Chromosomal Linkage Groups Research Articles

A chromosomal-scale reference genome of the New World Screwworm, Cochliomyia hominivorax.

The New World Screwworm, Cochliomyia hominivorax (Calliphoridae), is the most important myiasis-causing species in America. Screwworm myiasis is a zoonosis that can cause severe lesions in livestock, domesticated and wild animals, and occasionally in people. Beyond the sanitary problems associated with this species, these infestations negatively impact economic sectors, such as the cattle industry. Here, we present a chromosome-scale assembly of C. hominivorax's genome, organized in 6 chromosome-length and 515 unplaced scaffolds spanning 534 Mb. There was a clear correspondence between the D. melanogaster linkage groups A-E and the chromosomal-scale scaffolds. Chromosome quotient (CQ) analysis identified a single scaffold from the X chromosome that contains most of the orthologs of genes that are on the D. melanogaster fourth chromosome (linkage group F or dot chromosome). CQ analysis also identified potential X and Y unplaced scaffolds and genes. Y-linkage for selected regions was confirmed by PCR with male and female DNA. Some of the long chromosome-scale scaffolds include Y-linked sequences, suggesting misassembly of these regions. These resources will provide a basis for future studies aiming at understanding the biology and evolution of this devastating obligate parasite.

Remnant of Unrelated Amniote Sex Chromosomal Linkage Sharing on the Same Chromosome in House Gecko Lizards, Providing a Better Understanding of the Ancestral Super-Sex Chromosome.

Comparative chromosome maps investigating sex chromosomal linkage groups in amniotes and microsatellite repeat motifs of a male house gecko lizard (Hemidactylus frenatus, HFR) and a flat-tailed house gecko lizard (H. platyurus, HPL) of unknown sex were examined using 75 bacterial artificial chromosomes (BACs) from chicken and zebra finch genomes. No massive accumulations of microsatellite repeat motifs were found in either of the gecko lizards, but 10 out of 13 BACs mapped on HPL chromosomes were associated with other amniote sex chromosomes. Hybridization of the same BACs onto multiple different chromosome pairs suggested transitions to sex chromosomes across amniotes. No BAC hybridization signals were found on HFR chromosomes. However, HFR diverged from HPL about 30 million years ago, possibly due to intrachromosomal rearrangements occurring in the HFR lineage. By contrast, heterochromatin likely reshuffled patterns between HPL and HFR, as observed from C-positive heterochromatin distribution. Six out of ten BACs showed partial homology with squamate reptile chromosome 2 (SR2) and snake Z and/or W sex chromosomes. The gecko lizard showed shared unrelated sex chromosomal linkages—the remnants of a super-sex chromosome. A large ancestral super-sex chromosome showed a correlation between SR2 and snake W sex chromosomes.

Dissecting Genetic Resistance to Fire Blight in Three Pear Populations.

Fire blight, caused by the bacterial pathogen Erwinia amylovora, is a persistent problem for pear (Pyrus spp.) growers in the United States. Growing resistant cultivars is one of the best options for managing fire blight. The cultivars Potomac and Old Home and the selection NJA2R59T69 display resistance to fire blight. As such, three mapping populations (El Dorado × Potomac, Old Home × Bartlett, and NJA2R59T69 × Bartlett) were developed to identify genomic regions associated with resistance to fire blight. Progeny were phenotyped during 2017 and 2018 by inoculating multiple actively growing shoots of field-grown seedling trees with E. amylovora isolate E153n via the cut-leaf method. Genotyping was conducted using the recently developed Axiom Pear 70 K Genotyping Array and chromosomal linkage groups were created for each population. An integrated two-way pseudo-testcross approach was used to map quantitative trait loci (QTLs). Resistance QTLs were identified on chromosome 2 for each population. The QTLs identified in the El Dorado × Potomac and Old Home × Bartlett populations are in the same region as QTLs that were previously identified in Harrow Sweet and Moonglow. The QTL in NJA2R59T69 mapped proximally to the previously identified QTLs and originated from an unknown Asian or occidental source. Future research will focus on further characterizing the resistance regions and developing tools for DNA-informed breeding.

