The magnitude of genetic differences among and the heterogeneity within cultivated and wild American ginseng populations is unknown. Variation among individual plants from 16 geographically separated, cultivated populations and 21 geographically separated, wild populations were evaluated using RAPD markers. Cultivated populations from the midwestern U.S., the southern U.S., and Canada were examined. Wild populations from the midwestern U.S., the southern U.S., and the eastern U.S. were examined. Polymorphic bands were observed for 15 RAPD primers, which resulted in 100 scored bands. Variation was found within and among populations, indicating that the selected populations are heterogeneous with respect to RAPD markers. The genetic relationships among individual genotypes were estimated using the ratio of discordant bands to total bands scored. Multidimensional scaling of the relationship matrix showed independent clusters corresponding to the geographical and cultural origins of the populations. The integrity of the clusters were confirmed using pooled chi-squares for fragment homogeneity. Average gene diversity (Hs) was calculated for each population sample, and a one-way analysis of variance showed significant differences among populations. Overall, the results demonstrate the usefulness of the RAPD procedure for evaluating genetic relationships and comparing levels of genetic diversity among populations of American ginseng genotypes.
C.L. Boehm, H.C. Harrison, G. Jung, and J. Nienhuis
A. Dursun, D.P. Coyne, M.F. Mohamed, and G. Jung
Common bacterial blight, incited by the bacterium Xanthomonas campestris pv. phaseoli (Xcp), is a serious disease of common beans [Phaseolus vulgaris (P. v.)]. Some tepary beans (P. acutifolius) are resistant (R) to Xcp and used to breed P. v. with R to Xcp. The objective was to determine the inheritance of the reaction to different strains of Xcp in crosses between susceptible (S) and R tepary lines. The parents, F2, and F3 populations from six tepary crosses involving 3 R × S, 1 R × moderately (M) R, and 2 R × R were inoculated with Xcp strains EK-11, LB-2, and SC-4A. Different single dominant genes controlled the reaction to different Xcp isolates in R × S crosses. Coupling linkage was detected between the genes controlling the reactions to each of the Xcp strains in the crosses NE #4B(s) × NE #19(R) and NE #4B(S) × CIAT-640005(R), except for NE #8A(MR) × NE #4B(S) with strains EK-11 and LB-2 and EK-11 and SC-4A. Transgressive segregation for S was observed in the F2 and F3 NE #8A × NE #8B(R), indicating that the parents possessed different genes for R. No segregation for reactions occurred n the F2 NE #8B × NE #19 and NE #19 × CIAT-640005, indicating that these parents possessed the same genes for R to the three strains.
S.O. Park, A. Dursun, D.P. Coyne, and G. Jung
Common bacterial blight (CBB), incited by Xanthomonas campestris pv. phaseoli (Xcp), an important disease in common bean (Phaseolus vulgaris L.) Tepary bean (P. acutifolius A. Gray) is of interest to bean breeders because of resistance to CBB. Our objective was to identify RAPD markers linked to major genes for CBB resistance using bulked segregant analysis in an F2 population from a tepary bean cross CIAT640005 (R) X Nebr#4B (S). A total of 57 RAPD primers (602 RAPD primers screened) showed polymorphisms between bulked DNA derived from R and S CBB plants. All markers showed coupling linkage with CBB resistance. A good fit to a 3:1 ratio of bands for presence and absence using 11 RAPD primers was observed in 77 F2 plants. Markers of U-15 and L-7 primers were 2.4 cM distant from the gene for resistance to Xcp strain LB-2. RAPD markers of U-10, U-20, S-12, Y-4, F-13, P-6, Q-1, and Q-ll primers were 2.4 cM distant from the gene for resistance to Xcp strain SC-4A. RAPD markers of IJ-15 and L-7 primers were 8.4 cM distant from the gene for resistance to Xcp strain EKl l. The tepary RAPD linkage group includes three molecular markers and three genes for resistance to Xcp strains EK-l l, LB-2, and SC-4A and spans a length of 19.2 cM. This data supports the presence of Xcp races.
