Search

Black walnut ( Juglans nigra L.) heartwood is characterised by the appearance of a dark-brown colour in the inner part of the trunk. At the cellular level, phenolic compounds and peroxidases (PODs, EC 1.11.1.7) are most likely involved in the generation of this colour. Ortho -diphenols are revealed by Arnow’s reagent and condensed tannins by vanillin-HCl or p -dimethylaminocinnamaldehyde. These reagents reveal that only a small percentage of the living sapwood parenchyma cells accumulate phenolic compounds in the vacuole. POD activity is revealed either with guaiacol or 3,3’-diaminobenzidine. The simultaneous disclosure of o -diphenols and POD in the same cross-section clearly indicates that POD activity and its phenolic substrates are localised in different ray parenchyma cells of walnut sapwood. None of the cells exhibiting POD activity contain o -diphenols and vice versa. This compartmentation of active PODs and of their potential substrates in different cells within the living woody tissue of walnut is discussed.

Zinc (Zn) is essential for the optimal growth of plants but is toxic if present in excess, so Zn homeostasis needs to be finely tuned. Understanding Zn homeostasis mechanisms in plants will help in the development of innovative approaches for the phytoremediation of Zn-contaminated sites. In this study, Zn tolerance quantitative trait loci (QTL) were identified by analyzing differences in the Bay-0 and Shahdara accessions of Arabidopsis thaliana. Fine-scale mapping showed that a variant of the Fe homeostasis-related FERRIC REDUCTASE DEFECTIVE3 (FRD3) gene, which encodes a multidrug and toxin efflux (MATE) transporter, is responsible for reduced Zn tolerance in A. thaliana. Allelic variation in FRD3 revealed which amino acids are necessary for FRD3 function. In addition, the results of allele-specific expression assays in F1 individuals provide evidence for the existence of at least one putative metal-responsive cis-regulatory element. Our results suggest that FRD3 works as a multimer and is involved in loading Zn into xylem. Cross-homeostasis between Fe and Zn therefore appears to be important for Zn tolerance in A. thaliana with FRD3 acting as an essential regulator., Author Summary Plants are adapted to soils in which the amounts of different nutrients vary widely, like Zn-deficient or Zn-contaminated soils. Exploring the molecular bases of plant adaptation to Zn-contaminated soils is important in determining strategies for phytoremediation. Here, we describe the mapping and characterization of a QTL for Zn tolerance in A. thaliana that underlies the natural variation of the root response to excess Zn. This physiological variation is controlled by different alleles of the AtFRD3 gene, which codes for a citrate transporter that uploads citrate into the xylem sap, hence playing a role in Fe homeostasis. In the Zn-sensitive accession Shahdara, the expression of AtFRD3 is drastically reduced and the protein encoded is unable to efflux citrate in vitro. Less Fe and Zn are found in Shahdara root exudates, and less Fe and Zn are translocated from root to shoot when Zn is in excess. We deduce that a fine-tuned Fe and Zn homeostasis is crucial for Zn tolerance in A. thaliana. Finally, as a range of alleles were identified, some rare, it was possible to define a sequence motif that is a putative metal-responsive cis-element and demonstrate that two amino acids are essential for the function of the FRD3 transporter.