异源精子诱导条斑星鲽雌核发育  

Study on gynogenesis induced by heterogenous sperms in barfin flounder Verasper moseri

作　　者：杨景峰[1,2] 陈松林[2] 苏鹏志[2] 邵长伟[2] 田永胜[2] 廖小林[2] 翟介明 李波 

机构地区：[1]中国海洋大学海洋生命学院,山东青岛266003 [2]中国水产科学研究院黄海水产研究所农业部海洋渔业资源可持续利用重点开放实验室,山东青岛266071 [3]山东莱州明波水产有限公司,山东莱州266031

出　　处：《水产学报》2009年第3期372-378,共7页Journal of Fisheries of China

基　　金：国家"八六三"高技术研究发展计划(2006AA10A403);山东省泰山学者工程专项

摘　　要：用冷冻保存的花鲈精子作为异源精子，采用紫外线（UV）对精子进行照射使其遗传物质失活，然后与卵子进行授精，可以刺激条斑星鲽鱼卵进行雌核发育，在受精后一定时间采用冷休克处理“受精”卵，抑制第二极体排出成功获得了条斑星鲽雌核发育二倍体鱼苗。实验表明，花鲈精子只有经过紫外线照射遗传失活后，才能诱导产生条斑星鲽雌核发育二倍体鱼苗。经过大量实验筛选出诱导条斑星鲽雌核发育的最佳条件为花鲈冷冻精子采用80 MJ/cm2 的紫外线照射，然后与条斑星鲽卵子进行授精，在受精后7~9 min，将受精卵放在-1.0~1.5 ℃海水中进行冷休克处理，持续时间为60~90 min，在此条件下，获得的雌核发育二倍体的相对诱导率达40.68%±7.24%。由于不失活精子与条斑星鲽卵形成的杂交胚只能存活到原肠期，而染色体未被成功加倍的胚胎会由于单倍体的致死效应在孵化前后死亡，所以存活的仔鱼全部为条斑星鲽雌核发育二倍体。采用微卫星标记技术对雌核发育鱼苗进行了分析，证明了雌核发育鱼苗为雌核发育二倍体。首次报道了采用异源冷冻精子诱导条斑星鲽鱼卵进行雌核发育的技术方法，为条斑星鲽性别控制和遗传改良提供了技术手段。Fish in which gynogenesis has been induced have all their chromosomes inherited from the mother and, if females are the homogametic sex, they usually are all females. Because barfin flounder Verasper moseri females grow faster than males, the production of allfemale populations is highly desirable. Due to the low volumes of semen produced by male flounder, and to eliminate any potential genetic contribution by homologous sperm, activation of flounder eggs with heterologous sperm was also necessary. To test methods for inducing diploid gynogenesis in barfin flounder using homologous sperm, the cryopreservative sperm of Lateolabrax japonicas was used to activate eggs and cool shock was used to prevent extrusion of the second polar body. Four treatments, named for their expected outcome, were employed： hybrid, haploid, triploid, and gynogenetic diploid to prove the ability of Lateolabrax japonicas sperm activating flounder eggs. Diploid gynogenesis was induced by activating egg development with UV irradiated sperm （80 MJ/cm2） for 7-9 min in seawater, and then subjecting the eggs to cold shock in -1-1.5 ℃ seawater for 60-90 min. The hybrid of could not survive to hatching, thus the offspring was allogynogenesis only and the result of microsatellite markers analysis proved this result. This work provides procedures for induction of diploid gynogenesis in barfin flounder using heterologous sperm.

关 键 词：条斑星鲽 花鲈 雌核发育 异源冷冻精子 微卫星 

分 类 号：Q343.34[生物学—遗传学] S917[农业科学—水产科学]