猪3号染色体九个微卫星位点的遗传多态性研究和遗传图谱构建

从已公布的猪 3号染色体连锁图谱中选取 9个微卫星位点 ,分析了这些位点在大白猪×梅山猪资源家系F2代 140头个体上的多态性 ,利用Crimap软件分别构建了猪 3号染色体两性平均连锁图谱以及公、母畜连锁图谱。结果表明 :各位点等位基因数为 2～ 4个 ,杂合度为 0 .436～ 0 .6 5 6 ,多态信息含量为 0 .35 1～ 0 .5 82 ;本研究所构建的平均连锁图谱全长为 16 1.1cM ,公、母畜连锁图谱全长分别为 135 .8cM和 188.7cM。与USDA所公布的连锁图谱相比 ,两者的标记顺序一致 ,但我们的图谱标记间隔偏大。     

猪1、3号染色体微卫星位点多态性及遗传连锁图谱的构建

利用 3头英系大白猪和 7头梅山猪为父母本建立了F2 参考家系。F1 公猪 5头 ,母猪 2 3头 ,随机交配产生 147个F2 个体。根据美国肉畜研究中心 (USDA MARC)公布的猪遗传连锁图谱 ,在 1号和 3号染色体上等间隔 (2 0cM)选择 8个和 9个微卫星标记 ,对参考家系全部的F0 、F1 和F2 个体进行扩增 ,获得各标记位点基因型。研究结果表明 ,等位基因数介于 2到 5之间 ,平均每位点 3.35个等位基因 ;部分等位基因片段长度超过USDA MARC所报道的结果。标记位点杂合度为 0 .385 3～ 0 .70 97,平均 0 .5 795。有信息的减数分裂数为 35～ 30 5 ,平均 2 40。利用此参考家系和CRI MAP软件包构建的 1号和 3号染色体图谱分别长 182 .3cM和 180 .2cM。 1号染色体的母畜图谱短于公畜图谱 ,而 3号染色体正好相反。与USDA MARC报道相比较 ,微卫星排列顺序与报道相同 ,但 1号染色体长 44 .8cM ,3号染色体长 6 3.3cM。此连锁图的构建为以后的数量性状基因位点 (quantitativetraitloci,QTLs)定位奠定了基础。     

利用两个测序水稻品种构建微卫星连锁图谱

利用已完成基因组测序的两个水稻品种日本晴和931l的数据库成功开发出水稻微卫星新标记,并利用由90个单株组成的日本晴×9311 F2作图群体,构建了一张包含152个SSR标记位点、覆盖基因组总长度2 455.7 cM的连锁图谱,有46个SSR新标记为自主开发,该图谱标记间的平均遗传距离为16.16 cM;并将未能在Temnykh等人(2001)构建的图谱上定位的微卫星标记RM345和RM494定位在第6染色体上.通过与Temnykh等人(2001)和兰涛等人(2003)所构建的图谱从作图群体的类型和大小、标记的类型和数量、标记在染色体上的线性排列顺序等几个方面进行比较,所绘制的图谱其标记在染色体线性排列上与Temnykh等人绘制的图谱具有很高的一致性,达93.81%.     

利用鸡F2资源群体构建1号染色体遗传连锁图谱

在鸡1号染色体上选取23个微卫星标记,利用东北农业大学鸡F2资源群体构建了遗传连锁图谱。选用369只F2个体用于基因型测定。结果表明23个微卫星位点除MCW0058为低度多态,其他位点均为中高度多态。构建的连锁图谱覆盖1号染色体全长,总共637.9 cM。MCW0115和ROS0025标记顺序与EL图谱不同,但与WAU图谱一致。其他标记顺序与3大参考家系标记顺序一致,图谱总长和标记间距离大于3大参考家系。此连锁图谱的构建为数量性状位点（QTL）定位奠定了良好的基础。     

