In October 2007, blighted shoots were observed on highbush blueberry (Vaccinium corymbosum L. cv. O'Neal) plants in La Plata, Buenos Aires Province, Argentina. Isolations from surface-disinfested shoots onto carrot agar and Spezieller Nahrstoffarmer Agar (SNA) consistently yielded white colonies that produced black sclerotia, mainly near the edge of the culture plates, after 7 days. Sclerotia were transferred to SNA tubes and kept at 5°C for several months. The germination of sclerotia produced numerous 6 mm long initials, stipitate pale brown cup-shaped apothecia (10 × 6 mm) with eight-spored asci (137 × 7 μm) at 18°C and continuous light conditions. Asci with uniseriate ascospores were cylindrical and narrow at the base. Ascospores (11 to 12 × 4 μm) were hyaline, unicellular, smooth, and ellipsoid. This isolated fungus was morphologically identified as Sclerotinia sclerotiorum (Lib.) de Bary (2,3). The isolate was deposited in the IMYZA Microbial Collection as INTA-IMC 87. Mycelium was cultured in 100 ml of Czapek's-Dox medium, supplemented with sucrose, peptone, yeast extract, sodium nitrate, and vitamins (1), for 3 days and fungal DNA was obtained using a DNA extraction kit. ITS1 and ITS2 of ribosomal genes were amplified by PCR using universal primers (4) and the PCR product was sequenced. A BLAST algorithm search revealed 100% identity of the sequence with 12 GenBank entries for S. sclerotiorum. The nucleotide sequence was deposited in the GenBank with Accession No. JF277567. Pathogenicity testing was achieved by placing detached leaves of cvs. Emerald, Misty, and Start on water agar (WA) plates, inoculating with 9-mm2 mycelial blocks, and incubating at 20°C with 12 h of light. Young shoots of highbush blueberry, Misty and O'Neal, were inoculated by the cut shoot method with micropipette tips filled with mycelium and kept under greenhouse conditions at 24°C and 14 h of light. On control plants, WA blocks or WA-filled micropipette tips were used. Leaf blight was observed after 5 to 6 days and sclerotia appeared after 7 days on inoculated tissues. Shoot blight was recorded on inoculated plants after 5 days. The fungus was reisolated from inoculated tissues, with no symptoms showing on controls. To our knowledge, this is the first report of Sclerotinia rot caused by S. sclerotiorum in blueberry in Argentina.
References: (1) J.F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Hoboken, NJ, 2006. (2). J. E. M. Mourde and P. Holliday. No. 513 in: CMI Descriptions of Pathogenic Fungi and Bacteria. Kew, Surrey, UK, 1976. (3) S. Umemoto et al. Gen. Plant Pathol. 73:290, 2007. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.
