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Annals of Botany 71: 161-167, 1993
© 1993 Annals of Botany Company

Visualization of Membrane Calcium and Calmodulin in Embryo Sacs in situ and Isolated from Petunia hybrida L. and Nicotiana tabacum L.

Uday K. Tirlapur, Jac L. Van Went and Mauro Cresti

Departimento di Biologia Ambientale, Universita di Siena, Via P.A.Mattioli 4, I-53100 Siena, Italy and Department of Plant Cytology and Morphology, Wageningen Agricultural University, Arboretumlaan 4, NL-6703 BD Wageningen, The Netherlands

We have used chlorotetracycline (CTC) and fluphenazine (FPZ) as fluorescent probes to visualize the distributional patterns of membrane calcium (mCa2+) and the Ca2+-receptor protein calmodulin (CaM) in various cell types of unfixed living isolated and unisolated embryo sacs of Petunia hydrida L. and Nicotiana tabacum L. Our results indicate that in the young embryo sacs of Petunia, both synergids and the central cell sequester relatively higher amounts of mCa2+ and CaM than the egg cell and the antipodals.

Much of the mCa2+ in the synergids is polarized in its distribution in that the mCa2+ is higher towards the micropylar end of the synergids. Interestingly, in the mature embryo sacs of Petunia only one of the two synergids and the egg cell proper manifest a higher level of mCa2+. In vivo only one of the synergids in the young as well as in the mature embryo sacs if Nicotiana consistently show higher mCa2+.

In the mature embryo sacs of Petunia the level of CaM is almost uniform in all the cell types except that one of the synergids and the three antipodal cells show a slightly higher level of CaM.

The possible implications of these findings in the late events of vectorial orientation of pollen tube tip, pollen-tube-synergid interactions and sperm delivery mechanism are discussed.Copyright 1993, 1999 Academic Press

Membrane-Ca2+, calmodulin, living embryo sacs, Petunia, Nicotiana, pollen tube-synergid interaction


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