基于整合图谱的鲤生长相关性状QTL的分布及变异规律

单个群体数量性状位点(QTL)的检测和识别效率通常是有限的,而多个群体能提高QTL的检测效率并能更好地了解其等位基因的变异和分布.本研究利用4个群体构建了鲤鱼的整合图谱,在此基础上比较分析了不同群体QTL的分布及变异.整合图谱长度为2371.6cM,包含257个微卫星(SSR)和421个SNP标记分布于42个连锁群,平均标记间隔为3.7cM.将4个群体的体重、体长、体高和体厚共67个QTL定位到整合图谱上进行比较分析,发现仅有1个QTL为3个群体共享,9个QTL为2群体共享,未发现4个群体均共享的QTL.QTL的比较分析结果表明,共享QTL可能在控制不同鲤鱼种质间生长性状中起主要作用.此外,探讨了QTL在不同群体间变化的原因和规律,同时揭示了主效和微效基因在不同群体中的遗传表现,为QTL定位策略和分子育种改良鲤生长性状提供理论基础.     

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。     

凉山半细毛羊1号染色体微卫星遗传连锁图谱的构建

实验选择绵羊1号条染色体上的9个微卫星标记,采用父系半同胞家系群体（共387个个体）构建凉山半细毛羊1号染色体遗传连锁图。建立的资源参考家系通过20个微卫星标记进行了系谱确证。试验结果表明,9个标记的等位基因数变化范围为5~15个,杂合度在0.202~0.831之间,平均杂合度为0.617,各标记的平均多态信息含量PIC=0.604。构建的凉山半细毛羊1号条染色体遗传连锁图总长度311.0 cM,与美国肉畜中心（USDA）和国际绵羊作图中心（IMF）构建的绵羊1号条染色遗传连锁图结果基本一致。可用于下一步的QTL定位研究。     

甘蓝型油菜千粒重性状的QTL初步定位研究

千粒重是油菜重要的产量相关性状之一,构建油菜遗传连锁图谱是研究其产量性状基因的前提。本研究利用小孢子培养技术,选育出了甘蓝型油菜大粒品系(G-42)和小粒品系(7-9)的纯合DH系DH-G-42和DH-7-9,其千粒重分别为6.24 g和2.42 g,二者比值达2.58。以DH-G-42为母本、DH-7-9为父本,构建了含190个单株的F2遗传作图群体,利用SSR和SRAP标记技术绘制遗传连锁图谱,该图谱共包含20个连锁群,涉及128个SSR标记和100个SRAP标记,图谱总长1546.6cM,标记间平均图距为6.78cM。本研究共检测到3个与千粒重性状相关的QTL,分别位于A9和C1连锁群,其中qSW-A9-1和qSW-A9-2贡献率分别达到10.98%和27.45%,均可视为控制粒重的主效QTL。本研究为后续进行油菜千粒重性状QTL的精细定位分析、分子标记辅助选择育种及新基因的克隆等奠定了基础。     

南瓜遗传连锁图谱的构建及粒宽性状的QTL定位

以印度南瓜纯系大粒材料‘0515-1’和小粒材料‘0460-1-1’为亲本,获得193个南瓜F2单株群体,应用AFLP和SSR分子标记技术进行多态性筛选,构建了含84个标记位点的遗传连锁图谱。结果表明,整个图谱包含12个连锁群,全长683.50cM,标记平均间距为8.13cM。采用复合区间定位分析,共检测到控制南瓜籽粒宽度的4个数量性状位点(QTL),分别位于3个连锁群上,各QTL的贡献率在2.87%～29.68%之间。     

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。     

黄瓜SRAP遗传连锁图的构建及侧枝基因定位

在由黄瓜的两个自交系S06与S52杂交产生的F₂群体中, 应用SRAP(sequence-related amplified polymorphism)标记构建遗传连锁图谱, 检测控制黄瓜侧枝数(lbn)和侧枝平均长度(lbl)的数量性状座位(QTL). 使用筛选出的64个多态性引物组合对F2群体进行分析, 得到108个多态性位点. 经MAPMAKER/EXP3.0分析(LOD≥3.0), 获得由92个标记座位组成、覆盖7个连锁群的遗传图谱, 总长1164.2 cM, 标记平均间距12.6 cM. 应用QTLMapper1.6, 各检测到4个控制lbn和lbl的QTL, 其中, 对lbn贡献率最大的QTL位于第二连锁群的ME11SA4B-ME5EM5区间, 其S06基因型具有增效作用; 对lbl贡献率最大的QTL位于第二连锁群的DC1OD3-DC1EM14区间, 其S06基因型具有增效作用.     

