7/14/2023 0 Comments Spore microscope![]() ![]() Using metagenomic and BLAST analysis, 150 cereal pathogen species (33 different genera) were recorded on the spore trap tapes. , Blumeria graminis, Cladosporium spp., Fusarium spp., Puccinia spp., and Zymoseptoria spp.) were found using these techniques at all sites. Six major cereal fungal pathogen genera ( Alternaria spp. Spore traps were set up in four geographically distinct UK wheat fields (Carnoustie, Angus Bishop Burton, Yorkshire Swindon, Wiltshire and Lenham, Kent). We aimed to investigate the incidence of spores from major fungal pathogens of cereals in the field by comparing microscopic and metagenomic based approaches for spore identification. Fungal diseases such as Fusarium head blight, Septoria tritici blotch, spot blotch, tan spot, stripe rust, leaf rust, and powdery mildew cause serious yield losses in wheat and can impact quality. Wheat is one of the main staple food crops, and 775 million tonnes of wheat were produced worldwide in 2022. 4Institute for Life and Earth Sciences, School of Energy, Geosciences, Infrastructure and Society, Heriot-Watt University, Edinburgh, United Kingdom.3Department of Agriculture, Food and the Marine, Celbridge, Ireland.2School of Biology and Environmental Science and UCD Earth Institute, University College Dublin, Belfield, Ireland. ![]() 1School of Agriculture & Food Science and UCD Earth Institute, University College Dublin, Belfield, Ireland.All rights reserved.Paola Pilo 1, Colleen Lawless 2, Anna M. Fast green Hoescht 34580 Nile red Propidium iodide Spore Staining Wheat germ agglutinin 488.Ĭopyright © 2018 Elsevier Ltd. The proposed staining method has broad applicability and can potentially be applied to follow Clostridium spore behaviour in a range of different environments.ģD-SIM CLSM Clostridium sp. ![]() Direct staining of a cheese specimen using Nile Red and Fast Green allowed in situ observation of spores within the cheese and their position within the cheese matrix. Super-resolution microscopy of spores probed by Hoechst 34580 also allowed chromatin to be visualised. Dual staining with Hoechst/Propidium iodide differentiated spores from vegetative cells, provided information on the viability of cells and was successfully applied to follow spore production induced by heating. Staining was conducted without drying the sample, preventing changes induced by dehydration and cells observed by confocal laser scanner microscopy or using a super-resolution microscope equipped with a 3D-structured illumination module. ![]() Hoechst 34580, Propidium iodide and wheat germ agglutinin WGA 488 were used in combination to stain four strains of Clostridia at different life cycle stages. We have developed a quick, simple and non-destructive fluorescent staining procedure that allows a clear differentiation between spores and vegetative cells and effectively stains spores, allowing recovery and tracking in subsequent experiments. Due to their size, spores can be investigated by a range of microscopic techniques, many of which involve sample pre-treatment. The life cycle for Clostridium includes a spore stage that can assist in survival under environmentally stressful conditions, such as extremes of temperature or pH. Non-pathogenic spore-forming Clostridia are of increasing interest due to their application in biogas production and their capability to spoil different food products. ![]()
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