广西师范大学学报（自然科学版） ›› 2014, Vol. 32 ›› Issue (4): 126-134.

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凡纳滨对虾精子超微结构及遗传失活的研究

张彬, 韦嫔媛, 熊建华, 谢达祥, 赵永贞, 陈晓汉   

  1. 广西水产科学研究院广西水产遗传育种与健康养殖重点实验室,广西南宁530021
  • 收稿日期:2014-06-29 发布日期:2018-09-26
  • 通讯作者: 陈晓汉(1962-),男,湖北武汉人,广西水产研究所研究员。E-mail: chnxhn@163.com
  • 基金资助:
    广西科学研究与技术开发计划项目(桂科攻11107012-1-1);现代农业产业技术体系广西南美白对虾创新团队建设专项资金(nycytxgxcxtd-03-14-2)资助;国家虾产业技术体系岗位科学家经费(CARS-47-03B, CARS-47-02B);广西农业科技成果转化资金资助项目(桂科转14125004-23);广西“八桂学者”岗位专项资金课题(BGXZ-NMBDX-03);广西直属公益性科研院所基本科研业务费专项资金项目(2060302GXIF-2011-03,2060302GXIF-2012-16)

Preliminary Studies on Ultrastructure and Genetic Inactivity of Spermatozoa in Ltiopenaeus vannamei

ZHANG Bin, WEI Pin-yuan, XIONG Jian-hua, XIE Da-xiang, ZHAO Yong-zhen, CHEN Xiao-han   

  1. Guangxi Key Laboratory of Aquatic Genetic Breeding and Healthy Aquaculture, Guangxi Academy of Fisheries Science, Nanning Guangxi 530021, China
  • Received:2014-06-29 Published:2018-09-26

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摘要： 本文研究使凡纳滨对虾精子遗传失活的方法,用波长365 nm和照射距离10.5 cm的不同剂量紫外线照射凡纳滨对虾精液稀释液或精荚,然后进行人工授精。结果表明,新鲜精液稀释液在0~20 s照射范围内,受精作用都仍能正常发生,与卵子人工体外授精仍可获得5%的平均受精率,但胚胎在孵化前死亡;新鲜精荚在5~10 min照射范围内,采用精液移植授精法可获得20%的平均受精率和3%的平均幼体孵化率。精子在2×105个/cm2的条件下,精液稀释液厚度2 cm,经30 s 以上紫外线照射后,精子染色体可完全失活,而精荚经20 min以上紫外照射后,精子染色体亦可完全失活。应用透射电镜对正常和不同时间紫外辐射的凡纳滨对虾精子形态和超微结构进行观察,结果发现,凡纳滨对虾正常精子系单一棘突类型,主要由前端棘突、中间帽状体和后主体部构成,精子无尾部,不主动运动。紫外辐射凡纳滨对虾新鲜精液稀释液10~20 s的精子形态和超微结构与正常活精子无明显变化。紫外辐射30 s的精子表现为顶体和棘突形态正常,而染色质散乱,后主体部壁变薄;紫外辐射60 s时精子就出现明显的受损状态,主要表现为细胞核变形,核区缩小,核膜受损部分破裂,染色质散乱甚至溢出,后主体部或前部胞壁加厚、膜表面皱缩、底部凹陷,顶体部局部受损,细胞质带囊泡化,双层膜肿胀,膜间腔增大。

关键词: 凡纳滨对虾, 精子, 超微结构, 遗传失活

Abstract: The genetic inactivity of sperm in Litopenaeus vannamei by UV at 365 nm wavelength and 10.5 cm radiation distance were investigated. The inactivity sperm or spermatophore was used to fertilize by artificial insemination methods. The results indicated the fertilization of fresh semendiluter happened during 0~20 s radiation, and which could obtain 5% of average fertility rate by vitro fertilization method, yet all of embryos would not develop to nauplius. The average fertility rate and average hatching rate of fresh spermatophore by semen transplantation were 20% and 3% respectively during 0~10 min radiation. The sperm of fresh semendiluter radiated for more than 30 s would be inactive under the 2×105 sperm/cm2 with 2 mm thickness shaking in 90 time per minute, and the sperm of spermatophore radiated for more than 20 min would be inactive. The morphology and ultrastructure between fresh and different UV dosage radiated spermatozoa of Litopenaeus vannamei were observed by light and transmission electronic microscopy. The observation revealed that the spermatozoa of Litopenaeus vannamei was of the non-motile type. It consisted of three main parts, including the spike-anterior in position, the cap-shaped part at the middle and the main body part at the rear. The results showed that morphology and ultrastructure of sperm from fresh semendiluter UV radiated for 10~20 s didn’t change compared with normal fresh sperm. The acrosome and spike of sperm from fresh semendiluter UV radiated for 30 s were normal, but its chromatin dispersion and bottom wall became thinner. The sperm from fresh semendiluter UV radiated for 60 s was damaged obviously, mainly marked with cell nucleus deformed and membrane cracked, nucleus region diminished, chromatin dispersion and overflow, acrosome body impaired partly, membrane of posterior main body folded, and bottom hollowed, cytoplasmic banding vesiculated, nucleus membrane impaired and cracked, membrane swelled and inter-membrane space augmented.

Key words: Litopenaeus vannamei, spermatozoa, ultrastructure, genetic inactivity

中图分类号: 

  • Q248
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