Detection of linkage disequilibrium QTLs controlling low-temperature growth and metabolite accumulations in an admixed breeding population of <Emphasis Type="BoldItalic">Leymus</Emphasis> wildryes

Low-temperature soluble carbohydrate accumulations are commonly associated with anthocyanin coloration, attenuated growth, and cold adaptation of cool-season grasses. A total of 647 AFLP markers were tested for associations with anthocyanin coloration, tiller formation, leaf formation, cumulative leaf length, percent soluble carbohydrate, and dry matter regrowth among replicated clones of an admixed Leymus wildrye breeding population evaluated in low-temperature growth chambers. The admixed breeding population was derived from a heterogeneous population of L. cinereus × L. triticoides F₁ hybrids, with two additional generations of open pollination. Two AFLP linkage maps, constructed from two full-sib mapping populations derived from the same F₁ hybrid population, were integrated to produce a framework consensus map used to examine the distribution of marker-trait associations in the admixed F₁OP₂ population. Thirty-seven linkage blocks, spanning 258 cM (13.6%) of the 1895 cM consensus map, contained 119 (50%) of the 237 markers showing at least one possible trait association (P < 0.05). Moreover, 28 (68%) of the 41 most significant marker-trait associations (P < 0.005) were located in 15 QTL linkage blocks spanning 112.9 cM (6%) of the linkage map. The coincidence of these 28 significant marker-trait associations, and many less significant associations, in 15 relatively small linkage blocks (0.6 cM to 21.3 cM) provides evidence of admixture linkage disequilibrium QTLs (ALD QTLs) in this heterogeneous breeding population. At least four of the remaining 13 putative marker-trait associations (P < 0.005) were located in genetic map regions lacking other informative markers. The complexity of marker-trait associations results from heterogeneity within and substantial divergence among the parental accessions.     

Genetic linkage map construction for white jute (Corchorus capsularis L.) using SRAP,ISSR and RAPD markers

White jute (Corchorus capsularis) and dark jute (Corchorus olitorius) are two important cultivated crops that are used for natural fibre production. Some genetic maps have been developed for dark jute, but the genetic map information for white jute (C. capsularis) is limited. In this study, a linkage map comprising 44 sequence‐related amplified polymorphisms (SRAPs), 57 intersimple sequence repeats (ISSRs) and 18 randomly amplified polymorphic DNA (RAPD) covering 2185.7 cM with a mean density of 18.7 cM per locus was constructed in an F₂ population consisting of 185 individuals derived from a cross between two diverse genotypes of ‘Xinxuan No. 1’ and ‘Qiongyueqing’ in white jute. These markers were evenly distributed in the linkage groups without any clustering. This genetic linkage map construction will facilitate the mapping of agronomic traits and marker‐assisted selection breeding in white jute.     

Construction of a genetic linkage map with SSR,AFLP and morphological markers to locate QTLs controlling pathotype-specific powdery mildew resistance in diploid roses

Disease resistance is a sought-after trait in plant breeding programmes. One strategy to make resistance more durable is to increase the number of resistance genes, thereby increasing the number of pathotypes withstood. One of the most important diseases on roses is powdery mildew (PM) (Podosphaera pannosa). Recent studies show that pathotypes of PM and different types of resistances in roses exist. The results of this study aim to contribute to PM resistance in roses by the development of pathotype-specific markers on a genetic map. A diploid rose population (90 genotypes) derived from a cross between Rosa wichurana and Rosa ‘Yesterday’ was used to construct a genetic linkage map encompassing 20 AFLP primer combinations, 43 SSR, and 2 morphological markers. By applying the F1 pseudo test cross population strategy, two parental linkage maps were constructed (parent ‘Yesterday’ 536 cM; parent R. wichurana 526 cM). Both parental maps consisted of seven linkage groups with an average length of 70 cM (Kosambi) corresponding to the seven haploid rose chromosomes. These new maps were used to identify QTLs controlling disease resistance. The offspring population was screened for resistance to two PM pathotypes, R–E and R–P. QTLs for controlling pathotype-specific disease resistance were mapped by applying Kruskal–Wallis rank-sum tests and simple interval mapping. With two pathotypes analysed, nine QTL loci were detected on linkage groups 2, 3, 5 and 6, explaining 15–73% of the phenotypic variance for pathotype-specific disease response. The genetic maps developed here will be useful for future rose breeding, pathotype-specific resistance research and development of a consensus map for roses.     

