Pollen Wall Structure Using a New Stripping-Sputtering Device for Scanning Electron Microscopy

A new method has been developed to elucidate pollen wall architecture by the separation of wall layers and the use of scanning electron microscopy (SEM). Separation of wall layers at natural boundaries, breakage across the wall and metal-coating of the specimen have been achieved by controlled ramming of free scattered ions produced by a novel “ion separating and coating – model A” instrument. The stripping treatment reveals interfaces and cross profiles of pollen walls and the sputtering treatment results in metal coating for examination with SEM. An advantage of the method is that it provides intact interfaces that are not eroded or damaged. The application of the method is exemplified by SEM analyses of pollen grains of Gossypium hirsutum L., Zea mays L., Sesamum indicum L. and Brassica napus L. var. oleifera. Interfaces between the tectum, column, foot, nexine-2 and intine layers of the pollen wall were all portrayed in G. hirsutum and to a great part in the other species. In G. hirsutum, it was possible to document the attachment point of surface spines, the appearance of individual baculae and the irregular labrum-operculum but regular inner labrum-aperture structure. No tectum was found in S. indicum. In all four species it was not possible to separate the intine from the sporoplast. The numbers of apertures were 20, 1, 10–14 and 3 in G. hirsutum, Z. mays, S. indicum and B. napus, respectively. The dumbell-shaped arrangement of apertures in G. hirsutum, the gear-shaped oblate sporoplast of S. indicum and the abundance of micropores on the intine of B. napus are characteristic features.