RESUMEN
BACKGROUND: Endophytes are microbes that inhabit internal plant tissues without causing disease. Plant microbial communities consist of large numbers of endophyte species. Understanding the functions of these endophytes is a major challenge. An important function of some endophytes is to suppress fungal pathogens. Typically, plant associated microbes are screened for anti-fungal activities in vitro using the high-throughput dual culture screen, but it is not clear whether this method correlates with the activities of these microbes in planta. Furthermore, it is not clear whether in vitro screening captures all of the microbes that show this activity inside plants. The objective of this study was to evaluate the relevance of the in vitro dual culture method for screening endophytes with anti-fungal activity. RESULTS: In parallel, 190 bacterial endophytes from the corn grass family (Zea) were screened for suppression of two fungal pathogens (Sclerotinia homoeocarpa and Rhizoctonia solani) using the in vitro dual culture method, and in planta using the model plant, creeping bentgrass. All endophytes that showed anti-fungal activity in planta against Sclerotinia homoeocarpa and Rhizoctonia solani (3 or 4 strains, respectively, out of 190), were captured in vitro. The in vitro and in planta screening results strongly correlated (r = 0.81 and r = 0.94 for the two pathogens, respectively). CONCLUSIONS: Evidence was gained here that the in vitro dual culture method is a relevant method for high throughput screening of plant endophyte communities for anti-fungal activity. In our study, the method captured all of the microbes that suppressed the corresponding pathogens in planta.
Asunto(s)
Antibiosis , Bacterias/aislamiento & purificación , Fenómenos Fisiológicos Bacterianos , Endófitos/aislamiento & purificación , Endófitos/fisiología , Enfermedades de las Plantas/microbiología , Zea mays/microbiología , Ascomicetos/crecimiento & desarrollo , Ascomicetos/fisiología , Bacterias/clasificación , Bacterias/genética , Bacterias/crecimiento & desarrollo , Endófitos/clasificación , Endófitos/genética , Rhizoctonia/crecimiento & desarrollo , Rhizoctonia/fisiologíaRESUMEN
The Swede midge, Contarinia nasturtii Kieffer, is an economically significant pest of cruciferous crops in Canada and the northeastern United States. The effect of temperature on the virulence of three entomopathogenic nematode species, Heterorhabditis bacteriophora, Steinernema carpocapsae, and Steinernema feltiae, the entomopathogenic fungus Metarhizium brunneum, and a H. bacteriophora+M. brunneum combination treatment to C. nasturtii larvae, pupae, and cocoons was investigated. In the laboratory, all three nematode species successfully reproduced inside C. nasturtii larvae: H. bacteriophora produced the highest number of infective juveniles per larva, followed by S. carpocapsae and S. feltiae. H. bacteriophora and the H. bacteriophora+M. brunneum combination treatment generally caused the highest mortality levels to all C. nasturtii life stages at 20°C and 25°C, whereas S. feltiae caused the highest mortality to larvae and pupae at 16°C. No nematode species caused significant mortality when applied in foliar treatments to the infested host plant meristem and, in spite of high mortality, an antagonistic interaction was observed in the H. bacteriophora+M. brunneum combination treatment when compared with expected mortality. In trials conducted in broccoli fields in Elora, Ontario, M. brunneum suppressed adult emergence of C. nasturtii from infested soil in 2012 and all nematode treatments successfully suppressed adult emergence in 2013; however, no significant effects were observed in field trials conducted in Baden, Ontario.
Asunto(s)
Dípteros/parasitología , Interacciones Huésped-Parásitos , Metarhizium/fisiología , Nematodos/fisiología , Control Biológico de Vectores , Animales , Dípteros/crecimiento & desarrollo , Femenino , TemperaturaRESUMEN
A lysimeter-based field study was done to monitor the transfer of culturable Escherichia coli, general (ALLBAC), human (Hf183) and swine (PIG-BAC-1) specific 16S rRNA Bacteroides spp. markers, nutrients and metals through soils and leachate over time following land application of a CP1/Class A as well as two CP2/Class B municipal biosolids (MBs). Hf183 markers were detected up to six days following application in soils receiving dewatered and liquid MBs, but not in leachate, suggesting their use in source tracking is better suited for recent pollution events. The CP2/Class B biosolids and swine manure contributed the highest microbial load with E. coli loads (between 2.5 and 3.7 log CFU (100 mL)(-1)) being greater than North American concentration recommendations for safe recreational water. ALLBAC persisted in soils and leachate receiving all treatments and was detected prior to amendment application demonstrating its unsuitability for identifying the presence of fecal pollution. A significant increase in NO3-N (for Lystek and dewatered MBs) and total-P (for dewatered and liquid MBs) in leachate was observed in plots receiving the CP1/Class A and CP2/Class B type MBs which exceeded North American guidelines, suggesting impact to surface water. Metal (As, Cd, Cr, Co, Cu, Pb, Mo, Ni, Se, Zn and Hg) transfer was negligible in soil and leachate samples receiving all treatments. This study is one of the first to examine the fate of E. coli and Bacteroides spp. markers in situ following the land application of MBs where surface runoff does not apply.