Compression wood-responsive proteins in developing xylem of maritime pine (Pinus pinaster ait.).

Abstract

When a conifer shoot is displaced from its vertical position, compression wood (CW) is formed on the under side and can eventually return the shoot to its original position. Changes in cell wall structure and chemistry associated with CW are likely to result from differential gene/protein expression. Two-dimensional polyacrylamide gel electrophoresis of differentiating xylem proteins was combined with the physical characterization of wooden samples to identify and characterize CW-responsive proteins. Differentiating xylem was harvested from a 22-year-old crooked maritime pine (Pinus pinaster Ait.) tree. Protein extracted from different samples were revealed by high-resolution silver stained two-dimensional polyacrylamide gel electrophoresis and analyzed with a computer-assisted system for single spot quantification. Growth strain (GS) measurements allowed xylem samples to be classified quantitatively from normal wood to CW. Regression of lignin and cellulose content on GS showed that an increase in the percentage of lignin and a decrease of the percentage of cellulose corresponded to increasing GS values, i.e. CW. Of the 137 studied spots, 19% were significantly associated with GS effect. Up-regulated proteins included 1-aminocyclopropane-1-carboxylate oxidase (an ethylene forming enzyme), a putative transcription factor, two lignification genes (caffeic O-methyltransferase and caffeoyl CoA-O-methyltransferase), members of the S-adenosyl-L-methionine-synthase gene family, and enzymes involved in nitrogen and carbon assimilation (glutamine synthetase and fructokinase). A clustered correlation analysis was performed to study simultaneously protein expression along a gradient of gravistimulated stressed xylem tissue. Proteins were found to form "expression clusters" that could identify: (a) Gene product under similar control mechanisms, (b) partner proteins, or (c) functional groups corresponding to specialized pathways. The possibility of obtaining regulatory correlations and anticorrelations between proteins provide us with a new category of homology (regulatory homology) in tracing functional relationships.