Detection of Phytophthora ramorum Blight in Oregon Nurseries and Completion of Koch's Postulates on Pieris, Rhododendron, Viburnum, and Camellia.

Phytophthora ramorum, the cause of sudden oak death in California and Oregon coastal forests and ramorum blight in European nurseries and landscapes (1), was detected in six Oregon nurseries in Jackson, Clackamas, and Washington counties from May to June 2003. The pathogen was isolated from: Viburnum bodnantense 'Dawn', V. plicatum var. tomentosum 'Mariesii', Pieris japonica × formosa 'Forest Flame', P. japonica 'Variegata' and 'Flaming Silver', P. floribunda × japonica 'Brouwer's Beauty', Camellia sasanqua 'Bonanza' and other cultivars, C. japonica, and Rhododendron × 'Unique'. Samples of symptomatic tissues were plated on a Phytophthora-selective medium (PARP) and tested by polymerase chain reaction (PCR) (3). All samples positive for P. ramorum with PCR yielded P. ramorum isolates in culture. The isolates have the European genotype, mating type A1, except for the Camellia spp. isolates, which have the North American genotype, mating type A2 (2). Isolates are deposited in the American Type Culture Collection. Koch's postulates for this pathogen have been completed on V. bodnantense and C. japonica (1). To confirm pathogenicity on the new hosts, isolates from V. plicatum var. tomentosum 'Mariesii', Pieris × 'Forest Flame', Pieris × 'Brouwer's Beauty', and P. japonica 'Variegata' and 'Flaming Silver' were used to inoculate healthy plants of the same cultivars. For isolates from Rhododendron × 'Unique' and C. sasanqua 'Bonanza', pathogenicity was tested on Rhododendron × 'Nova Zembla' and C. sasanqua 'Sutsugekka' and 'Kanjiro'. Three to five plants of each cultivar were inoculated and three to five were noninoculated. Zoospore inoculum was prepared on dilute V8 agar for one isolate from each host. Foliage of plants growing in 10-cm pots was dipped for 5 sec in a zoospore suspension (3 × 104 zoospores per ml) or sprayed to runoff with a hand mister (6 × 104 zoospores per ml). Control plants were dipped in or sprayed with sterile water. C. sasanqua plants were also inoculated by placing 6-mm mycelial plugs on individual leaves that had been wounded by piercing with a pin. Control leaves were wounded but not inoculated. Foliage was enclosed in plastic bags to retain humidity and the pathogen, and plants were incubated in a locked growth chamber (21 to 23°C). After 21 days, plants were examined for symptoms, and isolations onto PARP were made. All inoculated plants showed foliar symptoms, and P. ramorum was consistently isolated from inoculated plants, but not from asymptomatic control plants. On Rhododendron × 'Nova Zembla', nearly all leaves were wilted and dead, as were terminal buds and stems. Pieris spp. cultivars exhibited leaf and stem necrosis and defoliation. On V. plicatum var. tomentosum 'Mariesii', necrotic leaf lesions and defoliation of the lower leaves were observed. On C. sasanqua, necrotic lesions developed only on wounded leaves inoculated with mycelial plugs; these leaves abscised. Our results confirm the pathogenicity of Oregon nursery isolates of P. ramorum on V. plicatum var. tomentosum 'Mariesii', P. japonica × formosa 'Forest Flame', P. japonica 'Variegata' and 'Flaming Silver', P. floribunda × japonica 'Brouwer's Beauty', C. sasanqua and Rhododendron and complete Koch's postulates for several new hosts. References: (1) J. M. Davidson et al. Online publication. doi:10.1094/PHP-2003-0707-01-DG. Plant Health Progress, 2003. (2) E. M. Hansen et al. Plant Dis. 87:1267, 2003. (3) L. M. Winton and E. M. Hansen. For. Pathol. 31:275, 2001.

First Report of A1 Mating Type of Phytophthora ramorum in North America.

