Huelva in southern Spain is a major production area for strawberry (Fragaria × ananassa). At the end of the 2006 season (May-June) collapsed and dying strawberry plants were observed on several cultivars in four fields. Cut crowns of affected plants revealed dark brown necrotic areas on the margins and along the woody vascular ring. Roots of these plants were also shown to be necrotic. Macrophomina-like isolates developed from surface-disinfested affected tissues plated on potato dextrose agar amended with 250 mg L-1 of chloramphenicol. Dark, oblong-shaped sclerotia were observed in affected crown tissue and in culture after 5 to 7 days incubation at 25 ° C. They had an average length of 107 (217 to 62) μ m and width of 71 (110 to 35) μ m. Sequenced rDNA fragments of a single sclerotium isolate CH 724 (Spanish Type Culture Collection, CECT 20715; GenBank Accession No. AM410964) presented a 99% identity with Macrophomina phaseolina. Morphological and molecular results confirmed this species as M. phaseolina (Holliday & Punithalingam, 1970). Six single sclerotium isolates of M. phaseolina from strawberry were used for pathogenicity tests. Each isolate was used to inoculate six strawberry runner plants (cv. Camarosa) growing in pots of coconut fibre substrate for 5 weeks. Plants were inoculated by inserting a fungal colonised toothpick into each crown (Mertely et al ., 2005). An equal number of uninoculated plants treated similarly were left as controls. After 58 days, the incidence of plant death ranged from 67 to 100% depending on isolate. Macrophomina phaseolina was reisolated from all plants showing symptoms. Uninoculated plants remained symptomless.
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