Mapping genes for grain protein concentration and grain yield on chromosome 5B of Triticum turgidum (L.) var. dicoccoides

Grain protein concentration (GPC) is an important quality factor in durum wheat [Triticum turgidum (L.) var. durum]. Due to the strong environmental influence on GPC, molecular markers linked to quantitative trait loci (QTL) affecting GPC have the potential to be valuable in wheat breeding programs. Various quantitative traits in a population of 133 recombinant inbred chromosome lines were studied in replicated trials at three locations in North Dakota. Segregation for GPC, 1000-kernel weight, gluten strength, heading date, and plant height was observed. By relating phenotypic data to a linkage map obtained from the same population, three QTL affecting GPC, and one affecting yield were identified. The genotypic coefficients of determination for both traits were high.     

甘蔗SSR和AFLP分子遗传连锁图谱构建

采用甘蔗商业品种Co419与野生种割手密Y75/1/2杂交,获得269个单株,组成F1群体,用F102/356与商业品种ROC25回交获得266个单株,组成BC1群体。利用筛选的多态性条带丰富的36对SSR引物和12对AFLP引物,对两个群体进行PCR扩增和分子遗传连锁分析,构建甘蔗分子遗传连锁图谱。用F1群体获得630个分离标记,经χ2检测,298个标记为单双剂量标记,占总标记数的47%;用BC1群体获得571个分离标记,有264个标记为单双剂量标记,占总标记数的46%;4个亲本获得单双剂量标记的数量依次为Co41902/356Y75/1/2ROC25。在LOD≥5.0,相邻标记遗传距离≤40cM的条件下,F1群体有134个单双剂量标记被纳入55个连锁群,其中39个连锁群归属8个同源组,16个未列入,总遗传距离为1458.3cM,标记间平均图距为10.9cM;BC1群体有133个单双剂量标记被纳入47个连锁群,其中34个连锁群归属于8个同源组,13个连锁群未列入,总遗传距离为1059.6cM,标记间平均图距为8.0cM。从4个亲本单双剂量标记进入的连锁群数来看,Co419最多,归入34个连锁群,其次为Y75/1/2,归入20个连锁群,第3为02/356和ROC25,归入19个连锁群。研究结果表明,从单双剂量标记比例、形成连锁群数量、总遗传距离来看,F1群体构图质量要优于BC1群体。     

The first genetic maps of cashew (Anacardium occidentale L.)

Cashew (Anacardium occidentale) is a widespread tropical tree crop that is grown primarily for its nuts and has a global production of over 2 million Mt. In spite of its economic importance to many countries, however, no linkage map containing STS anchor sites has yet been produced for this species. This is largely attributable to a prolonged juvenile phase of the tree (limiting mapping to F₁ progenies) and difficulty in effecting sufficient hand-pollinations to create mapping populations of effective size. Here, we produce an F₁ mapping population of 85 individuals from a cross between CP 1001 (dwarf commercial clone) and CP 96 (giant genotype), and use it to generate two linkage genetic maps comprising of 205 genetic markers (194 AFLP and 11 SSR markers). The female map (CP 1001) contains 122 markers over 19 linkage groups and the male map (CP 96) comprises 120 markers assembled over 23 linkage groups. The total map distance of the female map is 1050.7 cM representing around 68% genome coverage, whereas the male map spans 944.7 cM (64% coverage). The average map distance between markers is 8.6 cM in the female map and 7.9 cM in the male map. Homology between the two maps was established between 13 linkage groups of the female map and 14 of the male map using 46 bridging markers that include 11 SSR markers. These maps represent a platform from which to identify loci controlling economically important traits in this crop.     

An SSR-based molecular genetic map of cassava

Summary Microsatellites or simple sequence repeats (SSR) are the markers of choice for molecular genetic mapping and marker-assisted selection in many crop species. A microsatellite-based linkage map of cassava was drawn using SSR markers and a F₂ population consisting of 268 individuals. The F₂ population was derived from selfing the genotype K150, an early yielding genotype from an F₁ progeny from a cross between two non-inbred elite cassava varieties, TMS 30572 and CM 2177-2 from IITA and CIAT respectively. A set of 472 SSR markers, previously developed from cassava genomic and cDNA libraries, were screened for polymorphism in K150 and its parents TMS 30572 and CM 2177-2. One hundred and twenty two polymorphic SSR markers were identified and utilized for linkage analysis. The map has 100 markers spanning 1236.7 cM, distributed on 22 linkage groups with an average marker distance of 17.92 cM. Marker density across the genome was uniform. This is the first SSR based linkage map of cassava and represents an important step towards quantitative trait loci mapping and genetic analysis of complex traits in M. esculenta species in national research program and other institutes with minimal laboratory facilities. SSR markers reduce the time and cost of mapping quantitative trait loci (QTL) controlling traits of agronomic interest, and are of potential use for marker-assisted selection (MAS).     

