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      Microscopic Analyses of Latent and Visible Monilinia fructicola Infections in Nectarines

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          Abstract

          Little is known about the histologic features of a latent Monilinia fructicola infection and brown rot in infected fruit. This report informs on the results of an investigation whose aim was to analyze the microanatomy of nectarines with a latent and visible M. fructicola infection. Mature nectarines were inoculated with an M. fructicola isolate and incubated at 25°C for 0, 24, 48, 72, or 96 hours in the dark. For investigating the latent infection process, the inoculated nectarines were first incubated at 25°C for 24 hours in the dark and then incubated at 4°C for 72, 144, 216, and 288 hours in the dark. At the end of the incubation, samples of nectarine tissue were excised from the inoculation points and prepared for light and transmission electron microscopic examinations. No signs of disease were seen on the surface of nectarines with a latent infection over the 288-hour incubation period. When the tissue samples were microscopically examined, M. fructicola colonized the stomata and this stomatal colonization progressively increased over time and was associated with gradual collapse of the epidermal cells and colonization of the subepidermis. In nectarines with visible brown rot, the disease usually appeared after 24 hours on the surface and in the uppermost layers of epidermal cells, which began to collapse after 48 hours. Subsequently, the diseased tissues of the nectarines displayed (a) colonization of the epidermis and mesocarp by M. fructicola with thin and thick hyphae, (b) collapse and disruption of epidermal and mesocarpic cells, (c) lysogenic cavities in the subepidermis and mesocarp, (d) degradation of the cuticle and epidermis, and (e) M. fructicola sporulation. M. fructicola is active during latent infections because slow and progressive colonization of nectarine subcuticular cells by the fungus occurs.

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          Quiescent and necrotrophic lifestyle choice during postharvest disease development.

          Insidious fungal infections by postharvest pathogens remain quiescent during fruit growth until, at a particular phase during fruit ripening and senescence, the pathogens switch to the necrotrophic lifestyle and cause decay. During ripening, fruits undergo physiological processes, such as activation of ethylene biosynthesis, cuticular changes, and cell-wall loosening-changes that are accompanied by a decline of antifungal compounds, both those that are preformed and those that are inducible secondary metabolites. Pathogen infection of the unripe host fruit initiates defensive signal-transduction cascades, culminating in accumulation of antifungal proteins that limit fungal growth and development. In contrast, development of the same pathogens during fruit ripening and storage activates a substantially different signaling network, one that facilitates aggressive fungal colonization. This review focuses on responses induced by the quiescent pathogens of postharvest diseases in unripe host fruits. New genome-scale experimental approaches have begun to delineate the complex and multiple networks of host and pathogen responses activated to maintain or to facilitate the transition from the quiescent to the necrotrophic lifestyle.
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            Pathogen quiescence in postharvest diseases.

            D Prusky (1995)
            This chapter examines the quiescence period during different stages of fungal attack of postharvest pathogens: quiescence during spore germination and initial hyphal development, during and after appressorium formation, and quiescence of germinated appressorium and subcuticular hyphae. The different mechanisms for quiescence are reviewed: factors affecting quiescence of germinated spores, appressoria formation and germination, and fungal colonization. Special emphasis is given to mechanisms of quiescence involving fungal colonization: 1. the pathogen's nutritional requirements, 2. preformed antifungal compounds, 3. the elicitation of phytoalexins and preformed compounds, and 4. the activation of factors in fungal pathogenicity.
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              Early events in the Fusarium verticillioides-maize interaction characterized by using a green fluorescent protein-expressing transgenic isolate.

              The infection of maize by Fusarium verticillioides can result in highly variable disease symptoms ranging from asymptomatic plants to severe rotting and wilting. We produced F. verticillioides green fluorescent protein-expressing transgenic isolates and used them to characterize early events in the F. verticillioides-maize interaction that may affect later symptom appearance. Plants grown in F. verticillioides-infested soil were smaller and chlorotic. The fungus colonized all of the underground parts of a plant but was found primarily in lateral roots and mesocotyl tissue. In some mesocotyl cells, conidia were produced within 14 to 21 days after infection. Intercellular mycelium was detected, but additional cells were not infected until 21 days after planting. At 25 to 30 days after planting, the mesocotyl and main roots were heavily infected, and rotting developed in these tissues. Other tissues, including the adventitious roots and the stem, appeared to be healthy and contained only a small number of hyphae. These results imply that asymptomatic systemic infection is characterized by a mode of fungal development that includes infection of certain tissues, intercellular growth of a limited number of fungal hyphae, and reproduction of the fungus in a few cells without invasion of other cells. Development of visibly rotted tissue is associated with massive production of fungal mycelium and much less organized growth.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                5 August 2016
                2016
                : 11
                : 8
                : e0160675
                Affiliations
                [1 ]Department of Plant Protection, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain
                [2 ]Department of Vegetal Biology I, Faculty of Biology, Complutense University of Madrid, Madrid, Spain
                Fujian Agriculture and Forestry University, CHINA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: CGB PM ADC.

                • Data curation: CGB ADC BF.

                • Formal analysis: CGB ADC BF.

                • Funding acquisition: PM ADC.

                • Investigation: CGB BF.

                • Methodology: BF.

                • Project administration: PM ADC.

                • Resources: ADC BF.

                • Supervision: ADC.

                • Validation: CGB PM ADC BF.

                • Visualization: CGB ADC.

                • Writing - original draft: CGB ADC BF.

                • Writing - review & editing: CGB PM ADC BF.

                Article
                PONE-D-16-22333
                10.1371/journal.pone.0160675
                4975509
                27494620
                aff1894a-077a-465f-8fa1-d9b6453f27eb
                © 2016 Garcia-Benitez et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 3 June 2016
                : 23 July 2016
                Page count
                Figures: 4, Tables: 1, Pages: 16
                Funding
                Funded by: AGL2014-55287-CO2-01
                Funded by: AGL2011-30472-CO2-02
                Award ID: BES-2012-053796
                Award Recipient :
                This study was supported by grants AGL2011-30472-CO2-02 and AGL2014-55287-CO2-01 from the Ministry of Science and Innovation (Spain). C. García-Benitez received a scholarship BES-2012-053796 from the Ministry of Science and Innovation (Spain).
                Categories
                Research Article
                Biology and Life Sciences
                Agriculture
                Crop Science
                Crops
                Fruits
                Biology and Life Sciences
                Organisms
                Plants
                Fruits
                Research and Analysis Methods
                Microscopy
                Light Microscopy
                Biology and Life Sciences
                Cell Biology
                Cellular Structures and Organelles
                Cell Walls
                Biology and Life Sciences
                Anatomy
                Integumentary System
                Skin
                Epidermis
                Medicine and Health Sciences
                Anatomy
                Integumentary System
                Skin
                Epidermis
                Biology and Life Sciences
                Plant Science
                Plant Anatomy
                Leaves
                Stomata
                Biology and Life Sciences
                Plant Science
                Plant Anatomy
                Stem Anatomy
                Stomata
                Physical Sciences
                Physics
                Electromagnetic Radiation
                Light
                Visible Light
                Biology and Life Sciences
                Mycology
                Fungal Reproduction
                Fungal Spore Germination
                Research and Analysis Methods
                Microscopy
                Electron Microscopy
                Transmission Electron Microscopy
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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                Uncategorized

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