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。     

'百农64'×'京双16'小麦遗传连锁图谱构建

通过对小麦品种‘百农64’ב京双16’F3家系群体的SSR和AFLP分析,构建了含100个SSR标记(91个引物)和58个AFLP标记(12个引物)的小麦遗传连锁图,158个标记组成20个连锁群,覆盖小麦基因组3 114cM,标记间平均间距为19.7 cM.将前人未定位的12个SSR标记定位到了小麦遗传连锁图谱上.为小麦慢白粉病性等农艺性状的QTL分析打下了良好基础.     

微卫星标记对资源猪群的遗传分析和连锁图谱构建

在猪数量性状位点的定位研究中,标记的使用和图谱的构建是很重要的。本研究从猪的第4、6、7、8和13染色体上选取39个微卫星标记,在来源于约克夏和梅山214头猪组成的资源群中,分析了遗传特征并构建了图谱。研究表明,平均等位基因数、平均观察杂合度(Ho)和平均多态信息含量(PIC)在F1和F2代中分别为:3.2,0.528,0.463和3.2,0.496,0.447。结果表明大多数微卫星标记位点表现为中高度杂合性。在资源群体中,平均有信息减数分裂数是217.4(44-316),而各染色体上两性平均图谱的长度分别是:172.3cM(SSC4),168.7cM(SSC6),191.7cM(SSC7),197.3cM(SSC8),178.3cM(SSC13)。与USDA-MARC的参考图谱相比,标记位点的顺序相同,但长度均较长。雌雄两性图谱相比,第4和第6染色体上雌性图谱长于雄性图谱;而在另外3条染色体上,则雄性图谱长于雌性图谱。结果显示了标记位点在资源猪群的遗传特征和遗传关系,其连锁图谱可用于今后的QTL定位。     

茄子分子遗传图谱的构建及果实性状的QTL定位

以茄子(Solanum melongena)材料09-101-M和10TL-102-F4-1的重组自交系(RIL)为作图群体,构建总长度为991.7c M、共包含16个连锁群157个位点、平均图距为6.32 c M的遗传图谱。应用复合区间作图法(CIM),共检测到18个与茄子果实性状相关的QTLs,其中10个为主效QTLs,8个QTLs在两年两点的实验中能够被重复检测到。在所有QTLs中,控制果重的QTL fw1.1的效应值最大,为23.8%–31.6%,被定位在LG01(E09)上E25M34–E33M57b区域内;果长、果径与果重显著相关,且控制果长、果径与果重的QTL位于同一连锁群的相同区域。     

Genetic mapping of grapevine ( Vitis vinifera L.) applied to the detection of QTLs for seedlessness and berry weight

Parental and consensus genetic maps of Vitis vinifera L. (2n = 38) were constructed using a F₁ progeny of 139 individuals from a cross between two partially seedless genotypes. The consensus map contained 301 markers [250 amplification fragment length polymorphisms (AFLPs), 44 simple sequence repeats (SSRs), three isozymes, two random amplified polymorphic DNAs (RAPDs), one sequence-characterized amplified region (SCAR), and one phenotypic marker, berry color] mapped onto 20 linkage groups, and covered 1,002 cM. The maternal map consisted of 157 markers covering 767 cM (22 groups). The paternal map consisted of 144 markers covering 816 cM (23 groups). Differences in recombination rates between these maps and another unpublished map are discussed. The major gene for berry color was mapped on both the paternal and consensus maps. Quantitative trait loci (QTLs) for several quantitative subtraits of seedlessness in 3 successive years were searched for, based on parental maps: berry weight, seed number, seed total fresh and dry weights, seed percent dry matter, and seed mean fresh and dry weights. QTLs with large effects (R² up to 51%) were detected for all traits and years at the same location on one linkage group, with some evidence for the existence of a second linked major QTL for some of them. For these major QTLs, differences in relative parental effects were observed between traits. Three QTLs with small effects (R² from 6% to 11%) were also found on three other linkage groups, for berry weight and seed number in a single year, and for seed dry matter in 2 different years.     

Constructing a genetic linkage map and mapping quantitative trait loci for skeletal traits in Japanese flounder

A genetic linkage map of Japanese flounder was constructed using 165 doubled haploids (DHs) derived from a single female. A total of 574 genomic microsatellites (type II SSRs) and expressed sequence tag (EST)-derived markers (EST-SSRs) were mapped to 24 linkage groups. The length of linkage map was estimated as 1270.9 centiMorgans (cM), with an average distance between markers of 2.2 cM. The EST-SSRs were used together with type II SSR markers to construct the Japanese flounder genetic linkage map which will facilitate identify quantitative trait locus (QTL) controlling important economic traits in Japanese flounder. Thus, twelve skeletal traits at 2 years of age were measured for all DHs. Forty-one QTLs were detected on 14 linkage groups and totally account for a small proportion of phenotypic variation (4.5 to 17.3%). Most of QTLs detected distribute on linkage groups 5 (9 QTLs), 8 (9 QTLs), 9 (5 QTLs) and 20 (4 QTLs), in which, some QTLs perform the pleiotropy.     

