Phytophthora root rot, caused by many Phytophthora species, decreases the health of rhododendrons produced in nurseries. Optimizing nitrogen (N) fertilizer is often used to improve nursery stock quality, but there is little information on how N fertilizers influence root rot caused by these pathogens. To understand the impact of N fertilizer and pathogen species on root rot development, rhododendrons were grown with no (0 g N/pot), low (1.04 g N/pot) or high (3.12 g N/pot) rates of N and inoculated with either P. cinnamomi or P. plurivora. Noninoculated plants at low and high N rates had greater biomass, leaf greenness and enhanced N, potassium, magnesium, phosphorus, sulphur and manganese uptake compared to plants grown with no N. When either Phytophthora species was present, N application increased aboveground disease symptoms (wilting, chlorosis, reduced stomatal conductance and biomass), but had no effect on root rot severity belowground. In addition, P. cinnamomi restricted uptake of several nutrients while P. plurivora had less influence on nutrient uptake. Nurseries frequently apply high amounts of N to promote fast growth. However, our results show that this can exacerbate root rot when P. cinnamomi or P. plurivora is present. Although decreasing N can reduce the number of overtly symptomatic plants, this may conversely increase the risk for selling apparently asymptomatic plants with low levels of infection. Additional studies are needed to determine how N fertilization influences Phytophthora root rot for a broader range of rhododendron cultivars and nursery crop species.

The degree of flooding commonly used to induce disease in Phytophthora root rot studies rarely occurs in container nurseries. Instead, over-irrigation and poor drainage result in plants periodically sitting in shallow pools of water. Rhododendron plants were grown in a noninfested substrate or substrate infested with Phytophthora cinnamomi or P. plurivora to determine whether root rot induced by flooding represents disease that occurs under simulated nursery conditions when plants are in a shallow pool of water (saucers), or are allowed to freely drain and maintained at ∼75% container capacity (CC). Generally P. cinnamomi caused more disease than P. plurivora, and all water treatments were conducive to root rot. In experiment 1, the amount of disease caused by flooding was similar to that in the saucer treatment (75% CC not tested) while in experiment 2, flooding often caused more rapid and severe disease than the saucer or 75% CC treatment. Pathogens differed in their response to water treatments. P. cinnamomi caused more disease in treatments with >90% substrate moisture for either a short (flood) or long duration (saucer), while P. plurivora was less capable of causing disease when soil moisture was maintained >90% than when substrate moisture was maintained at a more moderate level (flood, 75% CC). Our results indicate that it is not necessary to flood plants to induce disease under experimental conditions and that disease induced by flooding can represent disease in container nurseries when containers are in pools of water or maintained at ∼75% CC. In addition, our results suggest that P. cinnamomi is a more aggressive pathogen than P. plurivora in nursery conditions where drainage is poor; however, both species are capable of causing a similar amount of disease under more typical irrigation management.

We evaluated whether reducing irrigation frequency and volume alters the ability of Phytophthora plurivora and P. cinnamomi to cause root rot on rhododendron grown in a noninfested potting medium or media infested with 1 or 100 propagules per gram (ppg) of pathogen. Plants were irrigated to maintain a substrate moisture of >70% container capacity (1.0X), one-half volume of 1.0X (0.5X), or two times the volume of 1.0X at each irrigation event for one week, followed by no irrigation, until soil moisture reached <50% container capacity. Aboveground disease symptoms (chlorosis, stomatal conductance, wilting, and plant death) were evaluated weekly and root rot, pathogen presence, plant biomass, and nutrient uptake were measured at the end of each trial. Both pathogens generally caused mild disease at 1 ppg and severe disease at 100 ppg. Reducing irrigation did little to lessen disease caused by either pathogen once infection had occurred. Instead, severe root infection often led to increased soil moisture and root rot across all irrigation treatments as roots became progressively compromised in their ability to take up water. Results show that reducing irrigation after infection has occurred is unlikely to effectively control root rot. Species used in this study: Phytophthora species (Phytophthora cinnamomi Rands; Phytophthora plurivora T. Jung and T.I. Burgess); rhododendron, Rhododendron catawbiense Michx. ‘Album', ‘Roseum Elegans', and ‘Roseum Pink'.

