First Report of Botryosphaeria dothidea Causing Fruit Rot of Plum in Spain.

Species in the Botryosphaeriaceae are known to produce cankers, dieback, blights, and leaf spots on many hosts, mainly under stress conditions. Several Botryosphaeria spp. may also cause pre- or post-harvest decay of stone fruit, such as peaches (2). In June 2012, fruit of plum (Prunus domestica cv. Golden Japan) showing soft, brown, and slightly sunken necrotic lesions were observed in several orchards affected by hail in Cordoba province (southern Spain). Symptomatic fruit were collected and incubated at 25°C and 100% relative humidity. Isolations were done on potato dextrose agar (PDA). Mycelium and black pycnidia developed on the surface of incubated fruit and on PDA plates. Conidia were hyaline, aseptate, and fusoid. According to morphological criteria, the fungus was identified as Fusicoccum aesculi, the anamorph of Botryosphaeria dothidea (3). The internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS4/ITS5 and sequenced. BLAST analysis of the 528-bp fragment showed 100% homology with the sequence of B. dothidea. Pathogenicity tests were performed on immature healthy fruit (2 weeks before harvest) of the same cultivar from the same orchards. Fruit were washed in deionized water with Tween 20 (Polyoxyethylene 20 sorbitan monolaureate 99%, 0.1 ml/liter) and surface sterilized in 10% sodium hypochlorite for 1 min. Twenty-four fruit were inoculated using mycelial-agar discs. Twelve fruit were previously wounded with a sterile 0.5-mm-diameter needle. The same number of fruit, wounded and unwounded, served as a control. All fruit were incubated at 25°C and 100% relative humidity. Seven days after inoculation, 83% of wounded inoculated fruit showed rot symptoms. After 9 days, fruit rot symptoms started to appear on unwounded inoculated fruit. Twenty days after inoculation, 100% of wounded and unwounded fruit showed rot symptoms that led to mummification of the fruit. Pycnidia developed on inoculated fruit and the fungus was reisolated. No symptoms developed on control fruit. These results demonstrate that B. dothidea is pathogenic on plum and that wounds favor infection, although they are not needed. To our knowledge this is the first report of B. dothidea causing fruit rot of plum in Spain. This pathogen is well known in southern Spain causing a serious fruit rot of olive (1) and could have a great impact on plum production in this region, especially when there is damage to the fruits as occurred this year with hail. References: (1) J. Moral et al. Phytopathology 100:1340, 2010. (2) J. M. Ogawa et al. Compendium of Stone Fruit Diseases. APS Press, St. Paul, MN, 1995. (3) B. Slippers et al. Mycologia 96:83, 2004.