Abstract
Programmed cell death (PCD) is a critical component of plant development, defense against invading pathogens, and response to environmental stresses. In this chapter, we provide detailed technical methods for studying PCD associated with plant development or induced by abiotic stress. A root hair assay or electrolyte leakage assay are excellent techniques for the quantitative determination of PCD and/or cellular injury induced in response to abiotic stress, whereas the lace plant provides a unique model that facilitates the study of genetically regulated PCD during leaf development.
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References
Kacprzyk J, Daly CT, McCabe PF (2011) The botanical dance of death: programmed cell death in plants. In: Kader JC, Delseny M (eds) Advances in botanical research, vol 60. Academic, UK, pp 169–261
Lockshin RA, Zakeri Z (2004) Apotosis, autophagy, and more. Int J Biochem Cell Biol 36:2405–2419
Fukuda H, Watanabe Y, Kuriyama H, Aoyagi S, Sugiyama M, Yamamoto R, Demura T, Minami A (1998) Programming of cell death during xylogenesis. J Plant Res 111:253–256
Gunawardena AHLAN, Greenwood JS, Dengler NG (2004) Programmed cell death remodels lace plant leaf shape during development. Plant Cell 16:60–73
Lombardi L, Ceccarelli N, Picciarelli P, Lorenzi R (2007) Caspase-like proteases involvement in programmed cell death of Phaseolus coccineus suspensor. Plant Sci 172:573–578
Simeonova E, Sikora A, Charzyńska M, Mostowska A (2000) Aspects of programmed cell death during leaf senescence of mono- and dicotyledonous plants. Protoplasma 214:93–101
Rogers HJ (2006) Programmed cell death in floral organs: how and why do flowers die? Ann Bot 97:309–315
Heath M (2000) Hypersensitive response-related death. Plant Mol Biol 44:321–334
Huh GH, Damsz B, Matsumoto TK, Reddy MP, Rus AM, Ibeas JI, Narasimhan ML, Bressan RA, Hasegawa PM (2002) Salt causes ion disequilibrium-induced programmed cell death in yeast and plants. Plant J 29:649–659
Jerome WG, Fuseler J, Price RL (2011) Specimen preparation. In: Jerome WG, Price RL (eds) Basic confocal microscopy. Springer, New York, pp 61–77
Lord CEN, Wertman JN, Lane S, Gunawardena AHLAN (2011) Do mitochondria play a role in remodelling lace plant leaves during programmed cell death? BMC Plant Biol 11:102
Gunawardena AHLAN, Navachandrabala C, Kane M, Dengler NG (2006) Lace plant: a novel system for studying developmental programmed cell death. In: da Silva JA T (ed) Floriculture, ornamental and plant biotechnology: advances and tropical issues. Global Science Books, Middlesex, pp 157–162
Wright H, van Doorn WG, Gunawardena AHLAN (2009) In vivo study of developmental programmed cell death using the lace plant (Aponogeton madagascariensis; Aponogetonaceae) leaf model system. Am J Bot 96:865–876
Wertman J, Lord CEN, Dauphinee AN, Gunawardena AHLAN (2012) The pathway of cell dismantling during programmed cell death in lace plant (Aponogeton madagascariensis) leaves. BMC Plant Biol 12:115
Hogg BV, Kacprzyk J, Molony EM, O’Reilly C, Gallagher TF, Gallois P, McCabe PF (2011) An in vivo root hair assay for determining rates of apoptotic-like programmed cell death in plants. Plant Methods 7:45
Kacprzyk J, McCabe PF (2015) A root hair assay to expedite cell death research. In: Estevez JM (ed) Plant cell expansion, vol 1242, Methods Mol Biol. Humana Press, New York, pp 73–82
Reape TJ, Molony EM, McCabe PF (2008) Programmed cell death in plants: distinguishing between different modes. J Exp Bot 59:435–444
McCabe PF, Leaver CJ (2000) Programmed cell death in cell cultures. Plant Mol Biol 44:359–368
He R, Drury GE, Rotari VI, Gordon A, Willer M, Farzaneh T, Woltering EJ, Gallois P (2008) Metacaspase-8 modulates programmed cell death induced by ultraviolet light and H2O2 in Arabidopsis. J Biol Chem 283:774–783
Mishiba KI, Nagashima Y, Suzuki E, Hayashi N, Ogata Y, Shimada Y, Koizumi N (2013) Defects in IRE1 enhance cell death and fail to degrade mRNAs encoding secretory pathway proteins in the Arabidopsis unfolded protein response. Proc Natl Acad Sci U S A 110:5713–5718
Acknowledgments
Department of Agriculture, Food & Marine (Ireland) is acknowledged for funding the work on ryegrass stress responses in the McCabe lab (VICCI grant. 14/S/819). The authors extend their thanks to the National Sciences and Engineering Research Council of Canada (NSERC) and the Killam Trusts for providing PhD funding to AND. AHLANG received funding from the Canadian Foundation for Innovation (CFI; Leaders Opportunity Fund) and the NSERC Discovery Grants Program.
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Time-lapse video of perforation formation via developmentally regulated programmed cell death (PCD) in a window stage lace plant (Aponogeton madagascariensis) leaf. Differential interference contrast (DIC) and dark field (DF) optics were used to monitor the same areole of a window stage leaf over a period of 48 h. PCD initiates in centralized cells and radiates outward before halting several cell layers before the longitudinal and transverse veins of the leaf (MOV 15,904 kb)
Time-lapse video of a lace plant (Aponogeton madagascariensis) late programmed cell death (LPCD) stage cell. The thin and transparent leaves of the lace plant make it an ideal tissue source for live cell imaging. In this fresh, unstained specimen the nuclei and nucleoli of cells are clearly visible. Chloroplasts can also be seen streaming within the cytoplasm or associated with the nuclei in a ring-like pattern. Aggregates of defunct organelles such as mitochondria and chloroplast can be seen undergoing Brownian motion in the vacuole. Nuclear displacement occurs late in the process, which is followed by collapse of the tonoplast and then plasma membrane. Following PCD, a packaged cell corpse is observed. 50× playback speed (MOV 73,470 kb)
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Kacprzyk, J., Dauphinee, A.N., Gallois, P., Gunawardena, A.H., McCabe, P.F. (2016). Methods to Study Plant Programmed Cell Death. In: Puthalakath, H., Hawkins, C. (eds) Programmed Cell Death. Methods in Molecular Biology, vol 1419. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3581-9_12
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DOI: https://doi.org/10.1007/978-1-4939-3581-9_12
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