Intraspecific Genetic Map Research Articles

A New Intra-Specific and High-Resolution Genetic Map of Eggplant Based on a RIL Population, and Location of QTLs Related to Plant Anthocyanin Pigmentation and Seed Vigour.

Eggplant is the second most important solanaceous berry-producing crop after tomato. Despite mapping studies based on bi-parental progenies and GWAS approaches having been performed, an eggplant intraspecific high-resolution map is still lacking. We developed a RIL population from the intraspecific cross ‘305E40’, (androgenetic introgressed line carrying the locus Rfo-Sa1 conferring Fusarium resistance) x ‘67/3’ (breeding line whose genome sequence was recently released). One hundred and sixty-three RILs were genotyped by a genotype-by-sequencing (GBS) approach, which allowed us to identify 10,361 polymorphic sites. Overall, 267 Gb of sequencing data were generated and ~773 M Illumina paired end (PE) reads were mapped against the reference sequence. A new linkage map was developed, including 7249 SNPs assigned to the 12 chromosomes and spanning 2169.23 cM, with iaci@liberoan average distance of 0.4 cM between adjacent markers. This was used to elucidate the genetic bases of seven traits related to anthocyanin content in different organs recorded in three locations as well as seed vigor. Overall, from 7 to 17 QTLs (at least one major QTL) were identified for each trait. These results demonstrate that our newly developed map supplies valuable information for QTL fine mapping, candidate gene identification, and the development of molecular markers for marker assisted selection (MAS) of favorable alleles.

Enriching an intraspecific genetic map and identifying QTL for fiber quality and yield component traits across multiple environments in Upland cotton (Gossypium hirsutum L.).

Cotton is a significant commercial crop that plays an indispensable role in many domains. Constructing high-density genetic maps and identifying stable quantitative trait locus (QTL) controlling agronomic traits are necessary prerequisites for marker-assisted selection (MAS). A total of 14,899 SSR primer pairs designed from the genome sequence of G. raimondii were screened for polymorphic markers between mapping parents CCRI 35 and Yumian 1, and 712 SSR markers showing polymorphism were used to genotype 180 lines from a (CCRI 35×Yumian 1) recombinant inbred line (RIL) population. Genetic linkage analysis was conducted on 726 loci obtained from the 712 polymorphic SSR markers, along with 1379 SSR loci obtained in our previous study, and a high-density genetic map with 2051 loci was constructed, which spanned 3508.29cM with an average distance of 1.71cM between adjacent markers. Marker orders on the linkage map are highly consistent with the corresponding physical orders on a G. hirsutum genome sequence. Based on fiber quality and yield component trait data collected from six environments, 113 QTLs were identified through two analytical methods. Among these 113 QTLs, 50 were considered stable (detected in multiple environments or for which phenotypic variance explained by additive effect was greater than environment effect), and 18 of these 50 were identified with stability by both methods. These 18 QTLs, including eleven for fiber quality and seven for yield component traits, could be priorities for MAS.

Molecular Mapping of Flowering Time Major Genes and QTLs in Chickpea (Cicer arietinum L.).

Flowering time is an important trait for adaptation and productivity of chickpea in the arid and the semi-arid environments. This study was conducted for molecular mapping of genes/quantitative trait loci (QTLs) controlling flowering time in chickpea using F2 populations derived from four crosses (ICCV 96029 × CDC Frontier, ICC 5810 × CDC Frontier, BGD 132 × CDC Frontier and ICC 16641 × CDC Frontier). Genetic studies revealed monogenic control of flowering time in the crosses ICCV 96029 × CDC Frontier, BGD 132 × CDC Frontier and ICC 16641 × CDC Frontier, while digenic control with complementary gene action in ICC 5810 × CDC Frontier. The intraspecific genetic maps developed from these crosses consisted 75, 75, 68 and 67 markers spanning 248.8 cM, 331.4 cM, 311.1 cM and 385.1 cM, respectively. A consensus map spanning 363.8 cM with 109 loci was constructed by integrating four genetic maps. Major QTLs corresponding to flowering time genes efl-1 from ICCV 96029, efl-3 from BGD 132 and efl-4 from ICC 16641 were mapped on CaLG04, CaLG08 and CaLG06, respectively. The QTLs and linked markers identified in this study can be used in marker-assisted breeding for developing early maturing chickpea.

