三倍体皱纹盘鲍活体倍性鉴定——用上足触手、外套膜活体组织制备染色体标本  

The Detection of Living Triploid of Haliotis discus hannai --Directly Making Chromosome Sample of Mantle and Epipedium Tentacle

作　　者：李雅娟[1] 毛连菊[1] 李霞[1] 王子臣[1] 宋坚[1] 

机构地区：[1]大连水产学院农业部海洋水产增养殖生态学重点开放实验室,辽宁大连116023

出　　处：《Acta Genetica Sinica》2002年第6期514-518,共5页

基　　金：国家 8 63高技术项目 (863 819 0 1 0 4)~~

摘　　要：以上足触手、外套膜活体组织为材料 ,采用PHA体外浸泡法 ,运用正交实验设计 ,探讨了暂养温度、PHA(植物血球凝集素 )浓度及取材时间对细胞分裂频度的影响。根据直观分析 ,得出用上足触手、外套膜活体组织直接制备染色体标本进行三倍体皱纹盘鲍活体倍性鉴定的最优组合是 :暂养温度 2 0℃ ;PHA浓度为 1.0 0 % ;取材时间17:0 0～ 18:0 0。 3因素的主次顺序 :暂养温度→PHA浓度→取材时间。并可获得大量图像清晰、长度适中。Ploidy detection method of living body is a very important component part in polyploid breeding There were few reports about chromosome ploidy detection of adult abalone ( Haliotis discus hannai ), except preparing adult abalone chromosome by means of chopping method using gill tissues and squashing method using gonad,also with these methods the abalone should be killed for getting these tissues. In polyploid breeding living triploid abalones are needed, so ploidy detection technique of living abalone is especially important. In this paper the orthogonal experiment of three factors and three levels L 9(3 4)using mantle and epipodium tentacle as material of living tissues which were vido immersing in PHA solution for preparing chromosome and count the frequence of cell division was first reported. Three factors and three levels were raising temperature: 16℃,18℃ ,20℃; PHA concentration:0.5%,l.0%,1.5%; sampling time:5:00～6:00, 17:00～18:00,23:00～24:00,respectively. The tests were repeated two times. The test results showed that the optimal patterns effecting the mitotic frequency of three factors and three levels in triploid abalone detection of chromosome from mantles and epipodium of living bodies were: raising temperature 20℃,PHA concentration 1.0%, and sampling time 17:00～18:00. The order of three factors was:raising temperature → PHA concentration → sampling time. Under optimal pattern of three factors and three levels many clear, proper in length and well scattered metaphase chromosomes could be obtained. The advantages of preparing chromosome using mantles and epipodium tentacle were sampling convenient, no body size restriction and no influence of survival of abalone. After using PHA treated tissues, mitotic frequency could be improved and 1.00% PHA concentration was the best. The key of chromosome preparation was the cells in mitotic metaphase, so the time of sampling was important. During 17:00～18:00, sampling of abalone many mitotic cells could be obtained because

关 键 词：三倍体皱纹盘鲍 活体倍性鉴定 上足触手 外套膜 活体组织 制备 染色体标本 

分 类 号：S968.315[农业科学—水产养殖]