Morphological, pathogenic and molecular characterization of Phoma spp. isolated from onion field soils in Puerto Rico

Abstract

Thirteen Phoma spp. isolates collected during a survey conducted in onion field soils in Santa Isabel, Puerto Rico, were examined on the basis of morphology, pathogenicity and molecular characteristics. Twelve isolates were identified as Phoma putaminum Speg. and one isolate as an atypical Phoma macrostoma var. incolorata (section Phyllostictoides).This is the first report of P. putaminum and P. macrostoma var. incolorata for Puerto Rico and the Caribbean. In vitro, Phoma putaminum isolates were pathogenic to onion cvs. Mercedes and Excalibur, resulting in necrosis of young bulbs and roots seven days after inoculation. Disease incidence caused by P. putaminum was higher in cultivar Mercedes, ranging from 75 to 100%, than in P. macrostoma var. incolorata (0 to 25%). No symptoms were observed on cultivar Excalibur inoculated with P. macrostoma var. incolorata or on control plants. In vitro inoculations of commercial onion bulbs and field inoculations of roots failed to reproduce symptoms, thus showing that P. putaminum and P. macrostoma var. incolorata are weak pathogens. Sequence size of the nuclear internal transcribed spacer (ITS) of ribosomal DNA (rDNA) gene ranged from 458 to 610 base pairs (bp) for P. putaminum and was 456 bp for P. macrostoma var. incolorata isolate. Analysis of rDNA ITS region by PCR-RFLP showed that restriction enzyme, Hindlll, among other restriction enzymes evaluated (Alul, EcoRI, Clal and Seal), differentiate between P. putaminum and P. macrostoma var. incolorata isolates.