雌二醇刺激对菲律宾蛤仔Dmrt基因表达的影响  

Effect of estradiol stimulation on Dmrt gene expression in Manila clam Ruditapes philippinarum

作　　者：吴启迪 谭月 王靖天 谢欠影 霍忠明 方蕾 闫喜武 WU Qi-di;TAN Yue;WANG Jing-tian;XIE Qian-ying;HUO Zhong-ming;FANG Lei;YAN Xi-wu(College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China;College of Marine Technology and Environment, Dalian Ocean University, Dalian 116023, China;Engineering Research Center of Shellfish Culture and Breeding in Liaoning Province, Dalian 116023, China)

机构地区：[1]大连海洋大学水产与生命学院,辽宁大连116023 [2]大连海洋大学海洋科技与环境学院,辽宁大连116023 [3]辽宁省贝类良种繁育工程技术研究中心,辽宁大连116023

出　　处：《大连海洋大学学报》2019年第3期362-369,共8页Journal of Dalian Ocean University

基　　金：现代农业产业体系建设专项(CARS-49)

摘　　要：为研究菲律宾蛤仔Ruditapes philippinarum不同发育时期及雌二醇刺激下Dmrt基因组织表达的变化状况,在菲律宾蛤仔基因组数据中(未发表),运用BLASTe比对确定了菲律宾蛤仔的3个Dmrt基因,运用MEGA、expasy、SMART等软件进行基因结构和进化分析,根据系统发生树的聚类对3个Dmrt基因进行命名,分别为Dmrt3-like、Dmrt4-like-1、Dmrt4-like-2,并采用实时荧光定量PCR方法对Dmrt基因在菲律宾蛤仔中的时空表达及其对雌二醇处理的响应进行了研究。结果表明:菲律宾蛤仔Dmrt 3个基因的表达量从担轮幼虫到壳顶幼虫时期呈迅速增长趋势,其中Dmrt4-like-2在菲律宾蛤仔的不同发育时期均有表达;在未进行雌二醇激素处理的正常菲律宾蛤仔体内,Dmrt4-like-1和Dmrt4-like-2在鳃和外套膜中表达量最高,Dmrt3-like在鳃和内脏团中表达量较高,在外套膜和水管中不表达;短时期内(72 h)雌二醇浸泡处理使得这3个基因在菲律宾蛤仔性腺中的表达量均发生上调;长时期(60 d)雌二醇浸泡处理后,空白组的雄性性腺中3个基因的表达量皆高于雌性性腺,试验组雌雄同体性腺中的Dmrt4-like-1和Dmrt4-like-2表达量略高于雌性中的表达量(P>0.05),但显著低于雄性中的表达量(P<0.05)。研究表明,菲律宾蛤仔Dmrt基因家族参与性别分化、性腺发育过程。In the unpublished data of clam genome, three Dmrt(double-sex and mab-3 related transcription factor) genes playing an important role in the process of sexual differentiation, gametogenesis, gonad development, and connected with DNA by transcription factors were identified in Manila clam Ruditapes philippinarum by BLASTe comparison and genetic identification was carried out using MEGA, expasy, SMART and other analysis software to evaluate the evolutionary relationship. The Dmrt genes were known as Dmrt3-like, Dmrt4-like-1, and Dmrt4-like-2 by the clustering of genetic evolution tree and their spatiotemporal expressions and response to estradiol treatment were investigated in the Manila clam by qT-PCR. The results showed that there was an explosive increase in the expression levels of the three genes from trochophore to umbo larvae, especially expression of Dmrt4-like-2 in each developmental stage. In the control clam, there was the maximal expression levels of Dmrt4-like-1 and Dmrt4-like-2 in gills and mantle, while the maximal expression levels of Dmrt3-like was observed in gills and viscera, without expression in mantle and siphon. A short period of the up-regulated expression of these three genes was found in the gonads. In the long period of estradiol exposure, however, there were higher expression levels of the three genes in the male gonads than that in the female gonads in the blank group. In the experimental group, the expression levels of Dmrt4-like-1 and Dmrt4-like-2 in the androgynous gonads were found to be higher than that in female gonads and lower than that in male gonads. The findings idicated that Dmrt gene family was involved in sex differentiation and gonadal development, which provides reference for the research on the sex determination mechanism of Manila clam.

关 键 词：菲律宾蛤仔 雌二醇 性别分化 性别决定 DMRT基因 

分 类 号：S968.31[农业科学—水产养殖]