３种虹鳟养殖群体的遗传结构及遗传多样性分析

从40个随机引物中筛选出20个,采用随机扩增多态DNA(RAPD)技术,对甘肃金鳟、道氏虹鳟和美国金鳟3个群体进行了遗传结构和遗传多样性分析.结果表明,甘肃金鳟、道氏虹鳟、美国金鳟群体多态位点百分比分别为82.79%、73.33 %、77.87%;平均等位基因数分别为1.8239、1.7429、1.7887,有效等位基因数分别为1 4823、1 4281、1.4527;基因多样度分别为0.2820、0.2528 、0.2675,Shannon遗传多样性指数分别为0.4219、0 3801、0 4022;以上参数表明,3个群体的遗传多样性均处于偏高的水平,且以甘肃金鳟的遗传多样性最为丰富.甘肃金鳟与美国金鳟之间的遗传距离最大,道氏虹鳟与美国金鳟之间的遗传距离最小.遗传分化指数表明,种内群体间的遗传分化程度很低,而群体内个体间的分化程度很高,变异主要来源于群体内个体间.综合分析认为,甘肃金鳟遗传多样性偏高,有进一步选育的遗传潜力;同时,群体内遗传分化程度较大,说明品种的纯度可能较低,经济性状不够稳定,需进一步选育.     

3种虹鳟养殖群体的遗传结构及遗传多样性分析

从40个随机引物中筛选出20个,采用随机扩增多态DNA(RAPD)技术,对甘肃金鳟、道氏虹鳟和美国金鳟3个群体进行了遗传结构和遗传多样性分析。结果表明,甘肃金鳟、道氏虹鳟、美国金鳟群体多态位点百分比分别为82.79%、73.33%、77.87%;平均等位基因数分别为1.8239、1.7429、1.7887,有效等位基因数分别为1.4823、1.4281、1.4527;基因多样度分别为0.2820、0.2528、0.2675,Shannon遗传多样性指数分别为0.4219、0.3801、0.4022;以上参数表明,3个群体的遗传多样性均处于偏高的水平,且以甘肃金鳟的遗传多样性最为丰富。甘肃金鳟与美国金鳟之间的遗传距离最大,道氏虹鳟与美国金鳟之间的遗传距离最小。遗传分化指数表明,种内群体间的遗传分化程度很低,而群体内个体间的分化程度很高,变异主要来源于群体内个体间。综合分析认为,甘肃金鳟遗传多样性偏高,有进一步选育的遗传潜力;同时,群体内遗传分化程度较大,说明品种的纯度可能较低,经济性状不够稳定,需进一步选育。     

5个虹鳟（Oncorhynchus mykiss）群体的生化遗传分析

采用不连续聚丙烯酰胺凝胶垂直板电泳技术，测定和比较了道氏虹鳟、美国加州虹鳟、挪威虹鳟、丹麦虹鳟、芬兰虹鳟5个群体的15种同工酶约40个基因座位的遗传变异。结果表明，5个虹鳟群体的多态座位比例为43.24%~47.37%；种群平均预期杂合度为0.1917~0.2178，平均实际杂合度为0.2703~0.3291；Hardy-Weinberg 遗传偏离指数(d)为0.4100~0.5331，位点有效等位基因数(Ne)为1.4865~1.5385。χ2检验表明，5个虹鳟群体多态座位基因频率均符合Hardy-Weinberg平衡。与其它鱼类相比，5个虹鳟群体的遗传多样性均处于较高水平，表明本研究的5个虹鳟群体种质资源状况尚好。聚类分析表明，挪威虹鳟、丹麦虹鳟、美国加州虹鳟、道氏虹鳟为同一聚类支，芬兰虹鳟为另一聚类支。本研究对5个群体虹鳟养殖品种的同工酶分析，为其种质资源保护和开发以及遗传育种等方面提供了基础性资料。 关键词：虹鳟；群体；同工酶；遗传结构；遗传变异     

金鳟苗种常见病害防治技术

金鳟是日本长野县水产试验场从虹鳟群体中发现的身体呈金黄色的突变品系。1996年12月黑龙江水产研究所从日本引进同型结合纯系发眼卵1万粒,经三年多的试验养殖从发眼卵至亲鱼育成及人工繁殖均取得了成功。在北京和黑龙江经2年-3年的饲养成国内第1代亲鱼后进行了金鳟与虹鳟杂交育种试验,现已成为我国养鳟业的重要养殖品种。金鳟苗种培育期间病害防治是提高成活率的关键,主要养殖病害有烂鳃     

