BackgroundThe potato genome sequence derived from the Solanum tuberosum Group Phureja clone DM1-3 516 R44 provides unparalleled insight into the genome composition and organisation of this important crop. A key class of genes that comprises the vast majority of plant resistance (R) genes contains a nucleotide-binding and leucine-rich repeat domain, and is collectively known as NB-LRRs.ResultsAs part of an effort to accelerate the process of functional R gene isolation, we performed an amino acid motif based search of the annotated potato genome and identified 438 NB-LRR type genes among the ~39,000 potato gene models. Of the predicted genes, 77 contain an N-terminal toll/interleukin 1 receptor (TIR)-like domain, and 107 of the remaining 361 non-TIR genes contain an N-terminal coiled-coil (CC) domain. Physical map positions were established for 370 predicted NB-LRR genes across all 12 potato chromosomes. The majority of NB-LRRs are physically organised within 63 identified clusters, of which 50 are homogeneous in that they contain NB-LRRs derived from a recent common ancestor.ConclusionsBy establishing the phylogenetic and positional relationship of potato NB-LRRs, our analysis offers significant insight into the evolution of potato R genes. Furthermore, the data provide a blueprint for future efforts to identify and more rapidly clone functional NB-LRR genes from Solanum species.

Genomewide association studies depend on the extent of linkage disequilibrium (LD), the number and distribution of markers, and the underlying structure in populations under study. Outbreeding species generally exhibit limited LD, and consequently, a very large number of markers are required for effective whole-genome association genetic scans. In contrast, several of the world's major food crops are self-fertilizing inbreeding species with narrow genetic bases and theoretically extensive LD. Together these are predicted to result in a combination of low resolution and a high frequency of spurious associations in LD-based studies. However, inbred elite plant varieties represent a unique human-induced pseudooutbreeding population that has been subjected to strong selection for advantageous alleles. By assaying 1,524 genomewide SNPs we demonstrate that, after accounting for population substructure, the level of LD exhibited in elite northwest European barley, a typical inbred cereal crop, can be effectively exploited to map traits by using whole-genome association scans with several hundred to thousands of biallelic SNPs.barley ͉ linkage disequilibrium ͉ oligo pool assay L inkage disequilibrium (LD), the nonrandom association of alleles at distinct loci in a sample population, is now routinely exploited to map disease genes in humans (1-4). In crop plants, the potential of exploiting LD in population-based association mapping, with the objective of estimating the position of a gene conferring a specific trait or phenotype by using LD between alleles of genetically mapped markers, has become a focus of considerable interest. A major attraction of association mapping is the potential to locate genes responsible for a wide range of traits in sample populations by using preexisting phenotypic data collected during crop improvement and cultivar registration programs. In addition, as association mapping exploits all historical recombination events that have occurred during establishment of the sample population (2, 5, 6) mapping resolution may greatly exceed that possible in small biparental experimental crosses.As spurious associations between phenotypes and marker loci can be caused by population structure, the extent and structure of LD within a sample population must be known before selecting an appropriate association mapping strategy (1). This is particularly true when looking at an inbreeding crop species such as barley where such complicating factors are expected to be more prevalent. In maize, a naturally outcrossing species, LD decays rapidly over 1-2 kb (7) and in diverse populations of Arabidopsis, an inbreeding species, LD extends to Ϸ1 cM (250 kb), although isolated populations exhibit LD Ͼ50 cM (8). Barley in addition to being an inbreeder has, as well as other crop plants, undergone a severe population bottleneck during domestication (9). In addition, the progenitors of contemporary European spring and winter barley varieties were selected from a relatively small number of successful European landraces d...

