ABSTRACT
Infection by Phytophthora ramorum was associated with stem and leaf lesions of Pacific madrone (Arbutus menziesii) seedlings and saplings. In addition, a common and native pathogen, Botryosphaeria dothidea, caused similar leaf and stem lesions. When exposed to natural levels of inoculum in forests infested with P. ramorum, 50 to 66% of madrone saplings used as bait died. Recovery of P. ramorum from colonized plant tissue on culture media was generally low. From initial infection, P. ramorum was not culturable from leaf tissue after a mean of 3.5 weeks or from stem tissue after a mean of 8 weeks. Generally, B. dothidea was recovered more frequently from necrotic stems and leaves than was P. ramorum. Experimental inoculations of madrone seedlings showed that leaf and stem lesion lengths were, on average, greater on tree seedlings inoculated with P. ramorum than on those inoculated with B. dothidea. P. ramorum and B. dothidea appear to coexist in stem and leaf tissue, forming a novel pathogen complex, affecting growth and reproduction of Pacific madrone.
ABSTRACT
Phytophthora ramorum S. Werres & A.W.A.M. de Cock was isolated from discolored leaves and cankers on small branches (<0.5 cm in diameter) on 27 coast redwood (Sequoia sempervirens) saplings (2 to17 cm in diameter) at two locations in California (Jack London State Park, Sonoma County and Henry Cowell State Park, Santa Cruz County). Symptoms were observed on branches throughout the crowns of affected trees. Isolates were identified as P. ramorum by their abundant chlamydospores and caducous, semi-papillate sporangia (2) and internal transcribed spacer (ITS) rDNA sequences identical to those of P. ramorum from Quercus spp., Lithocarpus densiflorus, and Rhododendron (1,2). P. ramorum was also detected in dying basal sprouts on mature redwood trees from an additional five locations in coastal California by polymerase chain reaction (PCR) amplification of the ITS region using DNA extracted from symptomatic tissue and P. ramorum-specific PCR primers. To test for pathogenicity, foliage inoculations were conducted on redwood seedlings in two trials by misting 30 leaves per trial (five leaves per seedling plus controls) with sterile distilled water and then pinning inoculum plugs to the upper surface of leaves. Inoculation resulted in lesions of 1 to 20 mm on individual leaves, and P. ramorum was recovered from 43% of inoculated leaves. Symptoms were not restricted to inoculated leaves because 15 inoculations of individual leaves led to discoloration of two or more adjacent leaves. On one inoculation, 60 mm of the adjacent stem was killed. Stems of redwood seedling (approximately 1 cm in diameter) were wound inoculated (1) in two trials consisting of 10 inoculated seedlings per trial plus 10 controls. After 6 weeks, lesion lengths in the cambium caused by P. ramorum averaged 13.7 mm (range 4 to 21 mm). P. ramorum was recovered from 100% of inoculated stems. Entire branches near the inoculation point became chlorotic even though no direct connection was evident between the lesion and the branches. No chlorosis was observed among the control inoculations. Mean lesion lengths of inoculated stems were significantly greater in both trials than those of control inoculations (mean 6.2 mm) at P < 0.05 based on analysis of variance (ANOVA). Redwood saplings (2.5 to 4.5 cm in diameter) were also wound inoculated in a separate trial. No phloem or cambial discoloration was observed after 7 weeks, but necrotic lesions in the xylem had a mean length of 39 mm (range 12 to 73 mm). In addition, narrow streaks, 1 to 2 mm in diameter, were also noted in the xylem extending from the necrotic areas upward to 90 cm. P. ramorum was recovered from 70% of inoculated stems in this trial. Mean lesion lengths of P. ramorum were significantly greater in all trials than those of control inoculations (mean 20 mm) at P < 0.05 based on ANOVA. While P. ramorum causes a lethal canker on Quercus spp. and L. densiflorus (1), we have not observed unusual mortality or disease symptoms on overstory redwoods in natural forests. The impact of infection by P. ramorum on understory redwoods is also unclear. However, the pathogen appears to be able to kill sprouts. References: (1) D. M. Rizzo et al. Plant Dis. 86:205, 2002. (2) S. Werres et al. Mycol. Res. 105:1155, 2001.