Controlled environment experiments were conducted to evaluate the effects of temperature on Calonectria pseudonaviculata mycelial growth and the effects of temperature and infection period on boxwood blight severity. In experiment 1, 15 Oregon isolates (representing five genotypes) were grown on potato dextrose agar (PDA) and malt extract agar (MEA) at six temperatures from 5 to 30°C. Growth (culture diameter) was measured after 2 weeks. Optimal growth occurred at 25°C on PDA and 20°C on MEA. Isolates of genotype G1 also grew faster than genotype G2, but only on MEA at 25°C. In experiment 2, Buxus cultivars Green Velvet (GV, more susceptible) and Winter Gem (WG, more resistant) were inoculated and incubated in moist chambers for 9 or 24 h at 22°C (infection period), then moved into growth chambers at 15 or 25°C. After 4 weeks, chamber temperatures were switched, and plants were incubated for 4 more weeks. Disease severity was evaluated weekly. During the first 4 weeks, disease was generally more severe on GV than WG, on plants with a 24-h versus a 9-h infection period, and on plants incubated at 15°C versus 25°C. However, disease was just as severe on WG as GV when the 24-h infection period was followed by incubation at 15°C. After the temperatures were switched, disease increased only on WG that were cooled from 25 to 15°C. Results show that Oregon isolates of C. pseudonaviculata are capable of growing faster and causing more severe disease at temperatures cooler than those reported previously.

Phytophthora root rot, caused by many soilborne Phytophthora spp., is a significant disease affecting the $42 million rhododendron nursery industry. Rhododendron growers have increasingly reported failure by two systemic fungicides, phosphorous acid and mefenoxam, to adequately control root rot. Both fungicides may be applied as a foliar spray or soil drench but it is unknown how application method, fungicide chemistry, or pathogen diversity affects disease control. Therefore, two experiments were conducted to (i) determine whether differences in application method or fungicide chemistry affect control of root rot caused by P. cinnamomi and P. plurivora and (ii) evaluate the sensitivity of Phytophthora spp. and isolates from the rhododendron industry to each fungicide. Results demonstrated that soil drenches of either fungicide were more effective than foliar sprays for control of P. cinnamomi but were ineffective for P. plurivora. Furthermore, Phytophthora spp. and isolates varied in sensitivity to phosphorous acid and mefenoxam, and there were multiple fungicide-insensitive isolates, especially within P. plurivora. Differences in sensitivity were also observed among isolates from different nurseries and production systems, with some nurseries having less sensitive isolates than others and with container systems generally having less sensitive isolates than field systems. Our results provide three potential reasons for why fungicide control of Phytophthora root rot might fail: (i) the fungicide can be applied to the wrong portion of the plant for optimal control, (ii) there are differences in fungicide sensitivity among soilborne Phytophthora spp. and isolates infecting rhododendron, and (iii) fungicide-insensitive isolates are present in the rhododendron nursery industry.

Inoculum production is an important part of conducting research with soilborne Phytophthora species. One common method is to incubate Phytophthora cultures in nutrient-amended vermiculite. However, inoculum levels often vary among batches of inoculum, even when production methods remain the same, and incubation typically takes ≥6 weeks, increasing risks for delayed experiments if the resulting inoculum level is too low. A more reliable and rapid method is needed for future studies. Experiments were conducted to determine inoculum levels of Phytophthora cinnamomi and Phytophthora plurivora after incubation in V8 juice-amended vermiculite (standard method); evaluate how inoculum viability was affected by air-drying; develop a modified method that takes less time to produce a vermiculite-based inoculum; and evaluate the effect of storage on inoculum viability. Results showed that the standard method produced inoculum levels from 716 to 1,808 colony forming units (CFUs)/g and that drying to <78% moisture content significantly reduced viability. The modified method used 2-week-old Phytophthora cultures to infest vermiculite at 80% moisture content and produced inoculum levels from 214 to 525 CFU/g. Storage for >1 day generally reduced inoculum viability. Although inoculum levels from the modified method were lower than the standard method, inoculum levels for each isolate were more consistent between trials, and the modified method was 6 to 8 weeks faster. Production with the modified method can also be easily scaled up by infesting a greater volume of vermiculite with additional cultures of Phytophthora. These results are important because they help explain variability in soilborne Phytophthora inoculum production and storage and provide a new method for producing inoculum more quickly.

