Andean blackberry (Rubus glaucus Benth) plants from the provinces of Tungurahua and Bolivar (Ecuador) started showing symptoms of black foot disease since 2010. Wilted plants were sampled in both provinces from 2014 to 2017, and fungal isolates were obtained from tissues surrounding necrotic lesions in the cortex of the roots and crown. Based on morphological characteristics and DNA sequencing of histone 3 and the translation elongation factor 1α gene, isolates were identified as one of seven species, Ilyonectria vredehoekensis, Ilyonectria robusta, Ilyonectria venezuelensis, Ilyonectria europaea, Dactylonectria torresensis, or Dactylonectria novozelandica. Pathogenicity tests with isolates from each species, excluding I. europaea and D. novozelandica whose isolates were lost due to contamination, confirmed that the four species tested can produce black foot disease symptoms in Andean blackberry. This is the first report of Dactylonectria and Ilyonectria species causing black foot disease of Andean blackberry.
Twenty-eight isolates of Sclerotinia homoeocarpa, causal agent of dollar spot disease in turf, were assessed for fungicide hormesis at sublethal concentrations of thiophanate-methyl (T-methyl). Each isolate was grown in corn meal agar amended with 11 concentrations of T-methyl (30,500 to 0.047 µg/liter), and the area of mycelial growth was determined relative to the control. Three replicates were used per concentration, and the experiment was repeated three to five times for each isolate. Reference isolates (EC50 > 20 µg/liter), with no prior history of T-methyl exposure, were highly sensitive and not stimulated by low doses. Likewise, no stimulation was observed in two highly sensitive isolates (EC50 > 30 µg/liter) that had been preconditioned by exposure to T-methyl, or in four T-methyl-tolerant isolates. Seventeen (81%) preconditioned T-methyl-tolerant isolates (EC50 = 294 to1,550 µg/liter) had statistically significant growth stimulation, in the range of 2.8 to 19.7% relative to the control. These results support that hormesis (low-dose stimulation, high-dose inhibition) is a common dose response in preconditioned S. homoeocarpa, particularly in response to subtoxic doses of T-methyl.
A multiplex end-point polymerase chain reaction (PCR) assay was developed for identifying the three-fungal species in the genus Ophiosphaerella that cause spring dead spot (SDS), a devastating disease of bermudagrass. These fungi are difficult to identify by morphology because they seldom produce pseudothecia. To achieve species-specific diagnosis, three pairs of primers were designed to identify fungal isolates and detect the pathogen in infected roots. The internal transcribed spacer region, the translation elongation factor 1-α, and the RNA polymerase II second-largest subunit were selected as targets and served as templates for the design of each primer pair. To achieve uniform melting temperatures, three to five random nucleotide extensions (flaps) were added to the 5′ terminus of some of the designed specific primers. Temperature cycling conditions and PCR components were standardized to optimize specificity and sensitivity of the multiplex reaction. Primers were tested in multiplex on DNA extracted from axenic fungal cultures and from field-collected infected and uninfected roots. A distinct amplicon was produced for each Ophiosphaerella sp. tested. The DNA from Ophiosphaerella close relatives and other common bermudagrass pathogens did not amplify during the multiplex assay. Metagenomic DNA from infected bermudagrass produced species-specific amplicons while DNA extracted from noninfected roots did not. This multiplex end-point PCR approach is a sensitive and specific molecular technique that allows for correct identification of SDS-associated Ophiosphaerella spp. from field-collected roots.
Con la finalidad de contar con un tratamiento biológico alternativo al uso de los fertilizantes químicos se evaluó el potencial biofertilizantes de consorcios de cianobacterias asilados de tapetes bacterianos recolectados en el volcán Pasochoa. Se reportó la influencia del biofertilizante en el crecimiento y valor nutricional de Pasto Raygrass, empleado comúnmente en la alimentación de ganado.
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