剧毒鹅膏菌是毒蘑菇中毒事故的头号杀手，其所含环肽毒素既具强致死性，又有广阔应用前景，是医药业寻找新药物的重要资源。脯氨酰寡肽酶（POP）是鹅膏肽类毒素生物合成过程中一种关键酶，但目前对该酶基因的研究很少，尤其是对我国剧毒鹅膏菌相关酶基因还未见报道。本项目利用前期研究中致命鹅膏菌转录组测序获得的POP unigene基因序列设计引物，扩增和克隆我国剧毒鹅膏菌POP基因的全长cDNA和基因组DNA，解析该基因及其预测蛋白的序列特征，构建毒鹅膏菌与其他菌类、动植物等不同生物类群的POP系统进化树，探讨其进化关系；同时经DNA杂交研究POP基因的拷贝现象，以及在有毒和无毒鹅膏菌中的表达情况；并对该酶基因在同一剧毒鹅膏不同组织和生长时期的表达量进行分析，探索POP基因在子实体发育过程中的表达与毒素含量累积的关系。本研究将为突破鹅膏环肽毒素生物合成的科学难题奠定重要的分子生物学基础。

Lethal Amanita species, containing the potent toxic cyclopeptides, are responsible for the most mushroom poisonings, but which were also the important new drug resources for their broad application prospects in the life sciences. Prolyl oligopeptidase (POP) is known as the key enzyme in the biosynthesis of the Amanita toxins, however, few studies have been reported in this key enzyme, especially in the lethal Amanita species endemic in China. In this study, the primers will be designed basing on the POP unigene sequences to clone the full-length cDNA sequences of POP in those lethal Amanita species, then to analyze the characterizations of the cDNA and protein sequences, to construct the phylogenetic tree of the POP genes among the fungi, bacterials, animals and plants, and to investigate the phylogenetic relationships between the Amanita species and other species; the POP gene copy and expression patterns in the toxic and nontoxic Amanita species will be studied by DNA blotting; quantitative real-time PCR will be conducted to detect the transcription levels of the POP genes in the different parts and development stages of the lethal Amanita species, and to explore the correlation between the POP gene transcription levels and the Amanita toxins accumulation. This study will lay an important molecular foundation for breakthroughing the biosynthesis of Amanita toxin genes.