The number of Phaseolus vulgaris germplasm accessions numbers more than 30,000. While the large numbers of accessions increase the probability of preserving genetic variability they simultaneously limit the efficient and routine utilization of this resource. From the approximately 4000 P. vulgaris accessions in the C.I.A.T. whole collection that were collected in Mexico, a core collection of 400 accessions was developed based on variation for agronomic performance, ecological adaptation, and seed characteristics. Random samples of 90 accessions each were drawn from the core and whole collections and evaluated for 224 polymorphic RAPD bands. Based on analysis of the RAPD data there were no significant differences in genetic diversity between the two samples. The correlation of marker frequency for the two samples was 0.984 confirming that the two samples represent the same population.
J. Nienhuis, P. Skroch, M. Sass, S. Beebe, J. Tohme and F. Pedraza
G. Jung, J. Nienhuis, S. Hirano, C. Upper, H. Ariyarathne and D.P. Coyne
Bacterial brown spot (BBS), incited by the bacterial pathogen Pseodomonas syringae pv. syringae is important disease of common bean. Phenotypic visual readings of infected areas and a leaf freezing assay estimating the population size of Pss on leaf surface were used for disease assessment for 2 years using 78 RI lines derived from Belneb RR-1 x A55 population grown in Wisconsin. The objectives of this research were to determine the genomic regions of QTL affecting the genetic variation of bacterial brown spot resistance in both assays over 2 years (1996 and 1998) and to determine the size of their genetic effects. In addition, we examined the consistency of detected QTL over environments. Three chromosomal regions associated with QTL for BBS resistance were identified in both assays in 1996 and one chromosomal region was consistently detected over 2 years.
C.L. Boehm, I.K. Lee, G. Jung, H.C. Harrison, J. Nienhuis, M. Sass and Moore Hall
Random amplified polymorphic DNA (RAPD) may have utility as genetic markers facilitating selection in ginseng crop improvement. This experiment determined chemical buffer and root tissue-type combinations that yield repeatable bands. The results allow further experiments using RAPD markers for estimating the genetic distance between ginseng landraces, selection for crop improvement, and extensive fingerprinting for use in determining the origin of tissue samples. This experiment determined mean band yields for all combinations of dry, fresh, and powdered root with cetyltrimethylammonium bromide, potassium/sodium ethyl xanthogenate, and urea buffers. The buffers were applied in replication to the tissue-types with other extraction protocol factors constant. Replications were amplified four times with four different primers using constant PCR and agarose gel electrophoretic protocols. Distinct bands were counted in each replication, and the summation of the replication repeats considered an observation. Least squares means for several response variables were analyzed. The most significant difference found was between buffers. The buffers ctab and urea were productive, and the pex was not. Significant difference was found when buffers were crossed with tissue. The applications of urea to fresh root, ctab to dry root, urea to dry root, and ctab to powdered root were productive. Based on these results we conclude 1) urea and ctab are productive when applied to all tissue-types, 2) dry root, which is easily collected and stored, yields sufficient DNA for analysis, and 3) powdered root, often the form of commercial products that might be tested for genetic origin, will yield sufficient DNA for analysis.
Geunhwa Jung, Paul W. Skroch, Dermot P. Coyne, James Nienhuis, E. Arnaud-Santana, H.M. Ariyarathne, Shawn M. Kaeppler and Mark J. Bassett
Randomly amplified polymorphic DNA (RAPD) molecular markers were used to construct a partial genetic linkage map in a recombinant inbred population derived from the common bean (Phaseolus vulgaris L.) cross PC-50 × XAN-159 for studying the genetics of bacterial disease resistance in common bean. The linkage map spanned 426 cM and included 168 RAPD markers and 2 classical markers with 11 unassigned markers. The seventy recombinant inbred lines were evaluated for resistance to two strains of common bacterial blight [Xanthomonas campestris pv. phaseoli (Smith) Dye] (Xcp). Common bacterial blight (CBB) resistance was evaluated for Xcp strain EK-11 in later-developed trifoliolate leaves and for Xcp strains, DR-7 and EK-11, in first trifoliolate leaves, seeds, and pods. One to four quantitative trait loci (QTLs) accounted for 18% to 53% of the phenotypic variation for traits. Most significant effects for CBB resistance were associated with one chromosomal region on linkage group 5 and with two regions on linkage group 1, of the partial linkage map. The chromosomal region (a 13-cM interval) in linkage group 5 was significantly associated with resistance to Xcp strains DR-7 and EK-11 in leaves, pods, and seeds. The regions in linkage group 1 were also significantly associated with resistance to both Xcp strains in more than one plant organ. In addition, a seedcoat pattern gene (C) and a flower color gene (vlae) were mapped in linkage groups 1 and 5, respectively, of the partial linkage map. The V locus was found to be linked to a QTL with a major effect on CBB resistance.