Online citations, reference lists, and bibliographies.
← Back to Search

Detecting Autophagy In Arabidopsis Roots By Membrane-permeable Cysteine Protease Inhibitor E-64d And Endocytosis Tracer FM4–64

Yuumi Oh-ye, Y. Inoue, Y. Moriyasu
Published 2011 · Biology, Medicine

Cite This
Download PDF
Analyze on Scholarcy
Autophagy is the process by which cells degrade their own components in lysosomes or vacuoles. Autophagy in tobacco BY-2 cells cultured in sucrose-free medium takes place in formed, autolysosomes in the presence of a cysteine protease inhibitor. The autolysosomes in BY-2 cells are located in the endocytotic pathway and thus can be stained with fluorescent endocytosis marker FM4–64. In the present study, in order to detect autophagy in the root cells of Arabidopsis, we incubated root tips from Arabidopsis seedlings in culture medium containing the membrane-permeable cysteine protease inhibitor E-64d and FM4–64, and examined whether autolysosomes stained with FM4–64 are accumulated. The results suggest that autophagy accompanying the formation of autolysosomes also occurs in Arabidopsis root cells. Such autophagy appeared to occur constitutively in the root cells in nutrient-sufficient culture medium. Even in atg5 mutants in which an autophagy-related gene is disrupted, accumulation of the structures stained with FM4–64, which likely correspond to autolysosomes, was seen although at lower level than in wild type roots.
This paper references
AtATG genes, homologs of yeast autophagy genes, are involved in constitutive autophagy in Arabidopsis root tip cells.
Y. Inoue (2006)
Vacuolar import of proteins and organelles from the cytoplasm.
D. Klionsky (1999)
A complex and mobile structure forms a distinct subregion within the continuous vacuolar membrane in young cotyledons of Arabidopsis.
C. Saito (2002)
Zur Entwicklung der Vacuole in Testa-Zellun des Leinsamens [On the development of the vacuole in the testa cells on Linum seeds
F Amelunxen (1984)
Processing of ATG8s, Ubiquitin-Like Proteins, and Their Deconjugation by ATG4s Are Essential for Plant Autophagy
K. Yoshimoto (2004)
Zur Entwicklung der Vacuole in Testa-Zellun des Leinsamens
F Amelunxen
A Novel type of Autophagy Occurs together with Vacuole Genesis in Miniprotoplasts Prepared from Tobacco Culture Cells
Kanako Yano (2007)
The Autophagic and Endocytic Pathways Converge at the Nascent Autophagic Vacuoles
W. Liou (1997)
Use of protease inhibitors for detecting autophagy in plants.
Y. Moriyasu (2008)
Autophagy in Tobacco Suspension-Cultured Cells in Response to Sucrose Starvation
Y. Moriyasu (1996)
Constitutive Autophagy in Plant Root Cells
Kanako Yano (2007)
Autophagy in tobacco BY-2 cells cultured under sucrose starvation conditions: isolation of the autolysosome and its characterization.
Chihiro Takatsuka (2011)
Vacuole biogenesis and protein transport to the plant vacuole: A comparison with the yeast vacuole and the mammalian lysosome
D. Robinson (2005)
Endosomal proteases facilitate the fusion of endosomes with vacuoles at the final step of the endocytotic pathway.
Kenji Yamada (2005)
Peroxisome Degradation by Microautophagy in Pichia pastoris: Identification of Specific Steps and Morphological Intermediates
Y. Sakai (1998)
Formation Process of Autophagosome Is Traced with Apg8/Aut7p in Yeast
T. Kirisako (1999)
Autophagy in yeast demonstrated with proteinase-deficient mutants and conditions for its induction
K. Takeshige (1992)
Cytochemical studies on GERL, provacuoles, and vacuoles in root meristematic cells of Euphorbia.
F. Marty (1978)
Zur Entwicklung der Vacuole in Testa-Zellen des Leinsamens
F. Amelunxen (1984)
Autophagy is not a main contributor to the degradation of phospholipids in tobacco cells cultured under sucrose starvation conditions.
Y. Inoue (2006)
Autophagy in plants
S. Kwon (2009)
Contribution of the plasma membrane and central vacuole in the formation of autolysosomes in cultured tobacco cells.
Kanako Yano (2004)
Autophagy and related mechanisms of lysosome-mediated protein degradation.
W. Dunn (1994)
Autophagy in Development and Stress Responses of Plants
D. Bassham (2006)
Autophagy as a regulated pathway of cellular degradation.
D. Klionsky (2000)
Divergent modes of autophagy in the methylotrophic yeast Pichia pastoris.
D. Tuttle (1995)
Protein Storage Bodies and Vacuoles
E. Herman (1999)
Uptake--microautophagy--and degradation of exogenous proteins by isolated rat liver lysosomes. Effects of pH, ATP, and inhibitors of proteolysis.
J. Ahlberg (1985)
Plant autophagy--more than a starvation response.
D. Bassham (2007)
Sequestration of cytoplasmic enzymes in an autophagic vacuole-lysosomal system induced by injection of leupeptin.
E. Kominami (1983)

This paper is referenced by
Redox-Mediated Endocytosis of a Receptor-Like Kinase during Distal Stem Cell Differentiation Depends on Its Tumor Necrosis Factor Receptor Domain1[OPEN]
Yingying Qin (2019)
Correlation of Autophagosome Formation with Degradation and Endocytosis Arabidopsis Regulator of G-Protein Signaling (RGS1) through ATG8a
Y. Jiao (2019)
In vivo imaging and quantitative monitoring of autophagic flux in tobacco BY-2 cells
Shigeru Hanamata (2013)
Autophagy-related approaches for improving nutrient use efficiency and crop yield protection.
Tamar Avin-Wittenberg (2018)
Quantitative live cell imaging of autophagic flux and roles of autophagy in reproductive development in plants
T. Kurusu (2016)
Methods for analysis of autophagy in plants.
D. Bassham (2015)
C. elegans and Arabidopsis thaliana show similar behavior: ROS induce escape tropisms both in illuminated nematodes and roots
K. Yokawa (2015)
COP1 E3 ligase protects HYL1 to retain microRNA biogenesis.
S. K. Cho (2014)
Heterotrimeric G-proteins in unfolded protein response mediate plant growth-defense tradeoffs upstream of steroid and immune signaling
Jimi C. Miller (2018)
TWS1, a Novel Small Protein, Regulates Various Aspects of Seed and Plant Development1[OPEN]
E. Fiume (2016)
The roles of autophagy in development and stress responses in Arabidopsis thaliana
X. Lv (2014)
The TOR Complex: An Emergency Switch for Root Behavior.
K. Yokawa (2016)
Reactive oxygen species derived from NADPH oxidase regulate autophagy
H. Jing (2020)
Autophagy is required for gamete differentiation in the moss Physcomitrella patens
Victoria Sanchez-Vera (2017)
Semantic Scholar Logo Some data provided by SemanticScholar