Environmentally induced programmed cell death in leaf protoplasts of Aponogeton madagascariensis

Christina E.N. Lord; Arunika H.L.A.N. Gunawardena

doi:10.1007/s00425-010-1304-9

Environmentally induced programmed cell death in leaf protoplasts of Aponogeton madagascariensis

Christina E.N. Lord, Arunika H.L.A.N. Gunawardena

Medicine

Research output: Contribution to journal › Article › peer-review

25 Citations (Scopus)

Abstract

Within plant systems, two main forms of programmed cell death (PCD) exist: developmentally regulated and environmentally induced. The lace plant (Aponogeton madagascariensis) naturally undergoes developmentally regulated PCD to form perforations between longitudinal and transverse veins over its leaf surface. Developmental PCD in the lace plant has been well characterized; however, environmental PCD has never before been studied in this plant species. The results presented here portray heat shock (HS) treatment at 55°C for 20 min as a promising inducer of environmental PCD within lace plant protoplasts originally isolated from non-PCD areas of the plant. HS treatment produces cells displaying many characteristics of developmental PCD, including blebbing of the plasma membrane, increased number of hydrolytic vesicles and transvacuolar strands, nuclear condensation, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling positive nuclei, as well as increased Brownian motion within the vacuole. Results presented here for the first time provide evidence of chloroplasts in the vacuole of living protoplasts undergoing environmentally induced PCD. Findings suggest that the mitochondria play a critical role in the cell death process. Changes in mitochondrial dynamics were visualized in HS-treated cells, including loss of mitochondrial mobility, reduction in ΔΨ_m, as well as the proximal association with chloroplasts. The role of the mitochondrial permeability transition pore (PTP) was examined by pre-treatment with the PTP agonist cyclosporine A. Overall, HS is depicted as a reliable method to induce PCD within lace plant protoplasts, and proves to be a reliable technique to enable comparisons between environmentally induced and developmentally regulated PCD within one species of plant.

Original language	English
Pages (from-to)	407-421
Number of pages	15
Journal	Planta
Volume	233
Issue number	2
DOIs	https://doi.org/10.1007/s00425-010-1304-9
Publication status	Published - Feb 2011

Bibliographical note

Funding Information:
Acknowledgments The authors thank Dr. M. E. Kane (University of Florida, USA) for providing the original sterile lace plant cultures, Jaime Wertman (Dalhousie University, Canada) for critical review of this article, Bruno J. Roy (Dalhousie University, Canada) for assistance in formatting supplementary videos, Dr. Nancy Dengler (University of Toronto, Canada) and Dr. Wouter G. van Doorn (Wageningen University, The Netherlands) for help throughout the manuscript, and Dr. Jennifer Sheen (Mass General Hospital, Molecular Biology, Boston, MA, USA) for help regarding protoplast isolation. The authors also greatly acknowledge the Canadian Foundation for Innovation (CFI) for the Leaders Opportunity Fund, the Natural Sciences and Engineering Research Council (NSERC) for discovery and equipment grants for A.G., and Dalhousie University for partial doctoral funding for C.L.

ASJC Scopus Subject Areas

Genetics
Plant Science

PubMed: MeSH publication types

Journal Article
Research Support, Non-U.S. Gov't

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10.1007/s00425-010-1304-9

Cite this

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title = "Environmentally induced programmed cell death in leaf protoplasts of Aponogeton madagascariensis",

abstract = "Within plant systems, two main forms of programmed cell death (PCD) exist: developmentally regulated and environmentally induced. The lace plant (Aponogeton madagascariensis) naturally undergoes developmentally regulated PCD to form perforations between longitudinal and transverse veins over its leaf surface. Developmental PCD in the lace plant has been well characterized; however, environmental PCD has never before been studied in this plant species. The results presented here portray heat shock (HS) treatment at 55°C for 20 min as a promising inducer of environmental PCD within lace plant protoplasts originally isolated from non-PCD areas of the plant. HS treatment produces cells displaying many characteristics of developmental PCD, including blebbing of the plasma membrane, increased number of hydrolytic vesicles and transvacuolar strands, nuclear condensation, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling positive nuclei, as well as increased Brownian motion within the vacuole. Results presented here for the first time provide evidence of chloroplasts in the vacuole of living protoplasts undergoing environmentally induced PCD. Findings suggest that the mitochondria play a critical role in the cell death process. Changes in mitochondrial dynamics were visualized in HS-treated cells, including loss of mitochondrial mobility, reduction in ΔΨm, as well as the proximal association with chloroplasts. The role of the mitochondrial permeability transition pore (PTP) was examined by pre-treatment with the PTP agonist cyclosporine A. Overall, HS is depicted as a reliable method to induce PCD within lace plant protoplasts, and proves to be a reliable technique to enable comparisons between environmentally induced and developmentally regulated PCD within one species of plant.",

