Pulse crops (annual grain legumes such as field pea, lentil, dry bean, and chickpea) have become an important component of the cropping system in the northern Great Plains of North America over the last three decades. In many areas, the intensity of damping-off, seedling blight, root rot, and premature ripening of pulse crops is increasing, resulting in reduction in stand establishment and yield. This review provides a brief description of the important pathogens that make up the root rot complex and summarizes root rot management on pulses in the region. Initially, several specific Fusarium spp., a range of Pythium spp., and Rhizoctonia solani were identified as important components of the root rot disease complex. Molecular approaches have recently been used to identify the importance of Aphanomyces euteiches on pulses, and to demonstrate that year-to-year changes in precipitation and temperature have an important effect on pathogen prevalence. Progress has been made on management of root rot, but more IPM tools are required to provide effective disease management. Seed-treatment fungicides can reduce damping-off and seedling blight for many of the pathogens in this disease complex, but complex cocktails of active ingredients are required to protect seedlings from the pathogen complex present in most commercial fields. Partial resistance against many of the pathogens in the complex has been identified, but is not yet available in commercial cultivars. Cultural practices, especially diversified cropping rotations and early, shallow seeding, have been shown to have an important role in root rot management. Biocontrol agents may also have potential over the long term. Improved methods being developed to identify and quantify the pathogen inoculum in individual fields may help producers avoid high-risk fields and select IPM packages that enhance yield stability.

Clonostachys rosea f. catenulata (syn. Gliocladium catenulatum ) is an effective fungal biological agent against Fusarium root and stem rot and Pythium damping-off diseases on cucumber plants. Both chitinase and β-1,3-glucanase enzymes were produced when C. rosea was grown on a synthetic medium containing chitin or laminarin as a sole carbon source, respectively. Chitinase production was also induced by Fusarium cell walls, while β-1,3-glucanase activity was induced by both Fusarium and Pythium cell walls, as well as by growth on homogenized cucumber roots and on low-carbon media. Mycelial growth of Fusarium and Pythium, when exposed to C. rosea culture filtrates that contain glucanase activity, was significantly reduced compared with the controls, and cell walls of both pathogens were degraded. On excised cucumber roots, hyphae of C. rosea formed appressorium-like structures and coiled around hyphae of Pythium. In culture, C. rosea caused localized degradation of Fusarium hyphae. Cucumber root tissues colonized by C. rosea showed higher levels of β-1,3-glucanase activity at 7 days post-application compared with untreated controls. To determine if this activity was derived from C. rosea, glucanase isoforms were separated on activity gels. Fungal culture filtrates and root extracts contained the same predominant 20 kDa isoform. Reverse-transcription polymerase chain reaction (RT-PCR) using primers designed to amplify a β-1,3-glucanase gene in C. rosea confirmed glucanase expression on roots. These results show that C. rosea produces β-1,3-glucanase in situ, which can degrade hyphae of Fusarium and Pythium and contribute to biological control efficacy.

In recent years, root rots have severely impacted yields of field pea (Pisum sativum L.) in the Canadian province of Alberta. Above-normal precipitation levels in the springs of 2011 to 2013 led to the hypothesis that Aphanomyces euteiches Drechsler may play a role in root rot in water-saturated pea fields. To determine causal agent(s) of root rot, 145 pea fields were surveyed at flowering in July 2013 (1). Symptoms of root rot were abundant; the most prominent included red vascular streaking and dark brown rot of the tap root, indicative of Fusarium spp., but brown discoloration and cortical decay of lateral roots, indicative of A. euteiches, was also observed. Total genomic DNA was extracted from diseased root samples from each field, using the Qiagen DNeasy Plant kit, and amplified with species-specific primers for A. euteiches (2). Fusarium spp. were present in all fields, but seven fields located within a 200-km radius yielded a positive reaction for A. euteiches. Five fields were re-visited in May 2014 to collect soil for a bait test (3). Tests were performed using surface-sterilized pea seeds (cv. CDC Meadow) treated with Allegiance FL (Bayer, a.i. metalaxyl) at a rate of 110 ml/kg of seed. Five seeds per pot were planted into field soils in 10-cm pots with 12 replicate pots per field. Soils were irrigated as needed until the second-node stage and then kept at saturation for 14 days. Thirty day-old pea roots were evaluated for root rot symptoms; plated onto cornmeal agar amended with metalaxyl, benomyl, and vancomycin (MBV) without surface sterilization; and visualized microscopically for presence of oospores in the roots. Roots from three out of the five field soils showed symptoms typical of A. euteiches infection, including honey-brown discoloration, degradation of the root cortex, and presence of oospores. Root rot symptoms from the remaining fields were characteristic of Fusarium root rot, and oospores were not observed in roots. Fungal cultures with fast-growing, white, aerial mycelia characteristic of A. euteiches on MBV, were recovered from roots with Aphanomyces root rot symptoms, and transferred to PDA. To confirm pathogen identity, total DNA was extracted from 7-day-old cultures growing on PDA using the Qiagen DNeasy Plant Kit. The ribosomal DNA internal transcribed spacer (ITS) region was amplified using the primer pair ITS1 and ITS4 and sequenced (4). The sequences, deposited in GenBank with accession numbers KM486065, KM486066, and KM486067, were 100% identical to the ITS rDNA sequence of several isolates of A. euteiches using a BLASTn query. Fusarium spp. were also recovered from all root samples in the soil bait test. Total DNA extracted from roots was used in PCR assays with A. euteiches-specific primers as described above. PCR amplification of root DNA was successful only from the same three fields that showed Aphanomcyces root rot symptoms, further verifying presence of A. euteiches. The inability to detect or recover A. euteiches from two fields that had tested positive in the survey was likely due to patchy distribution of this pathogen and emphasizes the importance of rigorous soil collection methods to accurately detect pathogens. Although this is the first record of A. euteiches on field pea in Alberta, the distribution of A. euteiches within a 200-km radius in southern Alberta indicates that it has likely been present in soils for several years. The interaction between A. euteiches and Fusarium spp. infection in the root rot complex of field pea and their impact on field pea production in Alberta is currently being investigated. References: (1) S. Chatterton et al. Can. Plant Dis. Surv. 94:189, 2014. (2) C. Gangneux et al. Phytopathology 104:1138, 2014. (3) D. Malvick et al. Plant Dis. 78:361, 1994. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Soilborne fungal and oomycete pathogens are the causal agents of several important plant diseases. Infection frequently co-occurs with herbivory by root-feeding insects, facilitating tripartite interactions that modify plant performance and mortality. In an agricultural context, interactions between pathogens, herbivores and plants can have important consequences for yield protection. However, belowground interactions are inherently difficult to observe and are often overlooked. Here, we review the impact of direct and indirect interactions between root-associated insects, fungi, and oomycetes on the development of plant disease. We explore the relationship between insect feeding injury and pathogen infection, as well as the role of insects as vectors of fungal and oomycete pathogens. Synergistic interactions between insects and phytopathogens may be important in weed suppression, and we highlight several promising candidates for biocontrol. Bridging the gap between entomological and pathological research is a critical step in understanding how interactions between insects and microorganisms modify the community structure of the rhizosphere, and how this impacts plant functioning. Furthermore, the identification of belowground interactions is required to develop effective pest monitoring and management strategies.