• Fifteen poplar cDNA encoding fasciclin‐like arabinogalactan proteins (PopFLAs) were finely characterized, whereas the presence of arabinogalactan proteins (AGPs) was globally assessed during wood formation. • PopFLAs transcript accumulation was analysed through EST distribution in cDNA libraries, semi‐quantitative RT‐PCR, microarray experiment and Northern blot analysis. Similarly, AGPs contents were globally quantified by rocket electrophoresis. AGPs accumulation was further examined by Western blotting and immunocytolocalization. • Ten PopFLAs were specifically expressed in tension wood (TW) and not expressed in the cambial zone. Rocket electrophoresis revealed important AGPs accumulation in TW xylem. An anti‐AGPs specific antibody recognized two proteins preferentially present in the cell wall‐bound fraction from TW. Immunocytochemistry revealed a strong labelling close to the inner part of the G‐layer of TW fibres. • PopFLAs are expressed in xylem and many are up‐regulated in TW. It is suggested that some PopFLAs accumulating at the inner side of the G‐layer may have a specific function in the building of this layer. PopFLAs expression may therefore be linked to the specific mechanical properties of TW.
Xylogenesis involves successive developmental processes ‐ cambial division, cell expansion and differentiation, cell death ‐ each occurring along a gradient from the cambium to the pith of the stem. Taking advantage of the high level of organisation of wood tissues, we isolated cambial zone (CZ), differentiating xylem (DX) and mature xylem (MX) from both tension wood (TW) and opposite wood (OW) of bent poplars. Four different cDNA libraries were then constructed and used to generate 10 062 EST, reflecting the genes expressed in the different wood tissues. For the most abundant clusters, the EST distributions were compared between libraries in order to identify genes specific or over‐represented at some specific developmental stages. They clearly showed a developmental shift between CZ and DX, whereas there is a continuity of development between DX and MX. CZ was mainly characterized by clusters of genes involved in cell cycle, protein synthesis and fate. Interestingly, two clusters with no assigned function were found specific to the cambial zone. In DX and MX, clusters were mostly involved in methylation of lignin precursors and microtubule cytoskeleton. In addition, in DX, EST from TW and OW were compared: five clusters of arabinogalactan proteins, one for sucrose synthase and one for fructokinase were specific or over‐represented in TW. Moreover, a putative transcription factor and a cluster of unknown function were also identified in DX‐TW. The informative comparison of multiple libraries prepared from wood tissues led to the identification of genes ‐ some with still unknown functions ‐ putatively involved in xylogenesis and tension wood formation.
In functioning eucalypt ectomycorrhizas, biochemical alterations are accompanied by a differential accumulation of polypeptides including the synthesis of symbiosis-related proteins (JL Hilbert, Martin FM [1988] New Phytol 110: 339-346). In the present study, protein biosynthesis in the early stages of ectomycorrhiza formation on Eucalyptus globulus subsp. bicostata Kirkp. was examined using compatible and incompatible isolates of the basidiomycete Pisolithus tinctorius (Coker & Couch). Changes in polypeptide composition were observed within hours following contact of the compatible mycelium with the roots, well before the differentiation of typical symbiotic tissues. At this stage, at least seven symbiosis-related proteins (ectomycorrhizins) accumulated in root tissues. In vivo incorporation of [35S]methionine by ectomycorrhizas followed by electrophoresis of the labeled proteins revealed that most of these differences in polypeptide concentrations, including the ectomycorrhizin accumulation, are the result of differential protein biosynthesis rather than posttranslational modifications of the polypeptides. The initial development of eucalypt ectomycorrhizas, therefore, coincides with the synthesis of symbiosis-related proteins and the data presented here provide essential evidence to ascribe a functional developmental role to these proteins.
Cadmium (Cd) uptake by white lupin (Lupinus albus) was studied at low Cd concentrations (0.05nM to 5 μM) in hydroponic solution. Ten 12‐day old seedlings were pretreated in 0.5 mM CaCl2 solution in presence and absence of metabolic inhibitors (DCCD, DNP or NaN3). Cadmium solutions were labelled with carrier free 109CdCl2. Cadmium uptake was measured after a 2 h desorption in unlabelled CdCl2 solution. In the absence of any metabolic inhibitor and at 5 [μM Cd, roots absorbed 235.23 μg Cd/g root dry weight. Over the range of lnM to 5 μM Cd, exchangeable Cd represented approximately 5% of the absorbed fraction, and about 25 % of the total absorbed Cd was adsorbed to the root. Cadmium was passively absorbed to about 30% as observed in the presence of the inhibitor (DCCD). Ative absorption which represented 70% of Cd uptake involved H+‐ATPase carriers. Cadmium absorption was reduced to 30 and 20% in presence of lanthanum (La3+) and zinc (Zn2+), respectively which suggested that calcium (Ca), Cd, and Zn use the same carriers. Cadmium uptake in presence of DNP or NaN3 was approximately 4‐ fold that in control. Data showed presomption for an excretion of Cd out of root cells which could be the expression of a detoxification process limiting cell contamination.