With a typical Mediterranean climate, Aydin is the third largest strawberry-producing province, responsible for 13% of the overall strawberry production in Turkey. Strawberries (Fragaria × ananassa Duchesne) are mainly grown in raised, plastic-mulched beds under tunnels and soil solarization is the most effectively used management practice to control soil-borne pathogens. During October 2011 and 2012, 2 months after planting, wilting and collapse of plants were observed on commercial strawberry (cv. strawberry Festival) fields in Sultanhisar town of Aydin Province. Eleven percent of the plants were wilted and died. Symptomatic plants exhibited blackened necrotic discoloration of roots and in the cross section of crowns. A fungus was consistently isolated from pieces of infected tissue cut aseptically from the crowns and placed on potato dextrose agar. Fungus produced white colonies and later turned olivaecious black with dense aerial mycelium after 4 to 5 days incubation at 27°C. Dark brown to black pycnidia that formed on 20- to 30-day-old pure cultures under daylight conditions produced abundant conidia that were two-celled, thick-walled, and oval shaped with longitudinal striations. Single spore isolates from 12 samples were obtained and stored for further identification. The average size of 300 conidia was 25.42 ± 2.12 × 12.87 ± 1.08 μm. The morphology of the fungus was similar to Lasiodiplodia theobromae (Pat.) Griff. & Maubl. (syn. Botryodiplodia theobromae Pat.). To confirm the identity of the isolates, the internal transcribed spacer (ITS) region of ribosomal DNA and the elongation factor 1-alpha gene were amplified with the universal ITS1/ITS4 and EF1-688F/EF1-1251R (1) primers, respectively. The amplicons from 12 isolates were commercially sequenced at Macrogen (Korea) and were deposited in GenBank under consecutive accession numbers KF910369 to KF910380 and KJ641536 to KJ641547. Sequence comparison and phylogenetic analysis revealed that all 12 isolates were closely related and belonged to L. theobromae. Pathogenicity tests were performed by the toothpick technique (2) under greenhouse conditions (28°C, 14/10-h day/night, 70% RH) on potted strawberry plants (cv. strawberry Festival). Toothpicks carrying fungal growth taken from 1-week-old corn meal agar cultures of the tested isolates was placed into the basal crown tissue of the plants by piercing about 5 mm depth. Six plants were inoculated for each isolate and six were treated with sterile toothpick for control. All inoculated plants developed wilting and dieback symptoms resembling those of naturally infected plants within 2 to 3 weeks of incubation. All plants inoculated with the tested isolates collapsed after 4 weeks and showed discoloration of internal crown tissue. Control plants did not exhibit any disease symptoms, and crown tissue was symptomless. L. theobromae was successfully re-isolated from lesions of all inoculated plants. L. theobromae has been reported to cause cankers and dieback in a wide range of hosts in tropical and subtropical regions of the world (3). To the best of our knowledge, this is the first report of L. theobromae causing dieback on strawberry plants.
References: (1) A. Alves et al. Fungal Divers. 28:1, 2008. (2) M. E. A. El-Morsi and I. A. Ibrahim. Wudpecker J. Agric. Res. 1:215, 2012. (3) E. Punithalingam. Plant diseases attributed to Botryodiplodia theobromae Pat. J. Cramer, Vaduz, 1980.
