草鱼的微卫星亲权鉴定

【目的】从亲权排除率、鉴别准确率、鉴别成功率等方面评价微卫星标记鉴别草鱼亲权关系的可行性,为草鱼系谱信息的确定奠定基础。【方法】利用8个微卫星标记检测5个草鱼家系亲本（F0）及其子代（F1）的基因型,以Cervus3．0软件进行亲权鉴定,并统计微卫星位点的等位基因数（Na）、多态信息含量（PIC）、观测杂合度（H0）和期望杂合度（He）等;最后通过模拟分析和试验验证,评估8个微卫星位点最多可鉴定的亲本数及其与鉴定率、置信度之间的关系。【结果】8个微卫星位点中,当两个亲本基因型未知时,单个亲本排除率（E-1P）介于0．342-0．668,平均值为0．520;当一个亲本基因型已知时,另一个亲本的排除率（E-2P）介于0．521-0．802,平均值为0．681;一对亲本组合的亲权排除率（E-PP）介于0．711-0．937,平均值为0．847。以95％置信水平和100％鉴定率为标准,模拟分析及试验验证结果表明,所选择的8个微卫星位点最多可以鉴定598个已知性别的亲本或431个未知性别的亲本。【结论】所选的H118、H137、H165、H148、4703、C2489、H57、H81等8对微卫星标记可在生产及科研试验中用于获取草鱼系谱信息。     

应用6个STR基因座进行奶牛亲子鉴定

运用PCR技术用BM2113、BM1862、BMc701、BM2934、TGLA122、BM720共6个STR(短串联重复序列)基因座对7头奶牛DNA进行扩增，扩增产物分两组混合后经聚丙烯酰胺凝胶电泳和银染检测，扩增片段互不干扰。采用已确定母女关系的2头奶牛血液样本和5个嫌疑父亲的精液样本，对一头母本已知的奶牛在嫌疑父本中找出父本，并计算父权概率为99．993％。结果证明，银染法检测STR基因座扩增产物可用于奶牛亲子鉴定。     

Parentage testing for hatchery-produced abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> based on microsatellite markers: preliminary evaluation of early growth of selected strains in mixed family farming

This study dealt with the detection of parentage in a mixed family tank of abalone Haliotis discus hannai using microsatellite DNA markers. As a preliminary investigation, the early growth rate between selected and non-selected abalone families, which were reared together in the mixed family tank from settlement stage to juvenile stage (7 months of age), was also compared. The selected families were derived from seven parents (three females and four males) selected for larger size in shell length at about 1-year-old, and the non-selected families originated from five wild captives (three females and two males). Parentage analysis based on eight microsatellite markers unambiguously allocated the 170 juveniles sampled from the single tank to the 17 parental pairs. The family size was highly heterogeneous among families, as two males in the selected families and one male in the non-selected families dominated the contribution to the offspring pool (>80%). The mean shell length of the selected families was approximately 23% larger than that of the non-selected families (Student’s t-test, P<0.001). This study demonstrated that the use of microsatellite markers is effective for parentage determination in the mixed family farming, commonly used in abalone hatcheries, and selective operations for larger size could improve the growth of the next generation.     

柏氏鲤与荷包红鲤抗寒品系杂交子二代的RAPD分析

采用随机扩增多态性DNA（RAPD）分子标记技术，对荷包红鲤抗寒品系与柏氏鲤进行杂交所获得的F2代个体进行亲子鉴定分析。从180个随机引物中筛选出5个多态性较丰富的引物，并对其产生的多态性DNA片段进行统计分析，结果显示：双亲的RAPD谱带全部在F2代中表现出来，而子代中产生的所有谱带在双亲中的表现率在AB02、AB05、AC20、AI10、AI13几个引物中分别为87．0％、100．0％、92．1％、82．8％、92．6％。这也表明BAPD技术在对鲤的亲子鉴定中具有很强的可行性。     

应用微卫星对大足黑山羊进行亲缘关系鉴定

为了确定大足黑山羊的亲缘关系,本试验选用24个微卫星位点对133只完全没有血缘记录的大足黑山羊基因组DNA进行扩增,扩增产物用聚丙烯酰胺凝胶电泳进行分离,银染显色后获得个体的基因型.用亲缘分析软件Cervus2.0进行个体关系分析,最终将子代个体以父权分为了11个群体,并对11只种公羊进行聚类分析,得出整个群体的亲缘关系,本实验结果对大足黑山羊的亲缘关系进行了初步分析,可为该群体的选育选配提供了参考.     

