尼罗罗非鱼与萨罗罗非鱼回交一代与回交二代群体遗传特征的微卫星分析

利用尼罗罗非鱼(Oreochromis niloticus)第二代遗传连锁图谱中微卫星标记,对尼罗罗非鱼♀与萨罗罗非鱼(Sarotherodon melanotheron)回交一代(BC1)和回交二代(BC2)群体遗传特征进行了分析。结果表明,从28个微卫星位点中筛选出9个在尼萨回交一代和回交二代中存在差异的扩增位点,这些多态性位点可作为尼萨回交一代和回交二代群体遗传鉴别的重要依据。尼萨回交一代群体的平均观测杂合度为0.670,多态信息含量为0.423;回交二代群体的平均观测杂合度为0.460,多态信息含量为0.365,回交二代群体的遗传多态性低于回交一代群体。与回交一代相比,尼萨回交二代群体的平均有效等位基因数较为一致(BC1:Ne=2.17,BC2:Ne=2.20),基因型数较为一致,但纯合基因型比例、优势等位基因频率均有明显提高,表明连续回交导致后代遗传同质性提高。     

尼罗罗非鱼(♀)×萨罗罗非鱼(♂)杂交F_2与F_3群体遗传特征的微卫星分析

利用罗非鱼第二代遗传连锁图谱中微卫星标记对尼罗罗非鱼(Oreochromis niloticus)(♀)×萨罗罗非鱼(Sarotherodon melanotheron)(♂)杂交后代F_2、F_3群体遗传特征进行了初步比较分析。结果显示,28对引物中有22对引物能有效扩增,未观察到杂交后代F_2、F_3中有多倍体个体现象。筛选出7个在尼萨罗非鱼杂交后代F_2、F_3群体中存在差异的位点。杂交F_2群体平均等位基因(Na)为2.71,平均多态信息含量(PIC)为0.466,平均观测杂合度(Ho)为0.632;杂交F_3群体平均Na为2.14,平均PIC为0.370,平均Ho为0.432,杂交F_3遗传杂合性较杂交F_2降低。F_2自繁F_3过程中,F_3群体4个位点的等位基因与F_2完全相同,3个位点的等位基因少于F_2,且F_3群体在2个位点出现完全纯合,杂交F_3群体位点等位基因呈现纯合趋势。研究结果为尼萨罗非鱼杂交世代遗传变异与杂交利用积累基础资料。     

Mutualism between euryhaline tilapia Sarotherodon melanotheron heudelotii and Chlorella sp.—Implications for nano-algal production in warmwater phytoplankton-based recirculating systems

The West-African euryhaline tilapia, Sarotherodon melanotheron heudelotii shift from visually feeding on zooplankton when juveniles to mostly filter feeding on phytoplankton when adults. When reared using an appropriate ration in intensive aquaculture systems, S. m. heudelotii also consume algal-based detritus, and contribute to sediment mineralization, clean up their environment, and ultimately stimulate and sustain algal growth. We analysed such practical advantages for phytoplankton-based recirculating systems, using S. m. heudelotii and Chlorella sp. as biological material originating from the prototype of such a system operated in Senegal. We performed a 24-h factorial design experiment in 36 tubs, cross-classifying three levels of S. m. heudelotii (fishless control, unfed fish, and fed fish) with four levels of Chlorella initial density.Chlorella overall mean density increased significantly from fishless, to unfed fish, and fed fish treatments, and with Chlorella initial density. S. m. heudelotii did not alter nitrogen nor phosphorus concentrations, only affected by algal initial densities. Most ammonia excreted by fish was probably uptaken by Chlorella. Bacteria-mediated diurnal nitrification was possibly an alternative ammonium loss mechanism at highest oxygen concentrations. Algae were not limited by nitrogen or phosphorus but most likely by low dissolved organic carbon availability. Chlorella differential responses with fed vs. unfed Sarotherodon suggest that CO₂ supplied by heterotrophic S. m. heudelotii respiration played a key role. Observed Chlorella growth rates were similar to the highest rates obtained in algal mass cultures, enriched with CO₂, nitrate and phosphate, under artificial lighting.Our results suggest the existence of a Sarotherodon-Chlorella mutualism in our systems, where S. m. heudelotii provide CO₂, the major limiting factor of Chlorella growth, whereas Chlorella oxygenate and detoxify the water media from ammonia, promoting S. m. heudelotii production. This mutualism could be used to optimize photosynthetic suspended-growth aquaculture systems, particularly in the Tropics where light is abundant and temperature is continuously high.     

