贵州3个斑鳜野生种群形态学差异

为探究贵州境内沅江、柳江和乌江水系野生斑鳜外形差异,基于形态学和框架结构测量数据,运用多元分析法比较3条水系斑鳜种群的形态特征,经数据标准化和参数选择后12项比例性状和12项可数性状进行统计分析。研究结果显示,单因素分析中9项可数性状之间差异显著(P<0.05);主成分中提取了前6个主成分,其累计贡献率为72.668%,前3项主成分对种群间外形差异贡献较大,主要受其尾柄长、眼间距和体宽等性状影响。应用逐步判别方法建立了这3个种群的特征判别函数,其交互验证判别准确率为81.5%。形态学聚类图结果显示,沅江和柳江种群形态关系较近先聚为一支,之后再与乌江种群相聚。差异系数分析表明,乌江种群与沅江和柳江种群在尾柄长/体长性状上的差异达到亚种水平(差异性系数>1.28)。综上可知,贵州3个斑鳜种群在形态学上存在一定差异,其中柳江与沅江形态差异较小,乌江与其差异较大,建立的判别方程可对3个种群进行区分。本研究结果可用于3个水系斑鳜种群的识别,也可为种群保护和育种利用提供理论依据。     

FIASCO法筛选鳜鱼微卫星标记

通过FIASCO(Fast Isolation by AFLP Sequences Containing repeats)法构建鳜鱼(Siniperca chuatsi)基因组微卫星富集文库,分离微卫星DNA序列并对其特征进行分析。鳜鱼基因组DNA经MseⅠ限制性内切酶消化后,选取200~800 bp的片段与MseⅠ接头连接,用生物素标记的寡核苷酸探针(AC)8、(CT)8、(AT)7、(GATA)8、(GATT)7与其杂交,杂交复合物结合到包被有链霉亲和素的磁珠上,变性洗脱获得单链目的片段,经PCR扩增形成双链,然后克隆到pGEM-T载体上,转化至DH5α中,首次成功构建鳜鱼基因组微卫星富集文库。对其中100个阳性克隆进行测序,60个(60%)含有微卫星序列(GenBank Accession Number:DQ789247~DQ789306)。成功设计了47对鳜鱼微卫星引物,并合成21对引物进行PCR扩增,结果筛选出18个多态性微卫星标记。结果表明:FIASCO法能有效提高筛选微卫星标记的效率。本研究筛选的微卫星标记可以用于鳜鱼遗传背景分析和遗传连锁图谱构建,并将为鳜鱼基因组结构分析、标记辅助育种以及数量性状位点(QTL)基因的定位等研究提供候选微卫星标记。     

翘嘴鳜与斑鳜、花鲈、大口黑鲈杂交试验

本研究以翘嘴鳜(Siniperca chuatsi)为母本,分别以斑鳜(S.Scherzeri)、花鲈(Lateolabrax japonicas)、大口黑鲈(Micropterus salmoides)为父本进行杂交试验.结果显示:与对照组(翘嘴鳜自交)相比,杂交组的受精率明显偏低,远缘杂交(与花鲈、大口黑鲈)的受精...     

Impact of environmental and innate factors on the food habit of Chinese perch Siniperca chuatsi (Basilewsky) (Percichthyidae)

Laboratory and field investigations were conducted to study the food habit of Chinese perch Siniperca chuatsi (Basilewsky) from first feeding through adult stage. Only fish larvae were consumed by Chinese perch larvae (2–21 days from hatching), and the presence of zooplankton did not have any significant effect on their survival rate. The ability of Chinese perch to feed on zooplankton is clearly limited by some innate factor. Instead of gill rakers, Chinese perch larvae have well‐developed sharp teeth at the first feeding stage, and are well adapted to the piscivorous feeding habit unique to the larvae of Chinese perch, e.g. they bite and ingest the tails of other fish larvae. At the first feeding stage (2 days from hatching), daily rations were both very low, either in light or complete darkness. Although early‐staged Chinese perch larvae (7–17 days from hatching) could feed in complete darkness, their daily rations were always significantly higher in light than in complete darkness. Late‐staged Chinese perch larvae (21 days from hatching) were able to feed in complete darkness as well as in light, similar to the case of Chinese perch yearlings. Chinese perch yearlings (total length, 14–16 cm) consumed prey fish only and refused shrimp when visual cues were available (in light), but they consumed both prey when visual cues were not available (in complete darkness), suggesting that prey consumption by Chinese perch yearlings is affected by their sensory modality in predation. Both prey were found in the stomachs of similar‐sized Chinese perch (total length, 14–32 cm) from their natural habitat, suggesting that shrimp are consumed by Chinese perch at night. Prey selection of Chinese perch with a length >38 cm, which consumed only fish in the field, appears to be based upon prey size instead of prey type. These results suggest that although environmental factors (e.g. light intensity) affect prey detection by Chinese perch, this fish is anatomically and behaviourally predisposed to prey on live fish from first feeding. This makes it a difficult fish to cultivate using conventional feeds.     

