Newly developed microsatellite markers for the pan‐European duck mussel,Anodonta anatina: revisiting the main mitochondrial lineages

相似文献   

Identification,characterisation and discriminatory power of microsatellite markers in the parasitic weed Orobanche cumana

Orobanche cumana is an obligate root parasite of sunflower. It represents a major agricultural problem in many countries of southern and eastern Europe. Information on O. cumana population genetics, structure and dynamics is scarce, particularly due to the lack of suitable molecular markers for such studies. The objective of this study was to identify and characterise simple sequence repeat (SSR) markers for O. cumana. Four thousand two hundred SSR‐containing candidate sequences were obtained from O. cumana using next‐generation sequencing, from which 298 SSR primer pairs were designed and 217 of them used for validation. Seventy nine SSR primers produced reproducible, high quality amplicons of the expected size that were polymorphic among 18 O. cumana populations from different geographical locations and hosts (sunflower, wild hosts from the Compositae family). The number of alleles per locus ranged from 2 to 10, with an average polymorphism information content value of 0.37. The O. cumana SSR markers were highly transferable to the closely related species Orobanche cernua. SSR markers showed high resolving power; UPGMA cluster analysis allowed proper classification of Orobanche spp. samples into species (O. cumana and O. cernua), geographical origin and host. The functional SSR markers reported in this study constitute a valuable tool for genetic analyses in O. cumana and related species and will contribute insights into the biology and genetics of this parasitic weed.     

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.     

Isolation and characterization of microsatellite markers from guineagrass (Panicum maximum) for genetic diversity estimate and cross-species amplification

Guineagrass ( Panicum maximum Jacq.) is one of the major forage grasses in tropical and semitropical regions, largely apomictic and predominantly exist as tetraploid. Non-availability of polymorphic molecular markers has been a major limitation in its characterization and improvement. We report isolation and characterization of microsatellites in P. maximum and cross-species results with other five Panicum species. Based on microsatellite-motifs, 15 functional and polymorphic simple sequence repeat (SSR) primer-pairs were designed, validated and employed in estimating genetic relationship among 34 guineagrass accessions. Thirteen primer-pairs amplified single locus and remaining two generated more than two loci with an average of 3.57 bands per locus amounts to 63 bands with 34 guineagrass accessions. Average expected heterozygosity ( H _E) of 0.35 (maximum 0.97) and observed heterozygosity ( H _O) of 0.37 (maximum 0.91) established the efficiency of developed markers for discriminating guineagrass accessions. Dice's similarity coefficients-based unweighted pair group with arithmetic average method-clustering supported with high bootstrap values (≥40) indicated its significance and distinguished all accessions except IG97-93 and IG97-6. Utility of these new SSR loci in genetic diversity study of P. maximum and other cross–amplified species is discussed.     

滇东南濒危植物长梗杜鹃转录组微卫星特征分析

[目的]全面了解滇东南特有濒危植物长梗杜鹃转录组SSR位点的分布及序列特征,为长梗杜鹃的保护和合理开发利用提供遗传学资料,为同属植物及近缘种SSR标记的开发及遗传研究提供便利。[方法]利用Illumina Hiseq 4000高通量测序平台对长梗杜鹃叶片进行转录组测序,再通过MISA软件对测序所得Unigenes进行SSR位点的发掘和分析。[结果]发现含SSR的序列17 354条,共得到23 192个SSR,出现频率为31.30%,平均每3 kb出现1个SSR。二碱基和三碱基重复为长梗杜鹃SSR主要重复单元类型,分别占SSR总数的69.25%和15.07%,187种重复基元中,所占比例最高的是(AG/CT)n(62.01%),其次是(A/T)n(12.34%)、(AC/GT)n(4.52%)和(AAG/CTT)n(4.23%)。在SSR和CDS的交集基因中,共发现15 908个SSR位点,其中2 792个位于编码区,出现频率为0.076 SSR/kb,而非编码区为0.344 SSR/kb,在基因编码区中出现频率最高的是三碱基重复(1 356,48.57%)。在不同长度重复单元中,二碱基重复SSR长度变异程度最高,其次是单碱基重复。长梗杜鹃SSR的频率和长度呈显著负相关(P0.01),相关系数为-0.566。[结论]长梗杜鹃转录组SSR位点的出现频率高、分布密度大、基元类型丰富、重复次数较高、长片段较多,具有较高的多态性潜能,用于遗传分析的潜力很大,能满足该物种的保护遗传学研究。     

皱纹盘鲍微卫星多重PCR体系构建及其在家系鉴定中的应用

为提高微卫星分型效率,从以往报道的皱纹盘鲍微卫星中筛选出易扩增、特异性好的微卫星位点进行组合扩增,并通过优化退火温度、反应体系、引物浓度等条件,开发了4组多重PCR扩增体系.运用CERVUS3.0软件对12个皱纹盘鲍全同胞家系的372个子代进行家系鉴定,验证了这4组多重PCR在家系鉴定中的效率.结果发现,仅用1组微卫星多重PCR模拟和实际家系鉴定的成功率分别为86％和90％,两组则达到100％.结果表明,微卫星多重PCR技术能准确地把任意子代鉴定至其所属家系,可以进行大批量家系材料分析,具有较好的应用价值.     

