Differentiation of the "Ascochyta complex" fungi of pea by biochemical and molecular markers.

Abstract

Isolates of three closely related pea pathogens, Ascochyta pisi, Ascochyta pinodes (teleomorph Mycosphaerella pinodes) and Phoma medicaginis var. pinodella, were compared by means of isozyme analysis and restriction fragment length polymorphisms (RFLP) of amplified rDNA spacers. Three enzyme systems differentiated A. pisi from M. pinodes and P. m. pinodella. The internal transcribed spacers (ITSs) of the three fungi showed no intraspecific and very little interspecific variation after digestion with 12 endonucleases. Digestion of the intergenic spacer (IGS) with HinfI, and Sau3A revealed uniformity in A. pisi patterns which consistently differed from those of M. pinodes and P. m. pinodella. No clear distinction could be made between the latter two fungi which both showed intraspecific variability. Both biochemical and molecular markers thus discriminated between two Ascochyta species. The results also indicated a closer relationship between two organisms belonging to different genera (Ascochyta and Phoma) than between two species of the same genus (Ascochyta).