扇贝异源三倍体诱导  

Induction of scallop allotriploid

作　　者：孟庆磊[1] 黄晓婷[1] 赵海波[1] 赵婷[2] 李宁[1] 王昭萍[2] 胡晓丽[1] 胡景杰[1] 包振民[1] 

机构地区：[1]中国海洋大学海洋生命学院,山东青岛266003 [2]中国海洋大学水产学院,山东青岛266003

出　　处：《中国水产科学》2009年第5期718-727,共10页Journal of Fishery Sciences of China

基　　金：国家863计划项目(2006AA10A408);农业部公益性行业科研专项(nyhyzx07-047);国家支撑计划项目(2006BAD01A13);国家科技基础条件平台建设项目(2005DKA30470)

摘　　要：荧光显微观察表明,20℃水温下,栉孔扇贝(Chlamys farreri)的卵与海湾扇贝(Argopecten irradians)的精子可以正常受精和发育,具备人工诱导三倍体的可行性。亲贝充分促熟后,分开催产,以20:1的精卵比授精;在50%的受精卵排出第1极体时,以60mg/L6-二甲基氨基嘌呤(6-DMAP)处理受精卵10-25min,可诱导75.23%-92.14%的三倍体;6-DMAP处理15min综合诱导效果最好,三倍体诱导率可达88.56%,孵化率可达53.52%。得到的三倍体幼虫经基因组原位杂交(Genomic in situ hybridization,GISH)验证,为含有2套栉孔扇贝染色体组和1套海湾扇贝染色体组的异源三倍体。孵化后诱导组与杂交对照组(未经6-DMAP处理)幼虫生长越来越缓慢,受精后14d其幼虫存活率分别下降到0.00067%和0.00224%,没有幼虫度过附着变态期。GISH分析显示,栉孔扇贝与海湾扇贝不同倍性的杂种后代早在担轮幼虫阶段就出现母本偏向性染色体转变。Induction of triploidy is of great importance in mollusk breeding,and can be modified further with interspecific hybridization techniques to create allotriploids which may benefit both from triploidization and hybridization. Prior to the allotriploid induction,the feasibility of fertilization between Zhikong scallop Chlamys farreri eggs and Bay scallop Argopecten irradians sperm was verified by epifluorescence microscopy. Parental scallops were fully induced mature separately and induced spawning. Insemination was carried out with the ratio of sperms and eggs being 20 ： 1. Triploidy rates of D-stage larvae ranging from 75.23% to 92.14%, were obtained by inhibiting the second polar body with 60 mg/L 6-deimethylaminopurine （6-DMAP） for 10,15,20 and 25 rain durations when 50% zygotes released the first polar body in 20℃ seawater. An optimum effect with triploidy rate of 88.56% and incubation rate of 53.52% was gained under 15 rain duration. With genomic in situ hybridization （GISH）method, the consequent larvae were validated to be real interspecies hybrids from C. farreri x A. irradians cross with two C. farreri genome and one A. irradiams genome. Overall, allotriploid scallop can be readily produced, but no statistical advantages of growth or survival was found in triploid induction groups compared with controls during larvae development. Larva survival rate of triploid and diploid hybrids were 0.000 67% and 0.002 24%, at the 14 th day after insemination. All larvae died before metamorphism without forming eyespot or foot. Furthermore, maternal biased chromosome conversion was revealed by GISH in both triploid and diploid hybrids, as early as at trophore-larvae stage.

关 键 词：扇贝 异源三倍体 基因组原位杂交 染色体转变 

分 类 号：S91[农业科学—水产科学]