安徽地区克氏原螯虾群体的遗传多样性和遗传结构

为了解安徽地区克氏原螯虾养殖群体的遗传多样性，实验以安徽芜湖 (WH)、宣城 (XC)、合肥 (HF) 3个地区克氏原螯虾人工养殖群体为研究对象，分别以安徽铜陵 (TL)、马鞍山 (MAS) 2个野生群体和监利 (JL)、建湖 (JH)、滆湖 (GH)、兴化 (XH) 4个人工养殖群体作为对照，选用10对克氏原螯虾微卫星引物对其进行微卫星遗传多样性和遗传结构研究。结果显示，安徽地区人工养殖克氏原螯虾群体的平均遗传多样性均高于2个野生群体和江苏、湖北4个地区的人工养殖群体，其中XC群体的遗传多样性最高。9个群体全部10个位点经Bonferroni法校正后均显著偏离Hardy-Weinberg平衡且绝大多数位点显示杂合不足。AMOVA分析表明遗传变异是由群体内部决定的；大多数组的F_st表现出中度分化 (0.05 < F_st < 0.15)。基因流表明不同群体之间存在着广泛的基因交换，尤其是GH和JH群体之间。基于群体间Nei氏遗传距离及UPGMA聚类树结果显示，WU、GH、MAS和JH群体聚为一组，XC和HF群体同属于一组，而JL、TL和XH群体分别自成一组。STRUCTUR结果显示，XC和HF群体的大多数个体被分配到相同的遗传群中，说明上述群体起源相同。研究表明，安徽地区克氏原螯虾人工养殖群体具有较高的遗传多样性。实验结果可为安徽地区克氏原螯虾种质资源的保护和改良提供参考资料。     

Mapping of quantitative trait loci controlling cotton leaf curl disease resistance in upland cotton

Cotton leaf curl disease (CLCuD) is a major threat to cotton production in Asia and Africa. Using marker-assisted breeding can be the best sustainable approach to tackle CLCuD. Identification of new QTLs in the indigenous cotton germplasm is necessary to combat CLCuD. The current study was designed to construct a genetic linkage map of bi-parental F2:F3 populations developed from highly tolerant MNH-886 and highly susceptible S-12 cotton cultivars. One hundred seven CLCuD-associated simple sequence repeat (SSR) marker alleles were identified as polymorphic and three new QTLs were found on chromosomes C11, C19 and C21. Two QTLs on chromosomes C11 and C19 were detected in both F2 and F3 populations in the region flanked by SSR markers CIR316 and BNL4094, and BNL285 and BNL3348, respectively. Whereas, one QTL on chromosome C21 was detected in the region flanked by SSR markers JESPR158 and JESPR135 in both F2 and F3 generations. The CLCuD-associated QTLs identified in this study can help fine-tune the molecular mapping of the QTLs on the cotton genome against CLCuD.     

棘头梅童鱼七个野生群体遗传多样性的微卫星分析

为研究中国沿海地区棘头梅童鱼群体的遗传多样性,利用微卫星标记技术,采用9对微卫星引物对中国连云港(LYG)、大丰(DF)、崇明(CM)、舟山(ZS)、温州(WZ)、宁德(ND)、厦门(XM)棘头梅童鱼7个野生群体的遗传多样性和遗传结构进行了分析。结果显示,实验检测到63个等位基因,各位点等位基因数为3~13个,有效等位基因数为1.7510~8.0317;各位点的观测杂合度(Ho)为0.3596~0.7854,期望杂合度(He)为0.4300~0.8780;多态信息含量(PIC)为0.3604~0.8631,其中有2个位点表现为中度多态(0.25PIC0.5),7个位点表现为高度多态(PIC0.5),具有较丰富的遗传多样性水平。Hardy-Weinber平衡分析显示,7个群体的大部分位点未偏离平衡。基于群体间Nei氏标准遗传距离构建的7个野生群体UPGMA系统进化树结果显示,ND和WZ群体遗传关系最近,ZS和WZ群体遗传关系最远,WZ和ND聚为一支,但总体上没有显著的遗传分化。     

长江胭脂鱼人工放流子一代遗传多样性初步研究

运用微卫星分子标记对80尾胭脂鱼人工放流子一代个体的遗传多样性和种群遗传结构进行了分析。用磁珠富集法构建了胭脂鱼(AAAG)n富集文库，从中分离并鉴定得到的22个微卫星位点最终设计合成18对引物。对长江中、上游的80尾人工增殖放流的胭脂鱼子一代进行了遗传多样性的初步研究，其中5对引物呈现多态性，等位基因数目为 2～8个；多态信息含量0.2957～0.8038；Shannon多样性指数0.5466～1.8840；观测杂合度0.3056～0.7222；期望杂合度0.3658～0.8381；Hard-Weinberg遗传偏离指数（D）-0.1818～0.4287。结果初步表明，胭脂鱼人工放流子一代处于较高的遗传多样性水平，但很大程度上仍受到人类活动的影响。