The Genome of Caenorhabditis bovis.

The free-living nematode Caenorhabditis elegans is akey laboratory model for metazoan biology. C.elegans has also become a model for parasitic nematodes despite being only distantly related to most parasitic species. All of the ∼65 Caenorhabditis species currently in culture are free-living, with most having been isolated from decaying plant or fungal matter. Caenorhabditis bovis is a particularly unusual species that has been isolated several times from the inflamed ears of Zebu cattle in Eastern Africa, where it is associated with thedisease bovine parasitic otitis. C.bovis is therefore of particular interest to researchers interested in the evolution of nematode parasitism. However, as C.bovis is not in laboratory culture, it remains little studied. Here, by sampling livestock markets and slaughterhouses in Western Kenya, we successfully reisolated C.bovis from the ear of adult female Zebu. We sequenced the genome of C.bovis using the Oxford Nanopore MinION platform in a nearby field laboratory and used the data to generate a chromosome-scaledraft genome sequence. We exploited this draft genome sequence to reconstruct the phylogenetic relationships of C.bovis to other Caenorhabditis species and reveal the changes in genome size and content that have occurred during its evolution. We also identified expansions in several gene families that have been implicated in parasitism in other nematode species. The high-quality draft genome and our analyses thereof represent a significant advancement in our understanding of this unusual Caenorhabditis species.

Inheritance and mapping of drought tolerance in soybean at seedling stage using bulked segregant analysis

AbstractOccurrence of drought under rainfed conditions is the foremost factor responsible for yield reduction in soybean. Developing soybean cultivars with an inherent ability to withstand drought would immensely benefit the soybean production in rainfed areas. In the present study, F2 derived mapping populations were developed by crossing drought tolerant (PK 1180, SL 46) and susceptible (UPSL 298, PK 1169) genotypes to investigate the inheritance of seedling survival drought mechanisms and to identify simple-sequence repeat (SSR) markers associated with them, using bulked segregant analysis. Parents as well as a F2 derived mapping population were screened for drought tolerance based on seedling survivability under controlled conditions. Segregation analysis of F2 population derived from a cross between PK 1180 × UPSL 298 was previously shown to have a 3:1 tolerant to susceptible ratio and a probability of 0.61 at a χ2(3:1) value of 0.258. This was confirmed in another F2 population derived from a cross between PK 1169 × SL 46 with a χ2(3:1) value of 0.145 obtained at a probability of 0.70. One SSR marker Satt277 showed polymorphism between contracting bulks (tolerant and susceptible) out of 50 polymorphic markers identified during parental polymorphism. Single marker analysis suggested that the marker, Satt277 is linked to seedling survival drought tolerance and is located on chromosome linkage group C2 (chr 6) with a map distance of 3.40 cM. The tolerant genotypes identified could be used as a donor in soybean improvement programs. The marker identified can be used in marker-assisted selection while screening large collection of germplasm.

Genetic analysis and fine-mapping of Soybean mosaic virus SC7 and SC13 resistance genes in soybean (Glycine max)

