乌鳢和斑鳢微卫星制备及其对自交与杂交F1代的初步鉴定

通过磁珠富集法分离了乌鳢(Channa argus)和斑鳢(C.maculate)的微卫星DNA序列,根据这些微卫星DNA序列的两翼序列,采用Primer 3.0或Primer 5.O软件设计出乌鳢与斑鳢的微卫星引物各31对.用62对微卫星引物对乌鳢和斑鳢自交与杂交F1代的4个组合进行扩增,10对引物无扩增产物,其余5...     

一种乌鳢与斑鳢线粒体PCR-RFLP鉴定方法

为了高效的鉴别乌鳢与斑鳢,采用PCR-RFLP技术,对乌鳢与斑鳢开展分子生物学鉴定方法研究。通过比对乌鳢和斑鳢线粒体全序列,发现1处单核苷酸多态性位点,可以明确区分两个物种。利用1对引物对乌鳢与斑鳢线粒体基因组该区域进行PCR扩增,用限制性内切酶EcoR I分别对扩增产物进行酶切,并用1.5%的琼脂糖凝胶检测酶切结果。PCR-RFLP检测结果显示,斑鳢的PCR扩增产物被EcoR I酶切后生成两个不同大小的片段,分别为315 bp和875 bp,乌鳢则保持不变。由此可将乌鳢与斑鳢在酶切图谱上鉴别出来。     

基于ISSR分子标记的褐马鸡亲缘关系分析

褐马鸡(Crossoptilon mantchuricum)是中国特有濒危鸟类,国家一级保护动物。为了保护褐马鸡种质资源,保障驯养繁育种群和再引入种群的遗传基因结构优化,采用ISSR分子标记技术,对山西庞泉沟国家自然保护区和太原动物园两个种群35个褐马鸡进行了亲缘关系分析。从20条ISSR引物中筛选出10条扩增条带清晰稳定、重复性好的引物,共扩增出65条DNA条带,其中77%呈多态性。两个种群各个体之间的Nei's无偏差遗传距离和遗传相似度分析表明:个体间遗传相似性的平均值分别为0.5061与0.7591,遗传距离的平均值分别为0.4939与0.2409。用组平均法对35个褐马鸡个体进行聚类分析,结果显示:大多数同一种群的个体首先聚在一起,显示出生态地理条件相似的种群亲缘关系较近。     

日本白鲫(♀)×德国镜鲤(♂)F1代及双亲的RAPD分析

利用筛选的24个随机引物,对日本白鲫(Carassius Curatus Cuvieri)(♀)×德国镜鲤(Cyprinus carpio varspecularis Lacepede)(♂)杂种一代及双亲进行RAPD分析。结果表明:在德国镜鲤、日本白鲫、F1中检测到的位点数分别为200、207、218,其中多态性位点数分别为63、66、72,多态位点比例分别为31.5%、31.9%、33%。F1多态位点高于父母本,表明它的遗传多样性和变异水平最高。3个群体间的遗传距离分别为0.3068(德国镜鲤和日本白鲫)、0.2041(德国镜鲤和F1)、0.1961(日本白鲫和F1)。德国镜鲤和日本白鲫的遗传距离最大,表明它们之间具有最大的杂交潜力,F1与父本遗传距离大于F1与母本遗传距离,表明F1与母本亲缘关系更近。从扩增谱带中找到了1个引物(S21)的特异性带是德国镜鲤区分日本白鲫和F1的分子标记,找到4个引物(S79、S114、S331、S333)扩增出的特异性谱带是日本白鲫区分德国镜鲤和F1的特异标记,还有1个引物(S106)扩增的特异性谱带可以作为F1区分父母本的遗传标记。在3个群体所有个体的UPGMP分支图中,所有个体分别成3个群分支,清晰地反映出个体间的相互关系。     

长臀鮠的AFLP分析

采用 AFLP分子标记技术对珠江长臀鮠和海南长臀鮠共60个个体(每个群体30个)进行了遗传多样性研究。选取的18对引物组合均能扩增出清晰、可重复的扩增产物,扩增带型差异明显。两个群体均表现出较高的多态位点比例和特异性条带数。群体内个体间的遗传相似度(平均值)珠江长臀鮠种群为0.9462±0.0237, 海南长臀鮠为0.9465±0.0226,海南长臀鮠比珠江长臀鮠略高。群体间的遗传相似度是0.9367±0.0231,小于两个群体内的遗传相似度,两群体的遗传距离是0.0634±0.0230。根据遗传相似度绘制了UPGMA 聚类图。研究结果表明：珠江长臀鮠和海南长臀鮠间的遗传差异属于种内差异,两者同属一个有效种。     

