Dataset Information

Microfluidic device enabled quantitative time-lapse microscopic-photography for phenotyping vegetative and reproductive phases in Fusarium virguliforme, which is pathogenic to soybean.

ABSTRACT: Time-lapse microscopic-photography allows in-depth phenotyping of microorganisms. Here we report development of such a system using a microfluidic device, generated from polydimethylsiloxane and glass slide, placed on a motorized stage of a microscope for conducting time-lapse microphotography of multiple observations in 20 channels simultaneously. We have demonstrated the utility of the device in studying growth, germination and sporulation in Fusarium virguliforme that causes sudden death syndrome in soybean. To measure the growth differences, we developed a polyamine oxidase fvpo1 mutant in this fungus that fails to grow in minimal medium containing polyamines as the sole nitrogen source. Using this system, we demonstrated that the conidiospores of the pathogen take an average of five hours to germinate. During sporulation, it takes an average of 10.5?h for a conidiospore to mature and get detached from its conidiophore for the first time. Conidiospores are developed in a single conidiophore one after another. The microfluidic device enabled quantitative time-lapse microphotography reported here should be suitable for screening compounds, peptides, micro-organisms to identify fungitoxic or antimicrobial agents for controlling serious plant pathogens. The device could also be applied in identifying suitable target genes for host-induced gene silencing in pathogens for generating novel disease resistance in crop plants.

SUBMITTER: Marshall J

PROVIDER: S-EPMC5353701 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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Microfluidic device enabled quantitative time-lapse microscopic-photography for phenotyping vegetative and reproductive phases in Fusarium virguliforme, which is pathogenic to soybean.

Marshall Jill J Qiao Xuan X Baumbach Jordan J Xie Jingyu J Dong Liang L Bhattacharyya Madan K MK

Scientific reports 20170315

Time-lapse microscopic-photography allows in-depth phenotyping of microorganisms. Here we report development of such a system using a microfluidic device, generated from polydimethylsiloxane and glass slide, placed on a motorized stage of a microscope for conducting time-lapse microphotography of multiple observations in 20 channels simultaneously. We have demonstrated the utility of the device in studying growth, germination and sporulation in Fusarium virguliforme that causes sudden death synd ...[more]

PMID: 28295054

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Project description:Marine ecosystems are heavily influenced by a wide range of human-related impacts, and thus monitoring is essential to preserve and manage these sensitive habitats. Seabirds are considered important bioindicators of the oceans, but accessing breeding populations can be difficult, expensive and time consuming. New technologies have been employed to facilitate data collection on seabirds that can reduce costs and minimize disturbance. Among these, the use of time-lapse photography is a potentially effective way to reduce researcher effort, while collecting valuable information on key ecological parameters. However, the feasibility of this approach remains uncertain. Here, we assessed the use of time-lapse photography as a tool for estimating foraging behaviour from breeding seabirds, and evaluate ways forward for this method. We deployed cameras in front of active nests at a colony of black-legged kittiwakes (Rissa tridactyla) during two breeding seasons, 5 nests in 2013 and 5 in 2014, taking pictures every 4 minutes. A subsample of monitored individuals were also equipped with accelerometers. Approximately 100,000 frames, covering incubation and chick-rearing periods, were analysed. Estimates of foraging trip duration from images were positively correlated with accelerometry estimates (R2 = 0.967). Equal partitioning of effort between pairs, predation events, nest attendance patterns and variation in trip metrics with breeding stage were also identified. Our results suggest that time-lapse photography is potentially a useful tool for assessing foraging trip duration and other fine-scale nesting ecology parameters as well as for assessing the effect of bio-logging devices on seabird foraging behaviour. Nevertheless, the time investment to manually extract data from images was high, and the process to set up cameras was not straightforward. To encourage wide use of time-lapse photography in seabird ecology, we thus provide guidelines for camera deployment and we suggest a need for further development of automated approaches to allow data extraction.

Project description:BACKGROUND:Sudden death syndrome (SDS) caused by the ascomycete fungus, Fusarium virguliforme, exhibits root necrosis and leaf scorch or foliar SDS. The pathogen has never been identified from the above ground diseased foliar tissues. Foliar SDS is believed to be caused by host selective toxins, including FvTox1, secreted by the fungus. This study investigated if the xylem sap of F. virguliforme-infected soybean plants contains secreted F. virguliforme-proteins, some of which could cause foliar SDS development. RESULTS:Xylem sap samples were collected from five biological replications of F. virguliforme-infected and uninfected soybean plants under controlled conditions. We identified five F. virguliforme proteins from the xylem sap of the F. virguliforme-infected soybean plants by conducting LC-ESI-MS/MS analysis. These five proteins were also present in the excreted proteome of the pathogen in culture filtrates. One of these proteins showed high sequence identity to cerato-platanin, a phytotoxin produced by Ceratocystis fimbriata f. sp. platani to cause canker stain disease in the plane tree. Of over 500 soybean proteins identified in this study, 112 were present in at least 80% of the sap samples collected from F. virguliforme-infected and -uninfected control plants. We have identified four soybean defense proteins from the xylem sap of F. virguliforme-infected soybean plants. The data have been deposited to the ProteomeXchange with identifier PXD000873. CONCLUSION:This study confirms that a few F. virguliforme proteins travel through the xylem, some of which could be involved in foliar SDS development. We have identified five candidate proteinaceous toxins, one of which showed high similarity to a previously characterized phytotoxin. We have also shown the presence of four soybean defense proteins in the xylem sap of F. virguliforme-infected soybean plants. This study laid the foundation for studying the molecular basis of foliar SDS development in soybean and possible defense mechanisms that may be involved in conferring immunity against F. virguliforme and other soybean pathogens.

Dataset Information

Microfluidic device enabled quantitative time-lapse microscopic-photography for phenotyping vegetative and reproductive phases in Fusarium virguliforme, which is pathogenic to soybean.

Publications

Microfluidic device enabled quantitative time-lapse microscopic-photography for phenotyping vegetative and reproductive phases in Fusarium virguliforme, which is pathogenic to soybean.

Similar Datasets

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