Soybean mosaic virus (SMV) is one of the most destructive pathogens of soybean (Glycine max (L.) Merr.) worldwide. In this study, 184 F7:11 recombinant inbred line (RIL) populations derived from Kefeng No. 1 × Nannong 1138-2 were used to study the inheritance and linkage mapping of resistance genes against SMV strains SC7 and SC13 in Kefeng No. 1. Two independent dominant genes (designated Rsc7 and Rsc13) that control resistance to SC7 and SC13 were located on a molecular linkage group (MLG) of chromosome 2 (D1b). A mixed segregating population was developed by self-pollination of three heterozygous plants of residual heterozygous lines (RHL3-27, RHL3-30, RHL3-53) with five markers linked to the loci, and was used in fine-mapping of Rsc7 and Rsc13. In addition, Rsc7 was fine-mapped between BARCSOYSSR_02_0667 and BARCSOYSSR_02_0670 on MLG D1b. The genetic distance between the two closest markers was 0.7 cM and the physical distance of the interval was ~77 kb, which included one LRR gene and another gene containing an F-box region. Two SSR markers (BARCSOYSSR_02_0610 and BARCSOYSSR_02_0621) were closely linked to the SC13 resistance gene. The physical distance where Rsc13 was located was ~191 kb. Sequence analysis showed that there were two K-box region types of transcription factor genes; GmHSP40 and two serine/threonine protein kinase (STK) genes were the most likely candidate genes. These results will facilitate map-based cloning of the Rsc7 and Rsc13 genes and development of transgenic disease-resistant varieties, and will provide SMV-resistance breeding systems with excellent resistance germplasm.

The LsVe1L allele provides a molecular marker for resistance to Verticillium dahliae race 1 in lettuce.

BackgroundVerticillium wilt caused by the fungus Verticillium dahliae race 1 is among the top disease concerns for lettuce in the Salinas and Pajaro Valleys of coastal central California. Resistance of lettuce against V. dahliae race 1 was previously mapped to the single dominant Verticillium resistance 1 (Vr1) locus. Lines of tomato resistant to race 1 are known to contain the closely linked Ve1 and Ve2 genes that encode receptor-like proteins with extracellular leucine-rich repeats; the Ve1 and Ve2 proteins act antagonistically to provide resistance against V. dahliae race 1. The Vr1 locus in lettuce contains a cluster of several genes with sequence similarity to the tomato Ve genes. We used genome sequencing and/or PCR screening along with pathogenicity assays of 152 accessions of lettuce to investigate allelic diversity and its relationship to race 1 resistance in lettuce.ResultsThis approach identified a total of four Ve genes: LsVe1, LsVe2, LsVe3, and LsVe4. The majority of accessions, however, contained a combination of only three of these LsVe genes clustered on chromosomal linkage group 9 (within ~ 25 kb in the resistant cultivar La Brillante and within ~ 127 kb in the susceptible cultivar Salinas).ConclusionsA single allele, LsVe1L, was present in all resistant accessions and absent in all susceptible accessions. This allele can be used as a molecular marker for V. dahliae race 1 resistance in lettuce. A PCR assay for rapid detection of race 1 resistance in lettuce was designed based on nucleotide polymorphisms. Application of this assay allows identification of resistant genotypes in early stages of plant development or at seed-level without time- and labor-intensive testing in the field.

An ultra-dense integrated linkage map for hexaploid chrysanthemum enables multi-allelic QTL analysis

Key messageWe constructed the first integrated genetic linkage map in a polysomic hexaploid. This enabled us to estimate inheritance of parental haplotypes in the offspring and detect multi-allelic QTL.Construction and use of linkage maps are challenging in hexaploids with polysomic inheritance. Full map integration requires calculations of recombination frequency between markers with complex segregation types. In addition, detection of QTL in hexaploids requires information on all six alleles at one locus for each individual. We describe a method that we used to construct a fully integrated linkage map for chrysanthemum (Chrysanthemum × morifolium, 2n = 6x = 54). A bi-parental F1 population of 406 individuals was genotyped with an 183,000 SNP genotyping array. The resulting linkage map consisted of 30,312 segregating SNP markers of all possible marker dosage types, representing nine chromosomal linkage groups and 107 out of 108 expected homologues. Synteny with lettuce (Lactuca sativa) showed local colinearity. Overall, it was high enough to number the chrysanthemum chromosomal linkage groups according to those in lettuce. We used the integrated and phased linkage map to reconstruct inheritance of parental haplotypes in the F1 population. Estimated probabilities for the parental haplotypes were used for multi-allelic QTL analyses on four traits with different underlying genetic architectures. This resulted in the identification of major QTL that were affected by multiple alleles having a differential effect on the phenotype. The presented linkage map sets a standard for future genetic mapping analyses in chrysanthemum and closely related species. Moreover, the described methods are a major step forward for linkage mapping and QTL analysis in hexaploids.

