东昌湖野生鲤的线粒体DNA RFLP分析

利用差速离心法及RNase消化法制备并纯化了采自山东东昌湖野生鲤（Cyprinus carpio haematopterus）的肝脏及性腺线粒体DNA,用7种限制性内切酶对其mtDNA进行酶解,经琼脂糖凝胶电泳分离酶解片段,用Gel-5000凝胶图像分析系统进行采集和分析。结果显示,东昌湖野生鲤的mtDNA分子大小为（16.62±0.15）kb,BglⅠ、BglⅡ、EcoRⅠ、HindⅢ、PstⅠ、KpnⅠ和BamHⅠ的酶切点分别为3或2、1、3或4、4、1、0、3;其中,BglⅠ、EcoRⅠ和BamHⅠ     

乌鳢线粒体DNA限制性内切酶酶切分析

用BamHⅠ,EcoRⅠ,HindⅡ,HinfⅠ,MspⅠ,PstⅠ,SmalⅠ,XhoⅠ,DraⅠ,SalⅠ等十种限制内切酶对纯化后的乌鳢(Ophiocephalus argus)线粒体DNA(mtDNA)进行酶切,然后用电泳技术分离酶切片段。结果表明,十种限制性内切酶只有DraⅠ,SalⅠ两种限制性内切酶有多个酶切位点,且计算出乌鳢mtDNA大小约由17200个碱基对组成。     

草鱼线粒体DNA限制性内切酶分析

本文报道了用ＢａｍＨⅠ，ＢｇｌⅠ，ＢｇｌⅡ，ＣｌａＩ，ＥｃｏＲＩ，ＨｉｎｄⅢ，ＰｖｕⅡ，ＳａｃⅠ，ＸｂａⅠ，ＸｈｏⅠ等十种限制性内切酶酶切草鱼线粒体ＤＮＡ，计算出各酶切片段长度及草鱼ｍｔＤＮＡ大小。其中六种酶有多个切点，在所检测的样品中未发现碱基替换。     

鲇鱼线粒体DNA的酶切图谱

采用差速离心法及ＤＮａｓｅⅠ、ＲＮａｓｅ消化法制备并纯化了鲇（Ｓｉｌｕｒｕｓａｓｏｔｕｓ）肝脏线粒体ＤＮＡ（ｍｉｔｏｃｈｏｎｄｒｉａｌＤＮＡ，ｍｔＤＮＡ）。用８种限制性内切酶对ｍｔＤＮＡ进行了分析。ＢｇⅡ、ＥｃｏｒⅠ、ＰｓｔⅠ、ＢｇｌⅠ、ＢａｍＨⅠ、ＸｂａⅠ、ＨｉｎｄⅢ、ＸｈｏⅠ在鲇ｍｔＤＮＡ分子上分别具１、１、１、２、２、７、７和０个切点。ｍｔＤＮＡ分子量约１０．８４×１０￣６道尔顿，大小为１７．５４ｋｉｌｏｂａｓｅｐａｉｒｓ。根据单酶和双酶解片段的数目和分子量，建立了鲇ｍｔＤＮＡ的限制性酶切图谱。     

黑龙江野鲤线粒体DNA限制性内切酶酶切分析

本文运用HaeIII、HindIII、EcoRI、BamHI、PstI、ClaI、HpaI、SalI、SacI等 9种限制性内切酶对来自黑龙江抚远江段的 40个黑龙江野鲤样本进行线粒体DNA的酶切电泳分析 ,其切点分别为 7、6、4、3、1、1、1、1、1。分子量约为 1 0 1 2× 1 0 6 相当于 1 6 49Kb     

三倍体湘云鲫及其亲本线粒体DNA的比较研究

用人工选育的异源四倍体鲫鲤（♂）与白鲫（♀）杂交获得具有明显生长优势的三倍体湘云鲫。采用差速离心和核酸酶处理等方法，从三倍体、白鲫和异源四倍体鲫鲤肝组织中提取线粒体DNA，并用9种限制性内切酶进行单酶酶切分析。经琼脂糖凝胶电泳后计算出各酶切片段的大小，测得三倍体、白鲫和异源四倍体鲫鲤mtDNA的分子大小分别为16．24kb、16．60kb和16．20kb。根据各单倍型间的酶切片段共享度，估算出3个群体间的遗传距离，说明了mtDNA母系遗传的特性。     