美国金鳟的同工酶分析

本实验运用聚丙烯酰胺凝胶电泳法对养殖型美国金鳟眼(E)、心(H)、肾(K)、肝(L)、肌肉(M)五种组织的乙醇脱氢酶(ADH)、酯酶(EST)、葡萄糖-6-磷酸脱氢酶(G-6- PDH)、谷氨酸脱氢酶(GDH)、乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)和过氧化物酶(POD)等7种同工酶进行电泳研究。结果显示：美国金鳟中ADH、EST、G-6-PDH、GDH、LDH、MDH、POD、SOD 7种同工酶都有不同程度的组织特异性；美国金鳟群体同工酶分析共记录18个基因座位,其中有8个基因座位为多态,等位基因总数为26个,从而得出多态座位百分数P为44．44％,位点有效等位基因数Ne=1．4444,表明美国金鳟群体遗传多样性偏高。     

鲑鳟通用型低通量单核苷酸多态性芯片的开发

为开发常见鲑鳟养殖物种通用的遗传分析工具,本研究利用Affymetrix虹鳟(Oncorhynchus mykiss) 57K高通量单核苷酸多态性(Single nucleotide polymorphism,SNP)芯片,对国内代表性鲑鳟养殖群体开展了分型检测,包括山女鳟(Oncorhynchus masou masou)、银鲑(Oncorhynchus kisutch)、美洲红点鲑(Salvelinus fontinalis)、白斑红点鲑(Salvelinus leucomaenis) 4个物种,从57,501个SNP标记中筛选出96个共享多态性标记,应用Fluigidm 96.96动态芯片平台,构建了大麻哈鱼属(Oncorhynchus)和红点鲑属(Salvelinus)通用型低通量SNP芯片。该芯片分型结果准确性较高,与Affymetrix高通量芯片分型一致性达到96.55%。使用该芯片对来自6个家系的48尾银鲑个体及其候选亲本进行检测,应用Cervus 3.0.7软件进行亲权鉴定,结果能够准确重现复杂家系的真实系谱。在用于单亲本亲权鉴定时,第一亲本非排除率(Non-exclusion probability for first parent, NE-1P)为4.120×10~(–4);用于双亲本亲权鉴定时,双亲非排除率(Non-exclusion probability for parent pair, NE-PP)低至6.219×10~(–12),表明该芯片在鲑鳟养殖群体系谱鉴定应用中具有较高的准确性。使用该芯片开展4个鲑鳟养殖群体遗传结构分析,样本分群聚类结果与其所属的分类阶元相符,能够准确反映群体遗传组分构成和遗传关系。本研究构建的低通量SNP芯片在常见鲑鳟养殖物种中具有良好的通用性,将其应用于养殖群体遗传分析,能够为鲑鳟制种、育种和引种等科学决策提供基因组信息参考。     

金鳟的生物学特性及养殖

金鳟 (Oncorhynchus mykiss)是日本长野县水产试验场从虹鳟鱼群体中发现的身体呈金黄色的突变品系。 1996年 12月黑龙江水产研究所从日本引进同型结合纯系发眼卵 1万粒。经 3年多的试验养殖从发眼卵至亲鱼育成及人工繁殖均取得了成功。证明我国的冷水资源适于金鳟养殖，金鳟将成为我国养鳟业的重要养殖新品种。金鳟与虹鳟是同一种鱼类，引进后取名金鳟。 　　一、生物学特性 　　金鳟属鲑科鲑属鱼类。同型结合纯系金鳟发眼卵粒径 4.0～ 6.0毫米，因卵内有一直径约 1.5～ 2.5毫米的玫瑰红色油球而呈鲜艳的红色。金鳟体型与虹鳟相似，周身…     

