First Report of Crown Rot of Strawberry Caused by Macrophomina phaseolina in Chile.

Abstract

In recent years, an increase of collapsed and dead strawberry plants has been observed in several fields in central Chile, specifically in San Pedro, Melipilla, an important area for strawberry cultivation in the country. To determine the causal agent of the disease and the extent of the problem, 25 sample sites of 1 ha each, distributed in different San Pedro zones, were surveyed at the end of the 2011 season (from December 2010 to February 2011). Cross sections of the crowns of symptomatic strawberry plants showed necrotic tissue and brown-red to dark brown areas on the vascular ring. Samples of the affected crowns were superficially disinfested and plated on potato dextrose agar with 200 μg/ml of streptomycin sulfate. Dark gray colonies were observed after 7 days of incubation at 24°C. Pure cultures of the pathogen showed aerial mycelium and abundant dark oblong sclerotia. Fifty sclerotia were measured, averaging 120 × 74 μm. Twenty-one isolates were identified molecularly utilizing the species specific primers MpKFI and MpKRI (2) that yielded a 350-bp fragment. The amplified DNA fragments were sequenced and BLAST analysis showed a 99% nucleotide sequence identity with Macrophomina phaseolina (GeneBank Accession No JX535007.1). Both morphological and molecular analyses confirmed that the isolated species corresponded to M. phaseolina, causal agent of crown and root rot in strawberry. Four representative isolates were selected to conduct pathogenicity tests. Inoculum was prepared by incubating the pathogen for 28 days at 20°C in sterilized oat seeds. Pots of 1.5 liters were filled with a mixed substrate of peat and perlite (2:1), amended with inoculated oats at 9 g per liter of substrate. 'Camarosa' strawberry plants were planted and grown in a glasshouse for 1 month. Six replicated plants per isolate and six plants growing on non-inoculated substrate were left as controls. Ninety-five percent of the inoculated plants showed wilt and collapse symptoms 22 days after transplant, whereas no symptoms were observed in the control plants. M. phaseolina was reisolated from the crowns of symptomatic plants, fulfilling Koch's postulates. The pathogen was isolated from symptomatic strawberry plants in 14 of the 25 sampled sites. Although M. phaseolina was described previously on other crops in Chile, to our knowledge, this is the first report of M. phaseolina causing crown rot of strawberry. The disease has been recently reported in Spain, the United States, and Argentina (1,3,4).