鳜胚胎细胞系的建立与应用

为更好地开展鳜基因功能研究和药物筛选工作,提高基因转染效率,本研究以鳜囊胚期胚胎为材料,采用含有20%胎牛血清的DMEM培养基进行培养,建立了生长稳定的鳜胚胎细胞系MFE。在此基础上,采用绿色荧光蛋白(GFP)作为标记物,在HEK293T细胞中体外包装逆转录病毒,再感染MFE细胞系。MTT法分析表明传代后细胞培养96 h内,细胞生长率变化也经历增殖、降低到达稳定期。而且MFE细胞能稳定表达GFP基因,感染效率为20%±5%,而脂质体转染效率为3%±2%。可见包装病毒感染细胞不仅能获得稳转细胞系,效率也远高于脂质体瞬时转染。荧光定量PCR分析表明,MFE细胞系能表达Irf1、Irf3和Irf7基因,Irf1基因表达量最高。MFE细胞系受到poly I:C刺激后,Irf1、Irf3和Irf7的表达量分别升高3.5,2.3和2.1倍。因此,MFE细胞通过病毒感染可以获得较高的转染效率,该细胞可作为鳜免疫相关基因功能研究的工具。     

肖四海湖五种渔具的鳜渔获结构特征及其对鳜资源的影响

为评估不同渔具对鳜资源的影响,于2007年5月和12月对长江中游浅水湖泊肖四海湖刺网、延绳钓、网簖、电拖网和电捕仪5种渔具捕获的鳜渔获物结构特征进行了调查分析.结果发现,共采集鳜样本359尾,全长分布范围为92 ～600 mm,优势全长集中于251～350 mm;体质量分布范围为10～3 380 g,优势体质量集中于300 ～500 g.种群由5个年龄组构成,2～3龄为优势龄组,占总数的74.6％.刺网、延绳钓和网簖对鳜有较强的捕捞选择性,网目大小为80 mm和100 mm刺网的鳜渔获物中2龄及以上成熟个体占总数的93.3％,个体平均体质量466 g,“标鳜”(0.4 ～0.75 kg)个体占总数量的57.0％;延绳钓捕获的鳜渔获物中2龄及以上成熟个体占总数的86.9％,“标鳜”个体占总数量的43.5％;网簖捕获的鳜渔获物中90％以上为1龄的未成熟个体.电拖网和电捕仪捕获的鳜全长范围明显较大,其渔获物以1龄和2龄个体为主.综合分析表明,刺网适于作为鳜捕捞的主要渔具,延绳钓可以作为一种鳜捕捞的辅助渔具.网簖对鳜补充群体有较大危害,不适于作为鳜的捕捞网具.电拖网和电捕仪均属于违法渔具,对鳜资源危害巨大,应该加大监管力度,严禁使用.     

鳜鱼人工繁殖技术研究

专塘培育鳜鱼亲鱼，用ＨＣＧ＋ＬＲＨ－Ａ作为催产剂，胸腔２次注射，分４批催产，受精率９０￣９８％，将受精卵放入孵化环道流水孵化，孵化率３４．０￣３８．３％，以刚脱膜的团头鲂作为开口饵料，进行鱼苗环道培育，共育出２．５￣３．０ｃｍ夏花鳜种１２．５万尾，成活率３０￣４０％。     

鳜弹状病毒N蛋白与鳜c-Myc互作调控谷氨酰胺代谢的研究

为了研究鳜弹状病毒(Siniperca chuatsi rhabdovirus, SCRV)如何调控鳜c-Myc(Sc-c-Myc)进而调控谷氨酰胺代谢的分子机制,本研究通过免疫共沉淀联合蛋白质谱寻找可能与Sc-c-Myc互作的病毒蛋白,初步分析确定为核衣壳蛋白(N蛋白);Co-IP结果显示SCRV的N蛋白与Sc-c-Myc存在相互作用。通过PCR扩增获得了带有Flag标签序列的SCRV-N基因的ORF片段,并构建了SCRV-N蛋白过表达载体pcDNA-N-Flag;将pcDNA-N-Flag质粒转染CPB细胞,荧光观察发现Sc-c-Myc与SCRV-N在细胞质内存在共定位现象;并且通过qRT-PCR和Western blotting检测转染pcDNA-N-Flag的CPB细胞中Sc-c-Myc及谷氨酰胺代谢通路关键酶(GLS1,GDH,IDH2)的表达变化,发现Sc-c-Myc、GLS1的转录水平和蛋白水平均显著上调。以上结果表明SCRV的N蛋白与Sc-c-Myc互作促进Sc-c-Myc的表达,进而调控宿主细胞谷氨酰胺代谢途径,为SCRV防控提供了新的靶点。     

培养时间对柱状嗜纤维菌脂多糖蛋白质成分和免疫原性的影响

从培养不同时间后的柱状嗜纤维菌中提取菌体脂多糖 (LPS) ,测定了其中蛋白质含量及成分的变化 ,并分别作为免疫原接种翘嘴鳜对其免疫原性进行了比较。结果表明 ,培养时间对菌体LPS中蛋白质含量没有明显影响 ,对其蛋白质成分则有一定影响 ,而菌体LPS中蛋白质成分与其免疫原性有关 ,从培养 36h的菌体中提取的菌体LPS的免疫原性最强     

鸭绿江斑鳜胚胎及胚后发育观察

于2005、2006年,连续观察了斑鳜Siniperca scherzeri受精卵的胚胎及胚后发育全过程。结果表明:斑鳜受精卵在22.2~25.2℃水温条件下,孵化所需时间约为144 h,胚胎发育到111 h,口裂已经形成,张合自如;破膜后54 h即可捕食活体鱼苗。斑鳜胚胎期即形成口裂、心脏、胸鳍、鳃盖等器官,在破膜前具备了较其它鱼类先天功能更完善、更复杂的生理结构,这一生理特点为其胚后体质强壮,开口即可捕食,消化活体鱼苗奠定了基础。