Biochemical and tissue print analyses of hydroxyproline-rich glycoproteins in cell walls of sporophytic maize tissues

Abstract

In an effort to understand the role of hydroxyproline-rich glycoproteins (HRGPs) in plant cell wall structure, we studied the distribution and physical properties of PC-1-like proteins (PC-1 being the major pericarp HRGP) throughout sporophytic tissues of two maize (Zea mays L.) varieties. We determined total amounts of hydroxyproline, an indicator of HRGPs, and did tissue print and Western blot analysis. We found hydroxyproline in cell walls of stems, leaves, roots, tassels, and silks. We also observed reactivity of anti-PC-1 monoclonal antibodies with anatomical prints of these tissues on nitrocellulose paper. Stem nodes and silks contained the most hydroxyproline and exhibited the strongest reaction with the antibody. PC-1 was localized in vascular bundles and the epidermis of stem tissue. However, localization to a specific cell type in the silk could not be determined at the resolution of the tissue print. The stem node protein had the same electrophoretic mobility as the pericarp protein as determined on Western blots prepared from cationic neutral gels. Protein extracts from silk tissues of both varieties studied contained one protein of the same size/charge as that found in pericarp, as well as some minor variant bands. The data presented here document that cell wall proteins are present in many tissues of the maize plant, although they are primarily in cell types contributing to support.