条纹斑竹浙江省鲨线粒体DNA的研究

用６种限制性内切酶分析了４条条纹斑竹鲨的线粒体ＤＮＡ，ＰｓｔＩ，ＨｐａＩ，ＸｂａＩ，ＥｃｏＲＩ，ＥｃｏＲＶ，ＢｇｌⅡ在条纹斑竹鲨ｍｔＤＮＡ分子上分别具有０至２个切点，ｍｔＤＮＡ分子大小为１６．６ｋｂ，根据单酶和双酶完全酶解片段的大小，构建了条纹斑竹浙江省ｍｔＤＮＡ的限制性酶切图谱。     

建鲤线粒体DNA的遗传多态性初步分析

用16种限制性内切酶对建鲤mtDNA进行酶解分析,结果表明：建鲤的mtDNA分子大小约为16.6kb;除Bgl Ⅱ无切点,ClaⅠ、PstⅠ、SalⅠ和XhoⅠ有一个切点外,其余11种酶都有2个或2个以上酶切位点,其中DraⅠ、PvuⅡ检出mtDNA多态,将建鲤mtDNA分为两种不同的单倍型;计算建鲤种内（群体内）mtDNA多态值π＝0.00164。建鲤在分子水平显示出的遗传多样性,可能是建鲤能够适应全国各地的环境条件和养殖方式,并得以大规模推广的重要原因之一。     

鲇（Silurus asotus L．）线粒体DNA的初步研究

实验共用13种限制酶分别对鲇mtDNA作酶切分析。其中，酶BamHI、Bgll和Sacl在鲇mtDNA分子上产生2个切点；BglⅡ、EcoRⅠ、PstⅠ和ScaⅠ都产生1个切点；HindⅢ和PvuⅡ分别产生5个和3个切点；XbaⅠ的切点多于5个，而ClaⅠ、DraⅠ和KpnⅠ没有切点。根据酶解结果计算出鲇mtDNA的大小     

湛江沿海五种石斑鱼线粒体DNA的RFLP分析

In this paper, genetic diversity of intraspecies, and genetic relationship of interspecies in Epinephelus spp. (E. merra, E. fario, E. awoara, E. akaara and E. septemfasciatus) were assessed by using mitochondrial DNA restriction fragment length polymorphisms (mtDNA RFLPs).The samples were collected from the coastal area of Zhanjiang,Guangdong province. MtDNA was extracted from the fresh liver tissue by applying a difference centrifugation procedures. Using 17 restriction enzymes with 5-or 6-bp recognition sites, the purified mtDNA was cleaved by single enzymes. These enzymes included BamH Ⅰ,Bgl Ⅰ,Bgl Ⅱ,Dra Ⅰ,EcoR Ⅰ,EcoR Ⅴ,Hind Ⅲ,Kpn Ⅰ,Mlu Ⅰ,Pst Ⅰ,Pvu Ⅱ,Sal Ⅰ,Sca Ⅰ,Sma Ⅰ,Sty Ⅰ,Xba Ⅰ and Xho Ⅰ. The phylogenetic analysis was done using the Neighbor joining(NJ) method and Unweighted pairgroup method with arithmetic mean(UPGMA) method. Genetic diversity indices such as haplotype diversity (h), average genetic distance between haplotypes (P) and nucleotide diversity (π) were calculated using Nei and Li's segment method to quantify the genetic diversity within species. There were 8, 5, 8, 5 and 2 haplotypes detected within E. merra, E. fario, E.awoara, E.akaara and E. septemfasciatus, respectively. The haplotype diversity (h) was 0.8943, 0.6186, 0.9242, 0.6927 and 0.1820,respectively. The average genetic distance between haplotypes (P) was 0.62%±0.31%, 0.64%±0.37%, 1.12%±0.55%,0.72%±0.42% and 0.45%, respectively. And the nucleotide diversity (π) was 0.22%, 0.13%, 0.46%, 0.17% and 0.04%, respectively. The wild groupers in the Zhanjiang Coastal Area exhibited a relative higher level of genetic diversity. The net genetic distance between species (P_net) was 0.0694（E. merra - E. fario）,0.1337（E. merra - E. awoara）,0.1090 E. merra -E. akaara）, 0.1286（E. merra - E. septemfasciatus）,0.1590（E. fario -E. awoara）,0.0825（E. fario -E. akaara）,0.1153（E. fario - E. septemfasciatus）,0.1131 E. awoara - E. akaara）,0.0724（E. awoara -E. septemfasciatus） and 0.1336（E. akaara - E. septemfasciatus）. Both NJ and UPGMA methods yielded an identical phylogenetic tree for the five species. The E. merra and E. fario first clustered together, then joined with E. akaara, and finally clustered with E. awoara and E. septemfasciatus.     

