泰国斗鱼色氨酸羟化酶基因cDNA克隆及其在雄鱼好斗性中的作用  

cDNA Cloning and Expression of Tryptophan Hydroxylase Gene in Betta Splendens and Role of the Gene in Male Aggressiveness

作　　者：陈勇智 郑银娟 马细兰 CHEN Yongzhi;ZHENG Yinjuan;MA Xilan(School of Life Science,Huizhou University,Huizhou Guangdong 516007,China)

机构地区：[1]惠州学院生命科学学院,广东惠州516007

出　　处：《海南热带海洋学院学报》2022年第5期29-35,共7页Journal of Hainan Tropical Ocean University

基　　金：惠州学院科研创新群体培育项目(HZU201807)。

摘　　要：雄性泰国斗鱼(Betta splendens)表现出极强的好斗性,是研究鱼类好斗性的极佳模型生物。首先通过RT-PCR从泰国斗鱼的脑组织中克隆并鉴定出色氨酸羟化酶基因(Tryptophan hydroxylase gene,tph)的部分cDNA片段,进行进化树分析;然后通过配对斗争法和镜像斗争法观察记录泰国雄性斗鱼的打斗行为;最后用荧光实时定量PCR技术比较斗争前后个体或优势、劣势个体之间的tph mRNA表达差异。研究结果如下:(1)克隆得到253 bp泰国斗鱼tph部分核苷酸序列,编码84个氨基酸,与同属于鲈形目的军曹鱼、大黄鱼等的同源性高达97.59%,泰国斗鱼与哺乳纲、爬行纲的亲缘关系由近至远;(2)当把2尾雄鱼放入同一容器时,两者立即相互撕咬、撞击、追逐,直至分出胜负;当把镜子放入有1尾雄性斗鱼的容器后,斗鱼立即对镜像表现出张鳃、咬镜、撞镜等好斗性行为;(3)荧光实时定量PCR结果显示,斗争后个体tph的mRNA表达水平显著高于斗争前个体水平(P<0.05),优势个体tph的表达水平显著高于劣势个体水平(P<0.01)。由此推测泰国斗鱼tph与其好斗性相关。Male Betta splendens,which show strong aggressiveness, are an excellent model organism for studying fish aggressiveness.First, some cDNA fragments of tryptophan hydroxylase gene(tph) were cloned and identified from the brain tissue of Betta splendens by RT-PCR for evolutionary tree analysis.Then, the fighting behaviors of male Betta splendens were observed and recorded by paired struggle method and mirror struggle method.Finally, the differences of tph mRNA expression between individuals before and after the struggle or between superior and inferior individuals were compared by fluorescence real-time quantitative PCR.The results are as follows:(1) The 253 bp tph partial nucleotide sequence of Betta splendens,encoding 84 amino acids, was cloned and showed 97.59% homology with Rachycentron canadum and Larimichthys crocea.The tph of Betta splendens is related to mammalia and Reptilia from near to far.(2) When two male fish were put into the same container, they immediately bit, collided with and chased each other until the winner was determined;When the mirror was put into the container with a male Betta splendens,the Betta splendens immediately showed aggressive behavior towards the mirror, such as gill opening, biting and hitting the mirror and so on.(3) The results of fluorescence real-time quantitative PCR showed that the mRNA expression level of tph in individuals after struggle was significantly higher than that before struggle(P<0.05),and the expression level of tph in dominant individuals was significantly higher than that in disadvantaged individuals(P<0.01).Therefore, it is speculated that tph of Betta splendens is involved in its aggressive regulation.

关 键 词：泰国斗鱼 色氨酸羟化酶基因 好斗性 基因表达 

分 类 号：S917.4[农业科学—水产科学]