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A New Report of Phytophthora ramorum on Rhamnus
purshiana in Northern California. A. M. Vettraino, Department of
ESPM-ES, 137 Mulford Hall, University of California, Berkeley 94720 and
Department of Plant Pathology, University of Tuscia, Viterbo, Italy; and D.
Hüberli, S. Swain, A. Smith, and M. Garbelotto, Department of ESPM-ES, 137
Mulford Hall, University of California, Berkeley 94720. Plant Dis. 90:246, 2006;
published on-line as DOI: 10.1094/PD-90-0246C. Accepted for publication 28
November 2005.
Rhamnus purshiana, or cascara, is a deciduous tall shrub or small tree as
much as 9 m high with thin, smooth, silver-gray bark. It is often present in
shady sites in redwood and mixed evergreen forests of the North American west
coast, from British Columbia to northern California. In July 2005, symptomatic
leaves with irregular, black spots, 2 to 5 mm in diameter and concentrated
toward the tips, were collected from four cascara plants in the Samuel P. Taylor
State Park, Marin County, California. There was no evidence of defoliation.
Pieces of necrotic tissue were plated on selective medium (PARP) and maintained
at 19°C for 2 weeks. A Phytophthora sp. was consistently isolated and it
was identified as P. ramorum on the basis of morphological and molecular
traits published previously (3,4). The P. ramorum isolate Pr-418 has been
deposited in the American Type Culture Collection (ATCC MYA-3676) and a portion
of the internal transcribed sequence (ITS) of rDNA has been deposited in the
NCBI database (GenBank Accession No. DQ168874). Koch’s postulates were
completed using the leaf-dip method (2) on detached leaves collected from three
cascara plants growing at the University of California Botanical Garden at
Berkeley. Zoospore inoculum was prepared by flooding a 2-week-old culture
growing on V8 agar with sterile water for 4 days. The liquid was filtered after
cold shocking at 4°C for 30 min and incubated at room temperature for 1 h.
Fifteen leaves were dipped in the resulting zoospore suspension (1.6 × 10(^4)
zoospores per ml) for either 1 min (experiment 1) or overnight (experiment 2).
Leaves used as negative controls were dipped in sterile water. After removal
from the inoculum, excess liquid was allowed to drain. Leaves were maintained in
a moist chamber at 19°C with 13 h of natural light for 1 week. After 3 days of
incubation, necrotic spots similar to those observed in the field had developed
on leaves in experiment 2, while no symptoms were observed in experiment 1.
Necrotic lesions were observed on 12 and 15 of 15 leaves in experiments 1 and 2,
respectively, after 7 days of incubation. For each leaf, the necrotic area and
percent necrosis was determined by placing the leaves in a flatbed scanner and
processing the images with Assess (Version 1.01; The American Phytopathological
Society, St. Paul, MN). Lesions extended from the tip of the leaves and covered
3 ± 1% of the total leaf area in experiment 1 and 33 ± 3% in experiment 2.
Reisolation of P. ramorum on PARP was successful for all inoculated
leaves. P. ramorum was never isolated from negative controls and no
symptoms of infection were observed. The leaf-dip inoculation method is a rapid
and reliable indicator of host susceptibility to P. ramorum, with many
species developing necrosis when exposed to high concentrations of zoospores
(3). Our results show that exposure time to the pathogen can play an important
role in the development of symptoms. R. purshiana has been previously
reported as a host in Oregon (1,2), but to our knowledge, this is the first
report of cascara as a natural host of P. ramorum in the state of
California. Our results confirm those from Oregon (2). The impact of infection
by P. ramorum on cascara is unknown.
References: (1) J. M. Davidson et al. Plant Health Prog.
DOI:10.1094/PHP-2003-0707-01-DG, 2003. (2) E. Hansen et al. Plant Dis. 89:63,
2005. (3) D. M. Rizzo et al. Plant Dis. 86:205, 2002. (4) S. Werres et al. Mycol
Res. 105:1155, 2001.
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