The first genetic linkage map of crape myrtle (Lagerstroemia) based on amplification fragment length polymorphisms and simple sequence repeats markers

Lagerstroemia (crape myrtle) are famous ornamental plants with large pyramidal racemes, long flower duration and diverse colours. Genetic maps provide an important genomic resource of basic and applied significance. A genetic linkage map was developed by genotyping 192 F₁ progeny from a cross between L. caudata (female) and L. indica (‘Xiang Xue Yun’) (male) with a combination of amplification fragment length polymorphisms (AFLP) and simple sequence repeats (SSR) markers in a double pseudo‐testcross mapping strategy. A total of 330 polymorphic loci consisting of 284 AFLPs and 46 SSRs showing Mendelian segregation were generated from 383 AFLP primer combinations and 150 SSR primers. The data were analysed using JoinMap 4.0 (evaluation version) to construct the linkage map. The map consisted of 20 linkage groups of 173 loci (160 AFLPs and 13 SSRs) covering 1162.1 cM with a mean distance of 10.69 cM between adjacent markers. The 20 linkage groups contained 2–49 loci and ranged in length from 7.38 to 163.57 cM. This map will serve as a framework for mapping QTLs and provide reference information for future molecular breeding work.     

利用2个F_2群体整合中国豌豆高密度SSR遗传连锁图谱

豌豆(Pisum sativum L.)是一种重要的食用豆类作物,在全世界范围内广泛种植,既可作为人类食物,也可作为牲畜饲料。用SSR标记构建的遗传连锁图谱在豌豆和其他作物的标记辅助育种中发挥着重要的作用。尽管对豌豆遗传连锁作图的研究已有悠久历史,但公众可获得且可转移的SSR标记以及基于遗传独特的中国豌豆种质的高密度遗传连锁图谱仍然有限。为了获得更多可转移的SSR标记和中国豌豆的高密度遗传连锁图谱,本研究首先从自主开发和文献获取的12,491个全基因组SSR标记中筛选了617个多态性SSR标记,并用于G0003973×G0005527 F_2群体遗传连锁图谱的加密。加密后的图谱全长扩展到5330.6 cM,包含603个SSR标记,标记平均间距离8.8 cM,相比之前的图谱有明显改善。基于上述结果,我们又筛选了119个具有多态性的SSR标记,用于构建大样本W6-22600×W6-15174 F_2群体的遗传连锁图谱,新图谱累积长度为1127.1 cM,包含118个SSR标记,装配在7条连锁群上。最后,将来自以上2个遗传图谱的数据进行整合,得到了一张覆盖范围6592.6 cM的整合图谱,包含668个SSR标记,由509个基因组SSR、134个EST-SSR和25个锚定标记组成,分布在7条连锁群上。这些SSR标记和遗传连锁图谱将为豌豆的遗传研究和标记辅助育种提供有力工具。     

苦荞SSR分子遗传图谱的构建及分析

构建苦荞遗传连锁图谱,为今后有关苦荞基因组结构、重要农艺性状QTL定位、分子标记辅助育种和基因克隆等研究工作奠定基础。以栽培苦荞‘滇宁一号’和苦荞野生近缘种杂交产生的119份F4代分离材料为作图群体,利用SSR分子标记来构建苦荞的分子遗传连锁图谱。本研究构建的连锁图谱包含15个连锁群,由89个标记组成,其中偏分离的标记有22个,占24.7%,每条连锁群上的标记在2~16之间。连锁群长度在6.9~165.8 cM的范围,覆盖基因组860.2 cM,总平均长度9.7 cM。本研究构建了首张苦荞SSR遗传连锁图谱,为苦荞QTL定位、基因克隆、遗传选育等研究奠定了基础。     

A genetic linkage map in southern‐by‐spring oat identifies multiple quantitative trait loci for adaptation and rust resistance

We developed 178 recombinant inbred lines from a southern‐by‐spring oat population designated as “TxH.” These lines were genotyped to generate a high‐quality linkage map that resolved 6,902 markers into 21 linkage groups that matched closely with the latest hexaploid oat consensus map. Three major quantitative trait loci (QTLs) affecting heading date were found in locations that are consistent with known QTLs and candidate genes, and two other QTLs affecting heading date were found in novel locations. Five QTLs affecting plant height were found. Both sets of QTLs are responsible for transgressive segregation observed for these two traits. Four QTLs affecting resistance to crown rust, caused by the pathogen Puccinia coronata f. sp. avenae, were identified. Two of these QTLs are consistent with known clusters of rust resistance genes, while two may represent new locations of novel rust resistance genes. A complete set of SNP sequences suitable for generating markers for molecular selection is provided.     

