5个重庆地方猪种遗传多样性的微卫星分析

目的通过微卫星标记评估8个安徽地方猪品种的遗传多样性。方法采用PCR和聚丙烯酰胺凝胶电泳对FAO-ISAG联合推荐的15个微卫星标记进行分型,并计算有效等位基因数 (N_e)、期望杂合度 (H_e)、多态信息含量 (PIC)、F统计量和遗传距离等参数,探讨品种间的遗传多样性和遗传关系。结果15个微卫星标记共检测到272个等位基因,平均有效等位基因数9.561 2个。8个品种的有效等位基因数在2.445 1~3.786 6之间,期望杂合度在0.532 0~0.714 8之间,多态信息含量在0.369 8~0.890 6之间。F统计量和基因流 (N_m) 分析表明：安徽地方猪品种间存在一定程度的近交和基因流动。基于Nei氏遗传距离 (D_s) 构建系统发生树显示：8个地方猪种可分为两类：一类为皖南花猪;另一类又聚为两小支,一小支为安庆六白猪、圩猪和霍寿黑猪,另一小支为皖北猪、皖南黑猪、枞阳黑猪和定远猪。结论8个安徽地方猪种遗传多样性丰富,群体内存在一定程度的近交和基因流动。结果为安徽地方猪种的保护和利用提供了理论依据。     

ISSR分子标记在芦柑提纯复壮研究中的应用

采用ISSR(Inter-simple Sequence Repeat)分子标记技术,从随机引物中筛选出能稳定扩增的14条引物,利用2条引物对供试的10个芦柑样品进行PCR扩增,获得了与母本完全一致的条带,从而可以确定鉴定样本为珠心苗。     

Genetic Analysis and Molecular Tagging a Novel Yellow Rust Resistance Gene Derived from Synthetic Hexaploid Wheat Germplasm M08

Yellow rust of wheat （caused by Puccinia striiformis Westend. f. sp. tritici Eriks.） has been periodically epidemic and severely damaged wheat production in China. The development of resistant cultivars could be an effective way to reduce yield losses of wheat caused by yellow rust. Rust reaction tests and genetic analysis indicated that M08, the synthetic hexaploid wheat derived from hybridization between Triticum durum （2n = 6X = 28; genome AABB） and Aegilops tauschii （2n = 2X = 14; genome DD）, showed resistance to current prevailing yellow rust races at seedling stage, which was controlled by a single dominant gene, designated as YrAm. Bulked segregant analysis was used to identify microsatellite markers linked to gene YrAm in an F2 population derived from cross M08 （resistant） × Jinan 17 （susceptible）. Three microsatellite marker loci Xgwm77, Xgwm285, and Xgwml31 located on chromosome 3B were mapped to the YrAm locus. Xgwml31 was the closest marker locus and showed a linkage distance of 7.8 cM to the resistance locus. Thus, it is assumed that YrAm for resistance to yellow rust may be derived from Triticum durum and is located on the long arm of chromosome 3B.     

利用酵母磷酸甘露糖异构酶基因作为拟南芥遗传转化的筛选标记

利用PCR反应克隆来自酿酒酵母的磷酸甘露糖异构酶(PMI)基因pmi,通过农杆菌介导的花序侵染法将PMI的基因导入受体拟南芥中,将转化后的拟南芥种子放在质量浓度为1g/L的甘露糖MS培养基上进行筛选培养,共得到抗性拟南芥植株4株。PCR反应证明酵母PMI的基因pmi已经整合到拟南芥基因组中,RT-PCR检测证明该基因的内含子能够在拟南芥中被正确剪切,同时通过GUS化学组织染色方法证明了伴随转化的GUS基因gus在拟南芥中得以表达,以上研究结果说明酵母PMI/甘露糖系统可以作为拟南芥遗传转化的筛选标记。     

基于α-半乳糖苷酶基因为筛选标记的酿酒酵母诱导型表达载体的构建及应用

以α-半乳糖苷酶基因为筛选标记,构建GAL1诱导型启动子介导的酿酒酵母表达载体YGM-α-gal质粒,将枯草芽孢杆菌的β-1,3-1,4-葡聚糖酶基因克隆到此载体中,构建质粒YGMPA-α-gal,转化宿主酵母后,实现β-1,3-1,4-葡聚糖酶在酿酒酵母中的分泌表达.结果表明:在2%半乳糖诱导下,摇瓶发酵24h后分泌表达的β-葡聚糖酶活性达到411.9U爛mL-1,而在培养60h后,发酵液中α-半乳糖苷酶活性可达64.2U爛mL-1.说明α-半乳糖苷酶基因可用作酿酒酵母表达载体转化的有效筛选标记,为食品级酿酒酵母表达系统的构建提供了新选择.     

