Detection of Phytophthora ramorum in Nurseries and Forest Lands in California in 2004 to 2009

From December 2004 through May 2009, samples were collected from California nurseries and wild lands to survey for Phytophthora ramorum and comply with federal regulations of nursery stock. Samples were prescreened by an enzyme-linked immunosorbent assay (ELISA) that detects Phytophthora spp. and tested by culture, P. ramorum-specific real-time polymerase chain reaction (PCR), and nested PCR. Yearly percentages of infected samples ranged from 0.6 to 2.3%. Camellia spp., Rhododendron spp., Magnolia spp., Pieris spp., and Laurus nobilis tested positive the most frequently in the nurseries and Lithocarpus densiflorus, Umbellularia californica, and Quercus agrifolia tested positive most often from wild lands. Of the 118,410 samples isolated onto PARP media, 0.8% was identified as P. ramorum. Of 115,056 samples tested by ELISA, 5.9% tested positive for Phytophthora spp. Of the 6,520 samples tested by PCR, 12.4% tested positive for P. ramorum. The false-negative, positive, and internal control failure rates of the assays are discussed. After removing the seasonal effect of sampling strategy, isolation of the pathogen into culture was found to be seasonally dependent whereas detectability by PCR and ELISA was not. To our knowledge, this is the first evaluation of a regulatory testing program for a plant pathogen on this scale using standardized assays.

Productivity and Cost of Integrated Harvesting of Wood Chips and Sawlogs in Stand Conversion Operations

This study evaluated the operational performance and cost of an integrated harvesting system that harvested sawlogs and biomass (i.e., energy wood chips) in stand conversion clearcut operations. Douglas-fir (Pseudotsuga menziesii) trees were processed into sawlogs while whole trees of tanoak (Lithocarpus densiflorus), and sub-merchantable materials (small-diameter trees, tops and limbs) were fed directly into a chipper to produce biomass for energy production. A standard time study method was used to determine productivity and costs. Over 26 working days, the integrated system produced 1,316 bone-dry metric tonnes (BDTs) of sawlogs, and 5,415.89 BDT of chips, with an average moisture content of 43.2%. Using the joint products allocation costing method, the costs of the integrated system were $29.87/BDT for biomass and $4.26/BDT for sawlogs. Chipping utilization was as low as 41%, directly affecting production and cost of chipping operation. Single-lane, dirt, spur roads were the most costly road type to transport whole trees to a centralized processing site: transportation costs for biomass and sawlogs were increased by $0.08/BDT and $0.02/BDT, respectively, for every 50 meter increase in traveling distance. Diesel fuel price could raise total system cost for each product by $0.78/BDT and $0.08/BDT for each $0.10/liter increase.

Stream Monitoring for Detection of Phytophthora ramorum in Oregon Tanoak Forests

Stream monitoring using leaf baits for early detection of Phytophthora ramorum has been an important part of the Oregon Sudden Oak Death (SOD) program since 2002. Sixty-four streams in and near the Oregon quarantine area in the southwest corner of the state were monitored in 2008. Leaves of rhododendron (Rhododendron macrophyllum) and tanoak (Lithocarpus densiflorus) were placed in mesh bags, and bags were floated in streams. Leaf baits were exchanged every 2 weeks throughout the year. Leaves were assayed by isolation on selective medium and by multiplex rDNA internal transcribed spacer polymerase chain reaction (ITS PCR). The two methods gave comparable results, but multiplex PCR was more sensitive. P. ramorum was regularly recovered at all seasons of the year from streams draining infested sites 5 years after eradication treatment. In streams with lower inoculum densities, recovery was much higher in summer than in winter. P. ramorum was isolated from streams in 23 watersheds. When P. ramorum was detected, intensive ground surveys located infected tanoaks or other host plants an average of 306 m upstream from the bait station. P. ramorum was isolated from stream baits up to 1,091 m from the probable inoculum source.