Development of a large number of SSR and InDel markers and construction of a high-density genetic map based on a RIL population of pepper (Capsicum annuum L.)

Capsicum annuum, the most widely cultivated species of pepper, is used worldwide for its important nutritional and medicinal values. The construction of an intraspecific high-density genetic linkage map would be of practical value for pepper breeding. However, the numbers of PCR-based simple sequence repeat (SSR) and insertion/deletion (InDel) markers that are available are limited, and there is a need to develop a saturated, intraspecific linkage map. The non-redundant Capsicum species’ expressed sequence tag (EST) database from the National Center for Biotechnology Information was used in this study to develop a total of 902 usable EST-SSR markers. Additionally, 177,587 SSR loci were identified based on the pepper genomic information, including 9182 SSR loci 500 bp both upstream and downstream of coding regions. Another 4497 stable and reliable InDel loci were also developed. From 9182 SSR and 4497 InDel loci, 3356 pairs of genomic SSR primers and 1400 pairs of InDel primers that were evenly distributed in 12 chromosomes were selected. A high-density intraspecific genetic map of C. annuum was constructed using the F10-generation recombinant inbred line of parents PM702 and FS871 as the mapping population, screening the selected 3356 pairs of genomic SSR primers and 1400 pairs of InDel primers and the 902 EST-SSR markers developed earlier, and 524 published SSR markers and 299 orthologous markers (including 263 COSII markers and 36 tomato-derived markers) used previously to develop an interspecific genetic map (C. annuum × C. frutescens). Eventually, a high-density complete genetic intraspecific linkage map of C. annuum containing 12 linkage groups and 708 molecular markers with a length of 1260.00 cM and an average map distance of 1.78 cM was produced. This intraspecific, high-density, complete genetic linkage map of C. annuum contains the largest number of SSR and InDel markers and the highest amount of saturation so far, and it will be of considerable significance for the breeding of improved cultivars of this important field crop in the future.

Major Quantitative Trait Loci and Putative Candidate Genes for Powdery Mildew Resistance and Fruit-Related Traits Revealed by an Intraspecific Genetic Map for Watermelon (Citrullus lanatus var. lanatus).

An intraspecific genetic map for watermelon was constructed using an F2 population derived from ‘Arka Manik’ × ‘TS34’ and transcript sequence variants and quantitative trait loci (QTL) for resistance to powdery mildew (PMR), seed size (SS), and fruit shape (FS) were analyzed. The map consists of 14 linkage groups (LGs) defined by 174 cleaved amplified polymorphic sequences (CAPS), 2 derived-cleaved amplified polymorphic sequence markers, 20 sequence-characterized amplified regions, and 8 expressed sequence tag-simple sequence repeat markers spanning 1,404.3 cM, with a mean marker interval of 6.9 cM and an average of 14.6 markers per LG. Genetic inheritance and QTL analyses indicated that each of the PMR, SS, and FS traits is controlled by an incompletely dominant effect of major QTLs designated as pmr2.1, ss2.1, and fsi3.1, respectively. The pmr2.1, detected on chromosome 2 (Chr02), explained 80.0% of the phenotypic variation (LOD = 30.76). This QTL was flanked by two CAPS markers, wsb2-24 (4.00 cM) and wsb2-39 (13.97 cM). The ss2.1, located close to pmr2.1 and CAPS marker wsb2-13 (1.00 cM) on Chr02, explained 92.3% of the phenotypic variation (LOD = 68.78). The fsi3.1, detected on Chr03, explained 79.7% of the phenotypic variation (LOD = 31.37) and was flanked by two CAPS, wsb3-24 (1.91 cM) and wsb3-9 (7.00 cM). Candidate gene-based CAPS markers were developed from the disease resistance and fruit shape gene homologs located on Chr.02 and Chr03 and were mapped on the intraspecific map. Colocalization of these markers with the major QTLs indicated that watermelon orthologs of a nucleotide-binding site-leucine-rich repeat class gene containing an RPW8 domain and a member of SUN containing the IQ67 domain are candidate genes for pmr2.1 and fsi3.1, respectively. The results presented herein provide useful information for marker-assisted breeding and gene cloning for PMR and fruit-related traits.

