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Plant Biol (Stuttg) 2001; 3(5): 524-535
DOI: 10.1055/s-2001-17734
Original Paper
Georg Thieme Verlag Stuttgart ·New York

Deregulation of Light-Induced Plastidogenesis in Etiolated Euglena gracilis Klebs Treated with DNA Hypermethylating 3′-Azido-3′-deoxythymidine

S. Pancaldi 1 , A. Bonora 1 , C. Baldisserotto 1 , R. Gualandri 1 , L. M. Neri 2 , M. P. Fasulo 1
  • 1 Department of Biology, Section of Botany, University of Ferrara, Ferrara, Italy
  • 2 Department of Morphology and Embriology, Section of Human Anatomy, Ferrara, Italy
Further Information

Publication History

December 21, 2000

July 31, 2001

Publication Date:
11 October 2001 (online)

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Abstract

The effect of the DNA hypermethylating 3′-azido-3′-deoxythymidine (AZT) on plastidogenesis was investigated on asynchronous, etiolated Euglena gracilis exposed to continuous light for 72 h. Based on (i) fluorescence and electron microscope aspects of the plastids, (ii) cytochemical localizations of functional PSI and PSII, (iii) immunolocalization of LHCII apoproteins, and (iv) spectrophotometric and HPLC analyses of the photosynthetic pigments, it was found that AZT, at a dose of 5 μM, which does not encroach on cell viability and only slightly interferes with cell proliferation, causes profound alterations of plastids in 10 - 15 % of the cells, without apparently affecting the rest of the cell population. Proplastid-to-chloroplast transformation slowed in this cell sub-population and occurred without the normal co-regulation that characterizes the process in wild type Euglena. In particular, thylakoid assembly was partially prevented while building materials continued to be synthesized and accumulated in the stroma. Chlorophylls were over-synthesized, while carotenoids, phaeophytin and phylloquinone dramatically diminished. The assemblage of PSI and PSII was inhibited. The external envelopes and thylakoid membranes were frequently damaged. Plastids, whose total volume increased with respect to the controls, were often malformed and interconnected. Their fluorescence under UV light declined quickly. The fact that AZT acts specifically at the plastid level, causing effects that are greatly opposed to those previously observed in the alga treated with the DNA demethylating 5-azacytidine, points to a relationship between DNA methylation and plastid differentiation in Euglena.

Abbreviations

AZT: 3′-azido-3′-deoxythymidine

car(s): carotenoid(s)

Chl(s): chlorophyll(s)

CLSM: confocal laser scanning microscope

DAB: 3,3′-diaminobenzidine

HPLC: high-performance liquid chromatography

LHCII: light harvesting complex II

PSI: photosystem I

PSII: photosystem II

TCNBT: thiocarbamyl nitroblue tetrazolium

Key words

Euglena - DNA hypermethylating agent - 3′-azido-3′-deoxythymidine - plastid differentiation

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M. P. Fasulo

Department of Biology
Section of Botany
University of Ferrara

C.so Porta Mare, 2
44100 Ferrara
Italy

Email: fsm@dns.unife.it

Section Editor: A. M. C. Emons