Centre of Competence for Innovation in the Agro-Environmental Sector (AGROINNOVA), Via Leonardo da Vinci 44, 10095 Grugliasco, Italy
AGROINNOVA and DISAFA, Via Leonardo da Vinci, 44, 10095 Grugliasco, Italy
AGRIS Sardegna, Viale Trieste 11, 09123 Cagliari, Italy
Common bean (Phaseolus vulgaris L.) is grown worldwide for consumption of dry or green beans. During late spring of 2012, yellowing and wilting symptoms were observed in a commercial bean field cv. Lingua di fuoco in Cagliari Province (Sardinia, southern Italy) on 30% of plants 4 to 5 months after sowing. The first symptoms developed in May, when temperatures reached 18 to 30°C. Affected plants showed crown rot, necrosis of the cortex, and foliar chlorosis. As disease progressed, plants collapsed. In the presence of abundant moisture, white mycelium developed on the senescent tissue along with light to dark brown sclerotia (3.0 to 4.8 mm in diameter). Symptomatic tissue was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar (PDA) amended with 25 mg streptomycin sulfate/liter. The fungus that was isolated consistently from symptomatic plants onto PDA at 23°C grew rapidly in culture with silky-white, sterile mycelium, formed light to dark brown sclerotia (each 1.8 to 3.2 mm in diameter) after 7 days, and readily produced aerial hyphae. These morphological features are typical of Sclerotium rolfsii (2). The internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) was amplified for one isolate using ITS1/ITS4 primers (4), and sequenced (GenBank Accession No. KF002510). BLASTn analysis (1) of the 656-bp segment showed 87% homology with the ITS sequence of an S. rolfsii isolate (JF819727). Pathogenicity of one isolate was confirmed by inoculating healthy P. vulgaris plants cv. Lingua di fuoco grown in 2-liter pots in a steamed potting mix containing 50% Tecno2 (70% white peat and 30% clay) and 50% Tiesse 3 (60% white peat, 20% clay, and 20% perlite) (Turco Silvestro terricci, Bastia d'Albenga, SV, Italy). Inoculum consisting of mycelium and sclerotia of the pathogen produced from 10-day-old cultures on PDA was mixed in the soil at 0.5 g/liter substrate. Four 7-day-old plants per pot, with three replicate pots, were used for inoculation. The same number of control plants grown in the same substrate were inoculated with non-colonized PDA as a negative control treatment. The pathogenicity test was repeated. Plants were kept in a growth chamber at 30°C and 85% RH. Inoculated plants developed symptoms of leaf yellowing within 10 days, followed by crown rot, appearance of white mycelium and sclerotia, and eventual wilting. Control plants remained asymptomatic. Isolations from inoculated plants demonstrated the absence of latent infections by the fungus S. rolfsii, but the fungus was not reisolated from non-inoculated control plants. To our knowledge, this is the first report of S. rolfsii infecting P. vulgaris in Italy. Southern blight has been reported on common bean in sub-tropical and tropical areas of the world (3), where it can cause severe crop losses.
References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) J. E. M. Mordue. CMI Descriptions of Pathogenic Fungi and Bacteria No. 410, 1974. (3) H. F. Schwartz et al. Page 20 in: Compendium of Bean Diseases. American Phytopathological Society Press, St. Paul, MN, 2005. (4) T. J. White et al. PCR Protocols. Page 315 in: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.