High-density linkage map construction and mapping of seed trait QTLs in chickpea (Cicer arietinum L.) using Genotyping-by-Sequencing (GBS).

This study reports the use of Genotyping-by-Sequencing (GBS) for large-scale SNP discovery and simultaneous genotyping of recombinant inbred lines (RILs) of an intra-specific mapping population of chickpea contrasting for seed traits. A total of 119,672 raw SNPs were discovered, which after stringent filtering revealed 3,977 high quality SNPs of which 39.5% were present in genic regions. Comparative analysis using physically mapped marker loci revealed a higher degree of synteny with Medicago in comparison to soybean. The SNP genotyping data was utilized to construct one of the most saturated intra-specific genetic linkage maps of chickpea having 3,363 mapped positions including 3,228 SNPs on 8 linkage groups spanning 1006.98 cM at an average inter marker distance of 0.33 cM. The map was utilized to identify 20 quantitative trait loci (QTLs) associated with seed traits accounting for phenotypic variations ranging from 9.97% to 29.71%. Analysis of the genomic sequence corresponding to five robust QTLs led to the identification of 684 putative candidate genes whose expression profiling revealed that 101 genes exhibited seed specific expression. The integrated approach utilizing the identified QTLs along with the available genome and transcriptome could serve as a platform for candidate gene identification for molecular breeding of chickpea.

Ultra-High Density, Transcript-Based Genetic Maps of Pepper Define Recombination in the Genome and Synteny Among Related Species.

Our ability to assemble complex genomes and construct ultradense genetic maps now allows the determination of recombination rates, translocations, and the extent of genomic collinearity between populations, species, and genera. We developed two ultradense genetic linkage maps for pepper from single-position polymorphisms (SPPs) identified de novo with a 30,173 unigene pepper genotyping array. The Capsicum frutescens × C. annuum interspecific and the C. annuum intraspecific genetic maps were constructed comprising 16,167 and 3,878 unigene markers in 2108 and 783 genetic bins, respectively. Accuracies of marker groupings and orders are validated by the high degree of collinearity between the two maps. Marker density was sufficient to locate the chromosomal breakpoint resulting in the P1/P8 translocation between C. frutescens and C. annuum to a single bin. The two maps aligned to the pepper genome showed varying marker density along the chromosomes. There were extensive chromosomal regions with suppressed recombination and reduced intraspecific marker density. These regions corresponded to the pronounced nonrecombining pericentromeric regions in tomato, a related Solanaceous species. Similar to tomato, the extent of reduced recombination appears to be more pronounced in pepper than in other plant species. Alignment of maps with the tomato and potato genomes shows the presence of previously known translocations and a translocation event that was not observed in previous genetic maps of pepper.

Development of a 63K SNP Array for Cotton and High-Density Mapping of Intraspecific and Interspecific Populations of Gossypium spp.