Phytophthora ramorum is known in Europe and the western United States (1). In Europe, it is found in nurseries and landscape plantings. In the United States, it has been confined to coastal forests, and in California, it is found in a few horticultural nurseries. All European isolates tested have been A1 mating type, while all North American isolates were A2 mating type (2). Amplified fragment length polymorphism markers also indicated that the populations on the two continents are distinct, and nearly all North American isolates are from one clone (Kelly Ivors, unpublished). In June 2003, P. ramorum was isolated from diseased Viburnum and Pieris spp. cultivars from a Clackamas County nursery in northern Oregon and diseased Camellia sp. cultivar from a Jackson County nursery in southern Oregon. Representative isolates were submitted to the American Type Culture Collection, Manassas, VA. As part of the effort to determine the origin of these new infestations, we tested the nursery isolates for mating type. Seven Oregon nursery isolates, three Oregon forest isolates (from the predominant North American clone), and two European isolates were paired. Agar plugs from 3-day-old colonies were placed in close proximity on carrot agar plates, and then the plates were examined for oogonia after 3 and 10 days as advised by C. M. Brasier (personal communication). Oogonia and antheridia typical of P. ramorum (2) formed when isolates from the Clackamas County nursery were paired with the Oregon forest isolates and also when isolates from the Jackson County nursery were paired with the European isolates. Gametangia also formed in pairings between Oregon forest isolates and European isolates, but not in any other combinations. We developed polymerase chain reaction (PCR) primers for four microsatellite loci and determined allele sizes for the same set of isolates (unpublished). Microsatellite alleles of the Clackamas County isolates were identical to the European tester isolates, and alleles of the Jackson County isolates were identical to the Oregon forest isolates. These results indicate that the recent Oregon nursery infestations are of separate origins. The Clackamas County isolates are A1 mating type and have microsatellite alleles like the European testers, but according to shipping records, the nursery has received no host nursery stock directly from Europe. However, host nursery stock has been received from a Canadian nursery. The Jackson County isolates are of A2 mating type and have microsatellite alleles like the forest isolates of Oregon, which is consistent with the reported origin of these plants from a California nursery. These preliminary microsatellite results need to be validated against a larger isolate set but are congruent with the mating type results. The Oregon nursery infestations highlight the dangers of unregulated or underregulated transport of host nursery stock from infested areas to noninfested areas. All host plants from infested nursery blocks at the affected Oregon nurseries have been destroyed by incineration, and a monitoring program has been implemented. Other host nursery stock on site has been taken "off-sale" pending verification that it is disease free, per the United States Department of Agriculture, APHIS requirements. References: (1) J. M. Davidson et al. On-line publication. doi:10.1094/PHP-2003-0707-01-DG. Plant Health Progress, 2003. (2) S. Werres et al. Mycol. Res. 105:1155, 2001.

Sudden Oak Death Caused by Phytophthora ramorum in Oregon.

Sudden oak death, caused by Phytophthora ramorum (1,2), has been found for the first time in Oregon, killing tanoak, Lithocarpus densiflorus, trees. To our knowledge, this is the first report of the disease outside of the San Francisco to Monterey area in California, (300 km to the south). Nine areas of infestation, all within a 24-km2 area, were discovered on forest lands near Brookings, in southwest Oregon. Mortality centers ranged in size from 0.2 to 4.5 ha and included 5 to approximately 40 diseased trees. P. ramorum was isolated from stem cankers using Phytophthora-selective medium. Isolates had distinctive morphological features characteristic of P. ramorum, including abundant production of chlamydospores and caducous, semipapillate sporangia on solid media. Internal transcribed spacer (ITS) sequences of isolates of P. ramorum from Oregon were identical to ITS sequences of isolates from California (1). The pathogen also was isolated from necrotic lesions on leaves and stems of native Rhododendron macrophyllum and Vaccinium ovatum growing beneath diseased tanoaks. In July 2001, the disease was located by an aerial survey conducted cooperatively by the USDA Forest Service and Oregon Department of Forestry. All lands within 1.6 km (1 mile) of the mortality centers are subject to Oregon quarantine, which bars the transport of any host plant materials. An eradication effort is currently underway. Symptomatic plants and all known host plants within 15 to 30 m of symptomatic plants are being cut and burned in the first phase of this operation. The total treated area is approximately 16 ha. References: (1) D. M. Rizzo et al. Plant Dis. In press. (2) S. Werres et al. Mycol. Res. 105:1155, 2001.

Analysis of Resistance to Eastern Filbert Blight in Corylus avellana.

Inheritance of resistance to eastern filbert blight, caused by Anisogramma anomala, in European hazelnut (Corylus avellana) was evaluated in the progeny of seven cultivars crossed in 12 combinations. The progeny were subjected to inoculation with A. anomala in the greenhouse and in the field. Three disease responses were measured: disease incidence, number of cankers, and proportion of wood diseased. In both the greenhouse and the field, progeny produced by crossing VR6-28 with three susceptible cultivars segregated 1:1 for complete resistance to eastern filbert blight, confirming a previous report that VR6-28 is heterozygous for a single, dominant resistance gene. Histograms of disease responses in progeny of the remaining six parents showed continuous distributions for all crosses examined. Consequently, these parents were analyzed for general and specific combining abilities for each disease response. In the field, general and specific combining ability were both significant (P < 0.05) for all disease responses, with general combining ability having twice the magnitude of specific combining ability. These results suggest these disease responses are controlled by additive gene action in the cultivars examined, with nonadditive gene action being of some importance. Based on general combining ability values, high levels of partial resistance were transmitted by the pollen parents, Gem and Tonda di Giffoni, and the seed parent, Willamette. Heritability of disease incidence, number of cankers, and proportion of wood diseased were calculated to be 0.21, 0.39, and 0.47, respectively, for this set of nine crosses after the first exposure period in the field. This suggests that it will be possible to use partially resistant parents to breed for hazelnuts exhibiting fewer and smaller cankers.