Screening of micro-organisms for Venturia inaequalis control by means of DGGE

Venturia inaequalis causes apple scab. The control of the disease in organic apple production depends on the use of copper which will be restricted in future for environmental reasons. In the search for environmental friendly microbial biocontrol agents and stimulation of antagonistic populations in situ the molecular technique of DGGE fingerprinting can is applied. This fingerprinting technique allows to bypass culturing techniques and even can identify not yet culturable organisms. Comparison of microbial fingerprints and pathogen development in the leaf samples during winter and spring has been performed to identify populations with antagonistic potential. Such potential antagonistic organisms can be further identified by their specific DNA sequence. Preliminary results identify possible antagonists. The identification based on the sequence can help to focus on specific isolation or stimulation of a possible antagonist or a consortium of antagonists

Deciphering the modes of action of Golubevia sp., an antagonist against the causal agent of powdery mildew in wheat, using an mRNA-based systems approach

Biocontrol agents are living organisms with the potential to suppress populations of plant pathogens or pests in a cropping system. The complex interplay between the different players and the changing environment, results in a combination of different modes of action. Here, we applied an mRNA-based systems approach to gain insight into the antagonist-pathogen-host interaction of Golubevia sp. isolates BC0812 and BC0850 with the causal agent of wheat powdery mildew, Blumeria graminis f. sp. tritici, in planta over time. Bioassays were performed on potted wheat plants (water-treated control, antagonist, pathogen, antagonist+pathogen) under controlled conditions. A significantly higher percentage of mildew conidia were parasitized after treatment with Golubevia sp. BC0812 with 26% and BC0850 with 16% compared to the water control with 1%. Differential gene expression analysis of antagonists, pathogen and host 5, 6, 7, and 11 days after inoculation (dai) with the antagonist pointed to a combination of different modes of action: An interplay of modulating plant defense responses, impairing conidiogenesis of the pathogen by scavenging H2O2, facultative hyperparasitism and nitrogen competition. Microscopic observations supported the suggested hyperparasitism as thin mycelium could be observed on Bgt conidia at 6 dai and later. Taken together the results allowed the formulation of new hypothesis regarding modes of action and the interplay between antagonist, pathogen and host. It showed that a solid molecular understanding of the antagonist-pathogen relationship over time is essential for less biased mode of action studies. Understanding this complex interplay is the basis for targeted optimization strategies and allows discovery of new potential targets and markers for future biocontrol development

Transmission of escherichia coli from manure to root zones of field-grown lettuce and leek plants

Pathogenic Escherichia coli strains are responsible for food-borne disease outbreaks upon consumption of fresh vegetables and fruits. The aim of this study was to establish the transmission route of E. coli strain 0611, as proxy for human pathogenic E. coli, via manure, soil and plant root zones to the above-soil plant compartments. The ecological behavior of the introduced strain was established by making use of a combination of cultivation-based and molecular targeted and untargeted approaches. Strain 0611 CFUs and specific molecular targets were detected in the root zones of lettuce and leek plants, even up to 272 days after planting in the case of leek plants. However, no strain 0611 colonies were detected in leek leaves, and only in one occasion a single colony was found in lettuce leaves. Therefore, it was concluded that transmission of E. coli via manure is not the principal contamination route to the edible parts of both plant species grown under field conditions in this study. Strain 0611 was shown to accumulate in root zones of both species and metagenomic reads of this strain were retrieved from the lettuce rhizosphere soil metagenome library at a level of Log 4.11 CFU per g dry soil

Draft genome sequences of three isolates of golubevia sp. basidiomycete fungi isolated from powdery mildew pustules

The genomes of three Golubevia isolates (BC0812, BC0850, and BC0902) that have been shown to reduce conidiation of Blumeria graminis f. sp. tritici were sequenced using a dual-platform approach. The assembled genomes will help to elucidate the molecular mechanisms underlying the biocontrol effect of this understudied group

Stepwise screening of candidate antagonists for biological control of Blumeria graminis f. sp. tritici

Antagonists for the biological control of Blumeria graminis f. sp. tritici were selected using a stepwise screening approach. Fungal colonizers of powdery mildew pustules were isolated from leaves of cereals and other plant species. Spore production, cold tolerance, drought tolerance and UV-B resistance as important characteristics for application of biocontrol candidates in the phyllosphere were tested in in vitro assays and preliminary risk assessments were conducted. Amongst 850 tested isolates 58% belonged to various taxonomical groups of Cladosporium. Only 3% belonged to species that have been reported in literature as antagonistic to powdery mildews. The stepwise screening approach allowed to reduce the number of candidate antagonists using screening criteria that can be tested reliably and cost-effectively in in vitro assays and by data mining from initially 1237 isolates down to 143 candidate antagonists belonging to 42 taxonomical groups. The potential of these isolates to reduce conidia production of B. graminis f. sp. tritici. in wheat was assessed in bioassays on potted winter wheat plants under controlled conditions. A set of ten superior isolates was subsequently tested in a series of trials on potted spring wheat plants under open field conditions. Isolates Tilletiopsis pallescens BC0441 and T. pallescens BC0850 significantly reduced the number of powdery mildew pustules per flag leaf by 30 to 62%. Slopes of the regression lines fitted to data on number of powdery mildew pustules during time were significantly reduced by the antagonists indicating that the powdery mildew epidemics were slowed down. Treatments with T. pallescens BC0441 and T. pallescens BC0850 also reduced leaf coverage with powdery mildew pustules in a small-scale field trial in spring wheat.</p