High-throughput genotyping arrays provide a standardized resource for plant breeding communities that are useful for a breadth of applications including high-density genetic mapping, genome-wide association studies (GWAS), genomic selection (GS), complex trait dissection, and studying patterns of genomic diversity among cultivars and wild accessions. We have developed the CottonSNP63K, an Illumina Infinium array containing assays for 45,104 putative intraspecific single nucleotide polymorphism (SNP) markers for use within the cultivated cotton species Gossypium hirsutum L. and 17,954 putative interspecific SNP markers for use with crosses of other cotton species with G. hirsutum. The SNPs on the array were developed from 13 different discovery sets that represent a diverse range of G. hirsutum germplasm and five other species: G. barbadense L., G. tomentosum Nuttal × Seemann, G. mustelinum Miers × Watt, G. armourianum Kearny, and G. longicalyx J.B. Hutchinson and Lee. The array was validated with 1,156 samples to generate cluster positions to facilitate automated analysis of 38,822 polymorphic markers. Two high-density genetic maps containing a total of 22,829 SNPs were generated for two F2 mapping populations, one intraspecific and one interspecific, and 3,533 SNP markers were co-occurring in both maps. The produced intraspecific genetic map is the first saturated map that associates into 26 linkage groups corresponding to the number of cotton chromosomes for a cross between two G. hirsutum lines. The linkage maps were shown to have high levels of collinearity to the JGI G. raimondii Ulbrich reference genome sequence. The CottonSNP63K array, cluster file and associated marker sequences constitute a major new resource for the global cotton research community.

A combinatorial approach of comprehensive QTL-based comparative genome mapping and transcript profiling identified a seed weight-regulating candidate gene in chickpea.

High experimental validation/genotyping success rate (94–96%) and intra-specific polymorphic potential (82–96%) of 1536 SNP and 472 SSR markers showing in silico polymorphism between desi ICC 4958 and kabuli ICC 12968 chickpea was obtained in a 190 mapping population (ICC 4958 × ICC 12968) and 92 diverse desi and kabuli genotypes. A high-density 2001 marker-based intra-specific genetic linkage map comprising of eight LGs constructed is comparatively much saturated (mean map-density: 0.94 cM) in contrast to existing intra-specific genetic maps in chickpea. Fifteen robust QTLs (PVE: 8.8–25.8% with LOD: 7.0–13.8) associated with pod and seed number/plant (PN and SN) and 100 seed weight (SW) were identified and mapped on 10 major genomic regions of eight LGs. One of 126.8 kb major genomic region harbouring a strong SW-associated robust QTL (Caq'SW1.1: 169.1–171.3 cM) has been delineated by integrating high-resolution QTL mapping with comprehensive marker-based comparative genome mapping and differential expression profiling. This identified one potential regulatory SNP (G/A) in the cis-acting element of candidate ERF (ethylene responsive factor) TF (transcription factor) gene governing seed weight in chickpea. The functionally relevant molecular tags identified have potential to be utilized for marker-assisted genetic improvement of chickpea.

Enrichment of an intraspecific genetic map of upland cotton by developing markers using parental RAD sequencing.

RAD sequencing was performed using DH962 and Jimian5 as upland cotton mapping parents. Sequencing data for DH962 and Jimian5 were assembled into the genome sequences of ≈55.27 and ≈57.06 Mb, respectively. Analysing genome sequences of the two parents, 1,323 SSR, 3,838 insertion/deletion (InDel), and 9,366 single-nucleotide polymorphism (SNP) primer pairs were developed. All of the SSRs, 121 InDels, 441 SNPs, and other 6,747 primer pairs were screened in the two parents, and a total of 535 new polymorphic loci were identified. A genetic map including 1,013 loci was constructed using these results and 506 loci previously published for this population. Twenty-seven new QTLs for yield and fibre quality were identified, indicating that the efficiency of QTL detection was greatly improved by the increase in map density. Comparative genomics showed there to be considerable homology and collinearity between the AT and A2 genomes and between the DT and D5 genomes, although there were a few exchanges and introgressions among the chromosomes of the A2 genome. Here, the development of markers using parental RAD sequencing was effective, and a high-density intraspecific genetic map was constructed. This map can be used for molecular marker-assisted selection in cotton.

Genetic map and QTL controlling fiber quality traits in upland cotton (Gossypium hirsutum L.)