Genomic regions and genes related to inter-population differences in body size in the ground beetle Carabus japonicus

Body size is a key trait in diversification among animal species, and revealing the gene regions responsible for body size diversification among populations or related species is important in evolutionary biology. We explored the genomic regions associated with body size differences in Carabus japonicus ground beetle populations by quantitative trait locus (QTL) mapping of F2 hybrids from differently sized parents from two populations using restriction site-associated DNA sequencing and de novo assembly of the beetle whole genome. The assembled genome had a total length of 191 Mb with a scaffold N50 of 0.73 Mb; 14,929 protein-coding genes were predicted. Three QTLs on different linkage groups had major effects on the overall size, which is composed chiefly of elytral length. In addition, we found QTLs on autosomal and X chromosomal linkage groups that affected head length and width, thoracic width, and elytral width. We determined the gene loci potentially related to control of body size in scaffolds of the genome sequence, which contained the QTL regions. The genetic basis of body size variation based on a small number of major loci would promote differentiation in body size in response to selection pressures related to variations in environmental conditions and inter-specific interactions.

Genetic mapping of 15 human X chromosomal forensic short tandem repeat (STR) loci by means of multi-core parallelization

Typing of X chromosomal short tandem repeat (X STR) markers has become a standard element of human forensic genetic analysis. Joint consideration of many X STR markers at a time increases their discriminatory power but, owing to physical linkage, requires inter-marker recombination rates to be accurately known. We estimated the recombination rates between 15 well established X STR markers using genotype data from 158 families (1041 individuals) and following a previously proposed likelihood-based approach that allows for single-step mutations. To meet the computational requirements of this family-based type of analysis, we modified a previous implementation so as to allow multi-core parallelization on a high-performance computing system. While we obtained recombination rate estimates larger than zero for all but one pair of adjacent markers within the four previously proposed linkage groups, none of the three X STR pairs defining the junctions of these groups yielded a recombination rate estimate of 0.50. Corroborating previous studies, our results therefore argue against a simple model of independent X chromosomal linkage groups. Moreover, the refined recombination fraction estimates obtained in our study will facilitate the appropriate joint consideration of all 15 investigated markers in forensic analysis.

Mapping of chromosome loci determined manifestation of morphological and biochemical traits of quality in Brassica rapa L. crops

Mapping chromosomal loci responsible for seven morphological and five biochemical traits of quality in populations of doubled haploid lines of leafy, rooted, and oilseed crops of Brassica rapa L. species was performed for the first time. In total, 140 quantitative trait loci (QTLs) were mapped determining manifestation of the studied morphological and biochemical economically important traits of quality under field and greenhouse conditions; the control of the same traits under different growth conditions or simultaneous control of several traits by one locus were noted. Molecular markers genetically linked with the selected QTLs were detected. The block genomic structure of the genetic components (chromosomal loci and linkage groups) determining manifestation of morphological and biochemical traits of quality that make B. rapa plants nutritionally valuable is discussed for the first time. The conclusion is drawn that the revealed QTLs and identified molecular markers may be of interest for the forthcoming study of genetic control by them of economically important characters and for the marker assisted selection in B. rapa.