Rhododendron root rot is a severe disease that causes significant mortality in rhododendrons. Information is needed about the incidence and identity of soilborne Phytophthora and Pythium species causing root rot in Pacific Northwest nurseries in order to better understand the disease etiology and to optimize disease control strategies. The last survey focusing solely on soilborne oomycete pathogens in rhododendron production was conducted in 1974. Since then, advances in pathogen identification have occurred, new species may have been introduced, pathogen communities may have shifted, and little is known about Pythium species affecting this crop. Therefore, a survey of root-infecting Phytophthora and Pythium species was conducted at seven nurseries from 2013 to 2017 to (i) document the incidence of root rot damage at each nursery and stage of production, (ii) identify soilborne oomycetes infecting rhododendron, and (iii) determine whether there are differences in pathogen diversity among nurseries and production systems. Rhododendrons from propagation, container, and field systems were sampled and Phytophthora and Pythium species were isolated from the roots and collar region. Root rot was rarely evident in propagation systems, which were dominated by Pythium species. However, severe root rot was much more common in container and field systems where the genus Phytophthora was also more prevalent, suggesting that Phytophthora species are the primary cause of severe root rot and that most contamination by these pathogens comes in after the propagation stage. In total, 20 Pythium species and 11 Phytophthora species were identified. Pythium cryptoirregulare, Pythium aff. macrosporum, Phytophthora plurivora, and Phytophthora cinnamomi were the most frequently isolated species and the results showed that Phytophthora plurivora has become much more common than in the past. Phytophthora diversity was also greater in field systems than in propagation or container systems. Risks for Phytophthora contamination were commonly observed during the survey and included placement of potting media in direct contact with field soil, the presence of dead plants that could serve as continuous sources of inoculum, and the presence of excess water as a result of poor drainage, overirrigation, or malfunctioning irrigation equipment. In the past, research on disease development and root rot disease control in rhododendron focused almost exclusively on Phytophthora cinnamomi. More research is needed on both of these topics for the other root-infecting species identified in this survey.

Rhododendrons are an important crop in the ornamental nursery industry, but are prone to Phytophthora root rot. Phytophthora root rot is a continuing issue on rhododendrons despite decades of research. Several Phytophthora species are known to cause root rot, but most research has focused on P. cinnamomi, and comparative information on pathogenicity is limited for other commonly encountered oomycetes, including Phytophthora plurivora and Pythium cryptoirregulare. In this study, three isolates each of P. cinnamomi, P. plurivora, and Py. cryptoirregulare were used to inoculate rhododendron cultivars Cunningham’s White and Yaku Princess at two different inoculum levels. All three species caused disease, especially at the higher inoculum level. P. cinnamomi and P. plurivora were the most aggressive pathogens, causing severe root rot, whereas Py. cryptoirregulare was a weak pathogen that only caused mild disease. Within each pathogen species, isolate had no influence on disease. Both P. cinnamomi and P. plurivora caused more severe disease on Cunningham’s White than on Yaku Princess, suggesting that the relative resistance and susceptibility among rhododendron cultivars might be similar for both pathogens. Reisolation of P. cinnamomi and P. plurivora was also greater from plants exhibiting aboveground symptoms of wilting and plant death and belowground symptoms of root rot than from those without symptoms. Results show that both P. cinnamomi and P. plurivora, but not Py. cryptoirregulare, are important pathogens causing severe root rot in rhododendron. This study establishes the risks for disease resulting from low and high levels of inoculum for each pathogen. Further research is needed to evaluate longer term risks associated with low inoculum levels on rhododendron health and to explore whether differences among pathogen species affect disease control.

