The genomes of most, if not all, flowering plants have undergone whole genome duplication events during their evolution. The impact of such polyploidy events is poorly understood, as is the fate of most duplicated genes. We sequenced an approximately 1 million-bp region in soybean (Glycine max) centered on the Rpg1-b disease resistance gene and compared this region with a region duplicated 10 to 14 million years ago. These two regions were also compared with homologous regions in several related legume species (a second soybean genotype, Glycine tomentella, Phaseolus vulgaris, and Medicago truncatula), which enabled us to determine how each of the duplicated regions (homoeologues) in soybean has changed following polyploidy. The biggest change was in retroelement content, with homoeologue 2 having expanded to 3-fold the size of homoeologue 1. Despite this accumulation of retroelements, over 77% of the duplicated low-copy genes have been retained in the same order and appear to be functional. This finding contrasts with recent analyses of the maize (Zea mays) genome, in which only about one-third of duplicated genes appear to have been retained over a similar time period. Fluorescent in situ hybridization revealed that the homoeologue 2 region is located very near a centromere. Thus, pericentromeric localization, per se, does not result in a high rate of gene inactivation, despite greatly accelerated retrotransposon accumulation. In contrast to low-copy genes, nucleotide-binding-leucine-rich repeat disease resistance gene clusters have undergone dramatic species/homoeologuespecific duplications and losses, with some evidence for partitioning of subfamilies between homoeologues.The comparative approach to studying genes and genomes is a powerful method for addressing both
Resistance to grapevine powdery mildew is controlled by Run1, a single dominant gene present in the wild grapevine species, Muscadinia rotundifolia, but absent from the cultivated species, Vitis vinifera. Run1 has been introgressed into V. vinifera using a pseudo-backcross strategy, and genetic markers have previously been identified that are linked to the resistance locus. Here we describe the construction of comprehensive genetic and physical maps spanning the resistance locus that will enable future positional cloning of the resistance gene. Physical mapping was performed using a bacterial artificial chromosome (BAC) library constructed using genomic DNA extracted from a resistant V. vinifera individual carrying Run1 within an introgression. BAC contig assembly has enabled 20 new genetic markers to be identified that are closely linked to Run1, and the position of the resistance locus has been refined, locating the gene between the simple sequence repeat (SSR) marker, VMC4f3.1, and the BAC end sequence-derived marker, CB292.294. This region contains two multigene families of resistance gene analogues (RGA). A comparison of physical and genetic mapping data indicates that recombination is severely repressed in the vicinity of Run1, possibly due to divergent sequence contained within the introgressed fragment from M. rotundifolia that carries the Run1 gene.
Ascochyta blight, caused by Ascochyta rabiei (Pass.) Lab., is a devastating disease of chickpea (Cicer arietinum L.) worldwide. Resistant germplasm has been identified and the genetics of resistance has been the subject of numerous studies. The objectives of the present study were to determine the genetics of resistance to ascochyta blight of chickpea and to map and tag the chromosomal regions involved using molecular markers. We used a set of 142 F5:6 recombinant inbred lines (RILs) obtained from an interspecific cross of C. arietinum (FLIP84‐92C, resistant parent) × C. reticulatum Lad. (PI 599072, susceptible parent). The RILs were scored for disease reactions in the field over 2 yr and were genotyped for polymorphic molecular markers [isozyme, random amplified polymorphic DNA (RAPD), and inter simple sequence repeat (ISSR)] in the laboratory. The disease was scored quantitatively and data were used for QTL analysis. A linkage map was established that comprised nine linkage groups containing 116 markers covering a map distance of 981.6 centimorgans (cM) with an average distance of 8.4 cM between markers. Two quantitative trait loci (QTLs), QTL‐1 and QTL‐2, conferring resistance to ascochyta blight, were identified which accounted for 50.3 and 45.0% of the estimated phenotypic variation in 1997 and 1998, respectively, and were mapped to linkage groups 6 and 1, respectively. Two RAPD markers flanked QTL‐1 and were 10.9 cM apart while one ISSR marker and an isozyme marker flanked QTL‐2 and were 5.9 cM apart. These markers can be used for marker‐assisted selection for ascochyta blight resistance in chickpea breeding programs, and to develop durable resistant cultivars through gene pyramiding.