微卫星QTL标记分析豫选黄河鲤群体遗传结构及生长性状相关性

用35个镜鲤微卫星QTL标记评估了豫选黄河鲤(Cyprinus carpio var.haematopterus)群体的遗传结构,并评估了不同基因型与生长性状(体重、体长、体高和体厚)的相关性,筛选了在群体中具有性状优势的基因型。35个微卫星标记在288个豫选黄河鲤个体中的扩增结果显示,各标记等位基因数为3~13,平均每个标记6.828 6个;平均有效等位基因数为4.520 8;平均观测杂合度为0.603 0;平均期望杂合度为0.701 9;平均多态信息含量为0.665 6,群体整体处于高度多态水平(PIC0.5)。利用SPSS19.0的GLM模型分析标记与性状的相关性,共17个微卫星标记与性状表现出不同程度的相关性,其中HLJ2456、HLJ2387和CA1677 3个标记与相应的性状相关性达极显著水平。用Duncan氏多重比较找到了每个微卫星标记具有生长性状优势的基因型,下一步可根据优势基因型指导豫选黄河鲤家系配组,开展基于QTL结果的分子聚合育种研究。     

中国红鲤四群体线粒体DNA遗传多样性、起源及分化

用13种限制性内切酶对兴国红鲤、玻璃红鲤、荷包红鲤及瓯江彩鲤4种红鲤的线粒体DNA(mtDNA)进行RFLP分析。共产生17种限制性态型,其中5种酶为限制性片段长度多态性(RFLPs),归结为5种单倍型;兴国红鲤、玻璃红鲤、荷包红鲤和瓯江彩鲤的核苷酸多样性(Ⅱ)分别为0．0087、0．0059、0．0094、0．0286,瓯江彩鲤的遗传多样性最为丰富;瓯江彩鲤起源于单倍型Ⅱ为主的母系祖先,推算分化时间分别约在11．5万年、9．5万年前;玻璃红鲤和荷包红鲤起源于单倍型I为主的母系祖先,推算分化时间分别约为5～5．6万年、2万年前。     

利用OneMap软件构建鲤遗传连锁图谱

首次使用R环境中的OneMap软件包,以荷包红鲤抗寒品系(♂)和云南大头鲤(♀)为祖父母本所培育的110个F2个体为作图群体,以荧光扩增片段长度多态性(fluorescent amplification fragment length polymorphism,fAFLP)为主要分子标记,采用远交全同胞家系模型构建鲤的遗传连锁图谱。结果显示,110个F2个体中共产生1513个清晰的fAFLP标记,其中多态性标记911个;另开发多态性的EST标记12个,最后总计923个标记用于构建遗传连锁图谱;采用OneMap软件包构建的遗传图谱含有238个fAFLP标记和8个EST标记分布在50个连锁群上,总图距为2876.64cM,标记间平均间距为14.68cM,图谱覆盖率为66.56%。     

Effects of graded levels of dietary vitamin C on the growth,digestive capacity and intestinal microflora of juvenile Jian carp (Cyprinus carpio var. Jian)

A 60‐day experiment was carried out to study the effects of vitamin C [ascorbic acid (AA)] on the growth, digestive enzyme activities and intestinal microbial population. Diets with six levels (0.0, 21.4, 45.1, 69.5, 93.6 and 142.1 mg AA kg^?1 diet) of supplemental ascorbyl polyphosphate were fed to juvenile Jian carp (Cyprinus carpio var. Jian) (12.63±0.02 g). The specific growth rate (SGR), feed efficiency and productive protein value were significantly improved with increasing AA levels up to 45.1 mg AA kg^?1 diet (P<0.05). The hepatopancreas and muscle AA concentrations were increased with increasing dietary AA levels up to 69.5 and 45.1 mg kg^?1 diet respectively (P<0.05). The activities of intestinal trypsin, chymotrypsin, lipase, α‐amylase, Na⁺, K⁺‐ATPase, alkaline phosphatase, gamma‐glutamyl transpeptidase and creatinkinase were all positively affected by the AA supplementation (P<0.05). Intestinal Lactobacillus and Bacillus were increased with increasing AA levels up to 45.1 mg AA kg^?1 diet (P<0.05), while intestinal Escherichia coli decreased with increasing AA levels up to 45.1 mg AA kg^?1 diet (P<0.05). In conclusion, AA improve the digestive capacity and intestinal microbial population of Jian carp, and the dietary AA requirement for SGR of juvenile Jian carp was 40.9 mg AA kg^?1 diet.     

