Jennifer L. Parke

Live plant imports: the major pathway for forest insect and pathogen invasions of the US

Trade in live plants has been recognized worldwide as an important invasion pathway for non-native plant pests. Such pests can have severe economic and ecological consequences. Nearly 70% of damaging forest insects and pathogens established in the US between 1860 and 2006 most likely entered on imported live plants. The current regulation of plant imports is outdated and needs to balance the impacts of pest damage, the expense of mitigation efforts, and the benefits of live plant importation. To inform these discussions, we document large increases in the volume and value of plant imports over the past five decades and explain recent and proposed changes to plant import regulations. Two data sources were used to estimate the infestation rate of regulated pests in live plant shipments entering the US, thus allowing evaluation of the efficacy of the current port inspection process.

Susceptibility of Oregon Forest Trees and Shrubs to Phytophthora ramorum: A Comparison of Artificial Inoculation and Natural Infection

Phytophthora ramorum is an invasive pathogen in some mixed-hardwood forests in California and southwestern Oregon, where it causes sudden oak death (SOD) on some members of Fagaceae, ramorum shoot dieback on some members of Ericaceae and conifers, and ramorum leaf blight on diverse hosts. We compared symptoms of P. ramorum infection resulting from four different artificial inoculation techniques with the symptoms of natural infection on 49 western forest trees and shrubs; 80% proved susceptible to one degree or another. No single inoculation method predicted the full range of symptoms observed in the field, but whole plant dip came closest. Detached-leaf-dip inoculation provided a rapid assay and permitted a reasonable assessment of susceptibility to leaf blight. Both leaf age and inoculum dose affected detached-leaf assays. SOD and dieback hosts often developed limited leaf symptoms, although the pattern of midrib and petiole necrosis was distinctive. Stem-wound inoculation of seedlings correlated with field symptoms for several hosts. The results suggested that additional conifer species may be damaged in the field. Log inoculation provided a realistic test of susceptibility to SOD, but was cumbersome and subject to seasonal variability. Pacific rhododendron, salmonberry, cascara, and poison oak were confirmed as hosts by completing Kochs postulates. Douglas-fir was most susceptible to shoot dieback shortly after budburst, with infection occurring at the bud.

Standardizing the Nomenclature for Clonal Lineages of the Sudden Oak Death Pathogen, Phytophthora ramorum

Phytophthora ramorum, the causal agent of sudden oak death and ramorum blight, is known to exist as three distinct clonal lineages which can only be distinguished by performing molecular marker-based analyses. However, in the recent literature there exists no consensus on naming of these lineages. Here we propose a system for naming clonal lineages of P. ramorum based on a consensus established by the P. ramorum research community. Clonal lineages are named with a two letter identifier for the continent on which they were first found (e.g., NA = North America; EU = Europe) followed by a number indicating order of appearance. Clonal lineages known to date are designated NA1 (mating type: A2; distribution: North America; environment: forest and nurseries), NA2 (A2; North America; nurseries), and EU1 (predominantly A1, rarely A2; Europe and North America; nurseries and gardens). It is expected that novel lineages or new variants within the existing three clonal lineages could in time emerge.

Culture-Based and Non-Growth-Dependent Detection of the Burkholderia cepacia Complex in Soil Environments

ABSTRACT Burkholderia cepacia complex (Bcc) bacteria reside in soil, plant rhizospheres, and water, but their prevalence and distribution in outdoor environments is not clear. We sampled a variety of soil and rhizosphere environments with which people may have contact: playgrounds, athletic fields, parks, hiking trails, residential yards, and gardens. A total of 91 sites was sampled in three large U.S. cities. In the first phase of the study, putative Bcc isolates were recovered on Burkholderia cepacia selective agar and trypan blue tetracycline medium and subsequently examined for biochemical reactivity and growth at 32 and 22°C. Isolates were further examined by PCR assays targeting Bcc-specific ribosomal DNA and recA gene sequences. Among the 1,013 bacterial isolates examined, 68 were identified as Bcc; 14 (15%) of 91 sampled sites yielded Bcc isolates. In the second phase, DNA was extracted directly from soil samples and examined with PCR assays targeting Bcc 16S rRNA gene sequences. Either 82 or 93% of the soil samples were positive for at least one Bcc genomovar, depending on the PCR assay system used. Cloning and sequencing were performed to check the specificity of the PCR assays. Sequence analysis of the 463-bp 16S rRNA inserts from eight clones indicated that all were from members of the Bcc. The four soil samples from which these clones were generated did not yield isolates identified as Bcc. Based on PCR detection, Bcc appears to be prevalent in soil from urban and suburban environments. Culture-based recovery of Bcc may underestimate environmental populations.

Phytophthora ramorum colonizes tanoak xylem and is associated with reduced stem water transport

ABSTRACT 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.

