Bremia lactucae is an obligate, oomycete pathogen of lettuce that causes leaf chlorosis and necrosis and adversely affects marketability. The disease has been managed with a combination of host resistance and fungicide applications with success over the years. Fungicide applications are routinely made under the assumption that inoculum is always present during favorable environmental conditions. This approach often leads to fungicide resistance in B. lactucae populations. Detection and quantification of airborne B. lactucae near lettuce crops provides an estimation of the inoculum load, enabling more judicious timing of fungicide applications. We developed a quantitative polymerase chain reaction (qPCR)-based assay using a target sequence in mitochondrial DNA for specific detection of B. lactucae. Validation using amplicon sequencing of DNA from 83 geographically diverse isolates, representing 14 Bremia spp., confirmed that the primers developed for the TaqMan assays are species specific and only amplify templates from B. lactucae. DNA from a single sporangium could be detected at a quantification cycle (C_q) value of 32, and C_q values >35 were considered to be nonspecific. The coefficient of determination (R²) for regression between sporangial density derived from flow cytometry and C_q values derived from the qPCR was 0.86. The assay was deployed using spore traps in the Salinas Valley, where nearly half of U.S. lettuce is produced. The deployment of this sensitive B. lactucae-specific assay resulted in the detection of the pathogen during the 2-week lettuce-free period as well as during the cropping season. These results demonstrate that this assay will be useful for quantifying inoculum load in and around the lettuce fields for the purpose of timing fungicide applications based on inoculum load.

Oomycetes are widely distributed in various environments, including desert and polar regions. Depending upon different habits and hosts, they have evolved with both saprophytic and pathogenic nutritional modes. Freshwater ecosystem is one of the most important habitats for members of oomycetes. Most studies on oomycete diversity, however, have been biased mostly towards terrestrial phytopathogenic species, rather than aquatic species, although their roles as saprophytes and parasites are essential for freshwater ecosystems. In this study, we isolated oomycete strains from soil sediment, algae, and decaying plant debris in freshwater streams of Korea. The strains were identified based on cultural and morphological characteristics, as well as molecular phylogenetic analyses of ITS rDNA, cox1, and cox2 mtDNA sequences. As a result, we discovered eight oomycete species previously unknown in Korea, namely Phytopythium chamaehyphon, Phytopythium litorale, Phytopythium vexans, Pythium diclinum, Pythium heterothallicum, Pythium inflatum, Pythium intermedium, and Pythium oopapillum. Diversity and ecology of freshwater oomycetes in Korea are poorly understood. This study could contribute to understand their distribution and ecological function in freshwater ecosystem.

In order to elucidate the fungal diversity and community structure in freshwater environments, numerous fungal strains were isolated from freshwater, submerged soils, twigs, dead insects, etc. Among them, the present study has focused specifically on Aspergillus, Penicillium, and Talaromyces species, which produce diverse useful metabolites in general. Twelve strains of Aspergillus isolated were identified as A. japonicus (n = 5), A. tubingensis (3), A. niger (2), and A. flavus (2), 10 strains of which belong to Aspergillus section Nigri, named black Aspergillus. Eight strains of Penicillium were identified as P. brasilianim (n = 3), P. oxalicum (2), P. crustosum (1), P. expansum (1), and P. piscarium (1). Two different strains of Talaromyces were identified as T. pinophilus and T. versatilis. Thus far, Penicillium piscarium and Talaromyces versatilis have been unrecorded in Korea, for which we provide detailed morphological and molecular characteristics.