Genetic diversity and cultivar variants in the NCGR cranberry (Vaccinium macrocarpon Aiton) collection.

The American cranberry (Vaccinium macrocarpon) is an endemic domesticated species that has become an economically important commercial fruit crop. The USDA-ARS National Clonal Germplasm Repository (NCGR) houses the national Vaccinium collection, which includes representatives of historical cranberry cultivars and wild-selected germplasm. The objective of this study wasto examine the genotypes of 271 cranberry plants from 77 accessions representing 66 named cultivars using 12 simple-sequence repeats to assess clonal purity and cultivar relatedness. Using principal components analysis and neighbour-joining based on estimated genetic distances between individuals, we identified 64 unique genotypes and observed that intracultivar variants (i.e. subclones) existed in the germplasm collection and in the commercial bogs where some accessions originated. Finally, through a comparison of the genotypes of this study with the previous studies, pedigree analysis and the study of the geographic distribution of cranberry diversity, we identified consensus genotypes for many accessions and cultivars. We highlight the important role that the NCGR collection playsfor ex situ conservation of cranberry germplasm for future breeders and researchers. The NCGR continues to search for historically relevant cultivars absent from the collection in an effort to preserve these genotypes before they are lost and no longer commercially grown.

Mining and validation of pyrosequenced simple sequence repeats (SSRs) from American cranberry (Vaccinium macrocarpon Ait.).

The American cranberry (Vaccinium macrocarpon Ait.) is a major commercial fruit crop in North America, but limited genetic resources have been developed for the species. Furthermore, the paucity of codominant DNA markers has hampered the advance of genetic research in cranberry and the Ericaceae family in general. Therefore, we used Roche 454 sequencing technology to perform low-coverage whole genome shotgun sequencing of the cranberry cultivar 'HyRed'. After de novo assembly, the obtained sequence covered 266.3 Mb of the estimated 540-590 Mb in cranberry genome. A total of 107,244 SSR loci were detected with an overall density across the genome of 403 SSR/Mb. The AG repeat was the most frequent motif in cranberry accounting for 35% of all SSRs and together with AAG and AAAT accounted for 46% of all loci discovered. To validate the SSR loci, we designed 96 primer-pairs using contig sequence data containing perfect SSR repeats, and studied the genetic diversity of 25 cranberry genotypes. We identified 48 polymorphic SSR loci with 2-15 alleles per locus for a total of 323 alleles in the 25 cranberry genotypes. Genetic clustering by principal coordinates and genetic structure analyzes confirmed the heterogeneous nature of cranberries. The parentage composition of several hybrid cultivars was evident from the structure analyzes. Whole genome shotgun 454 sequencing was a cost-effective and efficient way to identify numerous SSR repeats in the cranberry sequence for marker development.

Hierarchical classification of switchgrass genotypes using SSR and chloroplast sequences: ecotypes, ploidies, gene pools, and cultivars.

Switchgrass (Panicum virgatum L.) is an important crop for bioenergy feedstock development. Switchgrass has two main ecotypes: the lowland ecotype being exclusively tetraploid (2n = 4x = 36) and the upland ecotype being mainly tetraploid and octaploid (2n = 8x = 72). Because there is a significant difference in ploidy, morphology, growth pattern, and zone of adaptation between and within the upland and lowland ecotypes, it is important to discriminate switchgrass plants belonging to different genetic pools. We used 55 simple sequence repeats (SSR) loci and six chloroplast sequences to identify patterns of variation between and within 18 switchgrass cultivars representing seven lowland and 11 upland cultivars from different geographic regions and of varying ploidy levels. We report consistent discrimination of switchgrass cultivars into ecotype membership and demonstrate unambiguous molecular differentiation among switchgrass ploidy levels using genetic markers. Also, SSR and chloroplast markers identified genetic pools related to the geographic origin of the 18 cultivars with respect to ecotype, ploidy, and geographical, and cultivar sources. SSR loci were highly informative for cultivar fingerprinting and to classify plants of unknown origin. This classification system is the first step toward developing switchgrass complementary gene pools that can be expected to provide a significant heterotic increase in biomass yield.

A consensus linkage map identifies genomic regions controlling fruit maturity and beta-carotene-associated flesh color in melon (Cucumis melo L.).