A Systems Approach for Management of Pests and Pathogens of Nursery Crops

Horticultural nurseries are heterogeneous and spatially complex agricultural systems, which present formidable challenges to management of diseases and pests. Moreover, nursery plants shipped interstate and internationally can serve as important vectors for pathogens and pests that threaten both agriculture and forestry. Current regulatory strategies to prevent this movement of pathogens and pests with nursery plants are based on visual inspections of plants just before shipping, a process that is costly and inadequate. Here we propose the application of a systems approach for horticultural nurseries modeled after the Hazard Analysis of Critical Control Points (HACCP) approach widely used in the food processing industry. We evaluated aspects of the systems approach to analyze contamination hazards by Phytophthora species and then implemented management practices targeting specific critical control points. The systems approach for analyzing and correcting unsafe practices offers a potential alternative strategy for preventing plant contamination that could be broadly applied to many pests and pathogens. One of the hallmarks of this approach is its flexibility, including the ability to improve and adapt the approaches as new technologies are developed or improved and to provide the grower with management options.

Effects of Genetic Differences among Crop Species and Cultivars Upon the Arbuscular Mycorrhizal Symbiosis

Crop species and cultivars within species can differ markedly in their ability to respond to colonization by arbuscular mycorrhizal (AM) fungi. The variation in response to mycorrhizas has largely been overlooked by plant breeders, even when germplasm is screened for efficiency of nutrient uptake and efficiency of nutrient use, processes influenced greatly by mycorrhizal fungi. Host germplasm is frequently selected under conditions that would not favor, and indeed might prevent, a response to mycorrhizal fungi, i.e. high nutrient availability. Continued breeding of crop plants without regard to mycorrhiza responsiveness is irresponsible, as it could result not only in increased requirement for nutrient inputs in the short term, but also the inadvertent, permanent loss of these genes in crop germplasm. Loss of colonization by AM fungi could also result in the loss of other important benefits provided by mycorrhizal fungi (drought tolerance, resistance to disease, uptake of other nutrients, maintaining soil structure) and reduce the population of AM fungi able to colonize other hosts in the cropping system. Plant breeders should determine the contribution of mycorrhizal fungi to nutrient uptake by evaluating plant growth with and without mycorrhizas over a full range of nutrient levels; i. e. develop response curves for the variable of interest. Ultimately the goal should be to map genes responsible for mycorrhizal colonization and responsiveness, and to utilize them in developing cultivars which can more effectively acquire nutrients from input low input agricultural systems and take advantage of the other benefits provided by the mycorrhizal symbiosis.

Phytophthora Community Structure Analyses in Oregon Nurseries Inform Systems Approaches to Disease Management

Nursery plants are important vectors for plant pathogens. Understanding what pathogens occur in nurseries in different production stages can be useful to the development of integrated systems approaches. Four horticultural nurseries in Oregon were sampled every 2 months for 4 years to determine the identity and community structure of Phytophthora spp. associated with different sources and stages in the nursery production cycle. Plants, potting media, used containers, water, greenhouse soil, and container yard substrates were systematically sampled from propagation to the field. From 674 Phytophthora isolates recovered, 28 different species or taxa were identified. The most commonly isolated species from plants were Phytophthora plurivora (33%), P. cinnamomi (26%), P. syringae (19%), and P. citrophthora (11%). From soil and gravel substrates, P. plurivora accounted for 25% of the isolates, with P. taxon Pgchlamydo, P. cryptogea, and P. cinnamomi accounting for 18, 17, and 15%, respectively. Five species (P. plurivora, P. syringae, P. taxon Pgchlamydo, P. gonapodyides, and P. cryptogea) were found in all nurseries. The greatest diversity of taxa occurred in irrigation water reservoirs (20 taxa), with the majority of isolates belonging to internal transcribed spacer clade 6, typically including aquatic opportunists. Nurseries differed in composition of Phytophthora communities across years, seasons, and source within the nursery. These findings suggest likely contamination hazards and target critical control points for management of Phytophthora disease using a systems approach.

Effects of Soil Solarization and Trichoderma asperellum on Soilborne Inoculum of Phytophthora ramorum and Phytophthora pini in Container Nurseries

Infested container nursery beds are an important source of soilborne Phytophthora spp. for initiating disease through movement with surface water or splashing onto foliage. We investigated the effects of soil solarization, alone or with subsequent amendment with a Trichoderma asperellum biocontrol agent, on the survival of Phytophthora spp. inoculum. In field trials conducted with Phytophthora ramorum in San Rafael, CA and with P. pini in Corvallis, OR, infested rhododendron leaf inoculum was buried at 5, 15, and 30 cm below the soil surface. Solarization for 2 or 4 weeks during summer 2012 eliminated recovery of Phytophthora spp. buried at all depths in California trial 1, at 5 and 15 cm in California trial 2, but only at 5 cm in Oregon. There was no significant reduction of Phytophthora spp. recovery after T. asperellum application. Although the population densities of the introduced T. asperellum at the 5-cm depth were often two- to fourfold higher in solarized compared with nonsolarized plots, they were not significantly different (P = 0.052). Soil solarization appears to be a promising technique for disinfesting the upper layer of soil in container nurseries under certain conditions.