Barley powdery mildew invasion coincides with the dynamic accumulation of leaf apoplastic extracellular vesicles that are associated with host stress response proteins

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Апрель 19, 2024

Abstract The mutual exchange of extracellular vesicles across kingdom borders is a feature many plant-microbe interactions. occurrence and cargos has been studied in several instances, but their dynamics the course infection have remained elusive. Here we used two different procedures, differential high-speed centrifugation polymer-based enrichment, to collect from apoplastic wash fluid barley ( Hordeum vulgare ) leaves challenged by its fungal powdery mildew pathogen, Blumeria hordei . Both methods yielded similar quality morphological characteristics, though polymer approach was associated with higher reproducibility. We noted that derived constitute polydisperse populations are selectively responsive leaf B. Extracellular ∼100 nm – 300 diameter became progressively more abundant, particular 72 hours post inoculation onwards, resulting major peak late during infection. Vesicles ∼300 500 showed accumulation reached much lower levels, suggesting they might separate population. Proteome analysis uncovered an enrichment biotic stress response proteins vesicles. t-SNARE protein Ror2, ortholog PEN1 marker Arabidopsis thaliana , accumulates infection, hence also qualifying as potential protein. Our study serves starting point for investigating role at stages

Язык: Английский

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A pipeline for validation of Serendipita indica effector-like sRNA suggests cross-kingdom communication in the symbiosis with Arabidopsis

Journal of Experimental Botany, Год журнала: 2024, Номер unknown

Опубликована: Дек. 19, 2024

Abstract Bidirectional communication between pathogenic microbes and their plant hosts via small (s)RNA-mediated cross-kingdom RNA interference (ckRNAi) is a key element for successful host colonisation. Whether mutualistic fungi of the Serendipitaceae family, known extremely broad range, use sRNAs to colonize roots still under debate. To address this question, we developed pipeline validate accumulation, translocation, activity fungal in post-transcriptional silencing Arabidopsis thaliana genes. Using stem-loop RT-qPCR, detected expression specific set Serendipita indica (Si)sRNAs, targeting genes involved cell wall organization, hormonal signalling regulation, immunity, gene regulation. confirm these cells, SisRNAs were transiently expressed protoplasts. Stem-loop PCR confirmed while qPCR validated predicted target Furthermore, ARGONAUTE 1 immunoprecipitation (AtAGO1-IP) revealed loading into RNAi machinery, suggesting translocation SisRNA from fungus root cells. In conclusion, study provides blueprint rapid selection analysis sRNA effectors further supports model Sebacinoid symbiosis.

Язык: Английский

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Cross-kingdom RNAi adds a new layer of communication to the arbuscular mycorrhizal symbiosis

Research Square (Research Square), Год журнала: 2023, Номер unknown

Опубликована: Окт. 17, 2023

Abstract Cross-kingdom RNA interference (ckRNAi) is a mechanism of interspecies communication where small RNAs (sRNAs) are transported from one organism to another, they silence target genes in trans1-9. ckRNAi well established pathosystems1-9 and has also been observed number mutualistic interactions10,11. Fungal oomycete pathogens, as symbiotic bacterium, send sRNAs into host cells hijack RNAi machinery by loading AGO proteins order genes. Previous research our group suggested the possibility occurring Arbuscular Mycorrhizal Symbiosis (AMS)12, most widespread ancient symbiosis on Earth13. Here, we show that specific sRNA AM fungus Rhizophagus irregularis (Rir2216) loaded an AGO1 silencing complex plant Medicago truncatula, leading cleavage transcript encoding for MtWRKY69 transcription factor. downregulated arbusculated mycorrhizal roots overexpression led reduced colonization level, indicating its relevant AMS. Thus, present first experimental evidence molecular

Язык: Английский

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Interkingdom Communication via Extracellular Vesicles: Unraveling Plant and Pathogen Interactions and Its Potential for Next-Generation Crop Protection

Microorganisms, Год журнала: 2024, Номер 12(12), С. 2392 - 2392

Опубликована: Ноя. 22, 2024

Recent advancements in the field of plant–pathogen interactions have spotlighted role extracellular vesicles (EVs) as pivotal mediators cross-kingdom communication, offering new vistas for enhancing crop protection strategies. EVs are instrumental transport small regulatory RNAs (sRNAs) and other bioactive molecules across species boundaries, thus playing a critical molecular warfare between plants pathogens. This review elucidates sophisticated mechanisms by which utilize to dispatch sRNAs that silence pathogenic genes, fortifying defenses against microbial threats. Highlighting both eukaryotic prokaryotic systems, this delves into biogenesis, isolation, functional roles EVs, illustrating their importance not only fundamental biological processes but also potential therapeutic applications. studies illuminated significant facilitating communication pathogens, highlighting host-defense mechanisms. However, despite these advancements, challenges remain efficient isolation characterization plant-derived EVs. Overcoming is fully harnessing developing next-generation proposes innovative strategies utilizing RNA-based interventions delivered via bolster plant resilience diseases. By integrating latest scientific findings with practical applications agriculture, aims enhance connection biology development management technologies.

Язык: Английский

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Messenger and message: Uncovering the roles, rhythm and regulation of extracellular vesicles in plant biotic interactions

Current Opinion in Plant Biology, Год журнала: 2024, Номер 83, С. 102672 - 102672

Опубликована: Дек. 28, 2024

Extracellular vesicles (EVs) are membrane-delimited nanoparticles found in every kingdom of life and known to mediate cell-cell communication animal systems through the trafficking proteins nucleic acids. Research into plant microbial EVs suggests that these have similar transport capacity, moreover able signalling not only within an organism but also between organisms, acting plants their partners cross-kingdom signalling. Here, we review recent research exploring roles EVs, both microbial, highlighting emerging trends functional conservation species across kingdoms, complemented by heterogeneity EV subpopulations at level places as powerful regulatory mechanisms biotic interactions.

Язык: Английский

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A robust pipeline for the systematic validation of sRNA effectors suggests cross-kingdom communication in the symbiosis ofSerendipita indicawithArabidopsis thaliana

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Дек. 19, 2023

Abstract Bidirectional communication between pathogenic microbes and their plant hosts via small (s)RNA-mediated cross-kingdom RNA interference (ckRNAi) is one key element for successful host colonisation. However, whether mutualistic fungi from the Serendipitaceae family, known extremely broad range, employ sRNAs to colonize roots still under discussion. To address this question, we developed a pipeline validate accumulation, translocation, activity of fungal in post-transcriptional silencing Arabidopsis thaliana genes. Using stem-loop PCR, detected expression specific set Serendipita indica ( Si) sRNAs, targeting genes involved cell wall organization, hormonal signalling regulation, immunity, gene regulation. confirm these sRNA cells, Si were transiently expressed protoplasts. Stem-loop PCR proved while qPCR validated predicted target Furthermore, ARGONAUTE 1 immunoprecipitation At AGO1-IP) revealed loading into RNAi machinery, suggesting translocation fungus root cells. In conclusion, study provides blueprint rapid selection analysis effectors plant-microbe interactions further suggests Sebacinoid symbiosis. Highlight Small RNAs beneficial are translocated silence at onset interaction, revealing sebacinoid

Язык: Английский

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