栽培种花生遗传图谱的构建及主茎高和总分枝数QTL分析

栽培种花生是异源四倍体,基因组大,构建花生的分子遗传连锁图谱并对相关性状进行QTL定位研究的工作缓慢。本研究以遗传差异大的亲本组配杂交组合富川大花生×ICG6375构建F2作图群体,采用公开发表的2653对SSR引物,构建了一张含有234个SSR标记、分布于20个连锁群的栽培种花生遗传图谱。该图谱覆盖基因组的长度为1683.43c M,各个连锁群长度在36.11~131.48 c M之间,每个连锁群的标记数在6~15个之间,标记间的平均距离为7.19 c M。结合F3在湖北武汉和阳逻环境下的主茎高和总分枝数鉴定结果,应用Win QTLCart 2.5软件采用复合区间作图法进行了QTL定位和遗传效应分析。共检测到17个与主茎高和总分枝数相关的QTL位点,贡献率在0.10%~10.22%之间,分布于8个连锁群上。综合分析武汉和阳逻环境的鉴定结果,获得重复一致的与主茎高相关的6个QTL,其中q MHA061.1和q MHA062.1位于连锁群LG06上TC1A2~AHGS0153标记区间,贡献率为5.49%~8.95%;q MHA061.2和q MHA062.2位于LG06上AHGS1375~PM377标记区间,贡献率为2.93%~5.83%;q MHA092.2和q MHA091.1位于连锁群LG09上GM2839~EM87标记区间,贡献率为0.53%~9.43%。     

Detection and integration of gene mapping of downy mildew resistance in maize inbred lines though linkage and association

Association mapping relies on the variation and extent of linkage disequilibrium within a species to identify genes of interest; this is an alternative to linkage mapping in traditional biparental population, which exploits only the variation in the two parents of the mapping population. This study was designed to identify association between 48 SSR markers and downy mildew (DM) resistance using a set of 60 public and private maize inbred lines in Thailand. Genetic diversity and population structure in the set were calculated. A total of 489 alleles with an average gene diversity of 0.70 revealed two subpopulations among the 60 maize inbred lines. Analysis of variance resulted in significant effects of phenotypic values of tested entries, with significant effects of inbred lines, locations, and their interaction. In addition, the analysis of variance for broad sense heritability also indicated high heritability (0.97), and association analysis revealed three significant SSR-trait associations (P < 0.05). These three significant SSR loci have not been reported in previous linkage mapping studies. Our results suggest that new allelic variants associated with DM resistance in these germplasm collections should be useful to help identify new lines carrying alleles for DM resistance in breeding marker-assisted selection programs.     

Development of an interspecific Vigna linkage map between Vigna umbellata (Thunb.) Ohwi & Ohashi and V. nakashimae (Ohwi) Ohwi & Ohashi and its use in analysis of bruchid resistance and comparative genomics

To facilitate transfer of bruchid resistance to azuki bean (Vigna angularis) from its relatives an interspecific mapping population was made between rice bean, V. umbellata, and the related wild species V. nakashimae. The V. umbellata parent is completely resistant and V. nakashimae is completely susceptible to the bruchid beetle pests, azuki bean weevil (Callosobruchus chinensis) and cowpea weevil (C. maculatus). There is very low cross compatibility between V. umbellata and azuki bean. Therefore, V. nakashimae, that crosses with both V. umbellata and V. angularis without the need for embryo rescue, is used as a bridging species. A genetic linkage map was constructed based on an interspecific F₂ mapping population between V. umbellata and V. nakashimae consisting of 74 plants. A total of 175 DNA marker loci (74 RFLPs and 101 SSRs) were mapped on to 11 linkage groups spanning a total length of 652 cM. Segregation distortion was observed but only three markers were not linked to any linkage group due to severe segregation distortion. This interspecific genome map was compared with the genome map of azuki bean. Of 121 common markers on the two maps, 114 (94.2%) were located on the same linkage groups in both maps. The marker order was highly conserved between the two genome maps. Fifty F₂ plants that produced sufficient seeds were used for quantitative trait locus (QTL) analysis and locating gene(s) for C. chinensis and C. maculatus resistance in V. umbellata. The resistance reaction of these F₂ plants differed between C. chinensis and C. maculatus. Both resistances were quantitatively inherited with no F₂ plants completely susceptible to C. chinensis or C. maculatus. One putative QTL for resistance to each of these bruchid species was located on different linkage groups. Other putative QTLs associated with resistance to both C. chinensis and C. maculatus were localized on the same linkage group 1. Linked markers associated with the bruchid‐resistant QTL will facilitate their transfer to azuki bean breeding lines.     