Cotton is a leading natural fiber crop in the textile industry worldwide. The improvement of cotton fiber quality has become more important because of changes in spinning technology and ever-increasing demands. Mapping quantitative trait locus (QTL) for fiber quality traits will enable molecular marker-assisted selection (MAS) to improve fiber quality and provide an access to reveal the molecular mechanism of fiber development. A high-density intraspecific genetic map is constructed based on an upland cotton recombinant inbred line (RIL) population. A total of 25,313 SSR primer pairs were used and yielded 1,333 polymorphic markers, with a polymorphic ratio of 5.3 %, producing 1,382 polymorphic loci in the RIL population. The map comprised 1,274 loci and spanned 3,076.4 cM with an average distance of 2.41 cM between two adjacent markers. Based on the phenotypic data of fiber quality traits from five environments, a total of 59 QTL were detected. These QTL comprised 15 QTL for fiber upper half mean length, 10 QTL for fiber length uniformity, 9 QTL for fiber strength, 10 QTL for fiber elongation and 15 QTL for fiber micronaire, respectively. The genetic map constructed in this study is the most detailed upland cotton intraspecific map based on SSR markers to date, and could be used to construct consensus map or as reference genetic map for tetraploid cotton genome assembly. Stable QTL identified across multiple environments reflect some important and favorable alleles shaping fiber quality, and they are valuable candidate alleles for MAS breeding projects as well as for gene function research related to cotton fiber development and quality improvement.

Genotyping-by-sequencing based intra-specific genetic map refines a ''QTL-hotspot" region for drought tolerance in chickpea.

To enhance the marker density in the “QTL-hotspot” region, harboring several QTLs for drought tolerance-related traits identified on linkage group 04 (CaLG04) in chickpea recombinant inbred line (RIL) mapping population ICC 4958 × ICC 1882, a genotyping-by-sequencing approach was adopted. In total, 6.24 Gb data from ICC 4958, 5.65 Gb data from ICC 1882 and 59.03 Gb data from RILs were generated, which identified 828 novel single-nucleotide polymorphisms (SNPs) for genetic mapping. Together with these new markers, a high-density intra-specific genetic map was developed that comprised 1,007 marker loci spanning a distance of 727.29 cM. QTL analysis using the extended genetic map along with precise phenotyping data for 20 traits collected over one to seven seasons identified 49 SNP markers in the “QTL-hotspot” region. These efforts have refined the “QTL-hotspot” region to 14 cM. In total, 164 main-effect QTLs including 24 novel QTLs were identified. In addition, 49 SNPs integrated in the “QTL-hotspot” region were converted into cleaved amplified polymorphic sequence (CAPS) and derived CAPS (dCAPS) markers which can be used in marker-assisted breeding.Electronic supplementary materialThe online version of this article (doi:10.1007/s00438-014-0932-3) contains supplementary material, which is available to authorized users.

An intra-specific consensus genetic map of pigeonpea [Cajanus cajan (L.) Millspaugh] derived from six mapping populations

Pigeonpea (Cajanus cajan L.) is an important food legume crop of rainfed agriculture. Owing to exposure of the crop to a number of biotic and abiotic stresses, the crop productivity has remained stagnant for almost last five decades at ca. 750 kg/ha. The availability of a cytoplasmic male sterility (CMS) system has facilitated the development and release of hybrids which are expected to enhance the productivity of pigeonpea. Recent advances in genomics and molecular breeding such as marker-assisted selection (MAS) offer the possibility to accelerate hybrid breeding. Molecular markers and genetic maps are pre-requisites for deploying MAS in breeding. However, in the case of pigeonpea, only one inter- and two intra-specific genetic maps are available so far. Here, four new intra-specific genetic maps comprising 59–140 simple sequence repeat (SSR) loci with map lengths ranging from 586.9 to 881.6 cM have been constructed. Using these four genetic maps together with two recently published intra-specific genetic maps, a consensus map was constructed, comprising of 339 SSR loci spanning a distance of 1,059 cM. Furthermore, quantitative trait loci (QTL) analysis for fertility restoration (Rf) conducted in three mapping populations identified four major QTLs explaining phenotypic variances up to 24 %. To the best of our knowledge, this is the first report on construction of a consensus genetic map in pigeonpea and on the identification of QTLs for fertility restoration. The developed consensus genetic map should serve as a reference for developing new genetic maps as well as correlating with the physical map in pigeonpea to be developed in near future. The availability of more informative markers in the bins harbouring QTLs for sterility mosaic disease (SMD) and Rf will facilitate the selection of the most suitable markers for genetic analysis and molecular breeding applications in pigeonpea.Electronic supplementary materialThe online version of this article (doi:10.1007/s00122-012-1916-5) contains supplementary material, which is available to authorized users.

