Macrophomina phaseolina has been observed on alfalfa and white clover in North America, but its pathogenicity to mature plants of these species has not been adequately documented and Koch's postulates have not been fulfilled. Isolates of M. phaseolina from alfalfa and white clover were evaluated for pathogenicity by inoculating tissues of mature plants with infested toothpick pieces. Excised leaf tissues also were inoculated with mycelium. In stolons of white clover and stems of alfalfa, M. phaseolina caused a brown-black, basipetally progressive necrosis of vascular tissue with subsequent collapse of the surrounding pith and epidermis to produce radially constricted, expanding lesions. In taproots and crowns of alfalfa, M. phaseolina caused dark discoloration of vascular tissues in bands or streaks above and below inoculation points with subsequent invasion and death of cortical tissues, lateral roots, and stems. Sclerotia formed in all tissues of both species. Excised leaf tissues were rapidly parasitized, but significant differences in rates of parasitism between genotypes suggested that differences in host resistance to M. phaseolina may be present in both species. Pycnidia formed on leaves of bean, lima bean, and cotton. All isolates of M. phaseolina were reisolated from margins of necrosis in all types of inoculated tissues and regrown in pure culture. These results fulfill Koch's postulates for M. phaseolina as a pathogen of mature white clover and alfalfa in North America, and they demonstrate its capacity to parasitize a variety of tissues of both species in the absence of other pathogens. Results indicate that M. phaseolina should be considered a potential cause for lack of persistence of white clover and alfalfa during summer months in the southeastern United States.