The domestication of cereals has involved common changes in morphological features, such as seed size, seed retention and modification of vegetative and inflorescence architecture that ultimately contributed to an increase in harvested yield. In barley, this process has resulted in two different cultivated types, two-rowed and six-rowed forms, both derived from the wild two-rowed ancestor, with archaeo-botanical evidence indicating the origin of six-rowed barley early in the domestication of the species, some 8,600-8,000 years ago. Variation at SIX-ROWED SPIKE 1 (VRS1) is sufficient to control this phenotype. However, phenotypes imposed by VRS1 alleles are modified by alleles at the INTERMEDIUM-C (INT-C) locus. Here we show that INT-C is an ortholog of the maize domestication gene TEOSINTE BRANCHED 1 (TB1) and identify 17 coding mutations in barley TB1 correlated with lateral spikelet fertility phenotypes.

Breeding in raspberry is time-consuming due to the highly heterozygous nature of this perennial fruit crop, coupled with relatively long periods of juvenility. The speed and precision of raspberry breeding can be improved by genetic linkage maps, thus facilitating the development of diagnostic markers for polygenic traits and the identification of genes controlling complex phenotypes. A genetic linkage map (789 cM) of the red raspberry Rubus idaeus has been constructed from a cross between two phenotypically different cultivars; the recent European cultivar Glen Moy and the older North American cultivar Latham. SSR markers were developed from both genomic and cDNA libraries from Glen Moy. These SSRs, together with AFLP markers, were utilised to create a linkage map. In order to test the utility of the genetic linkage map for QTL analysis, morphological data based on easily scoreable phenotypic traits were collected. The segregation of cane spininess, and the root sucker traits of density and spread from the mother plant, was quantified in two different environments. These traits were analysed for significant linkages to mapped markers using MapQTL and were found to be located on linkage group 2 for spines and group 8 for density and diameter. The availability of co-dominant markers allowed heterozygosities to be calculated for both cultivars.

Fluorescence recovery after photobleaching (FRAP) was used to study the mechanism by which fluorescent-protein-tagged movement protein (MP) of tobacco mosaic virus (TMV) is targeted to plasmodesmata (PD). The data show that fluorescence recovery in PD at the leading edge of an infection requires elements of the cortical actin/ endoplasmic reticulum (ER) network and can occur in the absence of an intact microtubule (MT) cytoskeleton. Inhibitors of the actin cytoskeleton (latrunculin and cytochalasin) significantly inhibited MP targeting, while MT inhibitors (colchicine and oryzalin) did not. Application of sodium azide to infected cells implicated an active component of MP transfer to PD. Treatment of cells with Brefeldin A (BFA) at a concentration that caused reabsorption of the Golgi bodies into the ER (precluding secretion of viral MP) had no effect on MP targeting, while disruption of the cortical ER with higher concentrations of BFA caused significant inhibition. Our results support a model of TMV MP function in which targeting of MP to PD during infection is mediated by the actin/ER network.Key words: actin, endoplasmic reticulum, microtubule, plasmodesmata, targeting, tobacco mosaic virus Tobacco mosaic virus (TMV), type member of the tobamoviruses, is one of the most extensively studied viral pathogens of plants. Despite over 100 years of research into the interaction of TMV with its hosts, the mechanism by which TMV moves between plant cells remains uncertain (1-6). However, it has been speculated that during coevolution with host plants, viruses 'hijacked' one or more host proteins for trafficking molecules through plasmodesmata (PD). As a result, studies of the interaction of TMV with host plants are of particular relevance because they form the basis for models of macromolecular trafficking. TMV, in common with a wide range of viruses, encodes a viral movement protein (MP) that is involved in a number of interrelated functions. These include targeting and accumulation within PD (7-10), an increase in the size exclusion limit (SEL) of the PD pore (9,11,12) and the ability to bind to single-stranded RNA (13). Although the MP accumulates within PD (14), and increases the SEL of the PD pore in transgenic plants (11), it appears that during a natural viral infection, the 'gating' of PD is transitory and is restricted to the leading edge of infection (9), while the loss of gating function possibly involves the phosphorylation of the MP within the PD pore (15).In recent years, considerable attention has focused on the mechanism by which the TMV MP is targeted to PD, although data remain conflicting as to the precise intracellular pathway used by MP to reach the PD pore. Over the past decade, a series of reports have shown the association of the TMV MP with elements of the cytoskeleton, including actin (16), but more predominantly, microtubules (MTs) (17-23), although the latter is under spatiotemporal control, being more pronounced several cell layers behind the leading edge of the infection. In keeping with...

