Abstract
The classical Golgi pathway is not the only mechanism for vacuolar protein transport in plants because alternative transport mechanisms have been described. The existence of these alternative pathways can be demonstrated using several chemicals and here we describe the use of brefeldin A (BFA), endo-β-N-acetylglucosaminidase H (Endo-H), and tunicamycin, on isolated tobacco leaf protoplasts. Two main methods are illustrated in this chapter, protoplast pulse-chase followed by protein immunoprecipitation, and protoplast immunofluorescence.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Brandizzi F, Barlowe C (2013) Organization of the ER-Golgi interface for membrane traffic control. Nat Rev Mol Cell Biol 14:382–392
Hwang I (2008) Sorting and anterograde trafficking at the Golgi apparatus. Plant Physiol 148:673–683
Xiang L, Etxeberria E, Van den Ende W (2013) Vacuolar protein sorting mechanisms in plants. FEBS J 280:979–993
Robinson DG, Brandizzi F, Hawes C, Nakano A (2015) Vesicles versus tubes: Is endoplasmic reticulum-Golgi transport in plants fundamentally different from other Eukaryotes? Plant Physiol 168:393–406
Herman EM, Schmidt M (2004) Endoplasmic reticulum to vacuole trafficking of endoplasmic reticulum bodies provides an alternate pathway for protein transfer to the vacuole. Plant Physiol 136:3440–3446
Honig A, Avin-Wittenberg T, Ufaz S, Galili G (2012) A new type of compartment, defined by plant-specific Atg8-interacting proteins, is induced upon exposure of Arabidopsis plants to carbon starvation. Plant Cell 24:288–303
De Marchis F, Bellucci M, Pompa A (2013) Unconventional pathways of secretory plant proteins from the endoplasmic reticulum to the vacuole bypassing the Golgi complex. Plant Signal Behav 8, e25129
De Marchis F, Bellucci M, Pompa A (2013) Traffic of human α-mannosidase in plant cells suggests the presence of a new endoplasmic reticulum-to-vacuole pathway without involving the Golgi complex. Plant Physiol 161:1769–1782
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158
Watanabe N, Lam E (2007) BAX inhibitor-1 modulates endoplasmic reticulum stress-mediated programmed cell death in Arabidopsis. J Biol Chem 283:3200–3210
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
De Marchis, F., Pompa, A., Bellucci, M. (2016). Chemical Secretory Pathway Modulation in Plant Protoplasts. In: Pompa, A., De Marchis, F. (eds) Unconventional Protein Secretion. Methods in Molecular Biology, vol 1459. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3804-9_4
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3804-9_4
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3802-5
Online ISBN: 978-1-4939-3804-9
eBook Packages: Springer Protocols