author = "Lord, {Christina E.N.} and Gunawardena, {Arunika H.L.A.N.}",

note = "Funding Information: Acknowledgments The authors thank Dr. M. E. Kane (University of Florida, USA) for providing the original sterile lace plant cultures, Jaime Wertman (Dalhousie University, Canada) for critical review of this article, Bruno J. Roy (Dalhousie University, Canada) for assistance in formatting supplementary videos, Dr. Nancy Dengler (University of Toronto, Canada) and Dr. Wouter G. van Doorn (Wageningen University, The Netherlands) for help throughout the manuscript, and Dr. Jennifer Sheen (Mass General Hospital, Molecular Biology, Boston, MA, USA) for help regarding protoplast isolation. The authors also greatly acknowledge the Canadian Foundation for Innovation (CFI) for the Leaders Opportunity Fund, the Natural Sciences and Engineering Research Council (NSERC) for discovery and equipment grants for A.G., and Dalhousie University for partial doctoral funding for C.L.",

year = "2011",

month = feb,

doi = "10.1007/s00425-010-1304-9",

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N1 - Funding Information: Acknowledgments The authors thank Dr. M. E. Kane (University of Florida, USA) for providing the original sterile lace plant cultures, Jaime Wertman (Dalhousie University, Canada) for critical review of this article, Bruno J. Roy (Dalhousie University, Canada) for assistance in formatting supplementary videos, Dr. Nancy Dengler (University of Toronto, Canada) and Dr. Wouter G. van Doorn (Wageningen University, The Netherlands) for help throughout the manuscript, and Dr. Jennifer Sheen (Mass General Hospital, Molecular Biology, Boston, MA, USA) for help regarding protoplast isolation. The authors also greatly acknowledge the Canadian Foundation for Innovation (CFI) for the Leaders Opportunity Fund, the Natural Sciences and Engineering Research Council (NSERC) for discovery and equipment grants for A.G., and Dalhousie University for partial doctoral funding for C.L.

PY - 2011/2

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N2 - Within plant systems, two main forms of programmed cell death (PCD) exist: developmentally regulated and environmentally induced. The lace plant (Aponogeton madagascariensis) naturally undergoes developmentally regulated PCD to form perforations between longitudinal and transverse veins over its leaf surface. Developmental PCD in the lace plant has been well characterized; however, environmental PCD has never before been studied in this plant species. The results presented here portray heat shock (HS) treatment at 55°C for 20 min as a promising inducer of environmental PCD within lace plant protoplasts originally isolated from non-PCD areas of the plant. HS treatment produces cells displaying many characteristics of developmental PCD, including blebbing of the plasma membrane, increased number of hydrolytic vesicles and transvacuolar strands, nuclear condensation, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling positive nuclei, as well as increased Brownian motion within the vacuole. Results presented here for the first time provide evidence of chloroplasts in the vacuole of living protoplasts undergoing environmentally induced PCD. Findings suggest that the mitochondria play a critical role in the cell death process. Changes in mitochondrial dynamics were visualized in HS-treated cells, including loss of mitochondrial mobility, reduction in ΔΨm, as well as the proximal association with chloroplasts. The role of the mitochondrial permeability transition pore (PTP) was examined by pre-treatment with the PTP agonist cyclosporine A. Overall, HS is depicted as a reliable method to induce PCD within lace plant protoplasts, and proves to be a reliable technique to enable comparisons between environmentally induced and developmentally regulated PCD within one species of plant.

AB - Within plant systems, two main forms of programmed cell death (PCD) exist: developmentally regulated and environmentally induced. The lace plant (Aponogeton madagascariensis) naturally undergoes developmentally regulated PCD to form perforations between longitudinal and transverse veins over its leaf surface. Developmental PCD in the lace plant has been well characterized; however, environmental PCD has never before been studied in this plant species. The results presented here portray heat shock (HS) treatment at 55°C for 20 min as a promising inducer of environmental PCD within lace plant protoplasts originally isolated from non-PCD areas of the plant. HS treatment produces cells displaying many characteristics of developmental PCD, including blebbing of the plasma membrane, increased number of hydrolytic vesicles and transvacuolar strands, nuclear condensation, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling positive nuclei, as well as increased Brownian motion within the vacuole. Results presented here for the first time provide evidence of chloroplasts in the vacuole of living protoplasts undergoing environmentally induced PCD. Findings suggest that the mitochondria play a critical role in the cell death process. Changes in mitochondrial dynamics were visualized in HS-treated cells, including loss of mitochondrial mobility, reduction in ΔΨm, as well as the proximal association with chloroplasts. The role of the mitochondrial permeability transition pore (PTP) was examined by pre-treatment with the PTP agonist cyclosporine A. Overall, HS is depicted as a reliable method to induce PCD within lace plant protoplasts, and proves to be a reliable technique to enable comparisons between environmentally induced and developmentally regulated PCD within one species of plant.

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Environmentally induced programmed cell death in leaf protoplasts of Aponogeton madagascariensis

Abstract

Bibliographical note

ASJC Scopus Subject Areas

PubMed: MeSH publication types

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