用商品群作为参考系构建猪的微卫星连锁图谱

由 19头杂种公猪 [皮特兰× (皮特兰×汉普夏 ) ]、5 2头杂种母猪 [Leicoma× (大约克×长白 ) ]及其 332头后代组成的商品群作为参考系 ,选择 172个微卫星标记和 3个Ⅰ类标记 (RYR1、PIT1、PRKAG3)对参考系的个体进行遗传标记分型 ,应用CRIMAP(2 4 )构建猪的整个基因组微卫星连锁图谱。采用多重PCR方法对微卫星标记进行扩增 ,用ABI 377测序仪进行电泳分离 ,应用Genescan 3 0和Genotyper 2 0软件进行基因型检测。 3个Ⅰ类标记用PCR RFLP技术进行分型。CRIMAP程序分析表明 :所构建的猪常染色体性平均连锁图谱的总长度为 2 36 8 7cM ,X染色体的长度为 14 3 10cM ,遗传标记的平均间距为 16 3cM ,亲本的微卫星标记座位的杂合度平均为 0 70。此连锁图谱的构建将为商品猪群的生长、胴体、肉质以及繁殖性状的QTL扫描打下基础。     

RAPD和SSR两种标记构建的中国对虾遗传连锁图谱

利用RAPD和SSR分子标记结合拟测交策略,对中国对虾(Fenneropenaeuschinensis)“黄海1号”雌虾与野生雄虾作为亲本进行单对杂交产生的F1代,采用RAPD和SSR两种分子标记技术初步构建了中国对虾雌、雄遗传连锁图谱。对460个RAPD引物和44对SSR引物进行筛选,共选出61个RAPD引物和20对SSR引物,用于对父母本和82个F1个体进行遗传分析。共得到母本分离标记146个(RAPD标记128个,微卫星标记18个)和父本分离标记127个(RAPD标记109个,微卫星标记18个)。雌性图谱包括8个连锁群、9个三联体和14个连锁对,标记间平均间隔为11·28cM,图谱共覆盖1173cM,覆盖率为59·36%;雄性图谱包括10个连锁群、12个三联体和7个连锁对,标记间平均间隔为12·05cM,图谱共覆盖1144·6cM,覆盖率为62·01%。中国对虾遗传图谱的构建为其分子标记辅助育种、比较基因组作图及数量性状位点的定位与克隆奠定了基础。     

水稻籼粳交F2、F6群体遗传连锁图谱的比较分析

以部分基因组和全基因组测序水稻籼稻(O sativa L. indica)品种“培矮64S”(Pei’ai 64S♀)和粳稻(O sativa L. japonica)品种“日本晴”(Nipponbare♂)为构图亲本, 选取F2代180个株系为作图群体, 构建含138个微卫星位点的水稻遗传连锁图谱, 覆盖基因组2 046.2 cM, 平均图距17.1 cM, 即F2 图谱; 采用单粒传法获得F2:6 代330个株系, 用相同的多态性标记分析F6群体, 构建含92个标记连锁图谱, 覆盖基因组2 563.5 cM, 平均图距27.86 cM, 即F6图谱; F2、F6图谱在连锁群数、定位标记数、标记的位置顺序、遗传图距、平均图距等方面发生了较大变化, 并对产生这些差异的原因进行了初步分析。     

RAPD和SSR两种标记构建的中国对虾遗传连锁图谱

利用RAPD和SSR分子标记结合拟测交策略，对中国对虾(Fenneropenaeus chinensis)“黄海1号”雌虾与野生雄虾作为亲本进行单对杂交产生的F1代，采用RAPD和SSR 两种分子标记技术初步构建了中国对虾雌、雄遗传连锁图谱。对460个RAPD引物和44对SSR引物进行筛选，共选出61个RAPD引物和20对SSR引物，用于对父母本和82个F1个体进行遗传分析。共得到母本分离标记146个(RAPD标记128个，微卫星标记18个)和父本分离标记127个(RAPD标记109个，微卫星标记18个)。雌性图谱包括8个连锁群、9个三联体和14个连锁对，标记间平均间隔为11.28 cM，图谱共覆盖1 173 cM，覆盖率为59.36%；雄性图谱包括10个连锁群、12个三联体和7个连锁对，标记间平均间隔为12.05 cM，图谱共覆盖1 144.6 cM，覆盖率为62.01%。中国对虾遗传图谱的构建为其分子标记辅助育种、比较基因组作图及数量性状位点的定位与克隆奠定了基础。     

