文章摘要
二倍体泥鳅与大鳞副泥鳅及杂交F1代精子结构与活力研究
Studies on the structure and vitality of diploid M.anguillicaudatus, P.dabryanus and their hybrid F1 sperm
投稿时间:2020-10-02  修订日期:2021-03-24
DOI:
中文关键词: 二倍体泥鳅  大鳞副泥鳅  远缘杂交  CASA  流式细胞仪  扫描电镜  透射电镜
英文关键词: Diploid Misgurnus anguillicaudatus  Paramisgurnus dabryanus  Distant hybridization  CASA  Flow cytometry  Scanning electron microscope(SEM)  Transmission electron microscopy(TEM)
基金项目:国家自然科学基金31372180
作者单位邮编
李猛 华中农业大学水产学院 430070
虞炯莹 华中农业大学水产学院 
王卫民 华中农业大学水产学院 430070
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中文摘要:
      为了初步探究二倍体泥鳅(Misgurnus anguillicaudatus)和大鳞副泥鳅(Paramisgurnus dabryanus)杂交F1代雄性不育的原因,本文通过计算机辅助精子活力分析系统(CASA)、光学显微镜、扫描电镜、透射电镜、流式细胞技术对二倍体泥鳅(DD)、大鳞副泥鳅(PP)、其正反杂交F1代(DP)和(PD)四种泥鳅精子的形态结构与活力进行了分析。结果显示,激活30 s时,DD和PP的精子运动率分别为76.50 ± 0.70%和75.17 ± 8.60%,极显著高于DP(3.65 ± 1.75%)和PD(2.68 ± 0.63%)(P<0.01),且杂交泥鳅的其它相关运动参数也极显著低于DD和PP(P<0.01),应用流式细胞技术对四种泥鳅精巢内细胞进行倍性鉴定,发现DD和PP精巢内大多为1N精子,而DP和PD精巢内除了正常单倍体精子外,还有大量的2N和4N以及少量的8N细胞。通过扫描电镜和透射电镜发现,DD与PP精巢内精子细胞致密,细胞结构完整,大小均匀,其全长分别为(33.71 ± 2.12)和(34.28 ± 1.83)μm。而DP和PD精巢内出现了大量的异常精子,如多尾精子、双核精子以及大头精子,其头部大小表现出明显的多态性,根据头部体积大小测量估算其倍性,其中有正常的精子即单倍体精子,同时有二倍体精子、四倍体精子、八倍体精子甚至更高倍性的精子。这些杂交泥鳅的异常精子为其精子活力低下以及雄性不育提供了重要证据。
英文摘要:
      The loach M.anguillicaudatus (DD) (Cobitidae; Cobitinae; Misgurnus), and large scale loach P.dabryanus (PP) (Cobitidae; Cobitinae; Paramisgurnus), hybrid F1 generation (DP) and (PD) four loach sperm morphology and vitality are analyzed through computer assisted sperm analysis system (CASA), optical microscope, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and flow cytometry technology in order to explore the reason of male sterility in diploid loach and large scale in vice loach hybrid F1 generation, preliminarily. The results showed that the sperm motility rates of DD and PP were 76.50 ± 0.70% and 75.17 ± 8.60% respectively at 30s of activation, which were significantly higher than DP (3.65 ± 1.75%) and PD (2.68 ± 0.63%) (P<0.01), and the related motion parameters of hybrid loach were also significantly lower than DD and PP (P < 0.01). Ploidy identification was carried out on the sperms of four species of lodges in the sperms of DD and PP by flow cytometers. It was found that the sperms of DD and PP were mostly 1N cell, while there were a large number of 2N and 4N and a small number of 8N cells in the sperms of DP and PD except for the normal haploid sperm. The spermatozoa in DD and PP spermatozoa were dense, intact and uniform in length (33.71 ± 2.12) and (34.28 ± 1.83) m, respectively, by SEM and TEM. DP and PD in the testis appeared a large number of abnormal sperm, such as multitailed sperm, binuclear sperm and big-head sperm. Head size showed obvious polymorphism, according to the head size measurements to estimate its ploidy, including normal sperm haploid sperm, at the same time have a diploid and tetraploid sperm, or even higher haploid sperm. The abnormal sperm of these hybrid loach provided important evidence for its low sperm motility and male sterility.
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