家蚕二化性品种(品系)转基因用蚕卵的预处理技术研究  

Studies on Pretreatment of Eggs From Bivoltine Silkworm Varieties(Strains) for Micro-injection Transgene

作　　者：唐顺明[1,2] 沈兴家[1,2] 赵巧玲[1,2] 章扬[2] 张国政[1,2] 郭锡杰[1,2] 

机构地区：[1]中国农业科学院蚕业研究所,农业部蚕桑遗传改良重点开放实验室,江苏镇江212018 [2]江苏科技大学,江苏镇江212003

出　　处：《蚕业科学》2009年第4期872-876,共5页ACTA SERICOLOGICA SINICA

基　　金：国家重点基础研究发展计划“973”项目(编号2005CB12-1004);国家高技术研究与发展计划“863”项目(编号2007AA100504);江苏省高校自然科学基金项目(编号07KJD230052)

摘　　要：家蚕为卵滞育昆虫,二化性家蚕品种（品系）的化性由上代卵期温度与光照等调控。将若干个二化性家蚕品种（品系）以1718℃温度催青诱导子代蚕卵非滞育,筛选出1个经济性状较优的实用品种＂秋丰＂、1个突变品系P33,二者非滞育卵圈比例分别达到96.4%和89.6%,正常催青的良卵实用孵化率分别达到96.11%和98.58%,解决了采用显微注射法以家蚕实用品种构建转基因系统过程中因蚕卵滞育不能及时孵化的难题。另对利用20%盐酸-2%甲醛混合液刺激解除蚕卵滞育的方法以及非滞育蚕卵表面用70%乙醇消毒的方法进行了尝试,前者可以获得96%以上的孵化率,后者可以缩短蚕卵预处理时间。The silkworm,Bombyx mori,is a species of insect with diapause during egg stage.The voltinism of bivoltine silkworm variety（strain） is regulated by temperature and illumination in its parental egg stage.From a number of bivoltine varieties（strains） of which eggs were induced to become non-diapause by control of temperature at 17～18 ℃ during egg incubation,one practical silkworm variety Qiufeng with excellent economic characters and one mutant strain P33 were selected.Their non-diapause egg batch percentages were 96.4% and 89.6%,and practical hatching abilities of good eggs under normal incubation condition were 96.11% and 98.58%,respectively.These results have resolved the difficulty that diapause eggs can not hatch directly after micro-injection in transgenic studies.In addition,the efficiency of dipping eggs in mixture of 20% hydrochloric acid and 2% formaldehyde solutions for activating diapause eggs and the effect of surface disinfection with 70% ethanol for non-diapause eggs before micro-injection were investigated.The results showed that the former could provide a hatching ability higher than 96%,and the latter could shorten the egg pretreatment duration significantly.

关 键 词：家蚕 化性 催青温度 非滞育卵 转基因 

分 类 号：S882[农业科学—特种经济动物饲养]