Four Phytophthora Species Causing Foot and Root Rot of Apricot in Italy

In the summer of 2006, 1-year-old apricot (Prunus armeniaca L.) trees with leaf chlorosis, wilting, and defoliation associated with root and crown rot were observed in a nursery in Sicily (Italy). Of 3,000 plants, ~2% was affected. Four Phytophthora spp. (45, 25, 20, and 10% of the isolations of the first, second, third, and fourth species, respectively) were isolated from decayed roots and trunk bark on BNPRAH (3). Axenic cultures were obtained by single-hypha transfers. Isolates of the first species formed petaloid colonies on potato dextrose agar (PDA) and had an optimum growth temperature of 25°C. On V8 agar (VA), they produced persistent, papillate (often bipapillate), ovoid to limoniform sporangia (length/breadth ratio 1.4:1). They did not produce gametangia when paired with A1 and A2 isolates of Phytophthora nicotianae. The second species formed arachnoid colonies, had an optimum growth of 30°C, and produced uni- and bipapillate, ellipsoid, ovoid or pyriform sporangia (length/breadth ratio 1.3:1). All isolates were A2. The third species formed rosaceous colonies on PDA, had an optimum temperature of 28 to 30°C, and produced papillate (sometime bipapillate), ellipsoid or limoniform (length/breadth ratio 2:1), caducous sporangia with a tapered base and a long pedicel (as much as 150 μm). All isolates were A1 type. The fourth species formed petaloid-like colonies on PDA and had an optimum growth of 26 to 28°C. On VA, it produced papillate (sometimes bipapillate), ovoid (length/breadth ratio 1.3:1), and decidous sporangia with a short pedicel (<4 μm). The isolates were homothallic and produced oogonia (25 to 31 μm in diameter) with paragynous antheridia and aplerotic oospores. On the basis of morphological and cultural characters, the species were identified as P. citrophthora, P. nicotianae, P. tropicalis and P. cactorum. Identification was confirmed by the electrophoretic analysis of total mycelial proteins and four isozymes (acid and alkaline phosphatases, esterase, and malate dehydrogenase) on polyacrylamide gel (1). Analysis of internal transcribed spacer (ITS) regions of rDNA using the ITS 4 and ITS 6 primers for DNA amplification (2) revealed 99 to 100% similarity between apricot isolates of each species and reference isolates from GenBank (Nos. AF266785, AB367355, DQ118649, and AF266772). The ITS sequence of a P. citrophthora isolate from apricot (IMI 396200) was deposited in GenBank (No. FJ943417). In the summer of 2008, pathogenicity of apricot isolates IMI 396200 (P. citrophthora), IMI 396203 (P. nicotianae), IMI 396201 (P. tropicalis), and IMI 396202 (P. cactorum) was tested on 3-month-old apricot seedlings (10 plants for each isolate) that were transplanted into pots filled with soil prepared by mixing steam-sterilized sandy loam soil (4% vol/vol) with inoculum produced on autoclaved kernel seeds. Ten control seedlings were grown in autoclaved soil. Seedlings were maintained in a screenhouse and watered daily to field capacity. Within 40 days of the transplant, all inoculated seedlings showed leaf chlorosis, wilting, and root rot. Control seedlings remained healthy. All four Phytophthora spp. were reisolated solely from inoculated plants. To our knowledge, this is the first report of Phytophthora root and crown rot of apricot in Italy and of P. tropicalis on this host. References: (1) S. O. Cacciola et al. Plant Dis. 90:680, 2006. (2) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (3) H. Masago et al. Phytopathology 67:425, 1977.