Seven hundred forty-nine isolates of Phytophthora spp. were obtained from irrigation canals in eastern Washington State during the 1992 to 1995 and 1999 growing seasons. Isolates were retrieved using pear baiting techniques. All isolates were pathogenic to pear and were present in irrigation water beginning early in fruit development. Over the course of the 5 year study, 10 and 5% of isolates were identified as P. cactorum and P. citricola, respectively, using morphological criteria. The remaining isolates could not be identified using morphological criteria. Colony morphology of these isolates was characterized during all years of the study. In 1999, more detailed studies utilizing polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis of entire internal transcribed spacer (ITS) regions (ITS1, 5.8S, and ITS2) of ribosomal DNA for 180 isolates, and sequence analysis of ITS2 for 50 isolates, were used to investigate genetic variation and phylogenetic relationships among isolates. Isolates were divided into 12 groups based on their growth type on corn meal agar. Restriction digestion of the entire ITS region with three enzymes revealed 11 restriction digestion patterns among 180 isolates. PCR-RFLP and sequence data were obtained for 12 reference Phytophthora spp. (two species in each of Waterhouse's six morphological groups). Phylogenetic analysis of ITS2 regions revealed nine clades, each with strong bootstrap support. Molecular analyses revealed 23 isolates that were in the P. gonapodyides clade, 9 in the P. parasitica clade, 1 in the P. cactorum clade, 7 in the P. citricola/capsici clade, and 4 in the P. cambivora/pseudotsugae clade. The three isolates comprising clade 5 were significantly distinct from all other Phytophthora spp. in the databases and may represent a new Phytophthora sp. Colony morphology was not consistently correlated to PCR-RFLP pattern or ITS2 phylogeny, suggesting that the former criterion is insufficient for species identification. The results of this study indicate that at least nine phylogenetically distinct taxa of Phytophthora pathogenic to pear are present in irrigation water in North Central Washington.

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During March to July 2003, a postharvest fruit rot was observed on ‘Golden Delicious’, ‘Granny Smith’, and ‘Red Delicious’ apples (Malus × domestica Borkh.) sampled from commercial packinghouses in Washington State. Losses as high as 24% in storage bins were observed in July on ‘Red Delicious’. The disease started at the stem bowl area or the calyx end of the fruit. Decayed fruit was apparently not wounded. Decayed areas were brown and firm. Internal decayed flesh appeared yellowish brown. On ‘Red Delicious’ apples, decayed fruit was apparently discolored from red to brown. As the disease advanced, pycnidia of a fungus might form on the stem, sepals, or the surface of decayed fruit. Pycnidia were 0.3 to 0.7 mm in diameter, black, and partially immersed in decayed tissues. To isolate the causal agent, decayed fruit was lightly sprayed with 70% ethanol and air dried. Fragments of diseased tissue were removed from the margin of diseased and healthy tissue and plated on acidified potato dextrose agar (PDA). A fungus was consistently isolated from decayed fruit with the symptoms described above. On PDA, the colonies of the fungus first appeared with dense hyaline mycelium and later turned light yellow to yellow. Black pycnidia of the fungus formed on 2- to 3-week-old oatmeal agar cultures at 20°C under 12-h alternating cycles of fluorescent light and dark. The fungus was identified as Sphaeropsis pyriputrescens Xiao & J. D. Rogers, based on the description of the fungus (1). Voucher specimens were deposited at the WSU Mycological Herbarium. Two isolates of the fungus recovered from decayed apples were tested for pathogenicity on apple. Fruit of ‘Golden Delicious’ and ‘Gala’ were surface-disinfested for 5 min in 0.5% NaOCl, rinsed, and air dried. Fruit was wounded with a sterile 4-mm-diameter nail head. A 4-mm-diameter plug from the leading edge of a 3-day-old PDA culture or plain PDA (control) was placed in the wound of each of 10 replicate fruit for each isolate or control. Fruit was tray packed with polyethylene liners and stored in cardboard boxes in air at 3°C, and decay was evaluated 2 weeks after inoculation. Five decayed fruits from each treatment were selected for reisolation of the causal agent. The experiment was conducted twice. In a separate pathogenicity test, two isolates (one each from apple and pear) were included in the test. Fruit of ‘Red Delicious’ apple was prepared and inoculated as the same manner described above, but fruit was stored in air at 0°C. The experiment was conducted twice. All fruit that were inoculated with the fungus developed decay symptoms. No decay developed on fruit in the controls. The same fungus was reisolated from decayed fruit. This indicates that isolates from apple and pear were pathogenic to apple. S. pyriputrescens is the causal agent of a newly reported postharvest disease on ‘d'Anjou’ pears (1). To our knowledge, this is the first report of this fungus causing postharvest fruit rot on apple. We propose ‘Sphaeropsis rot’ as the name of this new disease on apple and pear. Preliminary evidence suggests that infection of fruit by this fungus occurred in the orchard prior to storage. Reference: (1) C. L. Xiao and J. D. Rogers. Plant Dis. 88:114, 2004.