Integration of EST-SSR markers of Medicago truncatula into intraspecific linkage map of lentil and identification of QTL conferring resistance to ascochyta blight at seedling and pod stages

Microsatellite markers have been extensively utilised in the leguminosae for genome mapping and identifying major loci governing traits of interest for eventual marker-assisted selection (MAS). The lack of available lentil-specific microsatellite sequences and gene-based markers instigated the mining and transfer of expressed sequence tag simple sequence repeat (EST-SSR)/SSR sequences from the model genome Medicago truncatula, to enrich an existing intraspecific lentil genetic map. A total of 196 markers, including new 15 M. truncatula EST-SSR/SSR, were mapped using a population of 94 F5 recombinant inbred lines produced from a cross between cv. Northfield (ILL5588) × cv. Digger (ILL5722) and clustered into 11 linkage groups (LG) covering 1156.4 cM. Subsequently, the size and effects of quantitative trait loci (QTL) conditioning Ascochyta lentis resistance at seedling and pod/maturity stages were characterised and compared. Three QTL were detected for seedling resistance on LG1 and LG9 and a further three were detected for pod/maturity resistance on LG1, LG4 and LG5. Together, these accounted for 34 and 61% of the total estimated phenotypic variation, respectively, and demonstrated that resistance at the different growth stages is potentially conditioned by different genomic regions. The flanking markers identified may be useful for MAS and for the future pyramiding of potentially different resistance genes into elite backgrounds that are resistant throughout the cropping season.

Genetic mapping and quantitative trait locus analysis of resistance to sterility mosaic disease in pigeonpea [ Cajanus cajan (L.) Millsp.

Sterility mosaic disease (SMD), considered as the “green plague of pigeonpea” and caused by pigeonpea sterility mosaic virus (PPSMV) is one of the major biotic factors, which leads to heavy yield losses and hence poses a big challenge for pigeonpea production in the Indian subcontinent. Variability in the sterility mosaic pathogen revealed the occurrence of five different isolates in India. Among them, three distinct SMD isolates have been characterized, viz., Patancheru, Bangalore and Coimbatore. Molecular tools offer a viable option to tackle these biotic stresses via identification of the genomic regions associated with the trait such as SMD resistance. With an aim of identifying the gene(s)/QTLs linked with SMD resistance, two F 2 populations, i.e. ICP 8863 × ICPL 20097 (segregating for Patancheru SMD isolate) and TTB 7 × ICP 7035 (segregating for both Patancheru and Bangalore SMD isolates) were developed and F 2:3 families were phenotyped for resistance to respective isolate(s) of SMD. After screening over 3000 SSR markers on parental genotypes of each mapping population, intra-specific genetic maps comprising of 11 linkage groups and 120 and 78 SSR loci were developed for ICP 8863 × ICPL 20097 and TTB 7 × ICP 7035 populations, respectively. Composite interval mapping (CIM) based QTL analysis by using genetic mapping and phenotyping data provided four QTLs for Patancheru SMD isolate and two QTLs for Bangalore SMD isolate. Identification of different QTLs for resistance to Patancheru and Bangalore SMD isolates is an indication of involvement of different genes conferring the resistance to these two SMD isolates. One QTL namely qSMD4 identified within an interval of 2.8 cM on LG 7 explaining 24.72% of phenotypic variance, once it is validated in other genetic background, seems to be a promising QTL for use in marker assisted selection. In summary, this is the first study on development of intra-specific genetic maps and identification of QTLs for SMD resistance in pigeonpea.

