马氏珠母贝胚胎和早期幼虫冷冻的研究.pdf

第 28 卷 第 1 期 广东海洋大学学报 Vol.28 No.1 2008 年 2 月 Journal of Guangdong Ocean University Feb 2008 马氏珠母贝胚胎和早期幼虫冷冻的研究 王鹏飞，王梅芳，余祥勇 （广东海洋大学水产学院 湛江 524025） 摘 要：0～4 ℃下，不同发育时期的马氏珠母贝(Pinctada martensii)胚胎和幼虫在不同浓度的 DMSO 抗冻保护液 中放置 30 min 后，置室温恢复，并以不同浓度的 DMSO 作为抗冻保护液，通过缓慢降温对不同发育时期的胚胎 和早期幼虫进行超低温（-196 ℃）冷冻、35 ℃水浴快速解冻。结果表明：低温下 DMSO 对胚胎和早期幼虫活力 影响较弱；马氏珠母贝的囊胚和担轮幼虫存活率较低。当 DMSO 体积分数为 14%时，原肠胚的存活率为 10%左 右，且部分胚胎能够继续发育至 D 形幼虫; 当 DMSO 体积分数为 14%、16%时，解冻后的 D 形幼虫存活率超过 49.9% ，其中大部分存活48 h 以上。 关键词：马氏珠母贝；胚胎；早期幼虫；超低温冷冻；存活率 中图分类号：S968.31+6.1 文献标志码：A 文章编号：1673-9159(2008)01-0025-04 Study on Cryopreservation of Embryo and Early Larvae of Pinctada martensii WANG Peng-fei, WANG Mei-fang, YU Xiang-yong ( Fisheries College, Guangdong Ocean University, Zhanjiang 524025, China ) Abstract: Embryos and early larvae of Pinctada martensii had been placed in 0 to 4 ℃ for 30min,then resumed at room temperature.When the embryos and early larvae at different developing stages were frozen(-196℃) in DMSO of different concentration by slow cooling, then thawed in 35 ℃ water quickly. The result proved that the embryos vitality was weaker in the DMSO solution at lower temperature .the survival rates of blastrulae and trochophore were lower. When the volume fraction of DMSO was 14% ,the survival rate of gastrulae was about 10% ,and part of the gastrulae could develop to D-shape larvae. When the volume fraction of DMSO was 14% and 16% , the survival rate of D-shape larvae wa