秋水仙素诱导青虾次级卵母细胞二倍体初探  

Studies on the Inducement on Macrobrachium Nipponens s Secondary Oocyte for Diploid Using COM

作　　者：陆全平[1] 吕佳[2] 潘建林[1] 宋胜磊[2] 黄成[2] 

机构地区：[1]江苏省淡水水产研究所,南京210017 [2]南京大学生命科学学院,南京210093

出　　处：《南京大学学报（自然科学版）》2004年第4期510-516,共7页Journal of Nanjing University（Natural Science）

基　　金：江苏省水产三项更新工程 (P2 0 0 1- 0 2 - 5 )

摘　　要：以青虾卵巢为材料 ,采用秋水仙素诱导多倍体次级卵母细胞 .经滴片试验 ,确立此实验最佳滴片高度为 1.5cm ,通过设置 5个秋水仙素浓度 (0 0 8～ 0 .4 0mg/mL)及 3个作用时间(6～ 2 4h) ,观察发生染色体加倍的细胞相对数目 .实验结果表明 :在使用秋水仙素的浓度为0 0 8～ 0 .4 0mg/mL ,处理 6～ 2 4h时都可以获得多倍体卵细胞 .在秋水仙素浓度是 0 .32mg/mL ,处理 6h可得到最高二倍体突变率 (32 .5 8% ) .还测试了 0 .32mg/mLCOM作用 6h时ATPase活性与生理盐水对照组的变化 ,结果显示COM会在一定程度上影响细胞的新陈代谢 .但是 5 0 %左右的成活率表明 ,这种诱导突变的方法仍具有一定的应用价值 .As it is always the case that polyploidy animals are bigger in size than common ones,we are now doing some research for the method to double Macrobrachium nipponens s secondary oocytes chromosomes.In this experiment,we inject COM nearby Macrobrachium nipponens s ovary,intending to get the diploid of Macrobrachium nipponens s secondary oocytes.The technique of chopping method is found for the making of the chromosome specimen,using the chopping method with the height of1.5cm.The study shows that we all can get the polyploidy secondary oocytes with the COM strength0.08～0.40mg/L and affecting time6～24h,but the highest mutation rate(32.58%)is under the6hours COM affecting with the strength of0.32mg/mL.During this experiments,we also get the number of chromosomes both before and after COM s effect as52and102.Concerning the reality of this method,we test ATPase s activity of Macrobrachium nipponens s livers under COM s effect(0.32mg/mL,6h),comparing with the ones’under the effect of physiological saline(0.65% NaCl,6h).The results are as different as0.897U/mgprot and4.609U/mgprot.According to statistical inference,we get the surviving rates under different concentrations of COM and different affecting time.The surviving rates are around50%.In this way,although Macrobrachium nipponens s metabolism will be disturbed in a certain level,the surviving rats are relatively high.So that this method is somewhat practicable. But if we use the COM affected female Macrobrachium nipponens to mate with common male ones and produce triploid prawns,the results may be disappointed,as a result of the DNA trauma caused by COM.So we can try the way of artificial insemination,using the affected secondary oocytes and common sperms instead.

关 键 词：青虾 秋水仙素 二倍体 多倍体诱导 

分 类 号：Q492.5[生物学—生理学]