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Journal of Electron Microscopy Advance Access published online on April 1, 2008
Journal of Electron Microscopy, doi:10.1093/jmicro/dfn005
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Surface replicas of normal and vitrified leaves of Datura insignis, Barb Rodr**

Flávio Costa Miguens*

Laboratório de Biologia Celular e Tecidual, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darci Ribeiro, Avenida Alberto Lamego 2000, Parque Califórnia, 28013-600, Campos dos Goytacazes, Rio de Janeiro, Brazil

* To whom correspondence should be addressed. E-mail: flavio.miguens{at}pq.cnpq.br

Vitrification is a morphological and physiological disorder affecting plants during their in vitro vegetative propagation. Vitrified plants have a poor survival rate when transferred from in vitro to greenhouse conditions, a fact mainly due to water loss and dissecation. It has been shown that normal and vitreous leaves of Datura insignis differ in the frequency of normal and abnormal stomata. The purpose of this work was to compare the surface of normal and vitreous leaves of D. insignis, using a modification of the platinum/carbon replica method. Adaxial and abaxial leaf surfaces of normal plantlets have a smooth and homogenous cuticle. A granular aspect, probably due to leaf age, rarely occurs at the periphery of the epidermal cells. Both adaxial and abaxial leaf surfaces of vitrified plantlets show discontinuities in the cuticle, occurring at several regions of the outer periclinal cell walls. However, such discontinuities are most noticeable in the region between adjacent epidermal cells. Fibrils 20–30 nm thick show a random arrangement or an oriented pattern in cuticular discontinuities. In D. insignis vitrified plantlets, adaxial and abaxial leaf cuticle has discontinuities or gaps which may cause an increase in cuticular transpiration contributing to the low survival rate of vitrified plantlets.

Keywords     plant cuticle gaps, platinum/carbon replica, scanning electron microscopy, transmission electron microscopy, plant vitrification

Received     28 December 2007, accepted 24 February 2008

** I dedicate this paper to honour the memory of Raul Dodsworth Machado (1917–1996) for his scientific merit and example of conduct which inspired us all.


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