Arabidopsis Apoplastic Fluid Contains sRNA- and Circular RNA-Protein Complexes that Are Located Outside Extracellular Vesicles

Karimi, Hana Zand; Baldrich, Patricia; Rutter,; Borniego,; Zajt, Kamil K; Innes, Roger W.

doi:10.1101/2021.10.02.462881

Cited by 5 publications

(5 citation statements)

References 86 publications

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“…Alterations to these epigenetic marks can result in changes to the plant phenotype including resistance to abiotic and biotic stress, as well as alteration in the plant architecture and vigour. apoplast (Yang et al, 2019;Karimi et al, 2022). These results suggest the existence of a specific mechanism for the systemic mobilization of sRNA molecules in plants, which is currently subject of research and discussion (Voinnet, 2022;Yan and Ham, 2022).…”

Section: Movement Of Small Non-coding Rna Moleculesmentioning

confidence: 75%

Long distance signalling and epigenetic changes in crop grafting

Jeynes-Cupper

Catoni

2023

Front. Plant Sci.

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Humans have used grafting for more than 4000 years to improve plant production, through physically joining two different plants, which can continue to grow as a single organism. Today, grafting is becoming increasingly more popular as a technique to increase the production of herbaceous horticultural crops, where rootstocks can introduce traits such as resistance to several pathogens and/or improving the plant vigour. Research in model plants have documented how long-distance signalling mechanisms across the graft junction, together with epigenetic regulation, can produce molecular and phenotypic changes in grafted plants. Yet, most of the studied examples rely on proof-of-concept experiments or on limited specific cases. This review explores the link between research findings in model plants and crop species. We analyse studies investigating the movement of signalling molecules across the graft junction and their implications on epigenetic regulation. The improvement of genomics analyses and the increased availability of genetic resources has allowed to collect more information on potential benefits of grafting in horticultural crop models. Ultimately, further research into this topic will enhance our ability to use the grafting technique to exploit genetic and epigenetic variation in crops, as an alternative to traditional breeding.

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Section: Movement Of Small Non-coding Rna Moleculesmentioning

confidence: 75%

Long distance signalling and epigenetic changes in crop grafting

Jeynes-Cupper

Catoni

2023

Front. Plant Sci.

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“…RNA-mediated intercellular communication has been almost exclusively studied in the context of extracellular vesicles (EVs) (O'Brien et al 2020), despite most exRNAs being present outside EVs in human biofluids (Arroyo et al 2011;Turchinovich et al 2011), cell-conditioned media (Tosar et al 2015(Tosar et al , 2020Sork et al 2021), and even in plants (Borniego and Innes 2023;Zand Karimi et al 2022). A bias against the study of nonvesicular exRNAs can be explained by the widespread assumption that exRNA is unstable unless protected inside EVs.…”

Section: Discussionmentioning

confidence: 99%

Ribonuclease activity undermines immune sensing of naked extracellular RNA

Castellano,

Blanco,

Calzi

et al. 2024

Preprint

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The plasma membrane and the membrane of endosomal vesicles are considered physical barriers preventing extracellular RNA uptake. While naked RNA can be spontaneously internalized by certain cells types, functional delivery of naked RNA into the cytosol has been rarely observed. Here we show that extracellular ribonucleases, mainly derived from cell culture supplements, have so far hindered the study of extracellular RNA functionality. In the presence of active ribonuclease inhibitors (RI), naked bacterial RNA is pro-inflammatory when spiked in the media of dendritic cells and macrophages. In murine cells, this response mainly depends on the action of endosomal Toll-like receptors. However, we also show that naked RNA can perform endosomal escape and engage with cytosolic RNA sensors and ribosomes. For example, naked mRNAs encoding reporter proteins can be spontaneously internalized and translated by a variety of cell types, in an RI-dependent manner. In vivo, RI co-injection enhances the activation induced by naked extracellular RNA on splenic lymphocytes and myeloid-derived leukocytes. Furthermore, naked extracellular RNA is inherently pro-inflammatory in ribonuclease-poor compartments such as the peritoneal cavity. Overall, these results demonstrate that naked RNA is bioactive and does not need encapsulation inside synthetic or biological lipid vesicles for functional uptake, making a case for nonvesicular extracellular RNA-mediated intercellular communication.

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“…These proteins are also attractive targets for genetic analysis, in order to determine how they impact the formation and contents of fungal EVs. Most excitingly, C. higginsianum is a pathogen of the model plant Arabidopsis thaliana, the EVs of which are currently the best-defined among plant species (Baldrich et al, 2019;Karimi et al, 2021;Rutter & Innes, 2017). c All the best blastp hits were found to be involved in the biosynthesis of a variety of biologically active molecules.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of Extracellular Vesicles from the Fungal PhytopathogenColletotrichum higginsianum

Rutter

Chu

Zajt

et al. 2022

Preprint

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Fungal phytopathogens secrete extracellular vesicles (EVs) associated with enzymes and phytotoxic metabolites. While these vesicles are thought to promote infection, defining the true contents and functions of fungal EVs, as well as suitable protein markers, is an ongoing process. To expand our understanding of fungal EVs and their possible roles during infection, we purified EVs from the hemibiotrophic phytopathogen Colletotrichum higginsianum, the causative agent of anthracnose disease in multiple plant species, including Arabidopsis thaliana. EVs were purified in large numbers from the supernatant of protoplasts but not the supernatant of intact mycelial cultures. We purified two separate populations of EVs, each associated with over 700 detected proteins, including proteins involved in vesicle transport, cell wall biogenesis and the synthesis of secondary metabolites. We selected two SNARE proteins (Snc1 and Sso2) and one 14-3-3 protein (Bmh1) as potential EV markers and generated transgenic lines expressing fluorescent fusions. Each marker was confirmed to be protected inside EVs. Fluorescence microscopy was used to examine the localization of each marker during infection on Arabidopsis leaves. These findings further our understanding of EVs in fungal phytopathogens and will help build an experimental system to study EV inter-kingdom communication between plants and fungi.

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Arabidopsis Apoplastic Fluid Contains sRNA- and Circular RNA-Protein Complexes that Are Located Outside Extracellular Vesicles

Cited by 5 publications

References 86 publications

Long distance signalling and epigenetic changes in crop grafting

Long distance signalling and epigenetic changes in crop grafting

Ribonuclease activity undermines immune sensing of naked extracellular RNA

Isolation and Characterization of Extracellular Vesicles from the Fungal PhytopathogenColletotrichum higginsianum

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