辐照花粉辅助授粉对百合远缘杂交结实的影响及杂种早期分子鉴定

研究了辐照花粉辅助授粉对百合‘pollyanna’(♀)×毛百合杂交结实的影响。结果表明:采用普通柱头授粉能获得许多膨大果实和饱满种子,表明该组合为亲合性组合。用经1000和2500Gy的60Coγ射线辐照的母本花粉进行辅助授粉明显提高了果实中饱满种子的结籽率,表明2种剂量的辐照花粉辅助授粉均能有效克服该组合的杂交障碍。结籽数的提高能获得更多的后代,增加了从F1代中选育新优单株的几率。随机选取辐照花粉辅助授粉获得的杂交子代15株,采用ISSR技术对其组培苗进行亲子鉴定,6条ISSR随机引物在15个子代中共扩增出372个条带,其中具有父本特征带114条,具有母本特征带67条,具有双亲共有带65条。ISSR鉴定结果证实了杂种的真实性。     

三角帆蚌紫色选育系1龄阶段内壳色及生长性状的遗传参数估计

采用群体选育辅助种内群体间杂交选育的方法,以内壳色、体质量作为选育性状,经过连续4代的选育获得了紫色选育系F4。本实验以F4为亲本进行繁殖,利用6对微卫星标记,对15个同批繁育的F4母蚌的1龄后代进行亲子鉴定,鉴别出了来自12个父本、15个母本的42个全同胞家系,使用ASREML软件的约束极大似然法对三角帆蚌内壳色及生长性状进行了遗传参数分析。结果显示,内壳色颜色参数L*、a*、b*、dE*的遗传力分别为0.31±0.22、0.11±0.08、0.36±0.18、0.29±0.19,L*、a*、b*之间的遗传相关和表型相关均较低,范围为0.08~0.47和0.04~0.32,L*与dE*相关性最大,遗传相关为-0.94±0.06,表型相关为-0.96±0.01;生长性状壳长、壳高、壳宽、体质量和壳重的遗传力分别为0.24±0.19、0.37±0.27、0.26±0.16、0.26±0.17、0.31±0.19,各性状间遗传相关和表型相关均为正相关,分别为0.71~0.92、0.66~0.94;颜色参数与生长性状的遗传相关和表型相关均很低,为0.02~0.18。三角帆蚌紫色选育系1龄阶段内壳色和生长性状的遗传力多为中高水平,对其继续进行遗传改良预期能够获得良好遗传进展。内壳色与生长性状的相关度很低,无法实现相互选择,体质量与其他生长性状相关均较紧密,表明将内壳色、体质量作为目标性状进行同步选育的方法合理,可实现同时改良壳色及生长性能的目的。     

多倍体中华鲟微卫星亲子鉴定体系的建立

应用荧光引物和自动测序技术,检测了10个自主研发的微卫星分子标记在68个中华鲟(Acipenser sinensis)个体中的遗传多态性。结果表明,10个微卫星标记共检测到124个等位基因,等位基因数为7~22,期望杂合度为0.730 26~0.873 95,Shannon-Wiener多样性指数为1.561 61~2.219 60。基于亲子鉴定排除率不低于0.99的标准,最终确定Asi-75067、Asi-67648、Asi-67123、Asi-73843、Asi-72040、Asi-70421和Asi-56700等7个微卫星标记为中华鲟亲子鉴定的核心体系,其单亲、父权和双亲的累积排除率分别为99.12%、99.95%和99.99%。该微卫星标记组合为中华鲟准确高效经济的亲子鉴定体系建立提供了科学依据。     

Heritability estimates for growth‐related traits in the Pacific oyster (Crassostrea gigas) using a molecular pedigree

Pacific oyster is one of the leading species in world aquaculture, but heritability estimation applying mixed‐family approach has not been actively pursued. In this study, heritability for growth‐related traits in the Pacific oyster was first estimated by creating a single cohort of 45 families in a full‐factorial mating design consisting of nine sires and five dams. A total of 270 offspring were analysed and parentage assignment inferred by six microsatellite markers achieved 100% success. All parents contributed to the spawn and a total of 42 full‐sib families were represented. Using an animal model, heritability estimates at 12 months of age were 0.49 ± 0.25 for shell height, 0.36 ± 0.19 for shell length, 0.45 ± 0.23 for shell width and 0.35 ± 0.17 for wet weight. Genetic correlation between shell height and wet weight was quite high (0.79 ± 0.25), suggesting that direct selection of shell height, which is an easily measurable trait, also improves wet weight. The results obtained in this study indicate that growth‐related traits could be improved by exploiting additive genetic effects through selective breeding.