C.L. Boehm, H.C. Harrison, G. Jung, and J. Nienhuis
Genetic differences among eleven cultivated and eight wild-type populations of North American ginseng (Panax quinquefolium L.) and four cultivated populations of South Korean ginseng (P. ginseng C.A. Meyer) were estimated using RAPD markers. Cultivated P. ginseng population samples were collected from four regions of S. Korea. Cultivated P. quinquefolium population samples were collected from three regions in North America: Wisconsin, the Southeastern Appalachian region of the United States, and Canada. Wild-type P. quinquefolium was collected from three states in the United States: Pennsylvania, Tennessee, and Wisconsin. Evaluation of germplasm with 10 decamer primers resulted in 100 polymorphic bands. Genetic differences among populations indicate heterogeneity. The genetic distance among individuals was estimated using the ratio of discordant bands to total bands scored. Multidimensional scaling of the relationship matrix showed independent clusters corresponding to the distinction of species, geographical region, and wild versus cultivated types. The integrity of the clusters was confirmed using pooled chi-square tests for fragment homogeneity.
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. G. Kuo, M. C. H. Jung, and S. C. S. Tsou
The distribution of 14C-photosynthates was examined in pot-grown Tainan-1 mung bean plants (Vigna radiata (L.) Wilczek var. radiata). Whole plants were assimilated with 14CO2 at anthesis, and at 7 and 17 days after anthesis. The 14C-photosynthate fixed at anthesis was retained mostly in the vegetative tissue. However, of the 14C-photosynthate fixed at early pod development stage (i.e. 7 days after anthesis), 15-26% of the assimilated 14C was detected in the reproductive tissue within 24 hours after exposure, whereas about 43% was detected at maturity (i.e. 38 days after anthesis). When plants with full grown pods (i.e. 17 days after anthesis) were treated, 70% of the 14C was detected in the reproductive tissue 24 hours after exposure and at maturity.
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.
H.M. Ariyarathne, D.P. Coyne, G. Jung, P.W. Skroch, A.K. Vidaver, J.R. Steadman, P.N. Miklas, and M.J. Bassett
Diseases of beans (Phaseolus vulgaris L.) are primary constraints affecting bean production. Information on tagging and mapping of genes for disease resistance is expected to be useful to breeders. The objectives of this study were to develop a random amplified polymorphic DNA (RAPD) marker linkage map using 78 F9 recombinant inbred (RI) lines derived from a Middle-American common bean cross Great Northern Belneb RR-1 [resistant to common bacterial blight (CBB) and halo blight (HB)] × black A 55 [dominant I gene resistance to bean common mosaic potyvirus] and to map genes or QTL (quantitative trait loci) for resistance to CBB, HB, BCMV (bean common mosaic virus), and BCMNV (bean common mosaic necrosis virus) diseases. The RI lines were evaluated for resistance to leaf and pod reactions to Xanthomonas campestris pv. phaseoli (Xcp) (Smith Dye) strain EK-11, leaf reactions to two Pseudomonas syringae pv. phaseolicola (Psp) (Burkholder) Young et al. (1978) strains HB16 and 83-Sc2A, and BCMV strain US-5 and BCMNV strain NL-3. The linkage map spanned 755 cM, including 90 markers consisting of 87 RAPD markers, one sequence characterized amplified region (SCAR), the I gene, and a gene for hypersensitive resistance to HB 83-Sc2A. These were grouped into 11 linkage groups (LG) corresponding to the 11 linkage groups in the common bean integrated genetic map. A major gene and QTL for leaf resistance to HB were mapped for the first time. Three QTL for leaf reactions to HB16 were found on linkage groups 3, 5, and 10. Four regions on linkage groups 2, 4, 5, and 9, were significantly associated with leaf reactions to HB strain 83-Sc2A. The gene controlling the hypersensitive reaction to HB 83-Sc2A mapped to the same region as the QTL on LG 4. The I locus for resistance to BCMV and BCMNV was mapped to LG 2 at about 1.4 cM from RAPD marker A10.1750. Five and four markers were significantly associated with QTL for resistance to CBB in leaves and pods, respectively, with four of them associated with resistance in both plant organs. A marker locus was discovered on LG 10, W10.550, which could account for 44% and 41% of the phenotypic variation for CBB resistance in leaves and pods, respectively. QTL for resistance in pod to CBB, leaf resistance to HB, and the I gene were linked on LG 2.