不同倍性泥鳅鳍细胞培养及染色体标本制备方法的研究  

The cell culture of fin tissue and preparation of chromosomes in different ploid loach Misgurnus anguillicaudatus

作　　者：刘博[1] 李雅娟[1] 李霞[1] 隋燚 高养春[1] 王玉生[1] 马辰[1] 

机构地区：[1]大连海洋大学农业部北方海水增养殖重点实验室,辽宁大连116023

出　　处：《大连海洋大学学报》2013年第6期557-562,共6页Journal of Dalian Ocean University

基　　金：国家自然科学基金资助项目(31272650);辽宁省科学技术基金资助项目(201102019);大连海洋大学博士启动基金资助项目(017208)

摘　　要：以天然二倍体、杂交三倍体(4n♀×2n♂)和天然四倍体泥鳅Misgurnus anguillicaudatus的鳍组织为材料,对短期培养的消毒方法、秋水仙素的浓度(1.0、1.5、2.0μg/mL)与处理时间(2、3、4 h)和低渗处理时间(30、40、50 min)等细胞培养技术,及染色体标本制备方法进行了研究。结果表明:选择100 g/L的碘伏溶液杀菌效果良好,对于细胞生长影响较小,尤以处理15 min时的效果最明显,细胞迁出迅速;在终止培养前3 h加入秋水仙素至终浓度为1.5μg/mL,天然二倍体、杂交三倍体和天然四倍体泥鳅鳍细胞中期分裂相染色体众数最高,分别为93.33%、90.00%、70.00%;25℃下低渗处理40 min时,可以获得大量图像清晰、长度适中、分散良好、染色体数目完整的染色体中期分裂相。本研究中初步建立了以鳍组织培养法作为材料来源快速制备不同倍性泥鳅染色体标本的方法。The effects of disinfection methods,concentrations( 1. 0,1. 5,and 2. 0 μg /mL) of colchicine,processing time( 2,3,and 4 h) of colchicine and hypotonic treatment( 30,40,and 50 min) on short-term cell culture of fins and the condition for chromosome preparation were studied in natural diploid,hybrid triploid( 4n♀×2n♂) and natural tetraploid loach Misgurnus anguillicaudatus. Results showed that the best disinfection was observed at 100 g / L lodophor with little influence on cell growth,especially rapid cell movement at processing time of 15 min. The maximal chromosome mode was frequencies of normal metaphase spread of natural diploid( 93. 33%),hybrid triploid( 90. 00%) and natural tetraploid( 70. 00%) when the colchicine was added 3 h before the termination of cell culture until to final concentration of 1. 5 μg / mL. The clear metaphase spreads with moderate length,excellent dispersion and complete number was observed under condition of hypotonic treatment at below 25 ℃ for 40 min. In summary,the chromosome preparation method was preliminarily established using cell culture of fins in loach.

关 键 词：泥鳅 天然四倍体 杂交三倍体 二倍体 鳍组织 染色体 

分 类 号：S917.4[农业科学—水产科学]