The effects of Phytophthora ramorum infection on hydraulic conductivity and tylosis formation in tanoak sapwood

Tanoak ( Lithocarpus densiflorus (Hook. and Arn.) Rehder) is highly susceptible to sudden oak death, a disease caused by the oomycete Phytophthora ramorum Werres, De Cock & Man in’t Veld. Symptoms include a dying crown, bleeding cankers, and, eventually, death of infected trees. The cause of mortality is not well understood, but recent research indicates that water transport is reduced in infected trees. One possible mechanism causing the reduction in hydraulic conductivity is the presence of tyloses in xylem vessels. The development of tyloses was studied in relation to hydraulic conductivity in P. ramorum-infected sapwood. Inoculated logs showed a greater abundance of tyloses than noninoculated logs after 4 weeks. Inoculated trees with xylem infections had significantly more tyloses than noninoculated trees. In addition, the increase in number of tyloses was associated with a decrease in specific conductivity, suggesting that tyloses induced by infection with P. ramorum may interfere with stem sap flow. Over time, tylosis development increased in tissues farther from the inoculation site, in advance of the vertical spread of infection. The results suggest that infected sapwood contains numerous tyloses, which could significantly impede stem water transport.

Long-term effects of tanoak competition on Douglas-fir stand development

In 1- to 2-year-old Douglas-fir ( Pseudotsuga menziesii (Mirb.) Franco var. menziesii) plantations near Cave Junction and Glendale, Oregon, sprout clumps of tanoak (Lithocarpus densiflorus (Hook. & Arn.) Rehd.) and other hardwoods were removed with herbicides in April 1983 to leave relative covers of 0%, 25%, 50%, or 100% of the nontreated cover, which averaged 15%. In 1996 (Cave Junction) and 1998 (Glendale), precommercial thinning (PCT) of Douglas-fir and cutting of nonconifer woody species were operationally applied across the four densities of tanoak. In 2005, Douglas-fir in 0% relative cover of tanoak averaged 5–8 cm larger at breast height and 3–6 m taller, and had two to four times the net stand volume of those growing in 100% relative cover. From 1999 to 2005, Douglas-fir stand growth accelerated more rapidly in tanoak relative covers of 0% and 25% than in covers of 50% and 100%. Differential development of Douglas-fir and hardwoods in relative covers of 0%, 25%, and 100%, followed by selection of crop trees via PCT, resulted in three distinct stand structures: pure stands of Douglas-fir with a single canopy layer 12–16 m tall, mixed stands with overstory Douglas-fir (12 m) and midstory hardwoods (7 m), and mixed stands with a single canopy layer (8–9 m).

Sources of Inoculum for Phytophthora ramorum in a Redwood Forest

Sources of inoculum were investigated for dominant hosts of Phytophthora ramorum in a redwood forest. Infected trunks, twigs, and/or leaves of bay laurel (Umbellularia californica), tanoak (Lithocarpus densiflorus), and redwood (Sequoia sempervirens) were tested in the laboratory for sporangia production. Sporangia occurred on all plant tissues with the highest percentage on bay laurel leaves and tanoak twigs. To further compare these two species, field measurements of inoculum production and infection were conducted during the rainy seasons of 2003-04 and 2004-05. Inoculum levels in throughfall rainwater and from individual infections were significantly higher for bay laurel as opposed to tanoak for both seasons. Both measurements of inoculum production from bay laurel were significantly greater during 2004-05 when rainfall extended longer into the spring, while inoculum quantities for tanoak were not significantly different between the 2 years. Tanoak twigs were more likely to be infected than bay laurel leaves in 2003-04, and equally likely to be infected in 2004-05. These results indicate that the majority of P. ramorum inoculum in redwood forest is produced from infections on bay laurel leaves. Years with extended rains pose an elevated risk for tanoak because inoculum levels are higher and infectious periods continue into late spring.

