 基因的克隆及其在两种产孢模式中的表达特征分析 1．克隆了 PP5 基因的 DNA 及 cDNA（Genebank 登录号：HQ317137），并 进行了生物信息分析。PP5 基因 DNA 长 2100 bp，含有 7 个外显子和 6 个内含子， 开放阅读框长 1428 bp。该基因编码蛋白大小为 38.1 KDa，等电点为 9.66，为碱性 蛋白，无信号肽序列。 2．PP5 编码产物蛋白结构分析。PP5 编码产物在一级结构上显示了第五类 Ser/Thr 保守的结构区域，如 TRP 序列及催化结构域，与所选 5 个亲近物种也具有 较高的相似性（73%-82%）。在二级结构上约含 46.32%的 α 螺旋， 7.16%的 β 转角， 14.74%的延伸串，31.79%的不规则卷曲。Swiss-Model 进行三级结构同源建模显示 该酶的核心结构类型为 α/β 型，Procheck 对建模结果可信度进行检测，算得出 Ramachandran 图，证实了该建模结果具有较强的可信度。 3．定量 PCR 检测该基因在两种产孢模式中的表达特征。结果显示，PP5 基 因在绿僵菌微循环产孢的不同阶段表达水平具有显著性差异， 特别是在孢子接种 16、24、32 小时高表达，而在正常产孢模式下表达量非常少。 关键词：绿僵菌，微循环产孢，RNAi 重庆大学硕士学位论文英文摘要 重庆大学硕士学位论文 英文摘要 ABSTRACT Metarhizium anisopliae is a significant filamentous pathogenic fungus, because it infects ca. 200 insect species from more than 50 taxonomic families, including termites, grasshoppers, thrips and mosquitoes. The mechanisms by which the insect pathogenic fungus M. anisopliae infects the host insect are relatively well understood. Conidiation, as the efficient propagule used in biocontrol programs, is a key step for survival and transmission in the disease cycle. Just like two representative conidiation elucidated in detail for two model organisms: Neuropora crassa and Aspergillus nidulans, our previous work found that M. anisopliae also exhibited two different conidiation programs: normal conidiation on non-inducing medium and microcycle conidiation on inducing medium. Compared with normal conidiation, microcycle conidiation showed remarkable advantages in the speed of conidia production and resistance to stress factors. Therefore, better understanding of the molecular mechanism of microcycle conidiation would improve conidial commercial production of M. anisopliae, which would greatly facilitate expansion of the scope of insect fungal pathogen applications in pest control. Previously, genes preferentially expressed during microcycle conidiation vs