Abstract
As a major intracellular degradation pathway, autophagy contributes to nutrient recycling and is indispensable during plant senescence. Here we describe methods used for investigating the autophagic process during leaf senescence. These include transcript analysis of core machinery autophagy genes, immunoblotting of ATG8, and microscopic observation of autophagosome formation.
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References
Mizushima N, Yoshimori T, Ohsumi Y (2011) The role of Atg proteins in autophagosome formation. Annu Rev Cell Dev Biol 27:107–132
Diaz C, Lemaitre T, Christ A et al (2008) Nitrogen recycling and remobilization are differentially controlled by leaf senescence and development stage in Arabidopsis under low nitrogen nutrition. Plant Physiol 147:1437–1449
Guiboileau A, Yoshimoto K, Soulay F et al (2012) Autophagy machinery controls nitrogen remobilization at the whole-plant level under both limiting and ample nitrate conditions in Arabidopsis. New Phytol 194:732–740
Hanaoka H, Noda T, Shirano Y et al (2002) Leaf senescence and starvation-induced chlorosis are accelerated by the disruption of an Arabidopsis autophagy gene. Plant Physiol 129:1181–1193
Thompson AR, Doelling JH, Suttangkakul A et al (2005) Autophagic nutrient recycling in Arabidopsis directed by the ATG8 and ATG12 conjugation pathways. Plant Physiol 138:2097–2110
Xiong Y, Contento AL, Bassham DC (2005) AtATG18a is required for the formation of autophagosomes during nutrient stress and senescence in Arabidopsis thaliana. Plant J 42:535–546
Phillips AR, Suttangkakul A, Vierstra RD (2008) The ATG12-conjugating enzyme ATG10 is essential for autophagic vesicle formation in Arabidopsis thaliana. Genetics 178:1339–1353
Shibuya K, Niki T, Ichimura K (2013) Pollination induces autophagy in petunia petals via ethylene. J Exp Bot 64:1111–1120
Van Der Graaff E, Schwacke R, Schneider A et al (2006) Transcription analysis of arabidopsis membrane transporters and hormone pathways during developmental and induced leaf senescence. Plant Physiol 141:776–792
Wada S, Ishida H, Izumi M et al (2009) Autophagy plays a role in chloroplast degradation during senescence in individually darkened leaves. Plant Physiol 149:885–893
Ishida H, Yoshimoto K, Izumi M et al (2008) Mobilization of rubisco and stroma-localized fluorescent proteins of chloroplasts to the vacuole by an ATG gene-dependent autophagic process. Plant Physiol 148:142–155
Ono Y, Wada S, Izumi M et al (2013) Evidence for contribution of autophagy to rubisco degradation during leaf senescence in Arabidopsis thaliana. Plant Cell Environ 36:1147–1159
Contento AL, Xiong Y, Bassham DC (2005) Visualization of autophagy in Arabidopsis using the fluorescent dye monodansylcadaverine and a GFP-AtATG8e fusion protein. Plant J 42:598–608
Yoshimoto K, Hanaoka H, Sato S et al (2004) Processing of ATG8s, ubiquitin-like proteins, and their deconjugation by ATG4s are essential for plant autophagy. Plant Cell 16:2967–2983
Xiong Y, Contento AL, Nguyen PQ et al (2007) Degradation of oxidized proteins by autophagy during oxidative stress in Arabidopsis. Plant Physiol 143:291–299
Xia T, Xiao D, Liu D et al (2012) Heterologous expression of ATG8c from soybean confers tolerance to nitrogen deficiency and increases yield in Arabidopsis. PLoS One 7:e37217
Voinnet O, Rivas S, Mestre P et al (2003) An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. Plant J 33:949–956
Acknowledgments
This work was supported by the National Natural Science Foundation of China (31401179) and the Fundamental Research Funds for the Central Universities to QG.
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Cui, X., Zheng, J., Zheng, J., Gong, Q. (2018). Study of Autophagy in Plant Senescence. In: Guo, Y. (eds) Plant Senescence. Methods in Molecular Biology, vol 1744. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7672-0_23
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DOI: https://doi.org/10.1007/978-1-4939-7672-0_23
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Online ISBN: 978-1-4939-7672-0
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