Construction of Genetic Linkage Map Based on SSR Markers in Peanut （Arachis hypogaea L.）

Molecular genetic maps of crop species can be used in a variety of ways in breeding and genomic research such as identification and mapping of genes and quantitative trait loci （QTLs） for morphological, physiological and economic traits of crop species. However, a comprehensive genetic linkage map for cultivated peanut has not yet been developed due to the extremely low frequency of DNA polymorphism in cultivated peanut. In this study, 142 recombinant inbred lines （RILs） derived from a cross between Yueyou 13 and Zhenzhuhei were used as mapping population in peanut （Arachis hypogaea L.）. A total 652 pairs of genomic-SSR primer and 392 pairs of EST-SSR primer were used to detect the polymorphisms between the two parents. 141 SSR primer pairs, 127 genomic-SSR and 14 EST-SSR ones, which can be used to detect polymorphisms between the two parents, were selected to analyze the RILs population. Thus, a linkage genetic map which consists of 131 SSR loci in 20 linkage groups, with a coverage of 679 cM and an average of 6.12 cM of inter-maker distance was constructed. The putative functions of 12 EST-SSR markers located on the map were analyzed. Eleven showed homology to gene sequences deposited in GenBank. This is the first report of construction of a comprehensive genetic map with SSR markers in peanut （Arachis hypogaea L.）. The map presented here will provide a genetic framework for mapping the qualitative and quantitative trait in peanut.     

Polymorphism analysis of microsatellites and construction of linkage map in part regions of four chromosomes in chicken

Based on chicken' consensus map issued in 2000,17 microsatellites near 4 candidate genes such as IGF2,OBR,GDF8 and APOA1 in 4 chromosomes(chromosome 5,7,8 and 24)were chosen for polymorphism analysis and construction of linkage map.Combining the technique of PCR and the fluorescent semi-automated detection,genome scanning was performed for 440 chickens,which was derived from China Agricultural University chicken resource families within three generations.The individuals of this resource families were genotyped.The results showed that the number of alleles ranged from 4 to 14;heterozygosity(H) of markers was between 0.3116 and 0.9148.Polymorphic information content(PIC)varied from 0.2672 to 0.8679.Microsatellites along with above-mentioned 4 candidate genes doing as general markers were used to construct linkage map.The spans of 4 linkage maps constructed in the part region of chromosome 5,7,8 and 24 were 263.5,79.9,206.2 and 104.2 cM,respectively.The order of markers was consistent with that of counterpart of reported consensus map.However,The spans of linkage map were larger than that of consensus map.The constructed linkage maps laid the foundation for mapping quantitative trait loci(QTL)responsible for economically important traits in chicken.     

A genetic linkage map with 178 SSR and 1901 SNP markers constructed using a RIL population in wheat (Triticum aestivum L.)

The construction of high density genetic linkage map provides a powerful tool to detect and map quantitative trait loci(QTLs) controlling agronomically important traits. In this study, simple sequence repeat(SSR) markers and Illumina 9K i Select single nucleotide polymorphism(SNP) genechip were employed to construct one genetic linkage map of common wheat(Triticum aestivum L.) using 191 recombinant inbred lines(RILs) derived from cross Yu 8679×Jing 411. This map included 1 901 SNP loci and 178 SSR loci, covering 1 659.9 c M and 1 000 marker bins, with an average interval distance of 1.66 c M. A, B and D genomes covered 719.1, 703.5 and 237.3 c M, with an average interval distance of 1.66, 1.45 and 2.9 c M, respectively. Notably, the genetic linkage map covered 20 chromosomes, with the exception of chromosome 5D. Bioinformatics analysis revealed that 1 754(92.27%) of 1 901 mapped SNP loci could be aligned to 1 215 distinct wheat unigenes, among which 1 184(97.4%) were located on o ne single chromosome, and the rest 31(2.6%) were located on 2 to 3 chromosomes. By performing in silico comparison, 214 chromosome deletion bin-mapped expressed sequence tags(ESTs), 1 043 Brachypodium genes and 1 033 rice genes were further added onto the genetic linkage map. This map not only integrated genetic and physical maps, SSR and SNP loci, respectively, but also provided the information of Brachypodium and rice genes corresponding to 1 754 SNP loci. Therefore, it will be a useful tool for comparative genomics analysis, fine mapping of QTL/gene controlling agronomically important traits and marker-assisted selection breeding in wheat.     