The nutritional value and yield potential of US Western Shipping melon (USWS; Cucumis melo L.) could be improved through the introgression of genes for early fruit maturity (FM) and the enhancement of the quantity of beta-carotene (QbetaC) in fruit mesocarp (i.e., flesh color). Therefore, a set of 116 F(3) families derived from the monoecious, early FM Chinese line 'Q 3-2-2' (no beta-carotene, white mesocarp) and the andromonoecious, late FM USWS line 'Top Mark' (possessing beta-carotene, orange mesocarp) were examined during 2 years in Wisconsin, USA to identify quantitative trait loci (QTL) associated with FM and QbetaC. A 171-point F(2-3) based map was constructed and used for QTL analysis. Three QTL associated with QbetaC were detected, which explained a significant portion of the observed phenotypic variation (flesh color; R (2) = 4.0-50.0%). The map position of one QTL (beta-carM.E.9.1) was uniformly aligned with one carotenoid-related gene (Orange gene), suggesting its likely role in QbetaC in this melon population and putative relationship with the melon white flesh (wf) gene. Two major (FM.6.1 and FM.11.1; R (2) >or= 20%) and one minor QTL (FM.2.1; R (2) = 8%) were found to be associated with FM. This map was then merged with a previous recombinant inbred line (RIL)-based map used to identify seven QTL associated with QbetaC in melon fruit. This consensus map [300 molecular markers (187 co-dominant melon and 14 interspecific; 10 LG)] provides a framework for the further dissection and cloning of published QTL, which will consequently lead to more effective trait introgression in melon.

Mapping of genetic loci that regulate quantity of beta-carotene in fruit of US Western Shipping melon (Cucumis melo L.).

Melon (Cucumis melo L.) is highly nutritious vegetable species and an important source of beta-carotene (Vitamin A), which is an important nutrient in the human diet. A previously developed set of 81 recombinant inbred lines (RIL) derived from Group Cantalupensis US Western Shipper market type germplasm was examined in two locations [Wisconsin (WI) and California (CA), USA] over 2 years to identify quantitative trait loci (QTL) associated with quantity of beta-carotene (QbetaC) in mature fruit. A moderately saturated 256-point RIL-based map [104 SSR, 7 CAPS, 4 SNP in putative carotenoid candidate genes, 140 dominant markers and one morphological trait (a) spanning 12 linkage groups (LG)] was used for QbetaC-QTL analysis. Eight QTL were detected in this evaluation that were distributed across four LG that explained a significant portion of the associated phenotypic variation for QbetaC (R (2) = 8 to 31.0%). Broad sense heritabilities for QbetaC obtained from RIL grown in WI. and CA were 0.56 and 0.68, respectively, and 0.62 over combined locations. The consistence of QbetaC in high/low RIL within location across years was confirmed in experiments conducted over 2 years. QTL map positions were not uniformly associated with putative carotenoid genes, although one QTL (beta-car6.1) interval was located 10 cM from a beta-carotene hydroxylase gene. These results suggest that accumulation of beta-carotene in melon is under complex genetic control. This study provides the initial step for defining the genetic control of QbetaC in melon leading to the development of varieties with enhanced beta-carotene content.

Isolation and characterization of microsatellite markers for red elm (Ulmus rubra Muhl.) and cross-species amplification with Siberian elm (Ulmus pumila L.).

Ulmus pumila is an elm species, non-native to the USA that hybridizes with Ulmus rubra. In order to study the genetic structure and hybridization patterns between these two elm species, we developed 15 primer pairs for microsatellite loci in U. rubra and tested their cross-amplification in U. pumila. All 15 primers amplified in both species, 11 of which possessed species-specific alleles. Eight loci were polymorphic in U. pumila and eight in U. rubra, each with two to eight alleles per locus. In addition, five primer pairs previously developed in U. laevis and U. carpinifolia (syn. U. minor) cross-amplified and showed polymorphic loci in U. pumila and/or U. rubra. These markers will facilitate the study of genetic structure and gene flow between U. rubra and exotic, invasive U. pumila.

Detection of QTL for yield-related traits using recombinant inbred lines derived from exotic and elite US Western Shipping melon germplasm.

The inheritance of yield-related traits in melon (Cucumis melo L.; 2n = 2x = 24) is poorly understood, and the mapping of quantitative trait loci (QTL) for such traits has not been reported. Therefore, a set of 81 recombinant inbred lines (RIL) was developed from a cross between the monoecious, highly branched line USDA 846-1 and a standard vining, andromonoecious cultivar, 'Top Mark'. The RIL, parental lines, and three control cultivars ('Esteem', 'Sol Dorado', and 'Hales Best Jumbo') were grown at Hancock, WI and El Centro, CA in 2002, and evaluated for primary branch number (PB), fruit number per plant (FN), fruit weight per plant (FW), average weight per fruit (AWF), and percentage of mature fruit per plot (PMF). A 190-point genetic map was constructed using 114 RAPD, 43 SSR, 32 AFLP markers, and one phenotypic trait. Fifteen linkage groups spanned 1,116 cM with a mean marker interval of 5.9 cM. A total of 37 QTL were detected in both locations (PB = 6, FN = 9, FW = 12, AWF = 5, and PMF = 5). QTL analyses revealed four location-independent factors for PB (pb1.1, pb1.2, pb2.3, and pb10.5), five for FN (fn1.1, fn1.2, fn1.3, fn2.4, and fn8.8), four for FW (fw5.8, fw6.10, fw8.11, and fw8.12), two for AWF (awf1.3 and awf8.5), and one for PMF (pmf10.4). The significant (P