Sixty percent of the $109 million processed red raspberry industry of the United States occurs in northern Washington State. In 2012, late-summer symptoms of vascular wilt and root disease were observed in many raspberry plantings. These symptoms were initially attributed to Verticillium dahliae. However, diagnostic tests for the pathogen were often contradictory and other soilborne pathogens (Phytophthora rubi and Pratylenchus penetrans) or Raspberry bushy dwarf virus (RBDV) might also have been involved. Therefore, a survey was conducted in 2013 and 2014 to (i) establish the incidence and soil population levels of V. dahliae in red raspberry production fields, (ii) compare among diagnostic methods and laboratories for detecting and quantifying V. dahliae from raspberry field soil, and (iii) assess which pathogens are associated with late-summer disease symptoms of raspberry. Plant and soil samples were collected from 51 disease sites and 20 healthy sites located in 24 production fields. Samples were analyzed for the presence and quantity of each pathogen using traditional plating and extraction methods (V. dahliae, P. rubi, and P. penetrans), quantitative polymerase chain reaction (qPCR) (V. dahliae and P. rubi), and enzyme-linked immunosorbent assay (RBDV). Results showed that V. dahliae was present in 88% of the production fields and that detection of the pathogen differed by method and by laboratory: qPCR detected V. dahliae in the soil from approximately three times as many sites (51 of 71 total sites) as by plating on NP10 semi-selective medium (15 of 71 total sites). Soil populations of V. dahliae were slightly greater at disease sites, but the pathogen was detected with similar frequency from healthy sites and it was rarely isolated from diseased plants (4%). P. rubi, P. penetrans, and RBDV were also common in production fields (79, 91, and 53% of fields, respectively). Both P. rubi (soil and root samples) and P. penetrans (root populations only), but not RBDV, were more frequently found at disease sites than healthy sites, and the amount of P. rubi detected by qPCR was greater from disease sites than healthy sites. In addition, P. rubi was isolated from 27% of the symptomatic plants located at disease sites. Regardless of detection method, V. dahliae, P. rubi, and P. penetrans, either with or without RBDV, were more likely to co-occur at disease sites (73%) than healthy sites (35%), suggesting that a soilborne disease complex is present in raspberry production fields. Results indicate that P. rubi is the primary pathogen most strongly associated with late-summer symptoms of disease, but root populations of P. penetrans and higher soil populations of V. dahliae may also be of concern. Therefore, disease control methods should focus on all three soilborne pathogens.

Preplant soil fumigation is commonly used to control soilborne pathogens and weeds in forest seedling nurseries of Oregon and Washington. However, lower chemical inputs are desired to meet state and federal application regulations, to minimize buffer zone size requirements, and to help protect the environment. Therefore, the objectives of this research were to evaluate the efficacy of three reduced rate soil fumigants under totally impermeable film (TIF) in managing soilborne diseases and weeds, and to determine if combined applications of up to four biocontrol agents improved soilborne disease control. Reduced rates of methyl bromide, metam sodium, and 1,3-dichloropropene, all applied in combination with chloropicrin, were effective in decreasing soil populations of Pythium and Fusarium as well as the presence of Pythium in root debris from the previous crop. The roots of Douglas-fir (Pseudotsuga menziesii) seedlings transplanted into each fumigant treatment were also colonized less by Pythium and Fusarium than those transplanted into nonfumigated control plots. However, biocontrol treatments were not effective against either pathogen. Weed biomass and weeding times were also significantly reduced by fumigation. Application costs were similar for all three fumigant treatments, but seedling size was largest from the methyl bromide and metam sodium treatments followed by the 1,3-dichloropropene treatment. Based on the results of this study, reduced rates of methyl bromide, metam sodium, and 1,3-dichloropropene show promise in managing soilborne diseases and weeds in forest nurseries.

