The density and utility of the molecular genetic linkage map of the widespread use of RFLP markers and maps in suncultivated sunflower (Helianthus annuus L.) has been greatly inflower has been restricted by a lack of public RFLP creased by the development and mapping of several hundred simple sequence repeat (SSR) markers. Of 1089 public SSR markers de-probes, consequent lack of a dense public RFLP map, scribed thus far, 408 have been mapped in a recombinant inbred line and low-throughput nature of RFLP markers. The diffi-(RIL) mapping population (RHA280 ϫ RHA801). The goal of the culties posed by the historic lack of public, single-copy present research was to increase the density of the sunflower map by DNA markers were only weakly offset by the emerconstructing a new RIL map (PHA ϫ PHB) based on SSRs, adding gence of facile, universal DNA markers, e.g., RAPDs loci for newly developed SSR markers to the RHA280 ϫ RHA801 RIL (Williams et al., 1990, 1993) and AFLPs (Vos et al., map, and integrating the restriction fragment length polymorphism 1995). RAPDs have primarily been used for tagging (RFLP) and SSR maps of sunflower. The latter was accomplished by phenotypic loci in sunflower, e.g., rust (Puccinia helianadding 120 SSR marker loci to a backbone of 80 RFLP marker loci thi Schw.) and Orobanche cumana Wallr. resistance on the HA370 ϫ HA372 F 2 map. The map spanned 1275.4 centimorgans (cM) and had a mean density of 6.3 cM per locus. The genes (Lawson et al., 1998; Lu et al., 2000). While RAPD PHA ϫ PHB SSR map was constructed from 264 SSR marker loci, and AFLP markers have a multitude of uses, both are spanned 1199.4 cM, and had a mean density of 4.5 cM per locus. The dominant, multicopy, and often nonspecific in nature RHA280 ϫ RHA801 map was constructed by adding 118 new SSR and, as a whole, unsatisfactory for establishing a geand insertion-deletion (INDEL) marker loci to 459 previously nome-wide framework of DNA markers for anchoring mapped SSR marker loci. The 577-locus map spanned 1423.0 cM and cross referencing genetic linkage maps. Single-copy, and had a mean density of 2.5 cM per locus. The three maps were codominant DNA markers, e.g., SSRs, are preferred for constructed from 1044 DNA marker loci (701 unique SSR and 89 such purposes and, until recently, have been lacking unique RFLP or INDEL marker loci) and supply a dense genomein sunflower. wide framework of sequence-based DNA markers for molecular breeding and genomics research in sunflower.
Comparative genetic linkage maps provide a powerful tool for the study of karyotypic evolution. We constructed a joint SSR/RAPD genetic linkage map of the Helianthus petiolaris genome and used it, along with an integrated SSR genetic linkage map derived from four independent H. annuus mapping populations, to examine the evolution of genome structure between these two annual sunflower species. The results of this work indicate the presence of 27 colinear segments resulting from a minimum of eight translocations and three inversions. These 11 rearrangements are more than previously suspected on the basis of either cytological or genetic map-based analyses. Taken together, these rearrangements required a minimum of 20 chromosomal breakages/fusions. On the basis of estimates of the time since divergence of these two species (750,000-1,000,000 years), this translates into an estimated rate of 5.5-7.3 chromosomal rearrangements per million years of evolution, the highest rate reported for any taxonomic group to date.
Wild populations of common sunflower (Helianthus annuus L.) are self-incompatible and have deep seed dormancy, whereas modern cultivars, inbreds, and hybrids are self-compatible and partially-to-strongly self-pollinated, and have shallow seed dormancy. Self-pollination (SP) and seed dormancy are genetically complex traits, the number of self-compatibility (S) loci has been disputed, and none of the putative S loci have been genetically mapped in sunflower. We genetically mapped quantitative trait loci (QTL) for self-incompatibility (SI), SP, and seed dormancy in a backcross population produced from a cross between an elite, self-pollinated, nondormant inbred line (NMS373) and a wild, self-incompatible, dormant population (ANN1811). A population consisting of 212 BC(1) progeny was subsequently produced by backcrossing a single hybrid individual to NMS373. BC(1) progeny produced 0-838 seeds per primary capitula when naturally selfed and 0-518 seeds per secondary capitula when manually selfed and segregated for a single S locus. The S locus mapped to linkage group 17 and was tightly linked to a cluster of previously identified QTL for several domestication and postdomestication traits. Two synergistically interacting QTL were identified for SP among self-compatible (ss) BC(1) progeny (R(2)=34.6%). NMS373 homozygotes produced 271.5 more seeds per secondary capitulum than heterozygotes. Germination percentages of seeds after-ripened for 4 weeks ranged from 0% to 100% among self-compatible BC(1)S(1) families. Three QTL for seed dormancy were identified (R(2)=38.3%). QTL effects were in the predicted direction (wild alleles decreased self-pollination and seed germination). The present analysis differentiated between loci governing SI and SP and identified DNA markers for bypassing SI and seed dormancy in elite x wild crosses through marker-assisted selection.
Three-fourths of the recognition-dependent disease resistance genes (R-genes) identified in plants encode nucleotide binding site (NBS) leucine-rich repeat (LRR) proteins. NBS-LRR homologs have only been isolated on a limited scale from sunflower (Helianthus annuus L.), and most of the previously identified homologs are members of two large NBS-LRR clusters harboring downy mildew R-genes. We mined the sunflower EST database and used comparative genomics approaches to develop a deeper understanding of the diversity and distribution of NBS-LRR homologs in the sunflower genome. Collectively, 630 NBS-LRR homologs were identified, 88 by mining a database of 284,241 sunflower ESTs and 542 by sequencing 1,248 genomic DNA amplicons isolated from common and wild sunflower species. DNA markers were developed from 196 unique NBS-LRR sequences and facilitated genetic mapping of 167 NBS-LRR loci. The latter were distributed throughout the sunflower genome in 44 clusters or singletons. Wild species ESTs were a particularly rich source of novel NBS-LRR homologs, many of which were tightly linked to previously mapped downy mildew, rust, and broomrape R-genes. The DNA sequence and mapping resources described here should facilitate the discovery and isolation of recognition-dependent R-genes guarding sunflower from a broad spectrum of economically important diseases. Sunflower nucleotide and amino acid sequences have been deposited in DDBJ/EMBL/GenBank under accession numbers EF 560168-EF 559378 and ABQ 58077-ABQ 57529.
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