奥利亚罗非鱼和尼罗罗非鱼核型及DNA含量的比较研究

采用ＰＨＡ体内培养法，取鱼体肾细胞用空气干燥法制备奥利亚罗非鱼和尼罗罗非鱼的染色体，经核型分析，奥利亚罗非鱼为２ｎ＝４４，６ｓｍ＋２４ｓｔ＋１４ｔ；ＮＦ＝５０。尼罗罗非鱼为２ｎ＝４４，６ｓｍ＋２４ｓｔ＋１４ｔ；ＮＦ＝５０。经流式细胞仪测量外周血红血球的ＤＮＡ含量，奥利亚罗非鱼为２Ｃ＝２．２２ｐｇ（２．２２ｐｇ／Ｎｕｃｌ）；尼罗罗非鱼为２Ｃ＝２．２７ｐｇ（２．２７ｐｇ／Ｎｕｃｌ）。经对比分析，两种鱼     

尼罗罗非鱼(♀)×萨罗罗非鱼(♂)杂交后代F1、F2形态性状的遗传与变异

通过6个可数性状、10个可量性状及24个框架性状, 比较分析了尼罗罗非鱼(Oreochromis niloticus ♀)×萨罗罗非鱼(Sarotherodon melanotheron ♂)杂交后代F₁、F₂形态性状的遗传与变异特征, 方差分析结果表明, 除臀鳍棘数相同外, 杂交F₁、F₂的可数性状数目相近, 位于尼罗罗非鱼与萨罗罗非鱼之间; 杂交F₁体长/全长、框架参数D_3-5/全长、D_6-8/全长、尾柄长/全长、D_5-7/全长、D_8-10/全长等参数显著大于萨罗罗非鱼, 体厚/全长、体高/全长、D_1-6/全长、D_3-4/全长、尾柄高/全长等参数显著大于尼罗罗非鱼; 杂交F₂的16个可量、框架性状与杂交F₁间无显著差异, 而头长/全长、D_5-6/全长、D_7-8/全长、D_9-10/全长等参数显著大于杂交F₁, 躯干长/全长、尾柄长/全长、D_1-2/全长、D_5-7/全长、D_7-9/全长等参数显著小于杂交F₁。聚类分析结果表明, 杂交F₂与杂交F₁先聚为一类, 再依次与尼罗罗非鱼、萨罗罗非鱼聚类。分别建立了不同群体的可数性状、可量与框架性状的判别公式, 判别正确率高低依次为尼罗罗非鱼、萨罗罗非鱼>杂交F₁>杂交F₂; 利用尼罗罗非鱼和萨罗罗非鱼的判别公式对杂交F₁进行判别, 被判入尼罗罗非鱼的比例较高(可数性状92.9%、可量与框架性状57.1%); 利用尼罗罗非鱼、萨罗罗非鱼及杂交F₁的判别公式对杂交F₂判别, 被判入杂交F₁的比例较高(可数性状68.6%、可量与框架性状77.2%)。主成分分布图显示, 杂交F₁、F₂均位于尼罗罗非鱼和萨罗罗非鱼之间, 杂交F₁与尼罗罗非鱼重叠分布较多, 杂交F₂分布重叠区域较F₁分散。以上结果表明, 尼罗罗非鱼♀×萨罗罗非鱼♂杂交F₁形态性状介于亲本之间, 遗传了双亲的形态特征, 且有一定的偏母遗传; 杂交F₂大部分形态特征遗传了杂交F₁, 但也存在一定程度的变异。本研究旨在通过分析尼罗罗非鱼、萨罗罗非鱼2个杂交世代中亲子间形态性状的遗传规律, 为罗非鱼杂交育种研究与生产利用提供基础资料和依据。

     

尼罗罗非鱼（ ♀） × 萨罗罗非鱼（ ♂） F1家系亲权关系微卫星分析

利用尼罗罗非鱼（Oreochromis niloticus）第2代遗传连锁图谱标记,对3组不同尼罗罗非鱼（♀）×萨罗罗非鱼（Sarotherodon melanotheron）（♂）杂交F1家系内亲权关系进行分析。结果显示,86个微卫星位点中共筛选出20个在尼罗罗非鱼、萨罗罗非鱼中存在差异的扩增位点,含13个种间特异性和7个共享带差异位点。尼萨杂交F1中,平均等位基因2.90,平均多态信息含量0.439,位点多态性较高。3个尼萨杂交F1家系组间遗传距离0.362~0.504,组内个体间遗传距离0.245~0.316,组内遗传距离明显小于组间。利用3个种间特异位点组合,可对3个不同家系组父、母本个体进行鉴别。通过对各组亲本与子代位点基因型分析,家系A、B和C组分别使用4、8和12个特异位点组合进行亲权鉴定,累积排除概率分别为99.99%、99.99%、99.91%,家系A、B组分别含3个半同胞家系,家系C组含2对非同胞或4个半同胞家系。     

