大珠母贝精子短期保藏条件的初步筛选  

Preliminary screening of the short-term preservation conditions for Pinctada maxima sperm

作　　者：黄星美 赵旺[1,2,3,4] 邓正华 温为庚[1,2,3,4] 陈明强 王雨[1,2,3,4] 张钰伟 于刚[1,2,3,4] HUANG Xing-mei;ZHAO Wang;DENG Zheng-hua;WEN Wei-geng;CHEN Ming-qiang;WANG Yu;ZHANG Yu-wei;YU Gang(Key Laboratory.of South China Sea Fishery Resources Exploitation&Utilization by the Ministry of Agriculture and Rural Affairs,South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510300,China;Tropical Fishery R&D Center,South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Sanya 572018,China;Sanya Tropical Fisheries Research Institute,Sanya 572018,China;Key Laboratory of Efficient Utilization and Processing of Marine Fishery Resources of Hainan Province,Lingshui 572426,China)

机构地区：[1]中国水产科学研究院南海水产研究所,农业农村部南海渔业资源开发利用重点实验室,广东广州510300 [2]中国水产科学研究院南海水产研究所热带水产研究开发中心,海南三亚572018 [3]三亚热带水产研究院,海南三亚572018 [4]海南省深远海渔业资源高效利用与加工重点实验室,海南陵水572426

出　　处：《海洋科学》2023年第7期122-129,共8页Marine Sciences

基　　金：国家重点研发计划(2019YFD0900905);海南省重点研发计划项目(ZDYF2021XDNY132);现代农业产业技术体系专项资金(CARS-49);中国水产科学研究院中央级公益性科研院所基本科研业务费专项资金(2020TD55);三亚市农业科技创新项目(2019NK13);中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项资金(2020CY01)。

摘　　要：为初步筛选不同基础液、保藏温度(4℃与23℃)、保存时间(0~120 h)、保护剂(5%DMSO、DMA、DMF、EG)对大珠母贝(Pinctada maxima)精子活力的影响,通过采用精子质量分析仪检测精子总运动率、直线运动速率、曲线运动速率、平均路径运动速率、头部位移距离和鞭毛摆动频率。结果表明,A、B、C三组基础液能不同程度上激活精子,D、E两组结果与之相反;精子被激活后,5组基础液中精子活力存在显著的差异(P<0.05),其中E组基础液的精子总运动率最高,达到80.79%;与室温保藏结果相比,低温(4℃)组精子总运动率显著(P<0.05)高于室温(23℃)组,精子活力的保持时间明显优于室温组;低温(4℃)保存120 h内,精子的运动呈先平稳后下降的趋势,精子的最佳保藏时间为24 h内,精子总运动率、直线运动速率、曲线运动速率、平均路径运动速率分别为82.27%、59.62μm/s、81.95μm/s和70μm/s,头部位移距离为2.56μm,鞭毛运动频率为2.93 Hz;低温(4℃)保存12 h后,对照组精子总运动率显著(P<0.05)高于保护剂组,精子总运动速率保持在80%以上,而保护剂组的精子活力呈现逐步下降趋势,精子总运动率显著性(P<0.05)下降。综上表明,将使用1x D-Hank’s溶液为基础液的大珠母贝精子保存在低温(4℃)条件下,24 h内可保持其精子活力,并延长保存时间。The effects of different storage conditions,such as the extender,storage temperature(4℃and 23℃),storage time(0-120 h),and protective agent(5%dimethyl sulfoxide,Dimethyl Acetamide,dimethyl formamide or ethylene glycol),were determined on Pinctada maxima sperm motility.Total motility(TM),curvilinear velocity(VCL),straight-line velocity(VSL),average path velocity(VAP),head displacement(ALH),and beat-cross frequency(BCF)were detected with a sperm quality analyzer.The results showed that the extenders in groups A,B,and C activated the spermatozoa to varying degrees but not those from groups D or E.After the sperm were activated,there were significant differences in sperm motility in the five groups of extenders(P<0.05),and the total sperm motility rate in the group E extender was the highest at 80.79%.Total sperm motility in the low temperature(4℃)group was significantly(P<0.05)higher than that in the room temperature(23℃)group,and the retention time of sperm motility was significantly better than that in the room temperature group.Sperm motility was steady initially and then decreased during 120 h of storage at low temperature.The optimal storage time of the sperm was 24 h,and the TM,VSL,VCL,and VAP were 82.27%,59.62μm/s,81.95μm/s,and 70μm/s,respectively.The ALH was 2.56μm,and the flagellar BCF was 2.93 Hz.After 12 h of storage at 4℃,the TM of sperm in the control group was significantly higher than that in the protectant group(P<0.05),and the TM of sperm in the protectant group was maintained above 80%,while the motility rate of sperm in the protectant group showed a trend of gradual decline,and the TM of sperm was significantly decreased(P<0.05).In conclusion,P.maxima sperm viability was maintained for 24 h and preservation time was extended using 1×D-Hank’s solution as the base solution at a low temperature(4℃).

关 键 词：大珠母贝 精子活力 保藏条件筛选 保存期 

分 类 号：S917.4[农业科学—水产科学]