文蛤养殖群体和野生群体遗传多样性的AFLP 分析

应用AFLP标记技术对辽宁和山东沿海文蛤（Meretrix meretrix）养殖群体和野生群体的遗传多样性进行分析。采用7对AFLP引物组合对5个群体（3个野生群体，2个养殖群体）150个个体进行扩增，共得到364个的位点。5个群体内的多态位点比例为84．3945％～87．6465％，总多态位点比例为99．6300％。群体的Nei’s基因多样性指数（H）为0．2301～0．2634；群体的Shannon多样性指数为0．3617～0．4248，群体间的遗传距离为0．0394～0．1609，群体内个体间的遗传距离为0．2592～0．5360。辽宁大洼野生群体和山东河口野生群体的多态位点比例、Shannon多样性指数和群体内遗传距离均高于辽宁盘山和辽宁庄河养殖群体，辽宁庄河野生群体的遗传多样性处于中等水平。用UPGMA方法构建的群体系统进化树显示，辽宁庄河野生群体单独成为一支，辽宁大洼野生群体与庄河养殖群体聚到一起，辽宁盘山养殖群体与山东野生群体聚到一起，但是用这5个群体的150个个体进行的聚类结果显示，所有个体基本是随机交叉聚类，不能形成明显的类群分支。分子变异分析（AMOVA）表明，文蛤群体94．04%的变异来源于群体内，群体间的变异仅占5．96％。以上结果表明，文蛤群体内遗传多样性非常丰富，群体间相似性较大，且存在较强的基因交流。[中国水产科学，2008，15（2）：215-221]     

基于ITS1基因分析亚东鲑(♂)、虹鳟(♀)及其F_1代遗传关系

虹鳟、棕鳟均为优质水产养殖品种,亚东鲑为棕鳟的同物异名,是19世纪西藏由欧洲引进。为探索杂种优势,对亚东鲑和虹鳟进行杂交试验,对其子一代幼鱼与虹鳟、棕鳟进行了体长和体质量测定。采用ITS1基因分析其遗传关系,并与巴基斯坦引进的棕鳟对比研究。结果发现F_1代体长、体质量增长快于虹鳟、棕鳟,其遗传多样性高于亚东鲑和虹鳟,略低于棕鳟,表明F1代具有一定杂种优势。此外,棕鳟较高的遗传多样性也反映出从巴基斯坦引进的发眼卵遗传背景较好,具有进一步养殖选育的潜力。     

长江、珠江、黑龙江三水系的鲢、镛、草鱼原种种群的生化遗传结构与变异

用LKB平板电泳仪聚丙烯酰胺凝胶电泳测定了长江、珠江及黑龙江的鲢、鳙及草鱼的原种8个种群16个酶位点的遗传变异。同种鱼不同水系种群之间存在着明显的生化遗传差异。长江、珠江、黑龙江鲢鱼种群的多态位点的比例分别是13.3％、26.7％、13.3％,平均杂合度分别是0.0493、0.0484、0.0511;长江、珠江鳙鱼种群的多态位点的比例都是31.3％,平均杂合度分别是0.1375,0.0977;长江、珠江、黑龙江草鱼种群的多态位点比例分别是30％,38％,23.1％,平均杂合度分别是0.1241,0.0961,0.0525。南方种群的多态位点比例有比北方的高的趋向。长江鲢—珠江鲢,长江鲢—黑龙江鲢,珠江鲢—黑龙江鲢的遗传相似度依次是0.9957,0.9955及0.9696;长江鳙—珠江鳙的遗传相似度是0.9955;长江草鱼—珠江草鱼,珠江草鱼—黑龙江草鱼,长江草鱼—黑龙江草鱼的遗传相似度依次是0.9679、0.9483及0.9324。长江种群与珠江种群这两个中央群体间的遗传差异较小,边缘群体黑龙江种群与中央群体长江、珠江种群间的遗传差异较大。黑龙江草鱼种群很小,其资源的保护工作应引起注意。     

大西洋鲑、三倍体虹鳟和金鳟的肌肉营养成分与品质特性

为了解大西洋鲑(Salmo salar)、三倍体虹鳟(Oncorhynchus mykiss)、金鳟(Oncorhynchus mykiss)3种鱼肌肉营养成分和品质特性,利用生化分析、物性分析方法分析3种鱼肌肉的营养成分、氨基酸和脂肪酸组成、肉色、系水力和质构特性。结果表明,大西洋鲑、三倍体虹鳟、金鳟肌肉的水分质量分数分别为62.91%、67.15%、73.02%,粗蛋白质量分数分别为22.39%、21.03%、22.11%,粗脂肪质量分数分别为14.64%、17.16%、5.11%。3种鱼肌肉的滴水损失、黄色值(b~*)、羟脯氨酸含量、内聚性均显著不差异(P0.05)。3种鱼肌肉的硬度和咀嚼性由低到高依次为大西洋鲑、三倍体虹鳟、金鳟,而pH值的结果则与之相反(P0.05)。大西洋鲑和三倍体虹鳟肌肉的灰分、解冻损失、蒸煮损失、回复性、弹性和红色值(a~*)差异不显著(P0.05),但灰分、蒸煮损失、回复性均小于金鳟肌肉的对应指标(P0.05),弹性和红色值(a~*)则均大于金鳟肌肉的对应指标(P0.05)。大西洋鲑、三倍体虹鳟、金鳟肌肉中必需氨基酸含量占氨基酸总量分别为42.28%、41.84%、41.63%(质量分数),必需氨基酸/非必需氨基酸(EAA/NEAA)比值分别为73.25%、71.94%、71.32%,均符合联合国粮农组织/世界卫生组织(FAO/WHO)对优质蛋白质的评价标准;3种鱼肌肉中均检测到22种脂肪酸,组成丰富,其中不饱和脂肪酸含量较高。综上所述,3种鱼的肌肉都是符合人体营养需求的优质水产品,其中大西洋鲑和三倍体虹鳟肉质接近,且都优于金鳟的肉质。     

