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Confocal image analysis of spatial variations in immunocytochemically identified calmodulin during pollen hydration, germination and pollen tube tip growth in Nicotiana tabacum L.

Published online by Cambridge University Press:  26 September 2008

Uday K. Tirlapur ,
Monica Scali ,
Alessandra Moscatelli ,
Cecilia Del Casino ,
Gianpiero Cai ,
Antonio Tiezzi  and
Mauro Cresti
Uday K. Tirlapur
Affiliation:
Dipartimento di Biologia Ambientale, Università di Siena, and Dipartimento Science Ambientale, Università Della Tuscia, Viterbo, Italy
Monica Scali
Affiliation:
Dipartimento di Biologia Ambientale, Università di Siena, and Dipartimento Science Ambientale, Università Della Tuscia, Viterbo, Italy
Alessandra Moscatelli
Affiliation:
Dipartimento di Biologia Ambientale, Università di Siena, and Dipartimento Science Ambientale, Università Della Tuscia, Viterbo, Italy
Cecilia Del Casino
Affiliation:
Dipartimento di Biologia Ambientale, Università di Siena, and Dipartimento Science Ambientale, Università Della Tuscia, Viterbo, Italy
Gianpiero Cai
Affiliation:
Dipartimento di Biologia Ambientale, Università di Siena, and Dipartimento Science Ambientale, Università Della Tuscia, Viterbo, Italy
Antonio Tiezzi
Affiliation:
Dipartimento di Biologia Ambientale, Università di Siena, and Dipartimento Science Ambientale, Università Della Tuscia, Viterbo, Italy
Mauro Cresti*
Affiliation:
Dipartimento di Biologia Ambientale, Università di Siena, and Dipartimento Science Ambientale, Università Della Tuscia, Viterbo, Italy
*
M. Cresti, Dipartimento di Biologia Ambientale, Università di Siena, Via P.A. Mattioli 4, Sienna, Italy. Telephone: 0039 577 298854. Fax: 0039 577 298860.
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Summary

Using monoclonal anti-calmodulin antibodies in conjunction with confocal scanning laser microscopy we have analysed the spatial variations in the distribution pattern of calmodulin (CaM) during the sequential events of pollen hydration, germination and tube growth in Nicotiana tabacum. These immunocytochemical observations have been complemented by immunochemical studies wherein the anti-calmodulin antibody raised against pea CaM recognises a polypeptide of c. 18 kDa in the pollen extracts. Digitisation of confocally acquired optical sections of immunofluorescence images reveals that in hydrated pollen a high level of CaM is consistently present in the region of the germinal apertures. Subsequently, with the onset of germination a high CaM concentration was found associated with the plasma membrane of the germination bubble and in the cytoplasm in its vicinity, while in the vegetative cytoplasm a weak diffuse and intense punctate signal was registered. CaM immunostain was also detected in association with the plasma membrane of the tube tips in both short and long pollen tubes. Furthermore, the cytosol of the tubes invariably manifested an apically focused CaM gradient. We were, however, unable to detect any vacuolar association of CaM in the older regions of the pollen tubes. Although punctate immunostain was obvious across the pollen tube numerous punctate structures were invariably present in the extreme tip. The possible implications of these findings in development of cell polarity, polarised growth, maintenance of calcium homeostasis and CaM interactions with other mechanochemical motor proteins in effecting propulsion of organelles during pollen hydration, germination and pollen tube growth are discussed.

Keywords

CalmodulinConfocal microscopyGradientNicotiana tabacumPolarized growthPollen germination
Type
Commentary
Information
Zygote , Volume 2 , Issue 1 , February 1994 , pp. 63 - 68
Copyright
Copyright © Cambridge University Press 1994

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