Ultrastructural observations of cell wall regeneration around isolated tobacco protoplasts

Abstract

The process of cell wall regeneration around cultured protoplasts isolated from tobacco mesophyll has been examined by electron microscopy. The initially formed wall contains 2 components which stain with conventional heavy metal stains. The first consists of unbranched fibres, at first oriented at right angles to the plasmalemma surface. As wall growth proceeds the fibres lengthen and assume an orientation parallel to the plasmalemma. It seems probable that this component is cellulose. The second component of the wall is more amorphous and more densely stained. It is most frequently seen in situations where leaching of materials into the medium would be expected to be minimal. The endoplasmic reticulum and the plasmalemma are the only membrane systems which appear to contribute towards wall formation. No pattern of structure has been detected to explain the orientation or method of synthesis of the microfibrillar part of the wall.