Isolation of Extracellular Vesicles from Phloem Sap by Size Exclusion Chromatography.
SEC
extracellular vesicles
phloem sap
stem incision
stylectomy
Journal
Current protocols
ISSN: 2691-1299
Titre abrégé: Curr Protoc
Pays: United States
ID NLM: 101773894
Informations de publication
Date de publication:
Oct 2023
Oct 2023
Historique:
medline:
10
10
2023
pubmed:
9
10
2023
entrez:
9
10
2023
Statut:
ppublish
Résumé
Extracellular vesicles (EVs) are nanoparticles that are released by cells and participate in the transfer of information. It is now known that EVs from mammalian cells are involved in different physiological and pathophysiological processes (antigen presentation, tissue regeneration, cancer, inflammation, diabetes, etc.). In the past few years, several studies on plants have demonstrated that EVs are also key tools for plant intercellular and cross-kingdom communications, suggesting that these nanostructures may contribute to distinct aspects of plant physiology such as development, defense, reproduction, symbiotic relationships, etc. These findings are challenging the traditional view of signaling in plants. EVs are probably involved in the phloem's transport system, since this vascular tissue plays a crucial role in translocating nutrients, defensive compounds, and informational signals throughout the plant. The collection of phloem is experimentally challenging because sap is under high turgor pressure inside the sieve elements, which have a small diameter and are hidden within the plant organs. The goals of this work are to develop new protocols that allow us to detect EVs for the first time in the phloem of the plants, and to isolate these nanovesicles for in-depth analysis and characterization. Our protocols describe two distinct methods to collect the phloem sap from rice and melon. The first method (Basic Protocol 1) involves 'Aphid stylectomy by radiofrequency microcautery' using rice plants and the aphid Sitobion avenae. This is considered the least invasive method for collecting phloem sap. The second method, 'Stem incision', involves cutting the stem of melon plants for collecting the exuded sap. Phloem sap EVs are then isolated by size exclusion chromatography. The results obtained in this study represent the first report on typical EVs isolated from in vivo-collected phloem sap. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Isolation of EVs from phloem sap: Aphid stylectomy by radiofrequency microcautery Basic Protocol 2: Isolation of EVs from phloem sap: Stem incision method.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e903Informations de copyright
© 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.
Références
Bachurski, D., Schuldner, M., Nguyen, P. H., Malz, A., Reiners, K. S., Grenzi, P. C., Babatz, F., Schauss, A. C., Hansen, H. P., Hallek, M., & Pogge von Strandmann, E. (2021). Extracellular vesicle measurements with nanoparticle tracking analysis-An accuracy and repeatability comparison between NanoSight NS300 and ZetaView. Journal of Extracellular Vesicles, 8(1), 1596016. https://doi.org/10.1080/20013078.2019.1596016
Benedikter, B. J., Bouwman, F. G., Vajen, T., Heinzmann, A. C. A., Grauls, G., Mariman, E. C., Wouters, E. F. M., Savelkoul, P. H., Lopez-Iglesias, C., Koenen, R. R., Rohde, G. G. U., & Stassen, F. R. M. (2017). Ultrafiltration combined with size exclusion chromatography efficiently isolates extracellular vesicles from cell culture media for compositional and functional studies. Science Reports, 7, 15297. https://doi.org/10.1038/s41598-017-15717-7