贵州省八个种群角倍蚜ISSR遗传多样性

基于ISSR分子标记技术对采自贵州省8个种群共139个角倍蚜样本进行遗传多样性及其与环境因子的相关性分析。筛选的12条引物共扩增出多态性条带133条,没有种群特异带出现,平均每个引物扩增的多态带数为11条,Popgene软件分析显示,总群体多态性条带比率为100％,尽管不是特意选择变异高的引物。种群间遗传相似系数变异范围介于0．899～0．955,遗传距离介于0．046～0．106,说明角倍蚜不同种群之间的遗传分化不大。聚类分析显示,8个种群共形成2个大的聚类簇,各种群之间没有明显的地域性分布规律,但遗传多样性与3月温度和3月降水量呈负相关。     

应用RAPD技术分析奥利亚罗非鱼和尼罗罗非鱼的基因多态性

以随机扩增多态DNA技术(RAPD）分析了奥利亚罗非鱼和尼罗罗非鱼两个养殖群体的群体内及群体间遗传关系,并探讨了该技术在种群鉴定中的应用。RAPD引物筛选结果表明,所测试的20个随机引物中(Table 1),除一个引物未扩增出任何片段外,其余19个引物均扩增出1～11个大小不等的片段,长度大部分在500—3000bp之间,共扩增出220个片段,平均每个引物产生55个片段。两群体间共有片段70条,大部分引物的扩增产物具有种间多态性,种群间相似系数为0.727。以筛选的引物对两种群不同个体(Fig．1,Table 2)及种群混合样品(Fig.2,Table 3)进行RAPD分析。结果表明,不同引物在扩增图谱上表现很大差异：奥利亚罗非鱼不同个体间表现为一致的扩增图谱,种内相似系数达1000,显示了其种群内遗传变异的缺乏;尼罗罗非鱼种内相似系数为0．827,个体间存在不同程度的多态性;两个种群间的相似系数分别为0．767和0.742,表明种间有较高的同源性,遗传距离为0．235,略低于国外的报道．此外,两个养殖群体间的扩增图谱比较也暗示了遗传渐渗现象的存在。实验表明,RAPD标记可以作为一种可靠的遗传标记,用于不同鱼类种群的鉴定,RAPD分析方法是一种快速,简便且行之有鼓的鉴定鱼类种群的方法。     

送春与多花兰种间杂交后代的ISSR分析

该文使用简单重复序列间( ISSR)分子标记,对送春与多花兰种间杂交后代进行了研究。结果表明：从80个ISSR引物中筛选出14个扩增效果稳定的ISSR引物,对两亲本和59个F1代个体进行了ISSR扩增,得到107个扩增位点,扩增的片段大小位于90~2100 bp之间,平均每个引物扩增7.64条条带,得到11种类型的带。 ISSR标记在送春×多花兰的F1代中表现出一定的多态性,分离频率为44.86％,分离位点有83.33％符合孟德尔1︰1或3︰1的分离规律,产生偏孟德尔分离的位点占12.50％,余下的4.17％属于特殊分离带型。可能导致后代变异的位点为偏孟德尔分离的6条带、缺失的8条带或新生成的2条带。聚类图中父本和母本与F1代个体间的遗传距离较远,59个杂交后代先聚集成一组,再同母本相聚为一组,最后才同父本聚在一起,59个杂种均偏母本型。送春与多花兰的杂交后代在植株形态、染色体、遗传物质方面都具备双亲特点,61个个体间的ISSR分子量标记结果和植株形态学特征都说明,59个F1代杂种包含送春和多花兰的遗传特性是真杂种;F1代杂种既有双亲的互补特征带,又有双亲的重组片断即产生新的特异带,这说明送春与多花兰的杂交后代具有遗传变异的特点。该研究结果可以有效地对杂交后代进行定向选择,为兰花的杂交育种提供了分子依据。     

