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Secretion marker proteins and cell-wall polysaccharides move through different secretory pathways

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Abstract

The building up of the cell wall is tightly dependent on the functionality of the secretory pathway. Syntaxins as well as other SNARE proteins play important roles during vesicle secretion and fusion. We have compared the secretion of newly synthesised cell-wall polysaccharides to that of secretory marker proteins such as secreted green-fluorescent protein (secGFP) and secreted rat preputial β-glucuronidase (secRGUS) in leaf protoplasts and roots of wild-type and transgenic Nicotiana tabacum plants, overexpressing a syntaxin homologue NtSyr1 (Sp1) and its soluble variant Sp2 that interferes specifically with Sp1 function, affecting post-Golgi transport. In protoplasts transiently transformed with secGFP and Sp1, no variation was observed in the pattern of fluorescence with respect to control; on the contrary, GFP fluorescence accumulate within the cells in protoplasts co-transformed with secGFP and Sp2. Sp2 reduced the percentage of marker protein secretion to 53% as quantified with secRGUS. In protoplasts obtained from leaves of wild-type and transformed tobacco plants expressing Sp1, Sp2 and Sp1 plus Sp2, no remarkable differences in the percentage of newly synthesised polysaccharides incorporated into the regenerating cell walls were observed. The same results were confirmed in roots of whole transformed seedlings. Tests with cytochalasin D (CD) showed a marked decrease in the amount of newly synthesised polysaccharides into the wall and a simultaneous sharp increase in membrane-associated polysaccharides. SecRGUS secretion was also inhibited by CD. The data indicate that marker proteins and matrix polysaccharides, as well as cellulose synthase complexes, are secreted through the involvement of different secretory machineries.

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Abbreviations

BFA:

Brefeldin A

CD:

Cytochalasin

dex:

Dexamethasone

GFP:

Green-fluorescent protein

RGUS:

Rat preputial β-glucuronidase

Sp1:

Syntaxin in plant 1; syntaxin homologue NtSyr1 (SYP121)

Sp2:

Soluble variant of Sp1

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Acknowledgments

We would like to thank Prof. M.R. Blatt (Glasgow University) for providing transgenic seeds. Special thanks to ISPA-CNR section of Lecce for the use of Zeiss confocal microscope. This work was supported by a grant from the Ministero dell’Università e della Ricerca Scientifica e Tecnologica (MURST- PRIN 2005).

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  1. Di.S.Te.B.A., Università di Lecce, via prov.le Lecce-Monteroni, 73100, Lecce, Italy

    Maria Rosaria Leucci, Gian-Pietro Di Sansebastiano, Massimiliano Gigante, Giuseppe Dalessandro & Gabriella Piro

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  1. Maria Rosaria Leucci
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  2. Gian-Pietro Di Sansebastiano
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  3. Massimiliano Gigante
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  4. Giuseppe Dalessandro
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Correspondence to Gabriella Piro.

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Maria Rosaria Leucci and Gian-Pietro Di Sansebastiano contribute equally to this work.

Giuseppe Dalessandro and Gabriella Piro wish to dedicate this work to the memory of Professor Don Northcote.

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Leucci, M.R., Di Sansebastiano, GP., Gigante, M. et al. Secretion marker proteins and cell-wall polysaccharides move through different secretory pathways. Planta 225, 1001–1017 (2007). https://doi.org/10.1007/s00425-006-0407-9

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