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Three-dimensional reconstruction of anomalous chloroplasts in transgenic ipt tobacco

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Abstract

Anomalies in the ultrastructure of chloroplasts, from transgenic ipt tobacco, overproducing endogenous cytokinins (CKs) were studied. Detailed analyses of CKs and their metabolites showed that Pssu-ipt tobacco contained enhanced contents of CKs both in leaves and in isolated chloroplasts. The role of CKs in the formation of anomalous structures is suggested. Pssu-ipt chloroplasts frequently formed the distinct peripheral reticulum with a system of caverns that often involved mitochondria and/or peroxisomes. Large crystalloids, which were found in chloroplasts of Pssu-ipt, occupied up to 16% of chloroplast volume. We suggested that the crystalloids were formed by LHC II aggregates. This was supported by analysis of the fluorescence emission spectra at 77°K, chlorophyll a/b ratio, immunogold staining of the structures, and crystallographic unit size analysis.

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Abbreviations

C:

Control type rooted plants

CKs:

Cytokinins

DHZ:

Dihydrozeatin

DHZOG:

Dihydrozeatin O-glucoside

DHZR:

Dihydrozeatin riboside

DHZ7G:

Dihydrozeatin-7-glucoside

iP:

Isopentenyladenine

iPR:

Isopentenyladenine riboside

iP7G:

Isopentenyladenine-7-glucoside

iP9G:

Isopentenyladenine-9-glucoside

ipt :

The gene for isopentenyl transferase

LHC II:

Light-harvesting chlorophyll a/b of photosystem II

Pssu :

Promoter sequence of the gene coding for small subunit of RuBPCO

T:

Transgenic rooted plants

T/C:

Transgenic grafts

Z:

Zeatin

ZMP:

Zeatin riboside monophosphate

ZOG:

Zeatin O-glucoside

ZR:

Zeatin riboside

ZROG:

Zeatin riboside O-glucoside

Z7G:

Zeatin-7-glucoside

Z9G:

Zeatin-9-glucoside

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Acknowledgements

We are very grateful to Prof. Roland Valcke for his kind donation of transgenic Pssu-ipt. We also thank Dr. Noemi Čeřovská for the preparation of the LHC II antibodies. The work was supported by grants of the Grant Agency of the Czech Republic No. 206/01/1061, 206/03/0369, 206/03/0365, and MSM6007665808, AV0Z50510513.

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Authors and Affiliations

  1. Academy of Sciences of the CR, Na Karlovce 1a, Institute of Experimental Botany, 160 00, Praha 6, CZ, Czech Republic

    Helena Synková & Renáta Schnablová

  2. Department of Plant Anatomy and Physiology, Faculty of Sciences, Charles University, Viničná 5, 128 44, Praha 2, CZ, Czech Republic

    Renáta Schnablová & Lenka Polanská

  3. Academy of Sciences of the CR, Institute of Experimental Botany, Rozvojová 135, 165 02, Praha 6, Lysolaje, CZ, Czech Republic

    Lenka Polanská, Jiří Malbeck & Ivana Macháčková

  4. Institute of Physical Biology, University of South Bohemia, Branišovská 31, 370 05, České Budějovice, CZ, Czech Republic

    Michal Hušák & František Vácha

  5. Institute of Plant Molecular Biology, Branišovská 31, 370 05, České Budějovice, CZ, Czech Republic

    Pavel Šiffel & František Vácha

  6. Laboratory of Electron Microscopy, Institute of Parasitology, Branišovská 31, 370 05, České Budějovice, CZ, Czech Republic

    Jana Nebesářová

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  1. Helena Synková
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  2. Renáta Schnablová
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Correspondence to Helena Synková.

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Synková, H., Schnablová, R., Polanská, L. et al. Three-dimensional reconstruction of anomalous chloroplasts in transgenic ipt tobacco. Planta 223, 659–671 (2006). https://doi.org/10.1007/s00425-005-0119-6

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  • DOI: https://doi.org/10.1007/s00425-005-0119-6

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