氨氮胁迫下凡纳滨对虾血细胞的凋亡通路研究

项目名称： 氨氮胁迫下凡纳滨对虾血细胞的凋亡通路研究

项目编号： No.31302164

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 冼健安

作者单位： 华南师范大学

项目金额： 25万元

中文摘要： 氨氮是虾类养殖过程中最常见的环境胁迫因子，是造成虾类免疫功能低下、甚至大量发病和死亡的主要原因之一。本项目组前期的研究发现，氨氮胁迫下凡纳滨对虾血细胞的活性氧含量显著上升，随后血细胞发生显著的凋亡，我们推测虾类可能通过血细胞凋亡来清除受到过度氧化损伤的血细胞，这可能是虾类抗氨氮胁迫的生理调节保护策略。本项目拟针对此前期发现，深入研究氨氮胁迫下凡纳滨对虾血细胞凋亡的调控机制。先确定血细胞凋亡与氨氮浓度和胁迫时间的关系；然后应用流式细胞术分析血细胞各类凋亡事件的发生；另一方面，应用RNA高通量测序技术构建氨氮胁迫的转录组和表达谱文库，比对筛选差异基因并分析凋亡相关基因的表达变化，综合细胞和分子水平的研究结果阐明氨氮胁迫下血细胞的凋亡通路。本研究成果对深入研究虾类抗胁迫防御机理具有重要的理论意义，对虾类养殖业的健康发展具有重要的现实意义！

中文关键词： 氨氮；凡纳滨对虾；血细胞；凋亡；环境胁迫

英文摘要： Ammonia is a common environmental stress factor during the culture of shrimp. It is one of the main reasons for reduced immunity,disease and death of shrimp. Our previous study found that reactive oxygen species (ROS) production of haemocyte and haemocyte apoptosis ratio significantly increased in Litopenaeus vannamei exposed to ammonia. We hypothesize that shrimp may eliminate the serious damaged haemocyte by cell apoptosis. This process may be a protection strategy via physiological accommodation for ammonia stress. The present project will investigate the regulatory mechanism of haemocyte apoptosis in shrimp L. vannamei exposed to ammonia-N, according to our previous finding. Firstly, we will analyze the relationship between haemocyte apoptosis and ammonia dose/exposure time. Then we will detemine the occurrence of various apoptotic events by flow cytometry. On the other hand, the cDNA ibrary and digital gene expression library in haemocyte of ammonia-exposed shrimp will be constructed using the next generation sequecing technology RNA-seq. Then the genes with changed expression levels will be identified, and the expression levels of apoptosis-related genes will be analyzed. The differences of genes expression during the occurrence of apoptosis will be analyzed. The apoptosis pathway of haemocyte in ammonia-e

英文关键词： ammonia；Litopenaeus vannamei；haemocyte；apoptosis；environmental stress