Pythium species are common soilborne oomycetes that occur in forest nursery soils throughout the United States. Numerous species have been described from nursery soils. However, with the exception of P. aphanidermatum, P irregulare, P. sylvaticum, and P ultimum, little is known about the potential for other Pythium species found in nursery soils to cause damping-off of tree seedlings. A greenhouse study was conducted to evaluate the pathogenicity and virulence of 44 Pythium isolates representing 16 species that were originally recovered from soil at three forest nurseries in Washington and Oregon. Seeds of Douglas-fir (Pseudotsuga menziesii) were planted into soil infested with each of the isolates. Seedling survival, the number of surviving seedlings with necrotic root lesions, and taproot length were evaluated 4 weeks later. Responses of Douglas-fir to inoculation varied significantly depending on Pythium species and isolate. Eight species (P dissotocum, P. irregulare, P aff. macrosporum, P. mamillatum, P aff oopapillum, P rostratifingens, P. sylvaticum, and P. ultimum var. ultimum) significantly reduced the number of surviving seedlings compared to the noninoculated treatment. However, all Pythium species caused a greater percentage of seedlings to develop root lesions (total mean 40%) than was observed from noninoculated seedlings (17%). Taproot length varied little among Pythium treatments and was not a useful character for evaluating pathogenicity. Results confirm the ability of P. irregulare, P. mamillatum, and P ultimum var. ultimum to cause damping-off of Douglas-fir seedlings, and are indicative that other species such as P. dissotocum, P. aff macrosporum, P. aff oopapillum, P rostratifingens, and P. sylvaticum may also be responsible for seedling loss.

Five fumigation treatments (a conventional methyl bromide – chloropicrin application and four reduced-rate alternative fumigant treatments) and a nonfumigated treatment were evaluated at two forest nurseries in Oregon and one forest nursery in Washington for their effects on soil pathogen populations, weeds, and seedling morphology during a 2-year study. The effect of plastic tarp composition on fumigant efficacy was also evaluated (virtually impermeable film versus high-density polyethylene). All fumigant treatments reduced soil populations of Fusarium and Pythium for up to 7 months after fumigation and resulted in seedlings with significantly less pathogen colonization than those from the nonfumigated treatment. All fumigant treatments were more effective against pathogen inoculum buried at 15 cm rather than at 30 cm. Fusarium commune Skovgaard, O’Donnell et Nirenberg, Fusarium oxysporum Schlect. emend. Snyder & Hansen, isolates from the Gibberella fujikuroi Saw. complex, Pythium irregulare Buisman, Pythium aff. spiculum B. Paul 2006, Pythium sylvaticum Campbell & Hendrix, and Pythium ‘vipa’ Hermansen & Klemsdal were the most commonly isolated pathogens. Weed biomass and weeding times were significantly reduced by fumigation, but only at the Washington nursery with high weed pressure. No significant differences were observed in efficacy between the conventional methyl bromide – chloropicrin treatment and any of the reduced rate fumigants or between the two types of plastic tarp. Conifer seedling height, diameter, shoot volume, and root volume were significantly greater in all fumigated treatments compared with the nonfumigated treatment.

Douglas-fir seedlings are susceptible to Fusarium or Pythium damping-off that currently is controlled by pre-plant soil fumigation in bareroot nurseries and steam pasteurization or chemical drenches of soilless media in container nurseries. However, because few chemical or biological agents are registered for use on conifer seedlings, we tested several on greenhouse-grown seedlings and found that over-seed applications of Cleary's 3336, Strike, Compass, Compass + Strike, Cygnus, Endura, Medallion, Medallion + Strike, Thiram, and Enzone effectively suppressed pre-emergence damping-off by Fusarium oxysporum, but only Cleary's 3336WP and Medallion + Strike also reduced post-emergence damping-off. Compass, Medallion, and Thiram reduced post-emergence damping-off, but not to a statistically significant level. Pre-emergence damping-off by Pythium irregulare was reduced only by Ranman, but post-emergence damping-off was reduced by Thiram, Hurricane, Ranman, and Subdue MAXX. Over-seed drenches of biological control agents Companion, Kodiak, Subtilex, Taegro, Primastop, SoilGard, Actinovate, Mycostop, RootShield, and Green-Releaf were ineffective in suppressing either Fusarium or Pythium diseases, and combining several agents with chemicals did not improve efficacy. These results provide data in support of registration of some of the effective chemicals, but no biological control agents, for the control of conifer seedling damping-off. Accepted for publication 14 January 2008. Published 17 March 2008.