Construction of the First High-Density Genetic Linkage Map and Analysis of Quantitative Trait Loci for Growth-Related Traits in <Emphasis Type="Italic">Sinonovacula constricta</Emphasis>

The razor clam (Sinonovacula constricta) is an important aquaculture species, for which a high-density genetic linkage map would play an important role in marker-assisted selection (MAS). In this study, we constructed a high-density genetic map and detected quantitative trait loci (QTLs) for Sinonovacula constricta with an F₁ cross population by using the specific locus amplified fragment sequencing (SLAF-seq) method. A total of 315,553 SLAF markers out of 467.71 Mreads were developed. The final linkage map was composed of 7516 SLAFs (156.60-fold in the parents and 20.80-fold in each F₁ population on average). The total distance of the linkage map was 2383.85 cM, covering 19 linkage groups with an average inter-marker distance of 0.32 cM. The proportion of gaps less than 5.0 cM was on average 96.90%. A total of 16 suggestive QTLs for five growth-related traits (five QTLs for shell height, six QTLs for shell length, three QTLs for shell width, one QTL for total body weight, and one QTL for soft body weight) were identified. These QTLs were distributed on five linkage groups, and the regions showed overlapping on LG9 and LG13. In conclusion, the high-density genetic map and QTLs for S. constricta provide a valuable genetic resource and a basis for MAS.     

A second-generation linkage map of the sheep genome

A genetic map of Ovis aries (haploid n = 27) was developed with 519 markers (504 microsatellites) spanning ∼3063 cM in 26 autosomal linkage groups and 127 cM (female specific) of the X Chromosome (Chr). Genotypic data were merged from the IMF flock (Crawford et al., Genetics 140, 703, 1995) and the USDA mapping flock. Seventy-three percent (370/504) of the microsatellite markers on the map are common to the USDA-ARS MARC cattle linkage map, with 27 of the common markers derived from sheep. The number of common markers per homologous linkage group ranges from 5 to 22 and spans a total of 2866 cM (sex average) in sheep and 2817 cM in cattle. Marker order within a linkage group was consistent between the two species with limited exceptions. The reported translocation between the telomeric end of bovine Chr 9 (BTA 9) and BTA 14 to form ovine Chr 9 is represented by a 15-cM region containing 5 common markers. The significant genomic conservation of marker order will allow use of linkage maps in both species to facilitate the search for quantitative trait loci (QTLs) in cattle and sheep. Received: 20 September 1992 / Accepted: 18 November 1997     

QTL analysis of flower and fruit traits in sour cherry

The map locations and effects of quantitative trait loci (QTLs) were estimated for eight flower and fruit traits in sour cherry (Prunus cerasus L.) using a restriction fragment length polymorphism (RFLP) genetic linkage map constructed from a double pseudo-testcross. The mapping population consisted of 86 progeny from the cross between two sour cherry cultivars, Rheinische Schattenmorelle (RS)×Erdi Botermo (EB). The genetic linkage maps for RS and EB were 398.2 cM and 222.2 cM, respectively, with an average interval length of 9.8 cM. The RS/EB linkage map that was generated with shared segregating markers consisted of 17 linkage groups covering 272.9 cM with an average interval length of 4.8 cM. Eleven putatively significant QTLs (LOD >2.4) were detected for six characters (bloom time, ripening time, % pistil death, % pollen germination, fruit weight, and soluble solids concentration). The percentage of phenotypic variation explained by a single QTL ranged from 12.9% to 25.9%. Of the QTLs identified for the traits in which the two parents differed significantly, 50% had allelic effects opposite to those predicted from the parental phenotype. Three QTLs affecting flower traits (bloom time, % pistil death, and % pollen germination) mapped to a single linkage group, EB 1. The RFLP closest to the bloom time QTL on EB 1 was detected by a sweet cherry cDNA clone pS141 whose partial amino acid sequence was 81% identical to that of a Japanese pear stylar RNase. Received: 4 March 1999 / Accepted: 27 August 1999     

Construction of a linkage map of the Rennell Island Tall coconut type (Cocos nucifera L.) and QTL analysis for yield characters.

AFLP and SSR DNA markers were used to construct a linkage map in the coconut (Cocos nucifera L.; 2n = 32) type Rennell Island Tall (RIT). A total of 227 markers were arranged into 16 linkage groups. The total genome length corresponded to 1971 cM for the RIT map, with 5-23 markers per linkage group. QTL analysis for yield characters in two consecutive sampling periods identified nine loci. Three and two QTLs were detected for number of bunches and one and three QTLs for number of nuts. The correlation of trait values between characters and evaluation periods is partially reflected in identical QTLs. The QTLs represent characters that are important in coconut breeding. The cosegregation of markers with these QTLs provides an opportunity for marker-assisted selection in coconut breeding programmes.     