伞裙追寄蝇不同地理种群遗传多样性的ISSＲ分析

为从分子水平探索不同地区伞裙追寄蝇种群间的内在联系,本文利用ISSR分子标记技术对8个不同地区伞裙追寄蝇自然种群进行遗传多样性、种群间分化程度以及聚类分析等研究。结果表明:筛选出11对多态性稳定的ISSR引物,对8个地区伞裙追寄蝇群体的80个个体进行PCR扩增,共获得166个重复性好且清晰可辨的ISSR条带,平均每条引物扩增出15.0909个片段,且均为多态性条带,多态信息含量(PIC)为0.8441-0.8653;香农信息指数(I)为0.1240-0.3455;Nei's遗传多样性指数(H)为0.0841-0.2285;基因分化率(Gst)为28.78%,基因流(Nm)的均值为1.5702,即遗传变异主要存在于个体之间,不同种群间基因交流处于中等水平;8个地区伞裙追寄蝇种群被划分为4个类群,种群间的遗传分化与地理距离呈正相关关系。     

斑鳢、乌鳢及其杂种细胞核DNA流式含量分析

以斑鳢(Channa maculata)、乌鳢(C.argus)及其正交杂种斑乌鳢(斑鳢♀×乌鳢♂)和反交杂种乌斑鳢(乌鳢♀×斑鳢♂)的红细胞为材料,以鸡(Gallus gallus)血细胞为DNA标准(2.5 pg/2c,2c指2倍体),采用流式细胞仪测定了这4种鱼的细胞核DNA含量。斑鳢、乌鳢、斑乌鳢及乌斑鳢这4种鱼血细胞DNA的绝对含量分别为(1.488±0.035)pg/2c、(1.489±0.034)pg/2c、(1.522±0.077)pg/2c和(1.520±0.033)pg/2c。斑鳢和乌鳢的细胞核DNA含量差异不显著(P0.05),斑鳢和乌鳢与两种杂交鳢的DNA含量差异显著(P0.05),两种杂交鳢之间的细胞核DNA含量差异不显著(P0.05)。杂交鳢细胞核DNA含量显著高于亲本,可以作为杂种鉴定的依据。     

Vitamin C deficiency in Channa punctatus Bloch

The results of a series of experiments on Channa punctatus previously conditioned to a complete synthetic diet and later fed vitamin C deficient diet for 210 days followed by 30 days of recovery with fortified vitamin C ration are reported. The survival, growth, morphological abnormalities and biochemical changes in tissue ascorbic acid and cholesterol levels were studied.
Deficiency of vitamin C resulted in a mortality of up to 25% and retarded growth accompanied by anorexia and loss of weight. Marked skeletal abnormalities, viz. , lordosis, scoliosis and degenerative changes in the thoracic vertebrae were revealed by X-ray studies. Biochemical investigations showed a significant decrease in blood and kidney ascorbic acid levels within a month of feeding the deficient diet. A prolonged deficiency beyond 150 days resulted in a rapid rise in the cholesterol content of the liver. These biochemical changes were reversed when vitamin C was added to the ration.     

斑鳢血细胞初步研究

对斑鳢的外周血细胞用常规Wright’s法染色并进行显微观察。结果显示,斑鳢外周血中具有红细胞、幼稚红细胞、小淋巴细胞、大淋巴细胞、血栓细胞、单核细胞和嗜中性粒细胞,未发现嗜酸性粒细胞和嗜碱性粒细胞。分别对各种血细胞的显微结构进行了描述。     

In silico analysis of Myoglobin in Channa striata

Myoglobin is a cytoplasmic hemoprotein, expressed solely in cardiac myocytes and oxidative skeletal muscle fibers, that reversibly binds O₂ by its heme residue. Myoglobin is an essential oxygen-storage hemoprotein capable of facilitating oxygen transport and modulating nitric oxide homeostasis within cardiac and skeletal myocytes. Functionally, myoglobin is well accepted as an O₂- storage protein in muscle, capable of releasing O₂ during periods of hypoxia or anoxia. There is no evidence available regarding active sites, ligand binding sites, antigenic determinants and the ASA value of myoglobin in Channa striata. We further document the predicted active sites in the structural model with solvent exposed ASA residues. During this study, the model was built by CPH program and validated through PROCHECK, Verify 3D, ERRAT and ProSA for reliability. The active sites were predicted in the model with further ASA analysis of active site residues. The discussed information thus provides the predicted active sites, ligand binding sites, antigenic determinants and ASA values of myoglobin model in Channa striata.     

