Identification of metalaxyl and ethaboxam insensitive Pythium sylvaticum pathogenic to pulse crops in Montana, USA.

Pythium root rot and damping-off caused by Pythium spp. are important diseases of pulse crops. In a 2016 pathogen survey from dry pea growing fields in Montana, along with commonly known causal agents P. ultimum and P. irregulare, an isolate identified as P. sylvaticum (LPPY17) was isolated from the rhizosphere of a diseased pea plant collected from Valley County, MT. Root rots and damping-off caused by P. sylvaticum have not previously been reported for chickpea, pea, and lentil crops. The isolate LPPY17 was tested for fungicide resistance in vitro, and results indicated a reduced sensitivity to metalaxyl and ethaboxam containing fungicides. LPPY17 was also tested for pathogenicity on chickpea, pea, and lentil seedlings in the greenhouse, and the results from the study revealed LPPY17 is capable of causing both root rots and damping off. Due to the potential pathogenicity and reduced fungicide sensitivity of this species, in the future it will be important to monitor for P. sylvaticum in pulse root rot surveys and diagnostics, as management options may be different from other common Pythium spp.

Resistance of the SoyNAM Parents to Seed and Root Rot Caused by Four Pythium Species

Some Pythium spp. cause damping off and root rot in soybeans and other crop species. One of the most effective management tools to reduce disease is host resistance; however, little is known about resistance in soybean to Pythium spp. The soybean nested associated mapping (SoyNAM) parent lines are a set of germplasms that were crossed to a single hub parent to create recombinant inbred line populations for the purpose of mapping agronomic traits. The SoyNAM parents were screened for resistance to Pythium lutarium, Pythium oopapillum, Pythium sylvaticum, and Pythium torulosum in separate assays to evaluate seed and root rot severity. Of the 40 SoyNAM parents, only ‘Maverick’ was resistant to the four species tested; however, 13 were resistant to three species. Other lines were resistant to two, one, or none of the species tested. Correlations between seed and root rot severity for the lines assessed were weak or insignificant. Results indicate that mechanisms of resistance to seed and root rot caused by Pythium spp. may not necessarily be the same.

The Effect of Cold Stress on Damping-Off of Soybean Caused by Pythium sylvaticum

To improve our understanding of the timing of cold stress and its effect on Pythium damping-off, we performed a factorial experiment with two cold stress temperatures (4 and 10°C); exposure to 96 h of cold stress at 0, 1, 2, 4, 6, and 8 days after planting; and inoculation with Pythium sylvaticum-infested millet or control. Increased susceptibility to damping-off resulting in reduced emergence was found in inoculated plants when the cold stress period began 2 or 4 days after planting. In the noninoculated controls, no effect of cold stress on emergence was observed. Slower seedling growth was observed during the cold stress period and in inoculated plants after exposure to cold stress. Seed exudation, mycelial growth, and sporangia germination of P. sylvaticum was evaluated at 4, 10, and 18°C. The greatest seed exudation was observed at 4°C. Low temperatures delayed mycelial growth of P. sylvaticum, although the pathogen was still able to grow at 4°C. Sporangia incubated for 3 h at 18°C in the presence of seed exudates had higher germination in comparison with sporangia incubated at 10 or 4°C. Moreover, more sporangia germinated in response to seed exudates that were previously collected from seed imbibed for 24 h at low temperatures (4°C). These results suggest that cold stress 2 to 4 days after planting increases soybean susceptibility to damping-off, presumably because of increased seed exudation and delayed seedling growth.

Seed Rot and Damping-off of Alfalfa in Minnesota Caused by Pythium and Fusarium Species

