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Mitochondrial bioenergetics linked to the manifestation of programmed cell death during somatic embryogenesis of Abies alba

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Abstract

The present work reports changes in bioenergetic parameters and mitochondrial activities during the manifestation of two events of programmed cell death (PCD), linked to Abies alba somatic embryogenesis. PCD, evidenced by in situ nuclear DNA fragmentation (TUNEL assay), DNA laddering and cytochrome c release, was decreased in maturing embryogenic tissue with respect to the proliferation stage. In addition, the major cellular energetic metabolites (ATP, NAD(P)H and glucose-6-phosphate) were highered during maturation. The main mitochondrial activities changed during two developmental stages. Mitochondria, isolated from maturing, with respect to proliferating cell masses, showed an increased activity of the alternative oxidase, external NADH dehydrogenase and fatty-acid mediated uncoupling. Conversely, a significant decrease of the mitochondrial K +ATP channel activity was observed. These results suggest a correlation between mitochondrial activities and the manifestation of PCD during the development of somatic embryos. In particular, it is suggested that the K +ATP channel activity could induce an entry of K+ into the matrix, followed by swelling and a release of cytochrome c during proliferation, whereas the alternative pathways, acting as anti-apoptotic factors, may partially counteract PCD events occurring during maturation of somatic embryos.

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Abbreviations

ANT:

Adenine nucleotide translocase

AOX:

Cyanide-resistant alternative oxidase

DAPI:

4′,6-Diamidino-2-phenylindole

∆F:

Variations of fluorescence

DTE:

Dithioerythritol

ΔΨ:

Transmembrane electrical potential

ETC:

Electron transport chain

FFA:

Free fatty acids

Glu-6-P:

Glucose-6-phosphate

IgG:

Immunoglobulins

PCD:

Programmed cell death

PEG:

Polyethylene glycol

PEMs:

Pro-embryogenic cell masses

PMSF:

Phenylmethanesulphonyl fluoride

PTP:

Permeability transition pore

PUMP:

Plant uncoupling mitochondrial protein

PVPP:

Polyvinylpolypyrrolidone

ROS:

Reactive oxygen species

SE:

Somatic embryogenesis

TMR:

Tetramethyl-rhodamine

TUNEL:

Terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick end labelling

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Acknowledgments

Dr. Thomas Elthon (University of Nebraska, USA) is greatly acknowledged for providing the monoclonal anti-AOX antibody. Ms. Carla Calligaro (Animal Science Dept. of University of Udine, Italy) is greatly acknowledged for technical support provided during TUNEL and histological assays. This research was supported by the University of Udine.

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Authors and Affiliations

  1. Section of Plant Biology, Department of Biology and Plant Protection, University of Udine, via delle Scienze 91, 33100, Udine, Italy

    Elisa Petrussa, Alberto Bertolini, Valentino Casolo, Francesco Macrì & Angelo Vianello

  2. National Forest Centre, Forest Research Institute, T. G. Masaryka 22, 96092, Zvolen, Slovakia

    Jana Krajňáková

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  1. Elisa Petrussa
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  2. Alberto Bertolini
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  3. Valentino Casolo
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  4. Jana Krajňáková
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  5. Francesco Macrì
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  6. Angelo Vianello
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Correspondence to Angelo Vianello.

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Petrussa, E., Bertolini, A., Casolo, V. et al. Mitochondrial bioenergetics linked to the manifestation of programmed cell death during somatic embryogenesis of Abies alba . Planta 231, 93–107 (2009). https://doi.org/10.1007/s00425-009-1028-x

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