Phytophthora diseases occur frequently in nurseries, and the recent incidence of ramorum blight, caused by P. ramorum, on nursery crops has underscored the need for improved management strategies against all Phytophthora diseases. We evaluated several chemicals that target Oomycete pathogens, inoculating detached rhododendron or lilac leaves removed from plants previously treated with various chemicals, or chemically-treated leaves on intact plants. Results indicated that Subdue MAXX (drench or foliar application) was the most effective chemical in suppressing infections caused by all species of Phytophthora tested (P. ramorum, P. citricola, P. citrophthora, and P. nicotianae) except P. citrophthora; with P. ramorum, it was active for at least 6 weeks after spray application. More chemicals were effective when sporangial rather than mycelial plug inoculum of P. ramorum was used, including Aliette, Ranman, Stature DM, and Fenamidone. All chemicals tested were fungistatic, not fungicidal. These tests indicate that several materials inhibit infection by Phytophthora species, and that the detached leaf test is effective in evaluating efficacy of chemical agents for the suppression of Phytophthora pathogens from nurseries. Accepted for publication 10 November 2007. Published 11 February 2008.

Phytophthora ramorum, causal agent of ramorum blight of woody shrub species, has caused serious damage to cultivars and species of camellia in commercial nurseries. Reports of relative susceptibility of camellia to P. ramorum have indicated a range from high to low susceptibility, both in nurseries and under experimental conditions. We inoculated a series of cultivars of camellia to determine their relative susceptibility to infection, and then compared lesion size to the capacity of the pathogen to produce sporangia on the lesions. We found, as did others, a wide range of susceptibility among cultivars, but lack of correlation between susceptibility (lesion size) and potential to produce sporangia that might spread the pathogen within the nursery. These results indicate that on some cultivars the pathogen might produce small or inconspicuous lesions, yet still produce copious numbers of sporangia that could spread the disease, both within the nursery and from nursery to nursery. Accepted for publication 10 April 2007. Published 22 August 2007.

Phytophthora ramorum, cause of Ramorum blight on numerous woody ornamental shrubs, is a regulated pathogen in the US and internationally. Currently, nurseries are inspected to detect infected plants; however, many plants are propagated by tissue culture nurseries and the behavior of P. ramorum in this system is unknown. Pathogen growth and sporulation in propagation vessels containing different multiplication and rooting media, with a range of plants and without plants, was evaluated with regard to pathogen visibility and induction of disease symptoms. Within 2 weeks, the pathogen colonies were visible to the naked eye on all 26 multiplication media and on 9 of 11 rooting media tested (without plants). The appearance of colonies on different media was variable and no sporangia but occasional chlamydospores were produced. The pathogen growth was very visible on multiplication media containing susceptible plants, inoculated plants exhibiting obvious discoloration and mortality. The pathogen was reisolated from terminal shoot tissue and roots of symptomatic plants. Variability occurred in susceptibility of different cultivars of a plant species, in virulence of the two isolates of the pathogen, and in recovery from shoot tissue. We conclude that fungal growth on the media, with or without plants, and symptoms of disease were apparent enough that contaminated vessels would be destroyed. Accepted for publication 9 April 2007. Published 22 August 2007.

Phytophthora ramorum, cause of sudden oak death of trees or ramorum blight of other plant species, has an ever-increasing host range. Some geographic regions are considered to be at high risk of becoming infested with the pathogen, possibly causing plant mortality such as seen in native habitats of California and Oregon. One such region is the Appalachian range of the eastern United States, where known susceptible plants occur and climatic characteristics appear favorable for infections by this pathogen. We collected foliage of a range of plant species native to Appalachia in Kentucky during two summer seasons, and the foliage was shipped to Oregon for inoculation with P. ramorum to determine relative susceptibility. Leaves were needle-wounded and inoculated with either mycelium agar plugs or sporangia of a North American (A2 mating type) or European (A1 mating type) isolate. After 14 days incubation at 20°C in moist boxes, lesions caused by either inoculum type or isolate generally were comparable using digital photos and ASSESS software. Some genera, species, and cultivars within species were highly susceptible, while others were moderately susceptible or not susceptible. These results provide a basis for regional surveyors to select target hosts and to generate survey and management practices for nursery and forest areas. Accepted for publication 24 April 2007. Published 17 September 2007.