我国泥鳅微卫星遗传多样性初步分析

用6个微卫星标记分析了安徽(AH)、甘肃(GS)、广东(GD)、广西(GX)、黑龙江(HLJ)、江苏(JS)和重庆(CQ)7个泥鳅(Misgurnus anguillicaudatus)群体的遗传多样性。这6个微卫星摘自泥鳅微卫星连锁图谱,不存在连锁不平衡,在HLJ群体中处于哈代-温伯格平衡,适用遗传多样性分析。六个微卫星共检测到69个等位基因,每位点7到16个不等,有效等位基因数(Ne)3.0-5.5个,观察杂合度在0.202和0.408之间,而期望杂合度在0.673和0.820之间,揭示的多样性信息含量(PIC)在0.643和0.796之间。这些数据显示7个泥鳅群体遗传多样性丰富。群体间遗传分化系数(Fst)达到0.499,49.9%的变异可归于群体间差异,而Fis在0.167和0.421之间,表明泥鳅群体存在较明显的近交或斑块化分布。聚类分析显示AH、GD、HLJ和JS聚成一个分支,而CQ、GS 和GX聚为另一个分支。这表明我国泥鳅呈现东西向梯度分布并在南北向可能存在扰动。这些发现将有助于我国泥鳅资源的有效管理和合理利用。     

镜鲤头长及头长体长比性状的主效QTL挖掘

为挖掘镜鲤头长及头长体长比性状的主效QTL区间,实验利用368个SSR、336个SNP标记对镜鲤良种后代杂交F1群体的68个个体进行基因型检测,运用JoinMap 4.0软件包构建遗传连锁图谱。该图谱包含535个分子标记并被分配到50个连锁群上,覆盖基因组总长度为2 244.66 cM,标记间平均距离为4.63 cM。利用MapQTL 5.0(interval mapping,IM)区间作图法进行QTL检测。结果显示,共得到2个与头长相关的QTL区间,分别分布在LG21和LG42,可解释型变异分别为28.2%、32.6%;6个与头长体长比性状相关的QTL位于LG8、LG15、LG18、LG21、LG39、LG40,可解释表型变异范围是16.4%~49.3%。全部QTL区间中贡献率大于20%的主效QTL有7个,HL-21和HL-42是头长性状的主效区间;HBR-8、HBR-15、HBR-21、HBR-39和HBR-40是头长体长比性状的主效QTL区间。利用SPSS的一般线性模型(GLM)针对另一群体进行验证,结果表明HLJ692与镜鲤头长体长比显著相关。     

Allozyme diversity in Australian rainbow trout, Oncorhynchus mykiss

Rainbow trout, Oncorhynchus mykiss (Walbaum), were first introduced into Australia over 100 years ago, and forms the basis of important recreational inland fisheries and an aquaculture industry in south‐eastern Australia. This paper investigates the genetic variation within and between samples of Australian rainbow trout using allozyme electrophoresis. The levels of genetic diversity within Australia do not show marked differences from those observed in hatchery and wild populations from throughout North America, New Zealand and South Africa, but there is evidence for the loss of some rare alleles during translocation from California to Australia via New Zealand. No appreciable difference in genetic diversity was apparent between hatchery and self‐sustaining wild populations of rainbow trout from mainland Australia. However, significant differences in allelic frequencies were observed, with consistent genetic differences between Victorian and New South Wales samples most likely reflecting state‐based hatchery and stocking policies.     