High density genetic map and quantitative trait loci(QTLs) associated with petal number and flower diameter identified in tetraploid rose

Rose is one of the most important ornamental and economic plants in the world. Modern rose cultivars are primarily tetraploid, and during meiosis, they may exhibit double reduction or preferential chromosome pairing. Therefore, the construction of a high density genetic map of tetraploid rose is both challenging and instructive. In this study, a tetraploid rose population was used to conduct a genetic analysis using genome sequencing. A total of 17 382 single nucleotide polymorphism(SNP) markers were selected from 2 308 042 detected SNPs. Combined with 440 previously developed simple sequence repeats(SSR) and amplified fragment length polymorphism(AFLP) markers, a marker dosage of 6 885 high quality markers was successfully assigned by GATK software in the tetraploid model. These markers were used in the construction of a high density genetic map, containing the expected seven linkage groups with 6 842 markers, a total map length of 1 158.9 c M, and an average inter-marker distance of 0.18 c M. Quantitative trait locus(QTL) analysis was subsequently performed to characterize the genetic architecture of petal number and flower diameter. One major QTL(qpnum-3-1) was detected for petal number in three consecutive years, which explained 20.18–22.11% of the variation in petal number. Four QTLs were detected for flower diameter; the main locus, qfdia-2-2, was identified in two consecutive years. Our results will benefit the molecular marker-assisted breeding of modern rose cultivars. In addition, this study provides a guide for the genetic and QTL analysis of autotetraploid plants using sequencing-based genotyping methods.     

Identification of QTLs Related to Bolting in Brassica rapa ssp. pekinensis （syn. Brassica campestris ssp. pekinensis）

A genetic linkage map of Brassica rapa ssp. pekinensis was constructed with 186 AFLP （amplified fragment length polymorphism） markers by using a doubled-haploid （DH） population with 183 individuals. The individuals were derived from F1 which was developed by crossing a bolting resistant DH line Y-177-12 and an easy bolting DH line Y195-93a. AFLPs were generated by the use of restriction enzymes EcoR Ⅰ and Mse Ⅰ . The segregation of each marker and linkage was analyzed by using JoinMap version 3.0. Mapped markers were aligned in ten linkage groups which covered 887.8 cM with an average marker interval of 4.47 cM. Markers showing skewed segregation ratio were clustered in six LGs. Quantitative trait loci （QTL） were mapped for bolting resistance by using MAPQTL 4.0 package. Four QTLs explaining from 7.0 to 9.4% of the total variation were detected, all of them increase bolting resistance. These mapped QTLs could be used to develop a marker assisted selection programme for bolting resistance breeding.     

Molecular Diversity and Association Analysis of Drought and Salt Tolerance in Gossypium hirsutum L. Germplasm

Association mapping is a useful tool for the detection of genes selected during plant domestication based on their linkage disequilibrium（LD）. This study was carried out to estimate genetic diversity, population structure and the extent of LD to develop an association framework in order to identify genetic variations associated with drought and salt tolerance traits. 106 microsatellite marker primer pairs were used in 323 Gossypium hirsutum germplasms which were grown in the drought shed and salt pond for evaluation. Polymorphism（PIC=0.53） was found, and three groups were detected（K=3） with the second likelihood ΔK using STRUCTURE software. LD decay rates were estimated to be 13-15 cM at r2 0.20. Significant associations between polymorphic markers and drought and salt tolerance traits were observed using the general linear model（GLM） and mixed linear model（MLM）（P 0.01）. The results also demonstrated that association mapping within the population structure as well as stratification existing in cotton germplasm resources could complement and enhance quantitative trait loci（QTLs） information for marker-assisted selection.     

QTL Mapping for Dough Mixing Characteristics in a Recombinant Inbred Population Derived from a Waxy×Strong Gluten Wheat （Triticum aestivum L.）