The inheritance of an inter-simple-sequence-repeat (ISSR) polymorphism was studied in a cross of cultivated chickpea (Cicer arietinum L.) and a closely related wild species (C. reticulatum Lad.) using primers that anneal to a simple repeat of various lengths, sequences and non-repetitive motifs. Dinucleotides were the majority of those tested, and provided all of the useful banding patterns. The ISSR loci showed virtually complete agreement with expected Mendelian ratios. Twenty two primers were used for analysis and yielded a total of 31 segregating loci. Primers based on (GA)n repeats were the most abundant while primers with a (TG)n repeat gave the largest number of polymorphic loci. Nucleotides at the 5' and 3' end of the primers played an important role in detecting polymorphism. All the markers showed dominance. We found an ISSR marker linked to the gene for resistance to fusarium wilt race 4. The marker concerned, UBC-855500, was found to be linked in repulsion with the fusarium wilt resistance gene at a distance of 5.2 cM. It co-segregated with CS-27700, a RAPD marker previously shown to be linked to the gene for resistance to fusarium wilt race 1, and was mapped to linkage group 6 of the Cicer genome. This indicated that genes for resistance to fusarium wilt races 1 and 4 are closely linked. The marker UBC-855500 is located 0.6 cM from CS-27700 and is present on the same side of the wilt resistance gene. To our knowledge this is the first report of the utility of an ISSR marker in gene tagging. These markers may provide valuable information for the development of sequence-tagged microsatellite sites (STMS) at a desired locus.
Randomly amplified polymorphic DNA (RAPD) markers were used for the identification of pigeonpea [Cajanus cajan (L.) Millsp.] cultivars and their related wild species. The use of single primers of arbitrary nucleotide sequence resulted in the selective amplification of DNA fragments that were unique to individual accessions. The level of polymorphism among the wild species was extremely high, while little polymorphism was detected within Cajanus cajan accessions. All of the cultivars and wild species under study could be easily distinguished with the help of different primers, thereby indicating the immense potential of RAPD in the genetic fingerprinting of pigeonpea. On the basis of our data the genetic relationship between pigeonpea cultivars and its wild species could be established.
Summary
Plant genomes contain numerous disease resistance genes (R genes) that play roles in defense against pathogens. Scarcity of genetic polymorphism makes peanut (Arachis hypogaea) especially vulnerable to a wide variety of pathogens.
Here, we isolated and characterized peanut bacterial artificial chromosomes (BACs) containing a high density of R genes. Analysis of two genomic regions identified several TIR‐NBS‐LRR (Toll‐interleukin‐1 receptor, nucleotide‐binding site, leucine‐rich repeat) resistance gene analogs or gene fragments. We reconstructed their evolutionary history characterized by tandem duplications, possibly facilitated by transposon activities. We found evidence of both intergenic and intragenic gene conversions and unequal crossing‐over, which may be driving forces underlying the functional evolution of resistance.
Analysis of the sequence mutations, protein secondary structure and three‐dimensional structures, all suggest that LRR domains are the primary contributor to the evolution of resistance genes. The central part of LRR regions, assumed to serve as the active core, may play a key role in the resistance function by having higher rates of duplication and DNA conversion than neighboring regions. The assumed active core is characterized by significantly enriched leucine residue composition, accumulation of positively selected sites, and shorter beta sheets.
Homologous resistance gene analog (RGA)‐containing regions in peanut, soybean, Medicago, Arabidopsis and grape have only limited gene synteny and microcollinearity.
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