Prediction of Genetic Variation in Stocks of Offspring and Its Correlation with Viability and Growth Rate

This study tested the accuracy of predictions of genetic variation with Genassemblage 1.0 software and investigated whether offspring with greater genetic variation have higher survivability in aquaculture conditions than offspring with less genetic variation. To achieve these aims, microsatellite polymorphism was used to determine genetic variation in rainbow trout, Oncorhynchus mykiss, spawners and progeny. Spawning pairs were assembled based on the estimated genetic variation of their offspring. The offspring were divided into two groups: (H) group had higher expected values of heterozygosity (0.93) and allelic richness (41), whereas (L) group had lower expected values (0.63 and 31). The heterozygosity, survivability, and growth rate of these groups were recorded. The observed heterozygosity was 0.93 in (H) group and 0.62 in (L) group. (H) group had slightly higher survivability (89.8%) than (L) group (87.6%), but this difference was not significant (p ≥ 0.05), possibly due to residual tetrasomy in the rainbow trout genome. In the offspring, predicted and observed variations were very similar, indicating that Genassemblage 1.0 accurately predicts genetic variation in progeny.     

Supplemental effect of bile salts to soybean meal-based diet on growth and feed utilization of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

A feeding experiment was conducted to evaluate the supplemental effect of bile salts to a defatted soybean meal-based non-fish meal diet for rainbow trout Oncorhynchus mykiss. A fish meal-based diet (FM) and two non-fish meal diets with and without bovine bile salts (SC+B and SC, respectively) were fed to fish (13 g initial weight) for 10 weeks. Fish fed diet SC showed inferior growth and feed efficiency, while bile salt supplementation improved the parameters to the same levels as fish fed diet FM. Crude fat and starch digestibility of diet SC-fed fish decreased after the 10-week feeding trial compared to the data obtained with fish that had no experience of the diet. Total biliary bile salt content and intestinal maltase activity of fish fed diet SC were the lowest among treatments, while these parameters were improved by bile salt supplementation. Morphological changes occurred in the distal intestine and liver of the diet SC group, although the histological features of fish fed diet SC+B were similar to those of fish fed diet FM. These results suggest that bile salt supplementation to a soybean meal-based diet improves the nutrient utilization by normalizing digestive processes in rainbow trout.     

Multilocus sequence typing (MLST) analysis of California Flavobacterium psychrophilum reveals novel genotypes and predominance of CC‐ST10 in California salmonid hatcheries

The Gram‐negative bacterium, Flavobacterium psychrophilum, is endemic to California, USA, where it is an important pathogen in salmonid aquaculture, especially in rainbow trout (Oncorhynchus mykiss). Disease outbreaks caused by F. psychrophilum in rainbow trout fingerlings can approach 90% mortality, resulting in millions of dollars of economic losses annually. The focus of this study was to investigate the genetic diversity of 49 F. psychrophilum isolates collected from disease outbreaks in 17 salmonid hatcheries in California, USA, from 2015 to 2018 using multilocus sequence typing. Results suggest California F. psychrophilum isolates are diverse, representing 11 distinct sequence types (STs), three of which were previously undescribed. Still, the majority of genotyped isolates (n = 41) belonged to a single clonal complex (CC), CC‐ST10, which is the largest CC worldwide and has been linked to disease outbreaks on several continents. Results of this study provide evidence of marked intraspecific genetic diversity of F. psychrophilum from California. The biological significance of this genetic variability is unclear but could have implications for future vaccine development and treatments. Further studies investigating the virulence, antigenic, and antimicrobial susceptibility profiles of F. psychrophilum are warranted to better understand the epizootiology of this pathogen in the Western United States.     

Growth in Arctic charr and rainbow trout fed temporally concentrated or spaced daily meals

The effect on growth of distributing feed over a few hours compared tomore frequent meals was tested on 1+ Arctic charr (Salvelinus alpinus L.) and 1+ rainbow trout (Oncorhynchus mykiss Walbaum). Triplicate hatchery groups for each treatment were fed at a ration level of 1%/dayeither with few meals (8 times per day divided into morning and evening)or with frequent meals (32 meals equally distributed during the day). Wefound an opposite effect of meal frequency on growth in the two species.Low feeding intensity (8 meals per day) had a significantly positive effecton growth in rainbow trout but a significantly negative effect on growth inArctic charr when compared to feeding the fish frequent meals. Theopposite response to meal frequency is likely to be an effect of thedifferences in activity during feeding. Rainbow trout feed much moreaggressively than charr which can result in feeding being a more stressfulevent. In this experiment, the specific growth rate was lower and the feedconversion ratio higher for Arctic charr compared to rainbow trout.