A comparative survey of proteins from recalcitrant tissues of a non‐model gymnosperm, D ouglas‐fir

Most research in plants and other organisms has, for the sake of convenience, focused on the use of model species to identify mechanisms that are conserved throughout the whole kingdom. Nevertheless, unique features and processes such as those related to plant cell wall and fiber formation, and to wood quality, sometimes need to be studied directly in the non‐model organism of interest. Such organisms, like the economically and ecologically important gymnosperm D ouglas‐fir ( P seudotsuga menziesii ), which is one of the crucial softwood timber species in Northern America, are often difficult to investigate. High phenolic, resin, and tannin contents in the woody tissues, as well as an incompletely sequenced genome, have contributed greatly to the species’ recalcitrance for molecular biology investigations. In this study, we present a complete procedure detailing protein sample preparation, separation, and proteomic analysis based on cross‐species identification of D ouglas‐fir. Proteins from the cambial zone, mature needles, and in vitro callus were extracted, purified, and separated via 1 D and 2 D SDS ‐ PAGE . One‐dimensional electrophoresis coupled with ESI ‐ MS / MS was used for cross‐species protein identification in order to evaluate the potential of this approach and reveal major differences in protein profiles among tested tissues. Identified proteins were functionally and developmentally compared. The likely contribution of these proteins to the properties of the cell wall and wood is indicated and discussed.