中国小麦抗穗发芽种质资源的挖掘与创制

根据4年的表型数据,并结合实验室开发和鉴定的13个分子标记,对我国833份小麦种质资源（主要包括278份小麦微核心种质、124份地方品种和431份现代推广品种及高代品系）穗发芽抗性进行鉴定。结果表明,13个分子标记鉴定的抗/感穗发芽等位类型间相对发芽指数（RGI）差异均达显著或极显著水平,其中TaMFT-222和Ta MFT-194标记鉴定的差异最大,U值分别为14.98**和11.30**,均达极显著水平,其优异等位类型可以降低相对发芽指数0.21～0.32。其次是Sdr2A、CNGC2AL、Vp1-b2、Ta MKK3-A、PM19、CAPS-2AL、A17-19和EX06323标记,其等位类型间穗发芽抗性差异也均达极显著水平;Qsd1和Barc321标记也能显著区分穗发芽抗性。共计鉴定出63份穗发芽抗性较好的种质资源,其中达到抗的有41份,多为红皮品种和地方品种;达到中抗的有22份,白皮半冬性居多。利用分子标记辅助选择,并结合杂交聚合,创制出12份穗发芽抗性水平达到中抗和抗的种质资源,至少携带3个抗穗发芽基因/位点。该结果为抗穗发芽新品种选育提供重要遗传资源。     

贵州21份玉米优质种质资源的SSR标记分析

为建立优质玉米种质资源的指纹图谱,给贵州玉米优良杂交组合的选配提供分子水平上的依据和基础,利用微卫星分子标记（SSR）技术对21份贵州玉米优质种质资源进行了遗传多样性分析。结果显示,共筛选出多态性丰富的20对 SSR 引物,在供试材料中检测出97位基因变异,每对引物检测到等位基因2-11个,平均4.85个。 SSR引物的多态性信息量（PIC）介于0.19-0.78,平均为0.53。UPGMA聚类分析表明,供试材料间遗传距离变幅为0.22-0.63,平均为0.43。     

花生晚斑病抗性AFLP标记

以抗感晚斑病组合“中花5号×ICGV 86699”的F2分离群体为材料，经田间抗性鉴定明确抗性亲本ICGV 86699的抗性受隐性单基因或主效基因控制；AFLP分析结合BSA法筛选到与晚斑病抗性连锁较紧密的AFLP标记3个，即E35/M51、E37/M48和E41/M47，它们与抗性间的图距分别为7.40cM、7.40cM和8.67cM；所获得的3个标记间连锁紧密，位于同一连锁群上。这3个标记在具有野生种亲缘的7个抗病花生品种或材料中均能检测到，而在栽培种以多粒型为代表的抗病材料中均未检测到，表明ICGV 86699的抗性基因与栽培种花生中的抗性基因不同。这是国内外有关花生晚斑病抗性分子标记的首例报道。     

生物大分子标记物检测在环境监测中的应用

生物大分子标记物是指生物体内的一些对外界环境变化敏感并能产生一些可检测变化的大分子物质,这些大分子物质能够反映环境变化对生物体的影响。随着社会对环境保护的日益重视和分子生物学技术的发展,将生物大分子标记物的检测应用到环境监测中已经成为一种趋势。生物大分子标记物检测由于其测定指标全面、准确、系统且具有特异性等优点,近十几年来作为污染物暴露和毒性效应的早期预警工具已被广泛应用于环境评价中。本文对一些主要的生物大分子标记物及其检测技术在环境监测中的应用状况及应用前景进行综述,旨在为生物大分子标记技术在环境检测实际操作中的应用提供参考依据。     

Accuracy of pairwise methods in the reconstruction of family relationships, using molecular information from turbot (Scophthalmus maximus)

Many estimators and algorithms have been developed to infer the genealogical relationships from molecular marker data when there is no pedigree information. Most pairwise methods provide estimates in a continuous range that needs to be converted into genealogical relationships (namely full-sibs, half-sibs and unrelated) if there is a previous knowledge of the population structure. Transformations are usually based on arbitrary thresholds, but the possibility of correcting the coancestry estimates via explicit pedigree reconstruction methods has been suggested. Using molecular data for ten highly polymorphic microsatellite loci on a population of turbot (Scophthalmus maximus) with a known genealogy, the behaviour of four pairwise marker-based coancestry estimators and the molecular coancestry has been evaluated. The population consisted on 138 families with 4 full-sib individuals each and one family with 8 full-sib individuals (up to 15 half-sib families were also present due to the sharing of parents between some of the full-sibs families). Our results suggested that transforming the pairwise estimators and the molecular coancestry to family relationships through the establishment of thresholds performs slightly better than the explicit pedigree reconstruction method, when accuracy is measured in a pairwise basis. However, if joint relationships between more than two individuals were tested at a time, the threshold methods led to a high percentage of incongruous triads of full-sib individuals, with Mendelian incompatibilities appearing in congruous full-sib families (more than 70% and 60% in our study, respectively). The explicit pedigree reconstruction approach, due to its nature, is free from such problems. Therefore, the pedigree reconstruction approach seems to be a valuable tool to provide a congruent and compatible pedigree when the pairwise marker-based coancestry matrices or the molecular coancestry need to be transformed.