A Carbohydrate Hydrolysis Enzymes encoding Genes in Neurospora crassa

Neurospora crassa, NCU05882.7 (423aa) and NCU09774.7 (303 aa) (NCU, Neurospora 7 crassa unit) genes encoding a Cellulase, which hydrolysis the Cellulose. In addition to that, 8 reporting here other 35 Carbohydrate hydrolysis enzymes encoding genes in N.crassa. A 9 metagenomic analysis for multiple sequences alignment and Phylogenetics analysis, the evaluated 10 result showed high sequence similarity and 99% homology to the other class of fungi; in the 11 bacterial species showed extremely very less sequence similarities and 100 % homology. 12 Where as in inter species between fungi and bacteria, the results showed extremely less sequence 13 similarities and 97 % homology. The studies on physiochemical properties of Cellulase using 14 GeneDoc, the evaluated results showed Cellulase was an amphoteric (polor), aromatic, aliphatic 15 and highly repeated amino acids of glycine and proline. These metagenomic studies could help 16 to straightforward isolation of Cellulase enzymes from NCU05882.7 (Chromosome/Linkage 17 Group-VII), NCU09774.7 (Chromosome Linkage Group- II) and other 35 Carbohydrate 18 hydrolysis enzymes encoding genes in N.crassa.

High-density sex-specific linkage maps of a European tree frog (Hyla arborea) identify the sex chromosome without information on offspring sex.

Identifying homology between sex chromosomes of different species is essential to understanding the evolution of sex determination. Here, we show that the identity of a homomorphic sex chromosome pair can be established using a linkage map, without information on offspring sex. By comparing sex-specific maps of the European tree frog Hyla arborea, we find that the sex chromosome (linkage group 1) shows a threefold difference in marker number between the male and female maps. In contrast, the number of markers on each autosome is similar between the two maps. We also find strongly conserved synteny between H. arborea and Xenopus tropicalis across 200 million years of evolution, suggesting that the rate of chromosomal rearrangement in anurans is low. Finally, we show that recombination in males is greatly reduced at the centers of large chromosomes, consistent with previous cytogenetic findings. Our research shows the importance of high-density linkage maps for studies of recombination, chromosomal rearrangement and the genetic architecture of ecologically or economically important traits.

An Ultra-High-Density, Transcript-Based, Genetic Map of Lettuce.

We have generated an ultra-high-density genetic map for lettuce, an economically important member of the Compositae, consisting of 12,842 unigenes (13,943 markers) mapped in 3696 genetic bins distributed over nine chromosomal linkage groups. Genomic DNA was hybridized to a custom Affymetrix oligonucleotide array containing 6.4 million features representing 35,628 unigenes of Lactuca spp. Segregation of single-position polymorphisms was analyzed using 213 F7:8 recombinant inbred lines that had been generated by crossing cultivated Lactuca sativa cv. Salinas and L. serriola acc. US96UC23, the wild progenitor species of L. sativa. The high level of replication of each allele in the recombinant inbred lines was exploited to identify single-position polymorphisms that were assigned to parental haplotypes. Marker information has been made available using GBrowse to facilitate access to the map. This map has been anchored to the previously published integrated map of lettuce providing candidate genes for multiple phenotypes. The high density of markers achieved in this ultradense map allowed syntenic studies between lettuce and Vitis vinifera as well as other plant species.

Use of Tyramide-Fluorescence in situ Hybridization and Chromosome Microdissection for Ascertaining Homology Relationships and Chromosome Linkage Group Associations in Oats

The physical mapping of single locus sequences by tyramide-fluorescence in situ hybridization (Tyr-FISH) and the analysis of sequences obtained from microdissected chromosomes were assayed as potential tools for (1) determining homology and homoeology among chromosome regions of Avena species, and (2) establishing associations between linkage groups and specific chromosomes. Low copy number probes, derived from resistance gene analogues (RGAs) and 2.8–4.5 kb long, successfully produced hybridization signals on specific chromosomes. Four sets of homoeologous chromosome regions were identified in the hexaploids using 3 probes that produced 4 single locus markers in A. strigosa and 2 in A. eriantha. Laser capture microdissection of metaphase I cells of A. sativa monosomic lines allowed the isolation of critical univalents. Sequences derived from 2 RGAs were successfully amplified in DNA extracted from univalents. In one instance, it was possible to map a nucleotide polymorphism specific for 1 chromosome. An association was established between this chromosome and its linkage groups in 2 hexaploid genetic maps. The results indicate that Tyr-FISH is useful in the characterization of homoeologous chromosome segments in hexaploids, whereas chromosome microdissection, as employed in this work, needs to be improved before it can routinely be used with meiotic chromosomes.

