A survey of soils associated with oak species was conducted in 2003 and 2004 in Indiana, Illinois, Maryland, Michigan, Minnesota, Pennsylvania, Ohio, West Virginia, and Wisconsin to investigate the occurrence of Phytophthora spp. Soils taken from around the base of healthy and declining oak trees were flooded with H2O and Quercus robur leaflets were used as bait for Phytophthora spp. From 829 soil samples collected near trees, 21% were positive for Phytophthora spp., with 55% of the 125 sites surveyed yielding a Phytophthora sp. Phytophthora cinnamomi was the most frequently isolated species, representing 69.4% of the Phytophthora-infested sites surveyed. Other species, in decreasing order of isolation frequency were Phytophthora sp. 2, P. citricola, P. europaea, P. cambivora, P. quercina-like isolates, and Phytophthora sp. 1. No significant association was found between the presence of Phytophthora organisms and site characteristics such as latitude, elevation, soil pH, or the crown condition of the trees. However, in P. cinnamomi-infested sites, a significant association was found with the deteriorating crown status of Q. alba and the presence of P. cinnamomi. The absence of P. cinnamomi above the 40°N latitude range also was noteworthy.
Isolates of Phytophthora cambivora, P. cinnamomi, P. citricola, P. europaea, P. quercetorum and two unidentified species were tested for their pathogenicity to eastern US oak species by root and stem inoculations. Experiments were conducted during two different periods and included 1-, 2-and 20-year-old oaks grown under greenhouse and field conditions. Species of Phytophthora were pathogenic in varying degrees to the oak species tested. All species were pathogenic to fine and taproots of at least one oak species. The fine root damage caused by the species of Phytophthora ranged from 9 to 55% when compared to the controls. Roots were more susceptible during the fall inoculation period than the summer. With exception of Phytophthora sp1 and P. quercina-like, all species of Phytophthora were pathogenic to oak stems with P. cinnamomi and P. citricola being the most aggressive. Quercus montana and Q. rubra were the most susceptible oak species to stem inoculation. Lesion sizes were considerably larger when 20-year-old trees were inoculated. Generally, no significant differences in lesion sizes were detected in greenhouse tests when the summer and fall inoculation periods were compared. However, on 2-year-old field-grown seedlings, lesion sizes were considerably smaller or not significantly different from controls during the fall inoculation period, suggesting lower, late season temperatures may restrict lesion development.
SummarySeven different Phytophthora species were used to test the foliar susceptibility of the common eastern US oak species and understory plants to Phytophthora infection. The Phytophthora species employed were Phytophthora cambivora, Phytophthora cinnamomi, Phytophthora citricola, Phytophthora europaea, Phytophthora quercetorum, Phytophthora quercina-like and Phytophthora sp1. Inoculation of detached-leaves with agar plugs containing mycelia of Phytophthora provided an estimate of their relative susceptibility. Lesions were always greater when foliage was wounded and young. On deciduous plants, lesion sizes were considerably reduced with the increasing foliar age, although with evergreen plants lesion sizes remained similar regardless of foliar age when more aggressive isolates were tested. Infections seldom resulted when foliage was not wounded. With young and mature foliage, P. citricola usually produced the largest lesions. Young foliage of Quercus rubra was the most susceptible to infection followed by Castanea dentata for both wounded and non-wounded inoculations. Mature foliage of Hamamelis virginiana, Kalmia latifolia and Quercus alba were the most susceptible to wound and non-wound inoculations.
In 2003 and 2004, soils in oak forest ecosystems in nine central and eastern states of the United States were surveyed for Phytophthora spp. Soil samples were collected around healthy and symptomatic trees. Symptoms included dieback of branches, gaps in lateral branch systems, yellowing of foliage, wilting and clustering of leaves, and the presence of epicormic shoots. Soil subsamples were collected in each of the four cardinal directions and at a distance of 1 to 2 m from the base of a tree. The four subsamples were bulked to produce a sample of approximately 2,000 ml. In the laboratory, each sample was mixed thoroughly and a single 250-g subsample was flooded with 500 ml of distilled water and baited with Quercus robur leaflets for 3 to 5 days at 17 to 20°C. Discolored leaflets were examined microscopically (×200) and those with sporangia typical of Phytophthora spp. were plated on PARPNH selective medium (1). Phytophthora europaea was recovered from soil samples collected from Q. alba in West Virginia, Q. rubra in Minnesota, West Virginia, and Wisconsin, Q. phellos in Ohio, and Q. velutina in Pennsylvania. Cultures were identified as P. europaea by their morphological, physiological, and molecular characteristics (4). Average dimensions of nine isolates were determined. Oogonia were 40 ± 3.9 μm in diameter and often had few bullet protuberances and tapered bases; oospores mostly filled the oogonia and averaged 36 ± 3.7 μm; sporangia dimensions averaged 42 ± 6.1 × 30 ± 4.1 μm with a length/width ratio of 1:4. Isolates produced larger oogonia and oospores but had similar sporangia length/width ratios comparable to the species description (4). Growth optimum (5.8 to 6.9 mm day-1) on V8 juice agar (V8A) occurred at 25°C. On potato dextrose agar, colonies produced dense, felt-like mycelia, often with a central mound of aerial hyphae. DNA also was extracted from eight representative isolates and the internal transcribed spacer (ITS) region of rDNA from each isolate was amplified and sequenced. ITS sequences were identical to those of P. europaea in the NCBI GenBank database (Accession No. DQ313222). Pathogenicity of six isolates (one from each site) was confirmed by wounding stems of 2-year-old Q. alba, Q. rubra, and Q. velutina seedlings and inoculating wounds with V8A plugs (6 mm) containing mycelia; V8A plugs without mycelia were used for controls. Two months after inoculation, P. europaea was reisolated on PARPNH medium from advancing lesions on all inoculated seedlings but was not isolated from control plants. Mean lesion lengths on seedlings inoculated with P. europaea were significantly greater (P < 0.05) than those on control plants; lesions averaged 0.46 cm on Q. alba, 1.38 cm on Q. rubra, and 1.01 cm on Q. velutina. Previously, P. europaea only was reported from oak trees and soil in forests of Austria, France, and Germany (1–4). These findings extend the current distribution of P. europaea and raise questions about its origin and role in the health of oak forests in eastern and north-central United States. Q. alba, Q. phellos, Q. rubra, and Q. velutina are new host associations for P. europaea. References: (1) Y. Balci and E. Halmschlager. For. Pathol. 33:157, 2003. (2) E. Hansen and C. Delatour. Ann. Sci. For. 56:539, 1999. (3) G. Hartmann and R. Blank. Forst Holz. 57:539, 2002. (4) T. Jung et al. Mycol. Res. 106:397, 2002.
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