Root dynamics in sprouting tanoak forests of southwestern Oregon

Root and shoot biomass were measured across an 8 year chronosequence in mature and regenerating stands of tanoak ( Lithocarpus densiflorus (Hook. & Arn.) Rehd.), following cutting and burning in mature tanoak forests. Tanoak stump sprouts rapidly replaced leaf biomass but did not maintain preexisting root systems. Rather, root biomass declined for several years, with the largest proportional decline in extra-fine roots. Four years after harvest, live root biomasses in diameter classes 0.25–2.00 and 0.25–5.00 mm were 25% and 30% of values in mature tanoak forests, respectively. The proportion of dead roots was strongly correlated with maximum summer soil temperature. Root/shoot ratios recovered to preharvest values by age 3–4 years, at which time the live biomass of fine roots and leaf biomass was approximately 30%–40% of values in mature forest. From age 4 to 8 years, stable root/shoot ratios were associated with a three- to four-fold reduction in growth rate of leaf biomass and a proportional increase in growth of fine roots. These findings support the general theory of a functional root–shoot balance in tanoak and suggest a possible role for soil temperature in postharvest root dynamics. Improved understanding of postdisturbance root and shoot dynamics in tanoak will help identify competitive interactions and priorities for vegetation management decisions in establishment of conifers following harvest of mixed conifer–hardwood forests of southwestern Oregon and northern California.

Epidemiology of Phytophthora ramorum in Oregon tanoak forests

We followed the local intensification and dispersal of Phytophthora ramorum Werres, De Cock, & Man In’t Veld in Oregon tanoak ( Lithocarpus densiflorus (Hook & Arn.) Rehd.) forests from its initial detection in 2001 through 2006, coincident with a continuing eradication effort. The initial infested area included nine scattered sites below 400 m elevation, close to the Pacific Ocean near Brookings, Oregon. In subsequent years, one-half of new infections were within 122 m of a previous infection, and 79% of the newly detected trees occurred within 300 m of a previously identified tree. Dispersal up to 4 km was occasionally recorded. Initial infection occurred in the upper crowns of tanoak trees. The pathogen was recovered in rainwater collected beneath diseased tanoak trees in every month from November 2006 through October 2007. Twenty-four multilocus microsatellite genotypes were identified among 272 P. ramorum isolates collected from Curry County. Genotypic analysis provided independent estimates of time of origin of the Oregon infestation, its clustered distribution, and dispersal distances. In all sampling years, 60%–71% of the isolates belonged to the same multilocus genotype. In 2001, 12 genotypes were detected and new genotypes were identified in each of the subsequent years, but all isolates belonged to the same clonal lineage. Knowledge of local intensification of the disease and long-distance dispersal should inform both Oregon eradication efforts and national quarantine regulations.

Phytophthora ramorum Colonizes Tanoak Xylem and Is Associated with Reduced Stem Water Transport

Isolation, detection with diagnostic polymerase chain reaction (PCR), and microscopy demonstrated the presence of Phytophthora ramorum in the sapwood of mature, naturally infected tanoak (Lithocarpus densiflorus) trees. The pathogen was strongly associated with discolored sapwood (P < 0.001), and was recovered or detected from 83% of discolored sapwood tissue samples. Hyphae were abundant in the xylem vessels, ray parenchyma, and fiber tracheids. Chlamydospores were observed in the vessels. Studies of log inoculation indicated that P. ramorum readily colonized sapwood from inoculum placed in the bark, cambium, or sapwood. After 8 weeks, radial spread of P. ramorum in sapwood averaged 3.0 to 3.3 cm and axial spread averaged 12.4 to 18.8 cm. A field study was conducted to determine if trees with infected xylem had reduced sap flux and reduced specific conductivity relative to noninfected control trees. Sap flux was monitored with heat-diffusion sensors and tissue samples near the sensors were subsequently tested for P. ramorum. Adjacent wood sections were excised and specific conductivity measured. Both midday sap flux and specific conductivity were significantly reduced in infected trees versus noninfected control trees. Vessel diameter distributions did not differ significantly among the two treatments, but tyloses were more abundant in infected than in noninfected trees. Implications for pathogenesis, symptomology, and epidemiology are discussed.