猪F2设计资源家系中4条染色体微卫星标记分析

利用中国地方猪种蓝塘猪(16头母猪)与外来品种长白猪(8头公猪)按F2设计建立资源家系, 根据美国肉畜中心(USDA-MARC 2.0)公布的猪连锁图谱, 在1、4、7和8号染色体上间隔10~20 cM选择一个微卫星标记, 共31个标记, 采用WAVEÒ核苷酸片段分析系统和ABI 377 DNA序列分析仪检测资源群体的P、F1和F2代个体微卫星的基因型, 对其基因频率、杂合度和多态信息含量等进行统计分析。结果发现: 利用ABI 377检测的猪1、4和8号染色体上的有效微卫星标记21个, 其中13个标记的18个等位基因片段大小超过了网上已报道的结果, 发现新等位基因的标记占62%; 在31个微卫星标记中, 杂合度(h)在0.043~0.7855之间, 总平均杂合度为0.6460, 其中70%座位的h>0.60; 总平均多态信息含量(PIC)为0.5949, 有77.4%位点的PIC>0.5。统计分析结果表明, 选用的微卫星标记能够较好地提供标记信息, 为进一步在该家系中进行猪重要性状的QTL定位打下了良好的基础。     

猪部分内脏器官性状和乳头数的QTL检测

对内脏器官重量性状的QTL定位研究,所见报道不多;对于猪的繁殖性状,尚需做进一步的探讨。本研究在总共214头(180头F2个体)组成的资源家系中,在猪的SSC4、SSC6、SSC7、SSC8 和 SSC13上共选取39个微卫星标记,检测了8种内脏器官的重量性状:心重 (HW)、肺重 (LW)、肝 胆重 (LGW)、脾重 (SPW)、胃重 (STW)、小肠重(SIW)、大肠重(LIW) 和肾重(KW);其他一些胴体性状:胴体长性状1(自第一颈椎,CL1)、胴体长性状2(自第一胸椎,CL2)、肋骨数(RNS)和繁殖性状乳头数(TNS)的QTL定位。结果表明,检测到3个染色体极显著水平的QTL(P≤0.01),它们是HW QTL定位在SSC6上30 cM处,RNS QTL定位在SSC7上115 cM处和TNS QTL定位在SSC7上 110 cM处;另外6个染色体显著水平的QTL(P≤0.05)是:LW(SSC13上119 cM处)、LGW(SSC6上94 cM处)、SPW(SSC8上106 cM处)、SIW(SSC 4上0 cM处)、LIW(SSC 4上170 cM 处)和TNS(SSC 6上95 cM处)。上述QTL解释的表型变异从 0.04% 到 14.06%,有些位点的 QTL 可以解释表型变异的 10%以上,如 HW 的 QTL 解释表型变异的9.52%、SIW的QTL解释表型变异的13.47%、定位在SSC6上的TNS QTL解释表型变异的14.06%,而定位在 SSC7上的TNS QTL解释表型变异的11.30%。多数内脏器官重量性状的QTL定位结果未见报道。胴体长未见显著水平的QTL,而在SSC7上定位染色体极显著水平的肋骨数QTL。     

A genetic linkage map of the model legume Lotus japonicus and strategies for fast mapping of new loci

A genetic map for the model legume Lotus japonicus has been developed. The F(2) mapping population was established from an interspecific cross between L. japonicus and L. filicaulis. A high level of DNA polymorphism between these parents was the source of markers for linkage analysis and the map is based on a framework of amplified fragment length polymorphism (AFLP) markers. Additional markers were generated by restriction fragment length polymorphism (RFLP) and sequence-specific PCR. A total of 524 AFLP markers, 3 RAPD markers, 39 gene-specific markers, 33 microsatellite markers, and six recessive symbiotic mutant loci were mapped. This genetic map consists of six linkage groups corresponding to the six chromosomes in L. japonicus. Fluorescent in situ hybridization (FISH) with selected markers aligned the linkage groups to chromosomes as described in the accompanying article by Pedrosa et al. 2002(this issue). The length of the linkage map is 367 cM and the average marker distance is 0.6 cM. Distorted segregation of markers was found in certain sections of the map and linkage group I could be assembled only by combining colormapping and cytogenetics (FISH). A fast method to position genetic loci employing three AFLP primer combinations yielding 89 markers was developed and evaluated by mapping three symbiotic loci, Ljsym1, Ljsym5, and Ljhar1-3.     