DEVELOPMENT OF MOLECULAR MARKERS FOR MARKER-ASSISTED SELECTION OF DIEBACK DISEASE RESISTANCE IN LETTUCE (LACTUCA SATIVA)

Lettuce dieback disease is widespread in commercially grown romaine and leaf-type lettuces, but not in iceberg-type cultivars. The disease is caused by two closely related Tombusviruses: Tomato bushy stunt virus (TBSV) and Lettuce necrotic stunt virus (LNSV). A single dominant gene (Tvr1) on chromosomal linkage group 2 has been found to be responsible for the resistance observed in modern iceberg lettuces. A population of 192 F 8 recombinant inbred lines (RILs) developed from crosses between the susceptible romaine cultivar 'Valmaine' and the resistant iceberg cultivars 'Salinas 88' and 'Salinas' was used to pinpoint the position of the resistance gene with EST-based molecular markers. Nine markers closely linked to the Tvrl gene were subsequently tested for association with resistance in a set of 68 accessions from all horticultural types of lettuce. Sequencing of the marker that matched precisely with resistance revealed the presence of three haplotypes in L. sativa, two of them associated with resistance. High-resolution DNA melting analysis, which allows detection of all three haplotypes in a single analysis, was successfully applied to study the marker-trait association. Results confirmed the usefulness of the marker for marker-assisted selection in all types of cultivated lettuce.

Gene Flow in the Face of Countervailing Selection: Adaptation to High-Altitude Hypoxia in the A Hemoglobin Subunit of Yellow-Billed Pintails in the Andes

When populations become locally adapted to contrasting environments, alleles that have high fitness in only one environment may be quickly eliminated in populations adapted to other environments, such that gene flow is partly restricted. The stronger the selection, the more rapidly immigrant alleles of lower fitness will be eliminated from the population. However, gene flow may continue to occur at unlinked loci, and adaptive divergence can proceed in the face of countervailing gene flow if selection is strong relative to migration (s > m). We studied the population genetics of the major hemoglobin genes in yellow-billed pintails (Anas georgica) experiencing contrasting partial pressures of oxygen in the Andes of South America. High gene flow and weak population subdivision were evident at seven putatively neutral loci in different chromosomal linkage groups. In contrast, amino acid replacements (Ser-beta13, Ser-beta116, and Met-beta133) in the betaA hemoglobin subunit segregated by elevation between lowland and highland populations with significantly elevated F(ST). Migration rates for the betaA subunit alleles were approximately 17-24 times smaller than for five unlinked reference loci, the alphaA hemoglobin subunit (which lacks amino acid replacements) and the mitochondrial DNA control region. The betaA subunit alleles of yellow-billed pintails were half as likely to be transferred downslope, from the highlands to the lowlands, than in the opposite direction upslope. We hypothesize that migration between the lowlands and highlands is restricted by local adaptation, and the betaA hemoglobin subunit is a likely target of selection related to high-altitude hypoxia; however, gene flow may be sufficiently high to retard divergence at most unlinked loci. Individuals homozygous for lowland alleles may have relatively little difficulty dispersing to the highlands initially but may experience long-term fitness reduction. Individuals homozygous for highland genotypes, in contrast, would be predicted to have difficulty dispersing to the lowlands if their hemoglobin alleles confer high oxygen affinity, predicted to result in chronic erythrocytosis at low elevation. Heterozygous individuals may have a dispersal advantage if their hemoglobin has a wider range of function due to the presence of multiple protein isoforms with a mixture of different oxygen affinities.

Association mapping and marker-assisted selection of the lettuce dieback resistance gene Tvr1.