Construction of high-density linkage map and identification of QTLs associated with resistance to black rot in radish (Raphanus sativus) by RAD sequencing

High-density marker-based QTL mapping can serve as an effective strategy to identify novel genomic information to facilitate crop improvement. In this study, we genotyped an F₂ population (KB12-1 × PP12-1) using a RAD-seq approach and constructed a high-density linkage map for radish. After a series of filtering procedures were performed, 17,124 SNPs and 3,336 indels with aa × bb genotyping were retained to obtain bin markers. Then, a linkage map comprising a total of 1,221 bin markers in nine linkage groups spanning 1,467.3 cM with an average marker interval of 1.2 cM was constructed. We evaluated the resistance of the F₂ mapping population to black rot using F₃ progeny, and two major QTLs related to black rot resistance were identified based on this map. Among these QTLs, qBRR2 on Chr.2 explained 26.97% of the phenotypic variation with a LOD score of 11.93, and qBRR7 on Chr.7 accounted for 27.06% of the phenotypic variation with a LOD score of 11.83. The additive effect of qBRR2 was positive (14.97); however, qBRR7 had the opposite effect (−11.99). The high-density linkage map and the major QTLs qBRR2 and qBRR7 provide new important information for disease resistance gene discovery and utilization in genetic improvement.     

QTL mapping for economic traits based on a dense genetic map of cotton with PCR-based markers using the interspecific cross of Gossypium hirsutum × Gossypium barbadense

A high-density molecular marker linkage map of cotton based entirely on polymerase chain reaction-based markers is useful for a marker-assisted breeding program. Four kinds of markers—simple sequence repeats (SSRs), sequence-related amplified polymorphism (SRAP), random amplified polymorphic DNA (RAPD), and retrotransposon-microsatellite amplified polymorphism (REMAP)—were used to assay an F₂ population from a cross between “Handan208” (Gossypium hirsutum) and “Pima90” (Gossypium barbadense). Sixty-nine F₂ plants were used for map construction using 834 SSRs, 437 SRAPs, 107 RAPDs, and 16 REMAPs. Linkage analysis revealed that 1,029 loci could be mapped to 26 linkage groups that extended for 5,472.3 cM, with an average distance between 2 loci of 5.32 cM. The corresponding 69 F_2:3 families were grown, arranged in two replicates, and scored for eight phenotypes. Quantitative trait loci (QTL) analysis was performed by means of composite interval mapping using WinQtlCart ver 2.0. A total of 52 distinct QTLs were detected: 4 QTLs for lint index, 8 for seed index, 11 for lint yield, 4 for seed cotton yield, 9 for number of seed per boll, 3 for fiber strength, 5 for fiber length, and 8 for micronaire value. The present map and QTL analysis may provide a useful tool for breeders to transfer desirable traits from G. barbadense to the mainly cultivated species, G. hirsutum.     

T1 locus in cotton is the candidate gene affecting lint percentage, fiber quality and spiny bollworm (Earias spp.) resistance

A genetic linkage map of chromosome 6 was constructed by using 270 recombinant inbred lines originated from an upland cotton cross (Yumian 1 × T586) F₂ population. The genetic map included one morphological (T₁) and 18 SSR loci, covering 96.2 cM with an average distance of 5.34 cM between two markers. Based on composite interval mapping (CIM), QTL(s) affecting lint percentage, fiber length, fiber length uniformity, fiber strength and spiny bollworm resistance (Earias spp.) were identified in the t₁ locus region on chromosome 6. The allele(s) originating from T586 of QTLs controlling lint percentage increased the trait phenotypic value while the alleles originating from Yumian 1 of QTLs affecting fiber length, fiber length uniformity, fiber strength and spiny bollworm resistance increased the trait phenotypic value.     

