First Report of Penicillium expansum Causing Postharvest Blue Mold of Fresh Date Palm Fruit (Phoenix dactylifera) in Spain

A survey of postharvest losses of commercially handled and cold-stored fruit of fresh date palm (Phoenix dactylifera L.), cvs. Medjool and Hayani, was conducted in the 2009 and 2010 seasons in the grove of Elx (Alacant Province, Southeast Spain). Disease symptoms consisting of circular, light brown, soft spots located in any part of the fruit skin were observed in 2 to 5% of the fruit. At room temperature, the lesions expanded rapidly and blue mold symptoms were apparent. The potential causal agent (isolate IVIA NiAA-2) was transferred to PDA and incubated at 25°C. The identification was performed at the Spanish Type Culture Collection (CECT, University of Valencia, Spain) based on colony morphology of the isolate grown on Czapeck yeast extract agar (CYA) and malt extract agar (MEA) at 26°C. Colonies were circular (average diameter of 40 mm at 7 days), radially sulcate, with dense velvety white mycelium, and very abundant, bluish green conidia. The underside of the plates showed light brown and pale green colonies on CYA and MEA, respectively. On CYA, but not on MEA, a light yellow exudate was produced and a brownish pigment diffused into the medium. At 5 and 37°C on CYA, white microcolonies and no colonies were observed, respectively. Conidia were ellipsoidal to subglobose, smooth and thin walled, measuring 3.0 to 3.5 × 2.5 to 3.0 μm (n = 50) (4). Based on these morphological characteristics, the isolate IVIA NiAA-2 was tentatively identified as Penicillium expansum L. To confirm the identity, we amplified and sequenced the rDNA internal transcribed spacer (ITS) region with primers ITS1 and ITS4 (GenBank Accession No. KC169942). A BLAST search showed 99% identity and 100% query coverage with P. expansum strain NRRL 6069 (DQ339562) (2). Selected healthy dates cv. Medjool were surface disinfected by dipping in 0.5% sodium hypochlorite for 2 min followed by thorough rinsing in deionized water. Pathogenicity was tested by pipetting 20 μl of a spore suspension (1 × 106 spores per ml), prepared from 7-day PDA cultures, onto fresh skin wounds, which were made on disinfected fruit using a sterile, stainless steel rod with a probe tip 1 mm in width × 2 mm in length (one wound on each of nine dates, incubated in one humid chamber). Disinfected, wounded, and non-inoculated dates were used as controls. The procedure was repeated three times. Disease symptoms were observed on all inoculated fruit (average lesion size of 6, 15, and 22 mm after 4, 7, and 10 days of incubation at 20°C, respectively) and P. expansum was consistently reisolated, thereby fulfilling Koch's postulates. No decay was observed on any of the non-inoculated fruit. Unidentified species of Penicillium have been reported to cause date palm fruit rot (1,3). To our knowledge, this is the first report of P. expansum causing postharvest decay of date palm fruit in Spain. References: (1) M. Djerbi. Diseases of the Date Palm. FAO Regional Project, Rome, 1983. (2) M. A. Dombrink-Kurtzman. Antonie Van Leeuwenhoek 91:179, 2007. (3) S. Ibrahim and M. A. Rahma. Bayero J. Pure Appl. Sci. 2:127, 2009. (4) R. A. Samson et al. Introduction to Food-Borne Fungi. Centraalbureau voor Schimmelcultures, Baarn, the Netherlands, 1995.