Mapping QTLs for phosphorus deficiency tolerance in rice (Oryza sativa L.)

 The amplified fragment length polymorphism (AFLP) technique combined with selective genotyping was used to map quantitative trait loci (QTLs) associated with tolerance for phosphorus (P) deficiency in rice. P deficiency tolerant cultivar IR20 was crossed to IR55178-3B-9-3 (sensitive to P-deficiency) and 285 recombinant inbred lines (RILs) were produced by single-seed descent. The RILs were phenotyped for the trait by growing them in P-sufficient (10.0 mg/l) and P-deficient (0.5 mg/l) nutrient solution and determining their relative tillering ability at 28 days after seeding, and relative shoot dry weight and relative root dry weight at 42 days after seeding. Forty two of each of the extreme RILs (sensitive and tolerant) and the parents were subjected to AFLP analysis. A map consisting of 217 AFLP markers was constructed. Its length was 1371.8 cM with an average interval size of 7.62 cM. To assign linkage groups to chromosomes, 30 AFLP and 26 RFLP markers distributed over the 12 chromosomes were employed as anchor markers. Based on the constructed map, a major QTL for P-deficiency tolerance, designated PHO, was located on chromosome 12 and confirmed by RFLP markers RG9 and RG241 on the same chromosome. Several minor QTLs were mapped on chromosomes 1, 6, and 9. Received: 21 April 1998 / Accepted: 9 June 1998     

Construction of a High-Density Microsatellite Genetic Linkage Map and Mapping of Sexual and Growth-Related Traits in Half-Smooth Tongue Sole (Cynoglossus semilaevis)

High-density genetic linkage maps of half-smooth tongue sole were developed with 1007 microsatellite markers, two SCAR markers and an F1 family containing 94. The female map was composed of 828 markers in 21 linkage groups, covering a total of 1447.3 cM, with an average interval 1.83 cM between markers. The male map consisted of 794 markers in 21 linkage groups, spanning 1497.5 cM, with an average interval of 1.96 cM. The female and male maps had 812 and 785 unique positions, respectively. The genome length of half-smooth tongue sole was estimated to be 1527.7 cM for the females and 1582.1 cM for the males. Based on estimations of the map lengths, the female and male maps covered 94.74 and 94.65% of the genome, respectively. The consensus map was composed of 1007 microsatellite markers and two SCAR markers in 21 linkage groups, covering a total of 1624 cM with an average interval of 1.67 cM. Furthermore, 159 sex-linked SSR markers were identified. Five sex-linked microsatellite markers were confirmed in their association with sex in a large number of individuals selected from different families. These sex-linked markers were mapped on the female map LG1f with zero recombination. Two QTLs that were identified for body weight, designated as We-1 and We-2, accounted for 26.39% and 10.60% of the phenotypic variation. Two QTLs for body width, designated Wi-1 and Wi-2, were mapped in LG4f and accounted for 14.33% and 12.83% of the phenotypic variation, respectively. Seven sex-related loci were mapped in LG1f, LG14f and LG1m by CIM, accounting for 12.5–25.2% of the trait variation. The results should prove to be very useful for improving growth traits using molecular MAS.     

A genetic linkage map of Guinea yam ( Dioscorea rotundata Poir.) based on AFLP markers

A genetic linkage map of the tetraploid white yam (Dioscorea rotundata Poir.) was constructed based on 341 co-dominantly scored amplified fragment length polymorphism (AFLP) markers segregating in an intraspecific F₁ cross. The F₁ mapping population was produced by crossing a landrace cultivar TDr 93-1 as female parent to a breeding line TDr 87/00211 as the male parent. The marker segregation data were split into maternal and paternal data sets, and separate genetic linkage maps were constructed since the mapping population was an F₁ cross between two presumed heterozygous parents. The markers segregated like a diploid cross-pollinator population suggesting that the D. rotundata genome is an allo-tetraploid (2n = 4x = 40). The maternal map comprised 155 markers mapped on 12 linkage groups with a total map length of 891 cM. Three linkage groups consisted of maternal parent markers only. The paternal map consisted of 157 markers mapped on 13 linkage groups with a total map length of 852 cM. Three and one quantitative trait loci (QTLs) with effects on resistance to Yam Mosaic Virus (YMV) were identified on the maternal and paternal linkage maps, respectively. Prospects for detecting more QTLs and using marker-assisted selection in white yam breeding appear good, but this is subject to the identification of additional molecular markers to cover more of the genome.