Name: Isolamento e purificazione delle vescicole extracellulari vegetali dalle foglie di Arabidopsis utilizzando un metodo ottimizzato di raccolta apoplastica (Video) | JoVE Journal
Uploaded: 2026-03-24T12:50:00.000+00:00
Duration: 9 min 38 s
Description: Impara a isolare e purificare vescicole extracellulari vegetali da Arabidopsis thaliana con un protocollo ottimizzato di raccolta apoplastica. Questo metodo standardizzato minimizza la contaminazione citoplasmatica e preserva l'integrità delle vescicole.

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</ol>

<table><tbody><tr><td>0,45 & micro; Filtri in nylon m</td><td>VWR</td><td>76479-032</td><td></td></tr><tr><td>1 ml di siringa inutile</td><td>BD in Biologia</td><td>309659</td><td></td></tr><tr><td>Filtro ultra centrifugo da 100 kDa</td><td>Amicon</td><td>UFC 910008</td><td></td></tr><tr><td>100 ml siringa inutile</td><td>BD in Biologia</td><td>551721050</td><td></td></tr><tr><td>Tubi conici da 15 ml</td><td>VWR</td><td>89039-664</td><td></td></tr><tr><td>Tubi conici da 50 ml</td><td>VWR</td><td>89039-656</td><td></td></tr><tr><td>Arabidopsis thaliana (Col-0)</td><td>Linee mantenute in laboratorio</td><td>Non applicabile</td><td></td></tr><tr><td>Griglie a 300 mesh carbonio-formvar</td><td>TED PELLA</td><td>01753-F</td><td></td></tr><tr><td>Nastro trasparente</td><td>Nastro magico scozzese</td><td>S-9783</td><td></td></tr><tr><td>Parafilm</td><td>Fisher Scientific</td><td>13-374-12</td><td></td></tr><tr><td>Buffer PBS</td><td>Realizzato in casa</td><td>Non applicabile</td><td></td></tr><tr><td>qEV colonna originale Gen2 35 nm</td><td>IZON Science</td><td>ICO-35</td><td></td></tr><tr><td>Centrifuga refrigerata</td><td>Eppendorf</td><td>5810R</td><td></td></tr><tr><td>Rotore a angolo fisso per centrifuga refrigerata</td><td>Eppendorf</td><td>F-34-6-38</td><td></td></tr><tr><td>Rotore a canna oscillante per centrifuga refrigerata</td><td>Eppendorf</td><td>A-4-62</td><td></td></tr><tr><td>Sucrosio</td><td>VWR</td><td>470302-810</td><td></td></tr><tr><td>Ultracentrifuga</td><td>Beckman Coulter</td><td>XPN-100</td><td></td></tr><tr><td>Tubi ultracentrifughe</td><td>Beckman Coulter</td><td>337986</td><td></td></tr><tr><td>Colorazione UranyLess</td><td>Scienze della microscopia elettronica</td><td>250218-01</td><td></td></tr></tbody></table>

isolamento e purificazione delle vescicole extracellulari vegetali dalle foglie di arabidopsis utilizzando un metodo ottimizzato di raccolta apoplastica

Isolation and Purification of Plant Extracellular Vesicles from Arabidopsis Leaves Using an Optimized Apoplastic Wash Collection Method

Isolamento e purificazione delle vescicole extracellulari vegetali dalle foglie di Arabidopsis utilizzando un metodo ottimizzato di raccolta apoplastica

Extracellular vesicles (EVs) are membrane-bound nanoparticles secreted by diverse organisms and cell types and are increasingly recognized as important ...

isolation-purification-plant-extracellular-vesicles-from-arabidopsis

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Questo video mostra una procedura per isolare protoplasti intatti da tessuti di 14-giorno-vecchie piantine di Arabidopsis. Dato che il protoplasti isolati rimangono intatte per almeno 96 ore e sono isolati da piantine invece di piante mature di un mese-vecchia, questa procedura accelera i test che richiedono protoplasti intatti.

Isolamento di protoplasti da tessuti di 14-giorni di età piantine di Arabidopsis thaliana</em

Fonte: Rufián, J. S., et al, Analisi a singola cellula dell'espressione dei geni di Pseudomonas syringae nel tessuto vegetale. J. Vis. Esp. (2022)Questo video mostra un metodo di pressione basato su siringa per estrarre batteri apoplastici, che proliferano e formano microcolonie all'interno dell'apoplasto, lo spazio intercellulare tra le cellule vegetali. Il metodo produce campioni batterici privi di detriti adatti all'analisi a valle di singola cellula.

