黄姑鱼三倍体的诱导、鉴定及其性腺发育特征  

THE INDUCTION AND IDENTIFICATION OF THE TRIPLOID YELLOW DRUM(NIBEA ALBIFLORA,SCIAENIDEA) AND CHARACTERIZATION OF THEIR GONADAL DEVELOPMENT

作　　者：卢磊 陈睿毅 杨阳 竺奇慧 胡伟华 李海东 徐冬冬[2,3] LU Lei;CHEN Rui-Yi;YANG Yang;ZHU Qi-Hui;HU Wei-Hua;LI Hai-Dong;XU Dong-Dong(College of Fisheries,Zhejiang Ocean University,Zhoushan 316022,China;Marine Fisheries Research Institute of Zhejiang,Zhoushan 316021,China;Key Laboratory of MaricuLture and Enhancement of Zhejiang Province,Zhoushan 316021,China)

机构地区：[1]浙江海洋大学水产学院,舟山316022 [2]浙江省海洋水产研究所,舟山316021 [3]浙江省海水增养殖重点实验室,舟山316021

出　　处：《水生生物学报》2023年第2期290-297,共8页Acta Hydrobiologica Sinica

基　　金：浙江省公益技术应用研究项目(LGN22C190018);国家自然科学基金(31972785);浙江省杰出青年基金(LR21C190001)资助。

摘　　要：研究采用冷休克方法诱导黄姑鱼(Nibea albiflora)三倍体并进行鉴定,进一步采用组织学和生殖相关基因表达分析比较了三倍体黄姑鱼的性腺发育特征。研究结果表明:(1)在受精后2.5min以3℃进行10min的冷休克处理,冷休克处理组的受精率和孵化率分别为(70.31±4.49)%和(21.5±6.63)%,其受精率和孵化率显著低于二倍体对照组;(2)经流式细胞仪倍性检测和染色体核型分析发现,三倍体的DNA含量为二倍体的1.5倍,染色体数目为72条,而二倍体的染色体数目为48条,三倍体的比例为100%;(3)三倍体的生殖腺指数显著低于二倍体,进一步通过组织切片观察发现三倍体的精巢和卵巢发育较二倍体滞后,在12月龄时,二倍体精巢和卵巢处于Ⅴ期,而三倍体精巢和卵巢分别处于Ⅲ期和Ⅰ期;(4)三倍体精巢的dmrt1和vasa基因及三倍体卵巢的cyp19a基因表达量均显著低于二倍体(P<0.05);三倍体卵巢的vasa基因表达量也比二倍体低(P>0.05)。综上结果表明:研究通过冷休克处理成功诱导了黄姑鱼三倍体;三倍体的性腺发育较二倍体滞后,育性明显降低。In this study, the cold shock treatment was used to induce triploid yellow drum(Nibea albiflora) and the ploidy was determined. The histology and gene expression analysis of reproduction-related genes were further used to characterize the gonadal development in triploid yellow drum. Firstly, the fertilized eggs were subjected to the cold shock treatment implemented at 3℃ for a period of 10 min commencing at post-fertilization 2.5min, and the fertilization rate and hatching rate in the cold shock treatment groups were(70.31±4.49)% and(21.5±6.63)%, respectively,which were significantly lower than that of diploid. Secondly, we found that the DNA relative content of triploid was1.5 times larger than that of diploid by flow cytometer analysis, and the number of the chromosome in the triploid was 72, whereas the number of the chromosome in the diploid was 48 shown by karyotype analysis. The rate of the triploid was 100% in the present study. Thirdly, the gonadosomatic indexes of the triploids were significantly lower than that of the diploids. Histological results showed that the gonadal development of the triploids was slower than that of the diploids. At the age of 12 months, the testis and ovary of the diploids were both at the stage Ⅴ, while the testis and ovary of the triploids were at stage Ⅲ and stage Ⅰ, respectively. Fourthly, the dmrt1 and vasa expression in the triploid testis and cyp19a expression in the triploid ovary were significantly lower than those in the diploid gonads(P<0.05), and the vasa expression in the triploid ovary was also lower than that in the diploids(P>0.05). The results indicated that the triploid yellow drum could be successfully induced by the cold shock treatment, and the gonadal development of the triploids was retarded. The results of present study will lay a basis for the application of the triploid in germ cell transplantation and the genetic improvement of yellow drum.

关 键 词：冷休克 染色体倍性 三倍体 性腺发育 黄姑鱼 

分 类 号：Q344.1[生物学—遗传学]