Plant anatomy, histochemistry, and gene expression as visualized by tissue printing

Abstract

Tissue printing on nitrocellulose membranes followed by localization with appropriate probes has been used to study the distribution of protein and mRNA as well as the timing of gene expression in a variety of plant tissues. Included in these studies are cell wall proteins in soybean and cellulase activity in relation to abscission in kidney bean. Our recent studies have focused on the distribution of enzymes involved in crassulacean acid metabolism in the leaves of various species of Peperomia, the localization of ubiquitinated protein in various plant tissues, and the development of a stain for total protein transferred to nitrocellulose by tissue printing.Tissue prints made on dry nitrocellulose membranes (0.45μm pore size) and viewed with side illumination at low (10X) magnification show the characteristic morphology and anatomical organization of the printed tissue (Figs. 1 and 2). Treatment of the membranes with 12 mM HC1 followed by a two minute incubation in an acidic 0.05% solution of copper phthalocyanine 3,4',4'',4'''-tetrasulfonic acid tetrasodium salt (CPTS) (Figs. 3 and 4) shows the distribution of total protein transferred to the membrane.