Estrazione di batteri apoplastici utilizzando un metodo di pressione a base di siringa

Conoscenza della composizione del floema nonché il meccanismo del suo carico e trasporto a lunga distanza è essenziale per la comprensione della segnalazione a lunga distanza in sviluppo delle piante e di stress / risposta patogeno e di assimilare trasporto. Questo manoscritto descrive la raccolta di essudati floema con il metodo EDTA-agevolata.

Raccolta e analisi dei<em&gt; Arabidopsis</em&gt; Floema Essudati Utilizzo del metodo EDTA-agevolata

Questo protocollo descrive in dettaglio l'estrazione ottimizzata del fluido di lavaggio dell'apoplast dalle foglie delle piante, utilizzando le piante di fagioli francesi (Phaseolus vulgaris) come esempio di modello.

La tecnica infiltrazione-centrifugazione per l'estrazione di apoplastico fluido da foglie delle piante Utilizzo Phaseolus vulgaris Come un esempio

Qui presentiamo un protocollo di cattura di interazione applicato ai protoplasti a foglia di Arabidopsis thaliana . Questo metodo si basa fondamentalmente sulla creazione di reticolazione UV in vivo e consente l'isolamento e l'identificazione delle proteine ​​legate alla mRNA da un ambiente fisiologico.

Cattura di Interactome mRNA da protoplasti vegetali

Qui, descriviamo un metodo per purificare i cloroplasti intatti da foglie di Arabidopsis e loro tre sub-scomparti principali (busta, lo stroma e tilacoidi), utilizzando una combinazione di centrifugazione differenziale, gradienti di Percoll continui, e pendenze del saccarosio discontinuo. Il metodo è utile per subplastidial e localizzazione subcellulare delle proteine mediante immunoblotting e proteomica.

Preparazione del cloroplasto Sub-compartimenti da Arabidopsis per l'analisi di localizzazione della proteina di Immunoblotting o proteomica

Questo protocollo isola le vesciche extracellulari (EV) lontano dai virioni con elevata efficienza e resa incorporando precipitazioni EV, ultracentrifugazione del gradiente di densità e cattura di particelle per consentire un flusso di lavoro semplificato e una riduzione dell'avvio volume, con conseguente preparazione riproducibile per l'uso in tutte le ricerche EV.

Purificazione dei preparati vescicoli extracellulari ad alto rendimento lontano dal virus

Viene presentato un protocollo rapido ed efficiente per l'isolamento delle goccioline lipidiche di plastoglobulo associate a vari organismi fotosintetici. Il successo della preparazione di plastoglobuli isolati è un primo passo cruciale che precede indagini molecolari dettagliate come analisi proteomiche e lipidomiche.

Isolamento di goccioline lipidiche plastoglobule dal tessuto fogliare vegetale e cianobatteri

La purificazione delle proteine ricombinanti dai sistemi vegetali è solitamente messa alla prova dalle proteine vegetali. Questo lavoro fornisce un metodo per estrarre e purificare efficacemente una proteina ricombinante secreta dall'apoplasto di Nicotiana benthamiana.

Metodo basato sull'estrazione di Apoplast per migliorare la purezza delle proteine ricombinanti prodotte dalle piante  

Descriviamo un metodo semplice ma altamente riproducibile per isolare l'RNA extracellulare dalla superficie fogliare e dall'apoplasto delle piante di Arabidopsis.

Una guida passo passo all'isolamento e all'analisi della superficie fogliare e dell'RNA apoplastico utilizzando le rosette di Arabidopsis

Impara a isolare e purificare vescicole extracellulari vegetali da Arabidopsis thaliana con un protocollo ottimizzato di raccolta apoplastica. Questo metodo standardizzato minimizza la contaminazione citoplasmatica e preserva l'integrità delle vescicole.

Isolamento e purificazione delle vescicole extracellulari vegetali dalle foglie di Arabidopsis utilizzando un metodo ottimizzato di raccolta apoplastica (Video) | JoVE Journal

Isolamento e purificazione delle vescicole extracellulari vegetali dalle foglie di Arabidopsis utilizzando un metodo ottimizzato di raccolta apoplastica

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