Macrophomina phaseolina is the cause of charcoal rot of soybean (Glycine max). Resistance to M. phaseolina in commercial soybean cultivars is not common but is needed in locations where the disease is chronic and severe. The objective of this study was to develop a reliable method to produce sufficient M. phaseolina conidia that can be used to inoculate soybean plants in a highthroughput resistance-screening program. Production of pycnidia is not common on most culture media, such as potato dextrose agar, but can be produced on media containing plant parts or oilseed extracts. Seven semi-defined media were tested to induce pycnidia production. Results indicated that the number of pycnidia that were produced by eight M. phaseolina isolates was dependent on induction medium; however, peanut butter extract-saturated filter paper placed over soynut butter extract agar (PESEA) allowed for greater pycnidia and conidia production than the other media tested. Production of pycnidia on PESEA ranged from 269 to 1,082 per plate. There were no differences among isolates in germination of conidia produced on PESEA, which averaged 83 ± 2% germination. A conidial suspension from one M. phaseolina isolate produced on PESEA and inoculated onto soybean radicles significantly distinguished (P < 0.01) ‘DT97-4290’, a soybean genotype with partial resistance to charcoal rot, from a susceptible genotype, ‘LS98-0358’. Results of this study indicated that multiple isolates of M. phaseolina from soybean produced sufficient amounts of conidia on PESEA to use as inoculum. This conidia inoculum production method will facilitate soybean charcoal rot resistance screening evaluation with different soybean isolates.

Charcoal rot, caused by the fungal pathogen Macrophomina phaseolina, is an economically important disease on soybean and other crops including maize (Zea mays L.), sorghum [Sorghum bicolor (L.) Moench], and sunflower (Helianthus annuus L.). Without effective cultural or chemical options to control charcoal rot in soybean [Glycine max (L.) Merr.], finding sources of genetic resistance is of high interest. In this study, 70 ancestral soybean genotypes were screened for resistance to M. phaseolina using a cut-stem inoculation technique under semi-controlled greenhouse conditions. Lesion progression on the stems in the first experiment was measured 7 to 15 d after inoculation. Three follow-up experiments were conducted to select and confirm the genotypes with the strongest partial resistance. Two experiments evaluated lesion lengths and the third experiment evaluated seedling survival. In the two experiments measuring lesion lengths, PI 548302 (42 and 38 mm) and PI 548414 (36 and 52 mm) had significantly shorter lesion lengths than the moderately resistant genotype, DT97-4290 (58 and 87 mm). In the fourth experiment, percentage survival of PI 548414 (88%), PI 548302 (81%), and PI 548178 (66%) were significantly higher than survival of DT97-4290 (32%). These three genotypes may be useful as parents for developing soybean cultivars with charcoal rot resistance.

Charcoal rot of soybean is caused by the fungal pathogen Macrophomina phaseolina. Effective and reliable techniques to evaluate soybean for resistance to this fungus are needed to work toward a management scheme that would utilize host resistance. Three experiments were conducted to investigate the use of a cut-stem inoculation technique to evaluate soybean genotypes for resistance to M. phaseolina. The first experiment compared aggressiveness of M. phaseolina isolates collected from soybean on different soybean genotypes. Significant (P < 0.05) differences among the isolates and genotypes for relative area under disease progress curve (RAUDPC) were found without a significant isolate–genotype interaction. The second experiment compared 14 soybean genotypes inoculated with M. phaseolina in multiple trials conducted in two environments, the greenhouse and growth chamber. Significant (P < 0.05) differences among environments and highly significant (P < 0.001) differences among soybean genotypes for RAUDPC were found. The environment–genotype interaction was nonsignificant (P > 0.05). Soybean genotypes DT97-4290, DT98- 7553, DT98-17554, and DT99-16864 had significantly (P < 0.05) lower RAUDPC than 7 of the 14 genotypes. The third experiment evaluated resistance in selected Phaseolus spp. and soybean genotypes. The range of RAUDPC for Phaseolus spp. was similar to that of soy- bean. The Phaseolus lunatus ‘Bush Baby Lima’ had significantly (P < 0.05) lower RAUDPC than P. vulgaris genotypes evaluated. The cut-stem inoculation technique, which has several advantages over field tests, successfully distinguished differences in aggressiveness among M. phaseolina isolates and relative differences among soybean geno- types for resistance to M. phaseolina comparable with results of field tests.