Mapping a new source of resistance to powdery mildew in mungbean

Both restriction fragment length polymorphism (RFLP) and amplified fragment length polymorphism (AFLP) analyses were employed to map a new source of resistance to powdery mildew in mungbean. Disease scores of an F₂ population derived from the cross between a moderately resistant breeding line VC1210A and a susceptible wild relative (Vigna radiata var. sublobata, accession TC1966) showed a continuous distribution and was treated as a quantitative trait. Although no significant quantitative trait loci (QTL) that can explain the variation was detected by QTL analysis based on the reconstructed RFLP linkage map, new marker loci associated with resistance were discovered by AFLP analysis. The RFLP loci detected by two of the cloned AFLP bands are associated with resistance and constitute a new linkage group. A major resistance quantitative trait locus was found on this linkage group that accounted for 64.9% of the variation in resistance to powdery mildew. One of the probes developed in this study has the potential to assist in breeding for powdery mildew resistance in mungbean.     

Quantitative analysis and QTL mapping for agronomic and fiber traits in an RI population of upland cotton

Genetic mapping is an essential tool for cotton (Gossypium hirsutum L.) molecular breeding and application of DNA markers for cotton improvement. In this present study, we evaluated an RI population including 188 RI lines developed from 94 F₂-derived families and their two parental lines, ‘HS 46’ and ‘MARCABUCAG8US-1-88’, at Mississippi State, MS, for two years. Fourteen agronomic and fiber traits were measured. One hundred forty one (141) polymorphic SSR markers were screened for this population and 125 markers were used to construct a linkage map. Twenty six linkage groups were constructed, covering 125 SSR loci and 965 cM of overall map distance. Twenty four linkage groups (115 SSR loci) were assigned to specific chromosomes. Quantitative genetic analysis showed that the genotypic effects accounted for more than 20% of the phenotypic variation for all traits except fiber perimeter (18%). Fifty six QTLs (LOD > 3.0) associated with 14 agronomic and fiber traits were located on 17 chromosomes. One QTL associated with fiber elongation was located on linkage group LGU01. Nine chromosomes in sub-A genome harbored 27 QTLs with 10 associated with agronomic traits and 17 with fiber traits. Eight chromosomes in D sub-genome harbored 29 QTLs with 13 associated with agronomic traits and 16 with fiber traits. Chromosomes 3, 5, 12, 13, 14, 16, 20, and 26 harbor important QTLs for both yield and fiber quality compared to other chromosomes. Since this RI population was developed from an intraspecific cross within upland cotton, these QTLs should be useful for marker assisted selection for improving breeding efficiency in cotton line development. Paper number J1116 of the Mississippi Agricultural and Forestry Experiment Station, Mississippi State University, Mississippi State, MS 39762. Mention of trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by USDA, ARS and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

Construction of a linkage map for watermelon (Citrullus lanatus (Thunb.) Matsum & Nakai) using random amplified polymorphic DNA (RAPD)

Summary A linkage map for watermelon (Citrullus lanatus) was constructed on the basis of RADP, ribosomal DNA restriction fragment length polymorphism (RFLP), isozyme, and morphological markers using F₁BC1. A segregating population of 78 individuals was the result of a backcross of a cultivated inbred line (H-7; Citrullus lanatus; 2n=22) and a wild form (SA-1; C. lanatus; 2n=22), in which the latter was the recurrent (male) parent. A total of 69 RAPD, one RFLP, one isozyme, and three morphological markers was found to segregate in the BC1 population. Linkage analysis revealed that 62 loci could be mapped to 11 linkage groups that extended more than 524 centimorgans (cM), while 12 loci segregated independently of all other markers. The locus for exocarp color was linked to two RAPD markers within a region of 5 cM on linkage group 4. The locus for flesh color was linked to a RAPD marker within a region of 30 cM on linkage group 6. The isozyme marker GOT was located on the linkage group 1. Linkage group 2 contained a locus for ribosomal DNA within 5 cM of a RAPD marker. Half of the RAPD markers on the linkage group 7 displayed severely distorted segregation. The construction of linkage map using molecular markers is necessary for the breeding of watermelon to introduce useful gene of wild watermelon efficiently. However the linkage map that was constructed for the most part on the basis of RAPD markers could not cover significant parts of the genome, the linkage map provides breeders of watermelons the possibility of tagging useful agronomic traits, as well as the gene for exocarp color.Abbreviations RAPD random amplified polymorphic DNA - RFLP restriction fragment length polymorphism - GOT glutamate oxaloacetate transaminase - MDH malate dehydrogenase - ACP acid phosphatase - 6PGH 6-phosphogluconate dehydrogenase