BackgroundLettuce (Lactuca saliva L.) is susceptible to dieback, a soilborne disease caused by two viruses from the family Tombusviridae. Susceptibility to dieback is widespread in romaine and leaf-type lettuce, while modern iceberg cultivars are resistant to this disease. Resistance in iceberg cultivars is conferred by Tvr1 - a single, dominant gene that provides durable resistance. This study describes fine mapping of the resistance gene, analysis of nucleotide polymorphism and linkage disequilibrium in the Tvr1 region, and development of molecular markers for marker-assisted selection.ResultsA combination of classical linkage mapping and association mapping allowed us to pinpoint the location of the Tvr1 resistance gene on chromosomal linkage group 2. Nine molecular markers, based on expressed sequence tags (EST), were closely linked to Tvr1 in the mapping population, developed from crosses between resistant (Salinas and Salinas 88) and susceptible (Valmaine) cultivars. Sequencing of these markers from a set of 68 cultivars revealed a relatively high level of nucleotide polymorphism (θ = 6.7 × 10-3) and extensive linkage disequilibrium (r2 = 0.124 at 8 cM) in this region. However, the extent of linkage disequilibrium was affected by population structure and the values were substantially larger when the analysis was performed only for romaine (r2 = 0.247) and crisphead (r2 = 0.345) accessions. The association mapping approach revealed that one of the nine markers (Cntg10192) in the Tvr1 region matched exactly with resistant and susceptible phenotypes when tested on a set of 200 L. sativa accessions from all horticultural types of lettuce. The marker-trait association was also confirmed on two accessions of Lactuca serriola - a wild relative of cultivated lettuce. The combination of three single-nucleotide polymorphisms (SNPs) at the Cntg10192 marker identified four haplotypes. Three of the haplotypes were associated with resistance and one of them was always associated with susceptibility to the disease.ConclusionWe have successfully applied high-resolution DNA melting (HRM) analysis to distinguish all four haplotypes of the Cntg10192 marker in a single analysis. Marker-assisted selection for dieback resistance with HRM is now an integral part of our breeding program that is focused on the development of improved lettuce cultivars.

First complete chromosomal organization of a protozoan plant parasite ( Phytomonas spp.)

Phytomonas spp. are members of the family Trypanosomatidae that parasitize plants and may cause lethal diseases in crops such as Coffee Phloem necrosis, Hartrot in coconut, and Marchitez sorpresiva in oil palm. In this study, the molecular karyotype of 6 isolates from latex plants has been entirely elucidated by pulsed-field gel electrophoresis and DNA hybridization. Twenty-one chromosomal linkage groups constituting heterologous chromosomes and sizing between 0.3 and 3 Mb could be physically defined by the use of 75 DNA markers (sequence-tagged sites and genes). From these data, the genome size can be estimated at 25.5 (± 2) Mb. The physical linkage groups were consistently conserved in all strains examined. Moreover, the finding of several pairs of different-sized homologous chromosomes strongly suggest diploidy for this organism. The definition of the complete molecular karyotype of Phytomonas represents an essential primary step toward sequencing the genome of this parasite of economical importance.

A high-density, integrated genetic linkage map of lettuce (Lactuca spp.).

An integrated map for lettuce comprising of 2,744 markers was developed from seven intra- and inter-specific mapping populations. A total of 560 markers that segregated in two or more populations were used to align the individual maps. 2,073 AFLP, 152 RFLP, 130 SSR, and 360 RAPD as well as 29 other markers were assigned to nine chromosomal linkage groups that spanned a total of 1,505 cM and ranged from 136 to 238 cM. The maximum interval between markers in the integrated map is 43 cM and the mean interval is 0.7 cM. The majority of markers segregated close to Mendelian expectations in the intra-specific crosses. In the two L. saligna x L. sativa inter-specific crosses, a total of 155 and 116 markers in 13 regions exhibited significant segregation distortion. Data visualization tools were developed to curate, display and query the data. The integrated map provides a framework for mapping ESTs in one core mapping population relative to phenotypes that segregate in other populations. It also provides large numbers of markers for marker assisted selection, candidate gene identification, and studies of genome evolution in the Compositae.