An intraspecific genetic map of water yam (Dioscorea alata L.) based on AFLP markers and QTL analysis for anthracnose resistance

Water yam (Dioscorea alata L.) is the most widely cultivated food yams. Despite its importance, its production is limited by anthracnose disease caused by Colletotrichum gloeosporioides (Penz.). The use of resistant yam varieties is the most reliable approach of management of this disease. The speed and precision of breeding can be improved by the development of genetic linkage maps which would provide the basis for locating and hence manipulating quantitative traits such as anthracnose resistance in breeding programmes. An F1 diploid population was developed by crossing ‘Boutou’ a female clone (with field resistance to anthracnose) with ‘Pyramide’ (susceptible). A linkage map was generated with 523 polymorphic markers from 26 AFLP primer combinations. The resulting map covered a total length of 1538 cM and included 20 linkage groups. It is the most saturated of all genetic linkage maps of yam to date. QTL analysis of anthracnose resistance was performed based on response to two isolates of C. gloeosporioides. Resistance to anthracnose appeared to be inherited quantitatively. Using a LOD significance threshold of 2.6 we identified a total of nine QTLs for anthracnose resistance. The phenotypic variance explained by each QTL ranged from 7.0 to 32.9% whereas the total amount of phenotypic variation for anthracnose resistance explained by all significant QTLs varied from 26.4 to 73.7% depending on the isolate and the variable considered. These QTLs displayed isolate-specific resistance as well as broad spectrum resistance. The availability of molecular markers linked to the QTLs of anthracnose resistance will facilitate marker-assisted selection in breeding programmes.

SNP Discovery and Linkage Map Construction in Cultivated Tomato

Few intraspecific genetic linkage maps have been reported for cultivated tomato, mainly because genetic diversity within Solanum lycopersicum is much less than that between tomato species. Single nucleotide polymorphisms (SNPs), the most abundant source of genomic variation, are the most promising source of polymorphisms for the construction of linkage maps for closely related intraspecific lines. In this study, we developed SNP markers based on expressed sequence tags for the construction of intraspecific linkage maps in tomato. Out of the 5607 SNP positions detected through in silico analysis, 1536 were selected for high-throughput genotyping of two mapping populations derived from crosses between ‘Micro-Tom’ and either ‘Ailsa Craig’ or ‘M82’. A total of 1137 markers, including 793 out of the 1338 successfully genotyped SNPs, along with 344 simple sequence repeat and intronic polymorphism markers, were mapped onto two linkage maps, which covered 1467.8 and 1422.7 cM, respectively. The SNP markers developed were then screened against cultivated tomato lines in order to estimate the transferability of these SNPs to other breeding materials. The molecular markers and linkage maps represent a milestone in the genomics and genetics, and are the first step toward molecular breeding of cultivated tomato. Information on the DNA markers, linkage maps, and SNP genotypes for these tomato lines is available at http://www.kazusa.or.jp/tomato/.

The first intraspecific genetic linkage maps of wintersweet [ Chimonanthus praecox (L.) Link] based on AFLP and ISSR markers

Wintersweet [ Chimonanthus praecox (L.) Link] ( n = 11), a dichogamous species, is a famous traditional fragrant flower plant in China. We used AFLPs and ISSR to construct a genetic linkage map on a pseudo-F 2 population of wintersweet derived from a cross between the individuals H4 and H29. A total of 370 polymorphism markers was generated from 12 AFLP primer combinations and 9 ISSR primers. Two separate female and male maps were constructed using 84 female-specific and 51 male-specific testcross markers, respectively. The maternal H4 map included 80 markers ordered in 12 linkage groups; while the paternal H29 map had 47 markers in 8 linkage groups. At a minimum logarithm of the odds (LOD) score of 3.0 and maximum map distance of 50 cM, the female map covered 2417.8 cM, with an average distance of 25.61 cM and maximum map distance of 48.2 cM between two loci. In contrast, the male map covered 1184.2 cM, with an average distance of 25.7 cM and maximum map distance of 49.0 cM between two loci. These genetic maps will serve as a framework for mapping QTLs and provide basic information for future molecular breeding studies.