A preliminary microsatellite genetic map of the ostrich (Struthio camelus)

Molecular genetic maps can provide information for the identification and localization of major genes associated with quantitative traits. However, there are currently no published genetic linkage maps for any ratites. Herein, a preliminary genetic map of ostrich was developed using a two-generation ostrich reference family by linkage analysis of 104 polymorphic microsatellite markers, including 40 novel markers reported in this study. A total of 35 microsatellite markers were placed into 13 linkage groups. Five linkage groups are composed of three or more loci, whereas the remaining eight groups each contained two markers. The sex-averaged map spans 365.4 cM. The marker interval of each linkage group ranges from 5.3 to 25.4 cM, and the average interval distance is 16.61 cM. The male map covers 342.7 cM, with an average intermarker distance of 15.58 cM, whereas the female map is 456.7 cM, with the average intermarker spacing of 20.76 cM. In order to screen the orthologous loci between ostrich and chicken, all of the flanking sequences of the 104 polymorphic loci, nine monomorphic loci and a further 12 reported microsatellite loci for ostrich were screened against the chicken genomic sequence using the BLAST algorithm (Altschul et al., 1990), and corresponding orthologs were found for 13 sequences. The microsatellite loci and genetic map developed in this study will be useful for QTL mapping, population genetics and phylogenetic studies in the ratite. In addition, the 13 orthologous loci identified in this study will be advantageous to the construction of a comparative genetic map between chicken and ostrich.     

High-density Linkage Map of Cultivated Allotetraploid Cotton Based on SSR, TRAP, SRAP and AFLP Markers

A high-density linkage map was constructed for an F2 population derived from an Interspecific cross of cultivated allotetraploid species between Gossypium hirsutum L. and G. barbadense L. A total of 186 F2 individuals from the Interspecific cross of ＂CRI 36 × Hal 7124＂ were genotyped at I 252 polymorphic loci Including a novel marker system, target region amplification polymorphism （TRAP）. The map consists of 1 097 markers, including 697 simple se- quence repeats （SSRs）, 171 TRAPs, 129 sequence-related amplified polymorphisms, 98 amplified fragment length polymorphisms, and two morphological markers, and spanned 4 536.7 cM with an average genetic distance of 4.1 cM per marker. Using 45 duplicated SSR loci among chromosomes, 11 of the 13 pairs of homologous chromosomes were Identified In tetraploid cotton. This map will provide an essential resource for high resolution mapping of quantitative trait loci and molecular breeding in cotton.     

A genetic and cytogenetic map for the duck (Anas platyrhynchos)

A genetic linkage map for the duck (Anas platyrhynchos) was developed within a cross between two extreme Peking duck lines by linkage analysis of 155 polymorphic microsatellite markers, including 84 novel markers reported in this study. A total of 115 microsatellite markers were placed into 19 linkage groups. The sex-averaged map spans 1353.3 cM, with an average interval distance of 15.04 cM. The male map covers 1415 cM, whereas the female map covers only 1387.6 cM. All of the flanking sequences of the 155 polymorphic loci--44 monomorphic loci and a further 41 reported microsatellite loci for duck--were blasted against the chicken genomic sequence, and corresponding orthologs were found for 49. To integrate the genetic and cytogenetic map of the duck genome, 28 BAC clones were screened from a chicken BAC library using the specific PCR primers and localized to duck chromosomes by FISH, respectively. Of 28 BAC clones, 24 were detected definitely on duck chromosomes. Thus, 11 of 19 linkage groups were localized to 10 duck chromosomes. This genetic and cytogenetic map will be helpful for the mapping QTL in duck for breeding applications and for conducting genomic comparisons between chicken and duck.     

A whole genome scan to detect quantitative trait loci for gestation length and sow maternal ability related traits in a White Duroc × Erhualian F2 resource population

Gestation length and maternal ability are important to improve the sow reproduction efficiency and their offspring survival. To map quantitative trait loci (QTL) for gestation length and maternal ability related traits including piglet survival rate and average body weight of piglets at weaning, more than 200 F2 sows from a White Duroc × Erhualian resource population were phenotyped. A genome-wide scan was performed with 194 microsatellite markers covering the whole pig genome. QTL analysis was carried out using a composite regression interval mapping method via QTL express. The results showed that total number of born piglets was significantly correlated with gestation length (r = -0.13, P < 0.05). Three QTL were detected on pig chromosome (SSC)2, 8 and 12 for gestation length. The QTL on SSC2 achieved the 5% genome-wide significant level and the QTL on SSC8 was consistent with previous reports. Four suggestive QTL were identified for maternal ability related traits including 1 QTL for survival rate of piglets at weaning on SSC8, 3 QTL for average body weight of piglet at weaning on SSC3, 11 and 13.     

