First Report of Pestalotiopsis clavispora Causing Dieback on Blueberry in Uruguay.

During the last 10 years, blueberry (Vaccinium corymbosum) production in Uruguay has increased to more than 850 ha. From 2005, symptoms of dieback characterized by the death of twigs and branches have been frequently observed in blueberry plants cv. O'Neal in orchards located in Uruguay. Symptomatic 4-year-old plants (cv. O'Neal) were collected and small pieces of necrotic tissues were surface disinfected and plated onto potato dextrose agar (PDA) with 0.2 g liter of streptomycin sulfate. Plates were incubated at 25°C in the dark. All affected tissues consistently developed colonies with white and cottony mycelium, turning slightly yellow after 7 to 10 days. Black acervuli distributed in concentric circles were observed after 10 days. Conidia were fusiform, straight, and had five cells. Basal and apical cells were colorless while the three median cells were dark brown. Conidia (n = 50) had an average of 22.1 (16.5 to 28.2) × 6.6 (5.6 to 7.7) μm. All conidia had one basal appendage of 6.1 (3.9 to 14.3) μm and two to four (usually three) apical appendages of 22.8 (17.4 to 42.9) μm. According to colony and conidia morphology, the isolates were initially identified as Pestalotiopsis clavispora (G.F. Atk.) Steyaert (1). To identify, the internal transcribed spacers (ITS1, 5.8S, ITS2) region of rDNA of a representative isolate (Ara-1) was amplified with ITS1/ITS4 primers (4), sequenced, and compared with those deposited in GenBank. The isolate Ara-1 (Accession No. JQ008944) had 100% sequence identity with P. clavispora (Accession Nos. FJ517545 and EU342214). To confirm pathogenicity, isolate Ara-1 was inoculated onto asymptomatic 1-year-old blueberry plants (cv. O'Neal). Mycelial plugs (4 mm in diameter) from an actively growing colony on PDA were applied to same-size bark wounds made with a cork borer in the center of the stems previously disinfected with 70% ethanol and covered with Parafilm. Control plants were inoculated with sterile PDA plugs. Inoculated plants (five per treatment) were randomly distributed in a greenhouse and watered as needed. After 2 weeks, all stems inoculated with P. clavispora showed brown necrotic lesions 2 to 3 cm in length and 1 to 2 mm deep. White mycelium was observed over lesions. Control plants remained symptomless. The pathogen was reisolated from all necrotic lesions, thus fulfilling Koch's postulates. P. clavispora has been reported as associated with blueberry in Hawaii (3) and Chile (2). To our knowledge, this is the first report of P. clavispora causing dieback disease on blueberry in Uruguay. References: (1) E. F. Guba, Monograph of Pestalotia and Monocheatia. Harvard University Press, Cambridge, MA, 1961. (2) J. G. Espinoza et al. Plant Dis. 92:1407, 2008. (3) L. M. Keith et al. Plant Dis. 90:16, 2006. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.