Protein and starch are the most important traits in determining processing quality in wheat. In order to understand the genetic basis of the influence of Waxy protein (Wx) and high molecular weight gluten subunit (HMW-GS) on processing quality, 256 recombinant inbred lines (RILs) derived from the cross of waxy wheat Nuomai 1 and Gaocheng 8901 were used as mapping population. DArT (diversity arrays technology), SSR (simple sequence repeat), HMW-GS, and Wx markers were used to construct the molecular genetic linkage map. QTLs for mixing peak time (MPT), mixing peak value (MPV), mixing peak width (MPW), and mixing peak integral (MPI) of Mixograph parameters were evaluated in three different environments. The genetic map comprised 498 markers, including 479 DArT, 14 SSR, 2 HMW-GS, and 3 Wx protein markers, covering 4 229.7 cM with an average distance of 9.77 cM. These markers were identified on 21 chromosomes. Eighteen additive QTLs were detected in three different environments, which were distributed on chromosomes 1A, 1B, 1D, 4A, 6A, and 7D. QMPT-1D.1 and QMPT-1D.2 were close to the Glu-D1 marker accounting for 35.2, 22.22 and 36.57% of the phenotypic variance in three environments, respectively. QMPV-1D and QMPV-4A were detected in all environments, and QMPV-4A was the nearest to Wx-B1. One minor QTL, QMPI-1A, was detected under three environments with the genetic distances of 0.9 cM from the nearest marker Glu-A1, explaining from 5.31 to 6.67% of the phenotypic variance. Three pairs of epistatic QTLs were identified on chromosomes 2D and 4A. Therefore, this genetic map is very important and useful for quality trait related QTL mapping in wheat. In addition, the finding of several major QTLs, based on the genetic analyses, further suggested the importance of Glu-1 loci on dough mixing characteristics.     

Genetic Analysis and Molecular Mapping of an All-Stage Stripe Rust Resistance Gene in Triticum aestivum-Haynaldia villosa Translocation Line V3

Triticum aestivum-Hayaldia villosa translocation line V3 has shown effective all-stage resistance to the seven dominant pathotypes of Puccinia striiforms f.sp.tritici prevalent in China.To elucidate the genetic basis of the resistance,the segregating populations were developed from the cross between V3 and susceptible genotype Mingxian 169,seedlings of the parents and F 2 progeny were tested with six prevalent pathotypes,including CYR29,CYR31,CYR32-6,CYR33,Sun11-4,and Sun11-11,F 1 plants and F 3 lines were also inoculated with Sun11-11 to confirm the result further.The genetic studied results showed that the resistance of V3 against CYR29 was conferred by two dominant genes,independently,one dominant gene and one recessive gene conferring independently or a single dominant gene to confer resistance to CYR31,two complementary dominant genes conferring resistance to both CYR32-6 and Sun11-4,two independently dominant genes or three dominant genes（two of the genes show cumulative effect） conferring resistance to CYR33,a single dominant gene for resistance to Sun11-11.Resistance gene analog polymorphism（RGAP） and simple-sequence repeat（SSR） techniques were used to identify molecular markers linked to the single dominant gene（temporarily designated as YrV3） for resistance to Sun11-11.A linkage map of 2 RGAP and 7 SSR markers was constructed for the dominant gene using data from 221 F 2 plants and their derived F 2：3 lines tested with Sun11-11 in the greenhouse.Amplification of the complete set of nulli-tetrasomic lines of Chinese Spring with a RGAP marker RG1 mapped the gene on the chromosome 1B,and then the linked 7 SSR markers located this gene on the long arm of chromosome 1B.The linkage map spanned a genetic distance of 25.0 cM,the SSR markers Xgwm124 and Xcfa2147 closely linked to YrV3 with genetic distances of 3.0 and 3.8 cM,respectively.Based on the linkage map,it concluded that the resistance gene YrV3 was located on chromosome arm 1BL.Given chromosomal location,the reaction patterns and pedigree analysis,YrV3 should be a novel gene for resistance to stripe rust in wheat.These closely linked markers should be useful in stacking genes from different sources for wheat breeding and diversification of resistance genes against stripe rust.     

QTL Analysis of the Oil Content and the Hull Content in Brassica napus L.

The QTLs of the oil content and the hull content were analyzed in Brassica napus L. By constructing the linkage map. The F26 RIL population with 188 lines, derived from the cross of GH06 × P147, was used as the mapping population. The SRAP, SSR, AFLP, and TRAP markers were used to construct the linkage map, and the composite interval mapping (CIM) to identify the quantitative trait loci associated with the oil content and the hull content. 300 markers were integrated into 19 linkage groups, covering 1 248.5 cM in total. Seven QTLs were found to be responsible for the oil content with the single contribution to phenotypic variance ranging from 3.73 to 10.46%; four QTLs were found for the hull content with the single contribution to phenotypic variance ranging from 4.89 to 6.84%. The yellow-seeded Brassica napus L. Has the advantage of higher oil content and the hull content has a significant effect on the oil content. In addition, the SRAP marker is good for detecting QTL.     