A new framework marker-based linkage map and SDPs for the Rat HXB/BXH strain set

A new contiguous genetic linkage map of the HXB/BXH set of rat recombinant inbred (RI) strains was constructed to enhance QTL mapping power and precision, and thereby make the RI strain set a better genomics resource. The HXB/BXH rat RI strains were developed from a cross between the hypertensive SHR/OlaIpcv and normotensive BN-Lx/Cub rat strains and have been shown useful for identifying quantitative trait loci (QTL) for a variety of cardiovascular, metabolic, and behavioral phenotypes. In the current analysis, the DNAs from 31 existing strains, 1 substrain, and 4 extinct strains were genotyped for a selection of polymorphic microsatellite marker loci, predominantly polymorphic framework markers from high-density integrated rat genome maps. The resulting linkage map consists of 245 microsatellite markers spanning a total length of 1789 cM with an average inter-marker distance of ~8.0 cM. This map covers the rat genome contiguously and completely with the exception of two locations on Chromosomes (Chrs) 11 and 16. The new genotypic information obtained also permitted further genetic characterization of the RI strain set including strain independence, genetic similarity among the individual strains, and non-syntenic associations between loci.     

A genetic linkage map based on AFLP and SSR markers and mapping of QTL for dry-matter content in sweetpotato

We developed a genetic linkage map of sweetpotato using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers and a mapping population consisting of 202 individuals derived from a broad cross between Xushu 18 and Xu 781, and mapped quantitative trait loci (QTL) for the storage root dry-matter content. The linkage map for Xushu 18 included 90 linkage groups with 2077 markers (1936 AFLP and 141 SSR) and covered 8,184.5 cM with an average marker distance of 3.9 cM, and the map for Xu 781 contained 90 linkage groups with 1954 markers (1824 AFLP and 130 SSR) and covered 8,151.7 cM with an average marker distance of 4.2 cM. The maps described herein have the best coverage of the sweetpotato genome and the highest marker density reported to date. These are the first maps developed that have 90 complete linkage groups, which is in agreement with the actual number of chromosomes. Duplex and triplex markers were used to detect the homologous groups, and 13 and 14 homologous groups were identified in Xushu 18 and Xu 781 maps, respectively. Interval mapping was performed first and, subsequently, a multiple QTL model was used to refine the position and magnitude of the QTL. A total of 27 QTL for dry-matter content were mapped, explaining 9.0–45.1 % of the variation; 77.8 % of the QTL had a positive effect on the variation. This work represents an important step forward in genomics and marker-assisted breeding of sweetpotato.     

A genetic map in the Mimulus guttatus species complex reveals transmission ratio distortion due to heterospecific interactions.

As part of a study of the genetics of floral adaptation and speciation in the Mimulus guttatus species complex, we constructed a genetic linkage map of an interspecific cross between M. guttatus and M. nasutus. We genotyped an F(2) mapping population (N = 526) at 255 AFLP, microsatellite, and gene-based markers and derived a framework map through repeated rounds of ordering and marker elimination. The final framework map consists of 174 marker loci on 14 linkage groups with a total map length of 1780 cM Kosambi. Genome length estimates (2011-2096 cM) indicate that this map provides thorough coverage of the hybrid genome, an important consideration for QTL mapping. Nearly half of the markers in the full data set (49%) and on the framework map (48%) exhibited significant transmission ratio distortion (alpha = 0.05). We localized a minimum of 11 transmission ratio distorting loci (TRDLs) throughout the genome, 9 of which generate an excess of M. guttatus alleles and a deficit of M. nasutus alleles. This pattern indicates that the transmission ratio distortion results from particular interactions between the heterospecific genomes and suggests that substantial genetic divergence has occurred between these Mimulus species. We discuss possible causes of the unequal representation of parental genomes in the F(2) generation.     

基于F2群体的香菇遗传图谱构建及其在QTL定位中的应用

以171个F2双核体菌株为作图群体,通过相互配对的2个单核体的基因型推断双核体基因型,构建了第一张基于双核体群体的香菇遗传图谱。该图谱包含分布于15个连锁群的459个标记,覆盖长度为989.7cM,平均标记间隔为2.2cM。此外,以此双核体群体作为表型分离群体,定位了6个与香菇双核体菌丝生长速度相关的QTLs,位于5个连锁群上。采用全同胞单核体随机交配策略,易于构建相对大的双核体群体,用于连锁图构建和QTL定位。研究表明,在食用菌连锁图谱构建及QTL定位研究中,利用F2群体,可能为提高遗传作图效率,解决作图群体与表型分离群体间不一致问题提供新的途径。