Food selection and growth of young snakehead Channa striatus

Food selection and growth of young snakehead Channa striatus were studied in the laboratory and in a field trial. In the laboratory, first-feeding snakehead larvae of 6–7 mm total length (TL) with a mouth opening of 0.55 mm selected for Artemia nauplii, and against formulated feed. Fish began feeding on formulated feed at 12mm TL when their mouth width reached 1.0mm. In both laboratory and field trials, snakehead diets changed as fish size increased. For fish 15–20 mm TL, cladocerans and copepods were 96.5% of their diet. With fish 30–40 mm TL, zooplankton consumption was greatly reduced while benthic organism consumption increased. Fish 45–50 mm TL fed exclusively on benthic invertebrates. Diet shift from zoo-plankton to benthic invertebrates was not due to reduced zoo-plankton availability, but was instead related to changes in gill raker structure. Low density of benthic invertebrates in the field trial caused reduced fish growth rates when fish switched diets from zooplankton to benthos. Our results indicate that snake-head can take Artemia nauplii as a larval starter food, then accept formulated feed at ≥12 mm TL. Zooplankton can serve as food for snakehead < 40 mm, but formulated feed should be provided for larger fish which are unable to catch zoo-plankton.     

Purification of C-reactive protein from Channa punctatus (Bloch).

C-reactive protein (CRP) is found to be a normal component of serum of freshwater air-breathing murrel Channa punctatus. Based on the property of binding with C-polysaccharide (CPS) of pneumococcus bacteria in presence of Ca2+, CRP was purified by phosphorylcholine-Sepharose affinity column chromatography. Molecular weight of the intact protein molecule was estimated to be approximately 141,000 by gel filtration. In non-reduced and reduced conditions the molecule showed molecular weight approximately 28,000 and 14,000 respectively in SDS-PAGE. Monospecific antisera was raised against the affinity purified CRP and used as a tool to detect CRP in the hepatic cytosol and egg extract. The level of CRP in the normal serum was estimated to be 220 micrograms/ml.     

鰤鱼诺卡氏菌对乌鳢血液指标的影响

应用浓度为106cfu/mL的鰤鱼诺卡氏菌(Nocardia seriolea)菌悬液腹腔注射乌鳢,人工感染诺卡氏菌病。在感染后3d、6d、9d、12d、18d和24d时抽取乌鳢血液,检测乌鳢的血细胞数、血细胞脆性、溶菌酶、血清总蛋白、碱性磷酸酶、乳酸脱氢酶和尿素氮等指标。结果表明,与对照组相比,感染鰤鱼诺卡氏菌后乌鳢血细胞数呈先升高后降低的趋势,而血细胞脆性在各实验时间点均高于对照组;血清碱性磷酸酶呈先降低后升高趋势,血清溶菌酶、血清总蛋白和血清尿素氮等均呈先降低后增高再降低的趋势,血清乳酸脱氢酶活力显著增加。说明鰤鱼诺卡氏菌感染会引起乌鳢血液指标明显变化。     

Dietary niacin requirement of fingerling Channa punctatus (Bloch)

A 16‐week experiment was accomplished to determine the dietary niacin requirement of fingerling Channa punctatus (6.8 ± 0.92 cm; 4.65 ± 0.46 g) by feeding seven casein‐gelatin based isonitrogenous (450 g/kg CP) and isoenergetic (18.39 kJ/g GE) diets with graded levels of niacin (0, 10, 20, 30, 40, 50 and 60 mg/kg diet) twice a day to apparent satiation to triplicate groups of fish. Significantly best absolute weight gain (AWG; 38.19 g/fish), feed conversion ratio (FCR; 1.42) and protein retention efficiency (PRE; 26.47%) were registered in fish fed 40 mg niacin/kg diet. Also, fish fed above diet exhibited maximum carcass protein. Hemoglobin (Hb), RBCs counts and hematocrit (Hct) were improved with the incremental levels of dietary niacin up to 40 mg/kg. However, liver niacin content showed the positive correlation up to 50 mg/kg niacin and then leveled off. Fingerling C. punctatus fed niacin‐free diet showed retarded growth, poor feed utilization, high mortality, difficult motion and skin haemorrhage. Broken‐line regression analysis of AWG, FCR and PRE indicated that fingerling C. punctatus require niacin in the range of 37.1–42.1, whereas that of liver niacin concentration indicated the niacin requirement at 52.3 mg/kg dry diet.     

Effect of alloxan administration upon Channa punctatus (Bloch)

Channa punctatus, a fresh water teleost, was administered intramuscularly with alloxan in doses varying between 200 and 600 mg/kg body weight. The course of cytological events occurring in the islets of alloxanized fish was followed over a period of 120 hours post injection. Alloxan caused mild degranulation of some of the beta cells in the beginning but later nuclear pycnosis, vacuolization and necrosis of the beta cells was observed. Like other vertebrates, Channa punctatus is alloxan-sensitive and its islet beta cells are susceptible to the cytotoxic effects of alloxan.