利用F2及其衍生群体定位陆地棉产量和纤维品质性状QTLs

以陆地棉(Gossypiumhirsutum L.)丰产品种中棉所35和优质品种渝棉1号杂交产生的F2群体为材料,利用SSR标记构建了包含46个连锁群和303个位点的连锁图谱.该图覆盖2 543.6 c M,约占四倍体棉花基因组的57.2%,标记间平均距离为8.4 c M.应用MQM作图法分析F2及其衍生群体家系的产量和纤维品质性状,共得到25个产量和23个纤维品质性状QTLs,其中1个QTL(qFS07-1)在3个环境下检测到,2个QTLs(qFS20-1和qFS21-1)在2个环境下检测到.在检测到的QTLs中,22个分布在A亚组,26个分布在D亚组.     

利用四交群体构建陆地棉栽培品种间的SSR标记遗传图谱

作物遗传作图与QTL定位常应用在源于两自交系的单交群体上,是利用2个亲本之间的遗传多态信息;而育种实践中经常用到的四交群体却很少用于遗传作图。本研究将异交物种中F1分离群体的作图方法应用于常异花授粉作物棉花上,以4个陆地棉栽培品种泗棉3号、苏棉12、中4133和8891为亲本,构建陆地棉品种间四交作图群体泗棉3号/苏棉12//中4133/8891,利用JoinMap3.0构建了1张陆地棉栽培品种间的SSR标记遗传图谱。该图谱由56个连锁群组成,总长为2113.3cM,含有286个SSR多态位点,覆盖率达42.3%;单个连锁群的标记数2~24个,平均5.2个;长度为0.37~125cM,平均38.4cM;标记间的平均距离为7.4cM。这是目前报道的第1张覆盖率达40%的陆地棉栽培品种间的分子标记遗传图谱。     

Construction of Genetic Linkage Map Based on SSR Markers in Peanut(Arachis hypogaea L.)

Molecular genetic maps of crop species can be used in a variety of ways in breeding and genomic research such as identification and mapping of genes and quantitative trait loci (QTLs) for morphological, physiological and economic traits of crop species. However, a comprehensive genetic linkage map for cultivated peanut has not yet been developed due to the extremely low frequency of DNA polymorphism in cultivated peanut. In this study, 142 recombinant inbred lines (RILs) derived from a cross between Yueyou 13 and Zhenzhuhei were used as mapping population in peanut (Arachis hypogaea L.). A total 652 pairs of genomic-SSR primer and 392 pairs of EST-SSR primer were used to detect the polymorphisms between the two parents. 141 SSR primer pairs, 127 genomic-SSR and 14 EST-SSR ones, which can be used to detect polymorphisms between the two parents, were selected to analyze the RILs population. Thus, a linkage genetic map which consists of 131 SSR loci in 20 linkage groups, with a coverage of 679 cM and an average of 6.12 cM of inter-maker distance was constructed. The putative functions of 12 EST-SSR markers located on the map were analyzed. Eleven showed homology to gene sequences deposited in GenBank. This is the first report of construction of a comprehensive genetic map with SSR markers in peanut (Arachis hypogaea L.). The map presented here will provide a genetic framework for mapping the qualitative and quantitative trait in peanut.     

玉米重组自交系群体遗传图谱的构建及标记

以黄早四和Mo 17为亲本,组建了含239份重组自交系的F9代分离群体.共选取了370对SSR引物用于亲本多态性筛选,结果有126对引物能扩增出清晰的多态性条带.用这些有多态性的引物进一步作群体分析,除有23对引物扩增效果较差外,其余引物在多态性、重复性方面均表现较好.最后,用该重组自交系群体构建了玉米分子标记遗传连锁图谱,该图谱拟合了103个分布于10个连锁群的微卫星标记位点,覆盖玉米整个基因组1 455.4 cM,标记间平均图距14.1 cM.     

小豆SSR分子标记遗传连锁图谱构建

【目的】以小豆SSR为锚定标记,将公开发表的豇豆SSR、普通菜豆SSR和EST-SSR标记定位整合到小豆遗传连锁群中,构建中国小豆遗传图谱,为小豆基因定位、图位克隆和分子标记辅助选择育种提供更多可用的分子标记。【方法】用1 473对SSR和EST-SSR引物进行PCR扩增,包括906对豇豆SSR、123对普通菜豆和196对小豆SSR引物及248对普通菜豆EST-SSR引物,筛选亲本间多态性标记,验证栽培小豆HB801×AG109及GM892×AG110的F2分离群体。【结果】整合和构建了含有145个SSR和EST-SSR标记小豆遗传连锁图谱,包括59个小豆SSR标记,新增63个豇豆SSR、9个普通菜豆SSR、14个普通菜豆EST-SSR标记和1个茎色标记。紫茎色性状被定位在第9连锁群,离CEDG022和cbess058标记的遗传距离分别为0.9 cM和0.1 cM。图谱全长823 cM,覆盖11个连锁群,每个标记间平均距离为5.64 cM。每个连锁群长度为49.1—125.6 cM,平均长度74.82 cM;每条染色体上的标记数7—26个,平均13.27个。【结论】率先把小豆近缘物种分子标记引入小豆,加密了小豆SSR分子标记遗传连锁图谱。     