QTL mapping in A-genome diploid Asiatic cotton and their congruence analysis with AD-genome tetraploid cotton in genus Gossypium

Asiatic cotton (Gossypium arboreum L.) is an Old World cultivated cotton species. The sinense race was planted extensively in China. Due to the advances in spinning technology during the last century, the species was replaced by the New World allotetraploid cotton G. hirsutum L. Gossypium arboreum is still grown in India and Pakistan and also used as an elite in current cotton breeding programs. In addition, G. arboreum serves as a model for genomic research in Gossypium. In the present study, we generated an A-genome diploid cotton intraspecific genetic map including 264 SSR loci with three morphological markers mapped to 13 linkage groups. The map spans 2,508.71 cM with an average distance of 9.4 cM between adjacent loci. A population containing 176 F(2:3) families was used to perform quantitative trait loci (QTL) mapping for 17 phenotypes using Multiple QTL Model (MQM) of MapQTL ver 5.0. Overall, 108 QTLs were detected on 13 chromosomes. Thirty-one QTLs for yield and its components were detected in the F2 population. Forty-one QTLs for yield and its components were detected in the F(2:3) families with a total of 43 QTLs for fiber qualities. Two QTLs for seed cotton weight/plant and lint index and three QTLs for seed index were consistently detected both in F2 and F(2:3). Most QTLs for fiber qualities and yields were located at the same interval or neighboring intervals. These results indicated that the negative correlation between fiber qualities and yield traits may result from either pleiotropic effect of one gene or linkage effects of multiple closely linked genes.

An intraspecific genetic map of velvetbean (Mucuna sp.) based on AFLP markers.

Velvetbean ( Mucuna sp., n=11), a self-pollinated species, is an important legume used in tropical agricultural systems in rotation with other crops for nematode management and/or soil improvement. A genetic map of velvetbean was constructed in order to identify potential molecular markers linked to important morphological and agronomic traits that would be particularly useful for developing and improving the species. Traits such as seed coat color, pod color, and pod pubescence were among the main parameters observed in a process of genetic diversity estimation. Two slightly divergent velvetbean accessions, PI364362 and 'Edgar Farm White', a land race from Alabama, were used to make an intraspecific F(1) hybrid. Amplified fragment length polymorphism analysis (AFLP) detected an average of six polymorphic fragments per primer pair between the two parents. As expected for dominant markers, the sum of all AFLP bands from both parents was generally observed to be present in the AFLP profiles of the F(1) progeny, indicating full penetrance and the dominant nature of AFLP markers. An F(2) population was generated by self-pollinating a single F(1) plant. Using 37 AFLP primer pairs, we detected 233 polymorphic markers of which 164 (70.4%) segregated in 3:1 Mendelian ratios, while the remaining 69 (29.6%) both segregated and were scorable. The genetic linkage map constructed from this population comprised 166 markers, including two morphological traits (pod color and pod pubescence). Twenty linkage groups were found with an average distance between markers of 34.4 cM, covering a total of 687.9 cM. The linkage groups contained from 2 to 12 loci each and the distance between two consecutive loci ranged from 0 to 21.8 cM. The newly designated morphological traits pod color ( pdc) and pod pubescence ( pdp) co-segregated with each other at a distance of 4.2 cM. Two DNA markers designated ACGCAG2 and ACTCTG1 were located in the same group as pdc and pdp. The AFLP linkage map provides opportunities for use in marker-assisted selection and in the detection of loci controlling morphologically important traits.