Böing, A. N., van der Pol, E., Grootemaat, A. E., Coumans, F. A., Sturk, A., & Nieuwland, R. (2014). Single-step isolation of extracellular vesicles by size-exclusion chromatography. Journal of Extracellular Vesicles, 3, 10. https://doi.org/10.3402/jev.v3.23430
Bokka, R., Ramos, A. P., Fiume, I., Manno, M., Raccosta, S., Turiák, L., Sugár, S., Adamo, G., Csizmadia, T., & Pocsfalvi, G. (2020). Biomanufacturing of tomato-derived nanovesicles. Foods, 9, 1852. https://doi.org/10.3390/foods9121852
Buhtz, A., Springer, F., Chappell, L., Baulcombe, D. C., & Kehr, J. (2008). Identification and characterization of small RNAs from the phloem of Brassica napus. The Plant Journal, 53, 739-749. https://doi.org/10.1111/j.1365-313X.2007.03368.x
Cai, Q., Qiao, L., Wang, M., He, B., Lin, F. M., Palmquist, J., Huang, S.-D., & Jin, H. (2018). Plants send small RNAs in extracellular vesicles to fungal pathogen to silence virulence genes. Science, 360, 1126-1129. https://doi.org/10.1126/science.aar4142
Chen, A., He, B., & Jin, H. (2022). Isolation of extracellular vesicles from Arabidopsis. Current Protocols, 2, e352. https://doi.org/10.1002/cpz1.352
Dinant, S., Bonnemain, J. L., Girouisse, C., & Kehr, J. (2010). Phloem sap intricacy and interplay with aphid feeding. Comptes Rendus Biologies, 333, 504-515. https://doi.org/10.1016/j.crvi.2010.03.008
Downing, N., & Unwin, D. M. (1977). A new method for cutting the mouth-parts of feeding aphids, and for collecting plant sap. Physiological Entomology, 2(4), 275-277. https://doi.org/10.1111/j.1365-3032.1977.tb00116.x
Fisher, D. B., & Frame, J. M. (1984). A guide to the use of the exuding stylet technique in phloem physiology. Planta, 161, 385-393. https://doi.org/10.1007/bf00394567
Garzo, E., Fernández-Pascual, M., Morcillo, C., Fereres, A., Gómez-Guillamón, M. L., & Tjallingii, W. F. (2018). Ultrastructure of compatible and incompatible interactions in phloem sieve elements during the stylet penetration by cotton aphids in melon. Insect Science, 26, 631-642. https://doi.org/10.1111/1744-7917.12447
He, B., Hamby, R., & Jin, H. (2021). Plant extracellular vesicles: Trojan horses of cross-kingdom warfare. FASEB BioAdvances, 3(9), 657-664. https://doi.org/10.1096/fba.2021-00040
Kehr, J., & Buhtz, A. (2008). Long distance transport and movement of RNA through the phloem. Journal of Experimental Botany, 59(1), 85-92. https://doi.org/10.1093/jxb/erm176
Kehr, J., & Kragler, F. (2018). Long distance RNA movement. New Phytologist, 218, 29-40. https://doi.org/10.1111/nph.15025
Killiny, N. (2019). Collection of the phloem sap, pros and cons. Plant Signaling and Behavior, 14(8), e1618181. https://doi.org/10.1111/j.1365-3032.1977.tb00116.x
Kim, D. K., & Rhee, W. J. (2021). Antioxidative effects of carrot-derived nanovesicles in cardiomyoblast and neuroblastoma cells. Pharmaceutics, 13, 1203. https://doi.org/10.3390/pharmaceutics13081203
Lohaus, G. (2022). Review primary and secondary metabolites in phloem sap collected with aphid stylectomy. Journal of Plant Physiology, 271, 153645. https://doi.org/10.1016/j.jplph.2022.153645
Lough, T. J., & Lucas, W. J. (2006). Integrative plant biology: Role of phloem long-distance macromolecular trafficking. Annual Review of Plant Biology, 57, 203-232. https://doi.org/10.1146/annurev.arplant.56.032604.144145
Lötvall, J., Hill, A. F., Hochberg, F., Buzás, E. I., Di Vizio, D., Gardiner, C., Gho, Y. S., Kurochkin, I. V., Mathivanan, S., Quesenberry, P., Sahoo, S., Tahara, H., Wauben, M. H., Witwer, K. W., & Thèry, C. (2014). Minimal experimental requirements for definition of extracellular vesicles and their functions: A position statement from the international society for extracellular vesicles. Journal of Extracellular Vesicles, 3, 26913. https://doi.org/10.3402/jev.v3.26913
