Ultrastructural effects of lysozymes on the cell wall of Caryophanon latum.

When Caryophanon latum was exposed to egg white lysozyme in isotonic sucrose and observed by phase-contrast microscopy, protoplasts emerged along the length of the trichomes, apparently at sites corresponding to cross septa. Electron microscopy of sections revealed that this enzyme initially attacked the core of the septal peptidoglycan and delamination of septa resulted. The inner densely staining layer of the lateral and polar wall (considered to contain peptidoglycan as the major component) remained intact except for destruction at the advancing tip of partial septa; protoplasts or cell debris could escape from the gaps formed at developing septa. Treatment of intact trichomes with pronase, a lipase - phospholipase C mixture, EDTA, glutaraldehyde, or heat, before exposure to egg white lysozyme did not alter this pattern nor did it render the remaining peptidoglycan more susceptible to attack. The wall material external to the peptidoglycan was solubilized by pronase. The peptidoglycan remaining after lysozyme treatment was not morphologically changed by treatment with pronase. Lysozyme derived from Chalaropsis hydrolyzed incomplete septa initially, while the lateral and polar wall and complete septa were degraded later. Therefore, it is most probable that the inner dense layer does contain the peptidoglycan component and that some biochemical maturation distinguishes the substrate for these enzymes in the lateral wall and septa.