Globally, 15 Pythium species have been found to cause damping-off and seed rot of alfalfa, although surveys of species causing disease on alfalfa in the midwestern United States are lacking. Pathogens were isolated by a seedling baiting technique from soil samples of five alfalfa fields in Minnesota with high levels of damping-off. Of the 149 organisms isolated, 93 (62%) were identified as Pythium spp. and 43 (29%) were identified as Fusarium species. Pythium sylvaticum, P. irregulare, and P. ultimum var. ultimum were aggressive pathogens on germinating alfalfa seedlings. Strains of seven Pythium spp. pathogenic on soybean and corn were also pathogenic on alfalfa. The majority of the Fusarium isolates were identified as F. solani and F. oxysporum with a low number of F. redolens and F. incarnatum-equiseti. The F. oxysporum and F. incarnatum-equiseti strains were the most aggressive in causing seed and root rot. Pythium strains were sensitive to Apron XL (mefenoxam) and pyraclostrobin in vitro but efficacy varied when the fungicides were applied as a seed treatment. Seed treatments with Apron XL were more effective than treatments with Stamina against Pythium. The presence of aggressive, broad-host-range pathogens causing seed rot and damping-off suggests that new strategies are needed for managing this disease in alfalfa production systems.

First Report of Basal Stem Rot and Foliar Blight Caused by Pythium sylvaticum on Miscanthus sinensis in Illinois

Miscanthus sinensis Anderss., a perennial grass, is native to eastern Asia. It has been widely grown as an ornamental in temperate regions of the world, including the United States, and recently has become an important component of public and private sector bioenergy feedstock Miscanthus selection programs. In August 2008, stem rot and blight was observed on M. sinensis plants in two irregular patches, ~2 to 2.5 × 1 to 1.5 m each in a trial plot that was preceded by corn, at the University of Illinois Energy Farm near Urbana, IL. At the time of the observation, most plants were dead and the wilted tillers had black, soft rotted basal stems. A few plants were stunted and the crowns of the tillers had black-to-brown soft rot. Some tillers' leaves were dead and others had turned light brown. Sample tissue fragments were surface disinfested in 0.5% NaOCl and plated on 1% water agar (WA). After 3 days of incubation in the dark at 23°C, colonies were transferred to corn meal agar (CMA), potato dextrose agar (PDA), or 10% V8 juice agar and incubated at 23°C under continuous white light for up to 2 weeks. Morphological characteristics of the isolates correspond to those originally described for Pythium sylvaticum W.A. Campb. & J.W. Hendrix (1). The mycelia grew and covered the 10-cm-diameter plates within 5 days. On PDA, the culture was a creamy white mycelial mat of coenocytic hyphae. The isolates produced only globose, terminal or intercalary hyphal swellings ranging from 28 to 48 μm in diameter, but no oogonia were produced on any of the three growth media. No zoospores were produced when agar blocks bearing mycelium were flooded with distilled water or 1% soil water. Sequence analysis was performed with the internal transcribed spacer (ITS) region of the rDNA amplified with primer pair ITS1/ITS4 (3) and the mitochondrially encoded cytochrome c oxydase subunit II (cox II) gene using primers FM58/FM66 (2). The resulting 871-bp ITS nucleotide sequence (Accession No. HM991706) was identical among all three isolates analyzed and 99% identical (100% coverage) to ITS sequences of multiple isolates of P. sylvaticum in GenBank. Likewise, the 544-bp cox II sequence (Accession No. HQ454429) was 99% identical (97% coverage) to cox II sequences of multiple isolates of P. sylvaticum. Six pots of M. sinensis seedlings were inoculated by placing two CMA plugs of a 2-week-old culture of isolate F71 at the crown. The control pots were mock inoculated with sterile CMA plugs. The plants were incubated at ~90% relative humidity (RH) and 25°C day and 22°C night for 3 days, and thereafter left on the greenhouse bench at ~65% RH with alternating 9 h of darkness and 15 h of light. Three weeks after inoculation, two of the inoculated seedlings wilted, others were stunted with leaves wilting from the tip downwards and the stems rotting from the crown upward. A thick mat of mycelia was seen on the rotted basal stems. No symptoms were observed in the control. P. sylvaticum was reisolated from both the rotted basal stems and the wilted foliage. To our knowledge, this is the first report of P. sylvaticum on M. sinensis. Infestation of farm soils with P. sylvaticum could limit M. sinensis biomass production significantly by limiting seedling establishment. References: (1) W. A. Campbell and F. F. Hendrix. Mycologia 59:274, 1967. (2) F. M. Martin. Mycologia 92:711, 2000. (3) T. J. White et al. Page 38 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.