To examine the performance and information content of different marker systems, comparative assessment of population genetic diversity was undertaken in nine populations of Athyrium distentifolium using nine genomic and 10 expressed sequence tag (EST) microsatellite (SSR) loci, and 265 amplified fragment length polymorphism (AFLP) loci from two primer combinations. In range-wide comparisons (European vs. North American populations), the EST-SSR loci showed more reliable amplification and produced more easily scorable bands than genomic simple sequence repeats (SSRs). Genomic SSRs showed significantly higher levels of allelic diversity than EST-SSRs, but there was a significant correlation in the rank order of population diversities revealed by both marker types. When AFLPs, genomic SSRs, and EST-SSRs are considered, comparisons of different population diversity metrics/markers revealed a mixture of significant and nonsignificant rank-order correlations. However, no hard incongruence was detected (in no pairwise comparison of populations did different marker systems or metrics detect opposingly significant different amounts of variation). Comparable population pairwise estimates of F(ST) were obtained for all marker types, but whilst absolute values for genomic and EST-SSRs were very similar (F(ST) = 0.355 and 0.342, respectively), differentiation was consistently higher for AFLPs in pairwise and global comparisons (global AFLP F(ST) = 0.496). The two AFLP primer combinations outperformed 18 SSR loci in assignment tests and discriminatory power in phenetic cluster analyses. The results from marker comparisons on A. distentifolium are discussed in the context of the few other studies on natural plant populations comparing microsatellite and AFLP variability.

Disease resistance is increasing in importance, as consumers require high-quality raspberry fruit at a time when chemical disease control is undesirable. Breeders have limited resources and rarely include a primary screen for each fungal disease. Marker-assisted breeding would facilitate the introduction of resistance into elite germplasm and breeding lines. An additional 20 simple sequence repeats have been added to the existing raspberry linkage map. Gene H, which determines cane pubescence (genotype HH or Hh), the recessive allele of which gives glabrous canes (genotype hh), has been mapped on to linkage group 2 and shown to be closely associated with resistance to cane botrytis and spur blight but not rust or cane spot. Other map regions on linkage groups 3, 5 and 6 associated with disease resistance are reported here.

Genome-wide association studies (GWAS) have become a powerful tool for analyzing complex traits in crop plants. The current study evaluates the efficacy of various GWAS models and methods for elucidating population structure in potato. The presence of significant population structure can lead to detection of spurious marker-trait associations, as well as mask true ones. While appropriate statistical models are needed to detect true marker-trait associations, in most published potato GWAS, a ‘one model fits all traits’ approach has been adopted. We have examined various GWAS models on a large association panel comprising diverse tetraploid potato cultivars and breeding lines, genotyped with single nucleotide polymorphism (SNP) markers. Phenotypic data were generated for 20 quantitative traits assessed in different environments. Best Linear Unbiased Estimates (BLUEs) for these traits were obtained for use in assessing GWAS models. Goodness of fit of GWAS models, derived using different combinations of kinship and population structure for all traits, was evaluated using Quantile-Quantile (Q-Q) plots and genomic control inflation factors (λGC). Kinship was found to play a major role in correcting population confounding effects and results advocate a ‘trait-specific’ fit of different GWAS models. A survey of genome-wide linkage disequilibrium (LD), one of the critical factors affecting GWAS, is also presented and our findings are compared to other recent studies in potato. The genetic material used here, and the outputs of this study represent a novel resource for genetic analysis in potato.