Aphanomyces root rot of pea (Pisum sativum) in many pea-growing regions. The genetic resistance to this fungal pathogen is quantitatively inherited and confers levels of tolerance to the disease. Genetic gains in selection have been hampered by the difficulty of differentiating the highly tolerant from tolerant lines in segregating populations. Reporter gene systems have been useful in studying genetic resistance to other soil-borne pathogens. We have transformed an isolate of Aphanomyces euteiches, the causal pathogen, with a reporter gene β-glucuronidase (GUS) and a selectable marker gene, hygromycin phosphotransferase or neomycin phosphotransferase. The transformed lines constitutively express GUS as determined fluorimetrically by measuring the conversion of 4-methylumbelliferyl glucuronide to 4-methlyumbelliferone. The efficacy of this GUS enzyme assay will be compared with an indirect enzyme linked immunosorbant assay (ELISA) and visual disease development ratings in inoculated seedlings of three populations recombinant inbred lines of pea segregating for tolerance.
Clarice J. Coyne and Fred J. Muehlbauer
Clarice J. Coyne, Shawn A. Mehlenbacher, and David C. Smith
Eastern filbert blight is an economically significant disease in European hazelnut (Corylus avellana L.) production in the United States. Since genetic resistance is the only viable disease control strategy to this fungal disease caused by Anisogramma anomala (Peck) E. Müller, greenhouse and field screening of germplasm was undertaken to study the inheritance from known resistant sources and to identify new sources for inclusion in the breeding program. We confirmed that `Gasaway' resistance to this disease is conferred by a single dominant gene. No major gen was identified in the field-resistant cultivar Gem. Representatives of six Corylus species were screened to identify new resistant germplasm. Corylus cornuta Marshall var. cornuta, C. cornuta var. californica (A.DC.) Sharp, C. heterophylla Fischer, and C. sieboldiana Blume were highly resistant, as were most C. americana Marshall genotypes and one C. colurna L. clone tested, but C. jacquenontii Decaisne was highly susceptible. In several cases, hybrids of these species with susceptible C. avellana were also resistant. These new sources of resisstance are being incorporated in the resistance breeding effort.
Melissa T. McClendon, Debra A. Inglis, Kevin E. McPhee, and Clarice J. Coyne
Dry pea (Pisum sativum L.) production in many areas of the world may be severely diminished by soil inhabiting pathogens such as Fusarium oxysporum f. sp. pisi race 1, the causal organism of fusarium wilt race 1. Our objective was to identify closely linked marker(s) to the fusarium wilt race 1 resistance gene (Fw) that could be used for marker assisted selection in applied pea breeding programs. Eighty recombinant inbred lines (RILs) from the cross of Green Arrow (resistant) and PI 179449 (susceptible) were developed through single-seed descent, and screened for disease reaction in race 1 infested field soil and the greenhouse using single-isolate inoculum. The RILs segregated 38 resistant and 42 susceptible fitting the expected 1:1 segregation ratio for a single dominant gene (χ2 = 0.200). Bulk segregant analysis (BSA) was used to screen 64 amplified fragment length polymorphism (AFLP) primer pairs and previously mapped random amplified polymorphic DNA (RAPD) primers to identify candidate markers. Eight AFLP primer pairs and 15 RAPD primers were used to screen the RIL mapping population and generate a linkage map. One AFLP marker, ACG:CAT_222, was within 1.4 cM of the Fw gene. Two other markers, AFLP marker ACC:CTG_159 at 2.6 cM linked to the susceptible allele, and RAPD marker Y15_1050 at 4.6 cM linked to the resistant allele, were also identified. The probability of correctly identifying resistant lines to fusarium wilt race 1, with DNA marker ACG:CAT_222, is 96% percent. These markers will be useful for marker assisted breeding in applied pea breeding programs.
Clarice J. Coyne, Shawn A. Mehlenbacher, Kenneth B. Johnson, John N. Pinkerton, and David C. Smith
A rapid and reliable assay for screening European hazelnut (Corylus avellana L.) genotypes for quantitative resistance to eastern filbert blight [Anisogramma anomala (Peck) E. Müller] was tested by comparing two methods using the same clones. In the first assay, disease spread was followed for five consecutive years (1992-96) in a field plot planted in 1990. Measured responses included disease incidence (the presence or absence of cankers) and total canker length, quantified as the length of perennially expanding cankers. The second assay consisted of annually exposing replicated sets of 2-year-old, potted trees to artificially high doses of pathogen inoculum and measuring incidence and canker lengths at the end of the next growing season. The potted trees were exposed to inoculum in 1990, 1992, 1993, and 1994. Compared to the field plot, disease incidence and total canker length were higher in all the potted-tree experiments. Nonetheless, disease responses of individual clones in the two screening methods were significantly correlated in some contrasts (rs = 0.97 between 1996 field and 1995 potted trees). However, for a few clones (`Camponica', `Tombul Ghiaghli', and `Tonda di Giffoni'), disease developed slowly in the field plot, but disease incidence on these clones averaged > 30% in most of the potted-tree studies. Disease responses also were significantly correlated among some of the potted-tree experiments (rs = 0.72 for the comparison of 1994 to 1995). Highly susceptible and highly resistant hazelnut clones were identified by both methods. However, the field plot method was superior to the potted-tree method for distinguishing among moderately resistant clones. `Bulgaria XI-8', `Gem', `Camponica', `Tombul Ghiaghli', and `Tonda di Giffoni' were identified as promising sources of quantitative resistance to eastern filbert blight.