棉花深棕色纤维基因Lc1的遗传定位

 【目的】研究棉花棕色纤维的遗传规律,寻找并定位与棕色纤维基因连锁的分子标记,为进一步在棉花基因组学水平上定位、克隆棕色纤维基因和棕色棉纤维品质改良奠定基础。【方法】基于陆地棉显性多基因标记系T586（具有深棕色纤维基因Lc1）与海岛棉新海16配制的海岛棉×陆地棉杂交F2群体,结合色彩色差仪对棕色纤维色泽的分类进行分析,并充分利用棉花基因组分子标记遗传连锁图谱信息、多态性分子标记筛选和图位克隆的方法,定位与棕色棉纤维基因Lc1连锁的分子标记。【结果】根据T586与新海16杂交后代F2群体（443个有效纤维单株）深棕色纤维、棕色（中间色）和洁白色纤维的分离比例,将Lc1定位于棉花基因组A亚组第7染色体微卫星标记NAU4030和CGR5119之间约8 cM的遗传距离内,其中,Lc1与标记CGR5119的遗传距离约为2.8 cM,与NAU4030之间的遗传交换距离约为5.1 cM,构建了Lc1位点的遗传连锁图谱。【结论】棉花深棕色纤维性状由单基因控制并呈现半显性遗传方式,Lc1位点附近的分子标记信息可在棕色棉分子标记辅助育种中得以利用。     

利用SSR和InDel标记构建白菜×芜菁分子遗传图谱

以结球白菜BY和芜菁MM为亲本建立的小孢子培养DH系作为图谱构建群体,利用筛选获得的86对SSR引物和66对InDel引物进行分离分析,并整合已有的55个SSR标记,构建包含10个连锁群、175个标记位点的分子连锁图谱,图谱总长度902.9cM,标记间的平均图距为5.7cM,每个连锁群上的标记数为9～28个,平均图距为3.2～8.8cM,连锁群长度为66.0～113.3cM。     

陆海杂交棉产量及重要农艺性状QTL定位

[目的]通过海7124和军棉1号构建的陆海杂交棉产量及重要农艺性状的QTL定位研究,筛选与产量及重要农艺性状紧密连锁的分子标记,提高棉花育种效率.[方法]选用陆地棉品种军棉1号和海岛棉品种海7124配制的一个包含148株F_2单株的群体,利用97对SSR标记构建了一个包含17个连锁群,长673.3 cM,标记间平均距离为17.3 cM的遗传图谱,覆盖整个棉花基因组12.2;.在此基础上对果枝始节、果枝台数、结铃数、单铃重、衣分和叶面积6个农艺性状进行了QTL定位研究.[结果]检测到3个稳定的QTL:1个与衣分相关的QTL,位于第4连锁群上;1个与果枝台数相关的QTL,位于第1连锁群上;1个与叶面积相关的QTL,位于第6连锁群上.[结论]这些QTL的效应值较大,对今后的棉花育种的分子辅助选择具有较大的意义.     

棉花Ligon lintless纤维突变体SSR的分子标记定位

陆地棉Ligon lintless是单基因显性突变体,其性状表现为成熟的长纤维极度缩短,一般在4~6 mm,植株的叶片和茎呈扭曲状。本研究用陆地棉遗传标准系TM-1与棉花纤维突变体Ligon lintless的杂交一代(由于材料的特殊性即相当于回交一代BC1,后文称为BC1)的246个单株为作图群体,利用4000对SSR引物筛选出在双亲间表现多态性的引物28对,多态频率为0.7%。江利用这28对引物对BC1群体进行分析,共检测到23个多态性位点。用Joinmap3.0软件绘制连锁图,该图谱共覆盖168 cM,相当于棉花总基因组的3.74%,标记间的平均遗传距离为7.3 cM,将棉花纤维突变体Ligon lintless的L i基因连锁定位在22号染色体上。