Margolis, L., & Sadovsky, Y. (2019). The biology of extracellular vesicles: The known unknowns. PLoS Biology, 17(7), e3000363. https://doi.org/10.1371/journal.pbio.3000363
Mol, E. A., Goumans, M. J., Doevendans, P. A., Sluijter, J. P. G., & Vader, P. (2017). Higher functionality of extracellular vesicles isolated using size-exclusion chromatography compared to ultracentrifugation. Nanomedicine, 13(6), 2061-2065. https://doi.org/10.1016/j.nano.2017.03.011
Pérez-Bermúdez, P., Blesa, J., Soriano, J. M., & Marcilla, A. (2017). Extracellular vesicles in food: Experimental evidence of their secretion in grape fruits. European Journal of Pharmaceutical Science, 98, 40-50. https://doi.org/10.1016/j.ejps.2016.09.022
Pinedo, M., de la Canal, L., & de Marcos Lousa, C. (2021). A call for Rigor and standardization in plant extracellular vesicle research. Journal of Extracellular Vesicles, 10(6), e12048. https://doi.org/10.1002/jev2.12048
Regente, M., Corti-Monzón, G., Maldonado, A. M., Pinedo, M., Jorrín, J., & de la Canal, L. (2009). Vesicular fractions of sunflower apoplastic fluids are associated with potential exosome marker proteins. FEBS Letters, 583, 3363-3366. https://doi.org/10.1016/j.febslet.2009.09.041
Regente, M., Pinedo, M., San Clemente, H., Balliau, T., Jamet, E., & de la Canal, L. (2017). Plant extracellular vesicles are incorporated by a fungal pathogen and inhibit its growth. Journal of Experimental Botany, 68, 5485-5495. https://doi.org/10.1093/jxb/erx355
Royo, F., Théry, C., Falcón-Pérez, J. M., Nieuwland, R., & Witwer, K. W. (2020). Methods for separation and characterization of extracellular vesicles: Results of a worldwide survey performed by the ISEV rigor and standardization subcommittee. Cells, 9(9), 1955. https://doi.org/10.3390/cells9091955
Rutter, B. D., & Innes, R. W. (2017). Extracellular vesicles isolated from the leaf apoplast carry stress-response proteins. Plant Physiology, 173(1), 728-741. https://doi.org/10.1104/pp.16.01253
Sánchez-López, C. M., Manzaneque-López, M. C., Pérez-Bermúdez, P., Soler, C., & Marcilla, A. (2022). Characterization and bioactivity of extracellular vesicles isolated from pomegranate. Food and Function, 13, 12870. https://doi.org/10.1039/d2fo01806c
Sidhom, K., Obi, P. O., & Saleem, A. (2020). A review of exosomal isolation methods: Is size exclusion chromatography the best option? International Journal of Molecular Science, 21, 6466. https://doi.org/10.3390/ijms21186466
Théry, C., Witwer, K. W., Aikawa, E., Alcaraz, M. J., Anderson, J. D., Andriantsitohaina, R., Antoniou, A., Arab, T., Archer, F., Atkin-Smith, G. K., Ayre, D. C., Bach, J. -. M., Bachurski, D., Baharvand, H., Balaj, L., Baldacchino, S., Bauer, N. N., Baxter, A. A., Bebawy, M., … Zuba-Surma, E. K. (2018). Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the international society for extracellular vesicles and update of the MISEV2014 guidelines. Journal of Extracellular Vesicles, 7(1), 1535750. https://doi.org/10.1080/20013078.2018.1535750
Tjallingii, W. F. (1978). Electronic recording of penetration behaviour by aphids. Entomologia Experimentalis et Applicata, 24, 721-730. https://doi.org/10.1111/j.1570-7458.1985.tb03516.x
Tjallingii, W. F. (1985). Electrical nature of recorded signals during stylet penetration by aphids. Entomologia Experimentalis et Applicata, 38, 177-186. https://doi.org/10.1111/j.1570-7458.1985.tb03516.x
You, J. Y., Kang, S. J., & Rhee, W. J. (2021). Isolation of cabbage exosome-like nanovesicles and investigation of their biological activities in human cells. Bioactive Materials, 6, 4321. https://doi.org/10.1016/j.bioactmat.2021.04.023