Matsukawa, S. | bioRxiv Science

bioRxiv2021

Chitin-induced systemic disease resistance in rice requires both OsCERK1 and OsCEBiP and is mediated via perturbation of cell-wall biogenesis in leaves

O_LIChitin is a well-known elicitor of disease resistance whose recognition by plants is crucial to perceive fungal infections. Chitin can induce both a local immune response and a systemic disease resistance when provided as a supplement in soils. Unlike local immune responses, how chitin-induced systemic disease resistance is deployed has not been studied in detail. C_LIO_LIIn this study, we evaluated systemic disease resistance against the fungal pathogen Bipolaris oryzae by performing a transcriptome analysis and monitoring cell-wall composition in rice plants grown in chitin-supplemented soils. We also examined the local immune response to chitin by measuring the production of reactive oxygen species in leaves. C_LIO_LIChitins induced both local immune response and systemic disease resistance with differing requirements for the receptors OsCERK1 and OsCEBiP. Transcriptome analysis suggested that a perturbation in cell-wall biogenesis is involved in the induction of systemic disease resistance, an idea which was supported by the induction of disease resistance by treatment with a cellulose biosynthesis inhibitor and alterations of cell-wall composition. C_LIO_LIThese findings suggest that chitin-induced systemic disease resistance in rice is caused by a perturbation of cell-wall biogenesis in leaves through long-distance signalling after recognition of chitins by OsCERK1 and OsCEBiP. C_LI

plant biology↗

bioRxiv2018

Direct PCR amplification of 16S rRNA genes offers accelerated bacterial identification using the MinION™ nanopore sequencer

Rapid identification of bacterial pathogens is crucial for appropriate and adequate antibiotic treatment, which significantly improves patient outcomes. 16S ribosomal RNA (rRNA) gene amplicon sequencing has proven to be a powerful strategy for diagnosing bacterial infections. We have recently established a sequencing method and bioinformatics pipeline for 16S rRNA gene analysis utilizing the Oxford Nanopore Technologies MinION sequencer. In combination with our taxonomy annotation analysis pipeline, the system enabled the molecular detection of bacterial DNA in a reasonable timeframe for diagnostic purposes. However, purification of bacterial DNA from specimens remains a rate-limiting step in the workflow. To further accelerate the process of sample preparation, we adopted a direct PCR strategy that amplifies 16S rRNA genes from bacterial cell suspensions without DNA purification. Our results indicate that differences in cell wall morphology significantly affect direct PCR efficiency and sequencing data. Notably, mechanical cell disruption preceding direct PCR was indispensable for obtaining an accurate representation of the specimen bacterial composition. Furthermore, 16S rRNA gene analysis of mock polymicrobial samples indicated that primer sequence optimization is required to avoid preferential detection of particular taxa and to cover a broad range of bacterial species. This study establishes a relatively simple workflow for rapid bacterial identification via MinIONTM sequencing, which reduces the turnaround time from sample to result, and provides a reliable method that may be applicable to clinical settings.

microbiology↗