No abstract

It is well known that in the rhizosphere soluble Fe sources available for plants are mainly a mixture of complexes between the micronutrient and organic ligands such as organic acids and phytosiderophores (PS) released by roots, microbial siderophores as well as fractions of humified organic matter. In the present work, mechanisms of Fe acquisition operating at the leaf level of plants fed with different Fe-complexes were investigated at the micro-analytical, physiological and molecular levels. Fe-deficient tomato plants (Solanum Lycopersicum L., cv. 'Marmande') were fed for 24 h with a solution (pH 7.5) containing 1 A mu M Fe as Fe-PS, Fe-citrate or Fe-WEHS. Thereafter, leaf tissue was used for the visualization of Fe distribution, measurements of Fe content, reduction and uptake, and evaluation of expression of Fe-chelate reductase (LeFRO1), Fe-transporter (LeIRT1) and Ferritin (Ferritin2) genes. Leaf discs isolated from Fe-deficient plants treated for 24 h with Fe-WEHS developed higher rates of translocation, Fe-chelate reduction and Fe-59 uptake as compared to plants supplied with Fe-citrate or Fe-PS. Leaves of plants treated with Fe-WEHS also showed higher transcript levels of LeFRO1, LeIRT1 and Ferritin2 genes with respect to plants fed with the other Fe-sources. Data obtained support the idea that the efficient use of Fe complexed to WEHS-like humic fractions involves, at least in part, also the activation of Fe-acquisition mechanisms operating at the leaf level

Iron (Fe) sources available for plants in the rhizospheric solution are mainly a mixture of complexes between Fe and organic ligands, including phytosiderophores (PS) and water-extractable humic substances (WEHS). In comparison with the other Fe sources, Fe-WEHS are more efficiently used by plants, and experimental evidences show that Fe translocation contributes to this better response. On the other hand, very little is known on the mechanisms involved in Fe allocation in leaves. In this work, physiological and molecular processes involved in Fe distribution in leaves of Fe-deficient Cucumis sativus supplied with Fe-PS or Fe-WEHS up to 5 days were studied combining different techniques, such as radiochemical experiments, synchrotron micro X-ray fluorescence, real-time reverse transcription polymerase chain reaction and in situ hybridization. In Fe-WEHS-fed plants, Fe was rapidly (1 day) allocated into the leaf veins, and after 5 days, Fe was completely transferred into interveinal cells; moreover, the amount of accumulated Fe was much higher than with Fe-PS. This redistribution in Fe-WEHS plants was associated with an upregulation of genes encoding a ferric(III) -chelate reductase (FRO), a Fe(2+) transporter (IRT1) and a natural resistance-associated macrophage protein (NRAMP). The localization of FRO and IRT1 transcripts next to the midveins, beside that of NRAMP in the interveinal area, may suggest a rapid and efficient response induced by the presence of Fe-WEHS in the extra-radical solution for the allocation in leaves of high amounts of Fe. In conclusion, Fe is more efficiently used when chelated to WEHS than PS and seems to involve Fe distribution and gene regulation of Fe acquisition mechanisms operating in leaves.

Cadmium (Cd) detoxification involves glutathione and phytochelatins biosynthesis: the higher need of nitrogen should require increased nitrate (NO(3)(-)) uptake and metabolism. We investigated inducible high-affinity NO(3)(-) uptake across the plasma membrane (PM) in maize seedlings roots upon short exposure (10 min to 24 h) to low Cd concentrations (0, 1 or 10 μM): the activity and gene transcript abundance of high-affinity NO(3)(-) transporters, NO(3)(-) reductases and PM H(+)-ATPases were analyzed. Exposure to 1 mM NO(3)(-) led to a peak in high-affinity (0.2 mM) NO(3)(-) uptake rate (induction), which was markedly lowered in Cd-treated roots. Plasma membrane H(+)-ATPase activity was also strongly limited, while internal NO(3)(-) accumulation and NO(3)(-) reductase activity in extracts of Cd treated roots were only slightly lowered. Kinetics of high- and low-affinity NO(3)(-) uptake showed that Cd rapidly (10 min) blocked the inducible high-affinity transport system; the constitutive high-affinity transport system appeared not vulnerable to Cd and the low-affinity transport system appeared to be less affected and only after a prolonged exposure (12 h). Cd-treatment also modified transcript levels of genes encoding high-affinity NO(3)(-) transporters (ZmNTR2.1, ZmNRT2.2), PM H(+)-ATPases (ZmMHA3, ZmMHA4) and NO(3)(-) reductases (ZmNR1, ZmNADH:NR). Despite an expectable increase in NO(3)(-) demand, a negative effect of Cd on NO(3)(-) nutrition is reported. Cd effect results in alterations at the physiological and transcriptional levels of NO(3)(-) uptake from the external solution and it is particularly severe on the inducible high-affinity anion transport system. Furthermore, Cd would limit the capacity of the plant to respond to changes in NO(3) (-) availability.