摘要
为研究B类Ⅰ型清道夫受体(SCARB1/SR-BI)在中华绒螯蟹代谢、生长以及免疫等生物学过程中的分子功能,实验对中华绒螯蟹Scarb1的克隆及时空表达进行了分析,观察了RNA干扰该基因后的相关组织结构和类胡萝卜素含量变化,筛选了该基因的SNP标记并与生长相关性状进行关联分析。结果显示,中华绒螯蟹的Scarb1由2个外显子和1个内含子组成,开放阅读框为2415 bp,编码805个氨基酸;系统进化分析显示中华绒螯蟹与三疣梭子蟹的氨基酸同源性高达80.51%,具有较高的物种间保守性。该基因在不同蜕壳时期的肝胰腺、肠道、血淋巴细胞、心脏、鳃和肌肉组织中均有表达,但在肝胰腺、肠道和血淋巴细胞中的表达量相对较高。RNA干扰Scarb1后,组织切片观察发现肝胰腺的肝小管管腔模糊并产生了部分空泡化结构,肠道内膜的肌肉层和黏膜下层处出现显著空洞现象;肝胰腺的颜色由黄色变成灰白色,其中的类胡萝卜素含量显著下降。在该基因的第一外显子筛选到1个SNP位点(C432T)与蜕壳后体重和壳长增长率存在显著相关性。本研究结果为Scarb1在中华绒螯蟹的遗传研究和育种应用提供参考。
In order to study the functions of class B type I scavenger receptor(SR-B1/SCARB1)in the metabolism,growth,immunity and other biological processes of Eriocheir sinensis,the cloning and spatiotemporal expression analysis of scarb1 gene in E.sinensis were analyzed,the changes of related tissue structure and carotenoid content after RNA interference were observed,the SNP markers of the gene were screened and their association with the growth related traits were evaluated.The results showed that the scarb1 gene in E.sinensis was composed of the two exons and one intron.The open reading frame(ORF)was 2415 bp and 805 amino acids were encoded;phylogenetic tree based on the amino acid sequences showed that the highest similarity with 80.51%were found between E.sinensis and Portunus trituberculatus,indicating its high conservation among species.Its expression was detected in all tissues encoded,including hepatopancreas,intestines,blood cells,heart,gills and muscle tissues at different molting stages,but the relatively high expression existed in hepatopancreas,intestines and blood cells.After RNA interference in scarb1 gene for 15 days and 30 days,histological observation showed that the lumen of the hepatic tubules in hepatopancreas was blurred and partially vacuolized,and significant cavities appeared in the muscular layer and submucosa of the intestinal intima.The color of hepatopancreas changed from yellow to gray white,and the content of carotenoids decreased significantly.One SNP locus(C432T)was screened in the first exon of the gene,which was significantly correlated with the weight gain rate and carapace length growth rate after molting(P<0.05).The results of this study provide useful guidance for genetic research of scarb1 gene in E.sinensis and its breeding.
作者
姜鹏飞
侯鑫
王军
陈晓雯
王成辉
JIANG Pengfei;HOU Xin;WANG Jun;CHEN Xiaowen;WANG Chenghui(Key Laboratory of Freshwater Aquatic Genetic Resources certificated by the Ministry of Agriculture and Rural Affairs/National Demonstration Centre for Experimental Fisheries Science Education/Shanghai Engineering Research Center of Aquaculture,Shanghai Ocean University,Shanghai 201306,China)
出处
《水产学报》
CAS
CSCD
北大核心
2023年第1期173-182,共10页
Journal of Fisheries of China
基金
上海市中华绒螯蟹现代农业产业技术体系建设项目(沪农科产字2022-4号)。