慢性盐度胁迫对吉丽罗非鱼(尼罗罗非鱼♀×萨罗罗非鱼♂)及其两亲本耐盐性的影响

为深入研究养殖新品种吉丽罗非鱼[尼罗罗非鱼(♀)×萨罗罗非鱼(♂)]的耐盐性能,对吉丽罗非鱼及其两个亲本尼罗罗非鱼和萨罗罗非鱼进行了慢性盐度胁迫实验,分析了3种罗非鱼耐盐性能的差异,建立了盐度胁迫过程中死亡率与致死盐度及时间的回归模型,结果显示:(1)3种罗非鱼的耐盐能力差异显著,吉丽罗非鱼的耐盐性能接近于萨罗罗非鱼,远高于尼罗罗非鱼,耐盐胁迫时3种罗非鱼的平均致死盐度分别为57.9、66.7和18.5.(2)盐度胁迫实验中尼罗罗非鱼个体间死亡时间差异最大,萨罗罗非鱼个体之间差异最小,吉丽罗非鱼介于其间;吉丽罗非鱼和萨罗罗非鱼死亡时间都有极显著的正态负偏移,离群值和极值较多,尼罗罗非鱼有较显著正态正偏移,只有离群值,没有极值.(3)3种罗非鱼盐度胁迫实验中死亡率与死亡时间及盐度之间的回归关系更适合一元回归,吉丽罗非鱼盐度胁迫实验中死亡率(y)与死亡时间(t)的回归模型为增长模型Y=e (-7.694+0.031t)(R2=0.979),死亡率(Y)与盐度(s)的回归模型为二次模型Y =0.542-0.037s +0.001s2(R2=0.950).     

A simple test to estimate the salinity resistance of fish with specific application to O. niloticus and S. melanotheron

In order to develop a simple and accurate index of the salinity resistance of tilapia, batches of 10 juveniles (5 to 20 g) of two different species Oreochromis niloticus and Sarotherodon melanotheron reared in freshwater were subjected to gradual increases in salinity until 100% mortality. Seven daily increments of salinity were tested with 4 replicates: 2, 4, 6, 8, 10, 12 and 14 g l⁻¹ day⁻¹, while control batches were kept in fresh water. The temperature was maintained at 27 °C. The concentration of oxygen, ammonia and the pH were not limiting factors. The mortality, monitored on a daily basis, appeared after 2–51 days and was spread out over 1–20 days, depending on the increment of salinity. The higher the daily rate in salinity increase, then the shorter the time lapse before total mortality occurred. The cumulative mortality as a function of salinity fit well with simple linear regressions. The criterion of the resistance to salinity was the index MLS (median lethal salinity) defined at each daily rate as the salinity at which 50% of fish died. For S. melanotheron, the mean MLS was 123.7±3.5 g l⁻¹ whatever the daily rate in salinity. For O. niloticus, the MLS was 46.3±3.4 g l⁻¹ for daily increases in salinity ranging from 2 to 8 g l⁻¹ day⁻¹ and decreased significantly (P<0.05) above this level. The MLS-8 g l⁻¹ day⁻¹ ,which takes into account the full capacity of the fish to adapt to the increasing salinity, appeared to be a simple, optimized and efficient criterion for assessing the resistance to salinity for O. niloticus and S. melanotheron. This criterion can be a useful tool for ranking the different parental strains and hybrids of different genus and species of tilapia used in programmes of genetic selection for growth and salinity tolerance.     

pH、盐度及K+、Ca2+和葡萄糖对萨罗罗非鱼精子活力的影响

[目的]确定萨罗罗非鱼精子活力最高时各因子的最佳参数,进一步提高尼罗罗非鱼♀×萨罗罗非鱼♂的受精率.[方法]利用显微镜观察对比不同梯度pH、盐度、离子及葡萄糖溶液中萨罗罗非鱼精子的活力状况,考察指标包括精子快速运动时间、寿命及激活率.[结果]pH 7.5～8.0时,萨罗罗非鱼精子激活率最高,达90％;在pH 7.5的条件下,精子快速运动时间和寿命最长,为66.33±2.25和933.17±25.79 s.在25‰～32‰的盐度范围内,萨罗罗非鱼精子活力较强,其中以30‰的精子活力最高,快速运动时间和寿命分别为63.50±2.35和402.50±7.31 s,激活率达90％.随K+浓度的增加,萨罗罗非鱼精子的快速运动时间、寿命及激活率均呈先升高后降低的变化趋势,其中以75mmol/L的精子活力最高,精子寿命为500.33±8.69 s,激活率为90％.Ca2+对萨罗罗非鱼精子活力有明显抑制作用,其浓度为37.8 mmol/L时精子快速运动时间和寿命最长,分别为38.50±2.43和117.50±3.21 s,精子激活率为80％;而后随Ca2+浓度的增加,精子开始出现聚集现象,活力明显下降,Ca2+浓度增至151.2mmol/L时,精子已完全失活.相对于K+和Ca2+,葡萄糖可适当延长萨罗罗非鱼精子寿命,其浓度为150mmol/L时,精子寿命最长,为281.00±5.90 s,激活率达90％.[结论]针对尼罗罗非鱼♀×萨罗罗非鱼♂人工繁殖中受精率低的问题,生产上除了选择优质尼罗罗非鱼卵细胞、优化孵化环境及人工繁殖技术外,还可通过适宜游动介质参数(pH 7.5～8.0、盐度30‰～32‰、75～100mmol/LK+、37.8 mmol/L Ca2+、100～125mmol/L葡萄糖)激活萨罗罗非鱼精子活力,进而提高其杂交受精率.