贵州喀斯特山区稻田养殖建鲤肌肉营养成分分析与健康评价

该研究分析了稻田养殖建鲤的肌肉营养成分,旨在了解建鲤在贵州省稻田养殖条件下的营养特点。试验采用常规营养成分分析方法测定了稻田养殖建鲤肌肉中的营养。结果表明:稻田养殖建鲤肌肉中水分含量为76.58%±0.31%,粗灰分含量2.20%±0.03%,粗脂肪含量为2.39%±0.06%,粗蛋白含量为21.02%±0.21%。稻田养殖建鲤肌肉中检测出18种氨基酸,总量为83.47%±1.17%,其中8种必需氨基酸占比34.29%±0.11%,非必需氨基酸占49.18%±0.23%,鲜味氨基酸占31.29%±0.64%,必需氨基酸和非必需氨基酸比值为69.12%,均符合FAO/WHO的评价标准;必需氨基酸指数为76.40%。稻田养殖建鲤肌肉中共检测出21种脂肪酸,饱和脂肪酸(SFA)、单不饱和脂肪酸(MUFA)、多不饱和脂肪酸(PUFA)分别占脂肪酸总量的30.65%、37.51%、31.87%;∑SFA:∑MUFA∶∑PUFA比例为1∶1.22∶1.04,ΣPUFA n-6∶ΣPUFA n-3为1.38。综上所述,相对于其他常见稻田养殖品种,稻田养殖的建鲤营养成分均衡,以期为稻田养殖建鲤以及居民...     

巢湖西半湖北岸湖滨带蓝藻堆积处细菌多样性初步研究

采用限制性片段长度多态性(restriction fragment length polymorphism,RFLP)方法,研究了巢湖西半湖北岸湖滨带蓝藻堆积处不同深度层次的细菌多样性.利用直接法提取了蓝藻堆积处3个不同深度层次样品(A层0～2 cm、B层2～10 cm、C层10 ～ 20 cm)中的细菌总DNA,并进一步构建各层细菌的16S rDNA克隆文库.结果表明,蓝藻堆积处各层细菌的Margalef丰富度指数(dMa)、Shannon-Wiener指数(H)和Simpson指数(D)均伴随深度的增加而减小.系统发育树分析显示,各层的细菌优势类群主要归属于厚壁菌门( Firmicutes)和变形菌门(Proteobacteria);且随深度层次的增加,归属于变形菌门的细菌比例减少,而归属于厚壁菌门的细菌比例增加;在A层样品中存在少量归属于蓝藻门( Cyanobacteria)、放线菌门(Actinobacteria)、浮霉菌门(Planctomycetes)、螺旋体门(Spirochaetes)类群的细菌;在C层样品中也存在少量归属于拟杆菌门(Bacteroidetes)类群的细菌.研究结果在一定程度上揭示了蓝藻堆积腐烂过程中细菌多样性和种类组成的变化.     

Effect of dietary iron levels on growth,body composition and intestinal enzyme activities of juvenile Jian carp (Cyprinus carpio var. Jian)

A 60‐day feeding trial was carried out to investigate the effect of iron on growth, body composition and digestive enzyme activities. Diets with seven levels of iron (53.9, 90.0, 115.6, 146.1, 176.0, 215.8 and 266.0 mg iron kg^?1 diet) were fed to Jian carp (initial weight 11.4 ± 0.0 g). Per cent weight gain (PWG), feed efficiency (FE) and protein efficiency ratio were the lowest in fish fed the basal diet (P < 0.05). Body protein content was increased with the increasing iron levels (P < 0.05), but moisture, lipid and ash of fish were not significantly affected by dietary iron levels (P > 0.05). Activities of trypsin, lipase, α‐amylase, Na⁺, K⁺‐ATPase, alkaline phosphatase and gamma‐glutamyl transpeptidase were improved with increasing dietary iron levels. Serum iron were significantly enhanced with dietary iron levels up to 146.1 mg iron kg^?1 diet, and plateaued. In conclusion, iron improved digestive enzyme activities of juvenile Jian carp and the dietary iron requirement for serum iron of juvenile Jian carp (11.4–64.0 g) was 147.4 mg iron kg^?1 diet with ferrous fumarate as the iron source.