Many studies have shown the capacity of soil humic substances (HS) to improve plant growth in natural ecosystems. This effect involves the activation of different processes within the plant at different coordinated molecular, biochemical, and physiological levels. However, the first event triggered by plant root-HS interaction remains unclear. Some studies suggest the hypothesis that the interaction of HS with root exudates involves relevant modification of the molecular conformation of humic self-assembled aggregates, including disaggregation, which might be directly involved in the activation of root responses. To investigate this hypothesis, we have prepared two humic acids. A natural humic acid (HA) and a transformed humic acid obtained from the treatment of HA with fungal laccase (HA enz). We have tested the capacity of the two humic acids to affect plant growth (cucumber and Arabidopsis) and complex Cu. Laccase-treatment did not change the molecular size but increased hydrophobicity, molecular compactness and stability, and rigidity of HA enz. Laccase-treatment avoided the ability of HA to promote shoot- and root-growth in cucumber and Arabidopsis. However, it does not modify Cu complexation features. There is no molecular disaggregation upon the interaction of HA and HA enz with plant roots. The results indicate that the interaction with plant roots induced in both HA and laccase-treated HA (HA enz), changes in their structural features that showed higher compactness and rigidity. These events might result from the interaction of HA and HA enz with specific root exudates that can promote intermolecular crosslinking. In summary, the results indicate that the weakly bond stabilized aggregated conformation (supramolecular-like) of HA plays a crucial role in its ability to promote root and shoot growth. The results also indicate the presence of two main types of HS in the rhizosphere corresponding to those non-interacting with plant roots (forming aggregated molecular assemblies) and those produced after interacting with plant root exudates (forming stable macromolecules).

Many studies have shown the close relationship between the beneficial action of soil and sedimentary humic acids on the growth of plants cultivated in calcareous soils and their ability to improve Fe plant nutrition. These results have been ascribed to the humic acid (HA) capability to improve Fe solubility and bioavailability. However, other effects more related to a humic acid action on the specific mechanisms activated in roots of plants under Fe deficiency cannot be ruled out. Although this question has been studied in dicotyledonous plants, in graminaceous plants there are no specific studies. Here we investigate the ability of a humic acid extracted from peat (HA) to improve Fe nutrition in wheat plants cultivated under Fe deficient and sufficient conditions. The results show that HA can improve the physiological status of Fe deficient wheat plants by alleviating some of the deleterious consequences of Fe deficiency on plant development and increasing the plant ability to secrete phytosiderophores to the nutrient solution. This action of HA is associated with increases in the Fe-active pool in leaves that might be related to the mobilization of the Fe complexed by HA resulting from the interaction of HA with the phytosiderophores in the nutrient solution. The Fe translocation from the root to the shoot may be favored by the action of trans-Zeatin Riboside (tZR) since the leaf concentration of this phytohormone was enhanced by HA in Fe deficient plants.

Although the ability of humic (HA) and fulvic acids (FA) to improve plant growth has been demonstrated, knowledge about the mechanisms responsible for the direct effects of HA and FA on the promotion of plant growth is scarce and fragmentary. Our study investigated the causal role of both root PM H+-ATPase activity and ABA in the SHA-promoting action on both root and shoot growth. The involvement of these processes in the regulation of shoot cytokinin concentration and activity was also studied. Our aim was to integrate such plant responses for providing new insights to the current model on the mode of action of HA for promoting root and shoot growth. Experiments employing specific inhibitors and using Cucumis sativus L. plants show that both the root PM H+-ATPase activity and root ABA play a crucial role in the root growth-promoting action of SHA. With regard to the HA-promoting effects on shoot growth, two pathways of events triggered by the interaction of SHA with plant roots are essential for the increase in root PM H+-ATPase activity-which also mediates an increase in cytokinin concentration and action in the shoot-and the ABA-mediated increase in hydraulic conductivity (Lp(r)).

The ability of rhizospheric humic substances to improve plant growth has been well established by many studies carried out using diverse plant species cultivated under many different conditions. These beneficial effects of humic substances on plant development are expressed in both root and shoot. However, the mechanisms responsible for this action of humic substances are only partially known and poorly integrated. In fact, although the studies focused on plant root development are numerous, those dealing with plant shoot development are scarce. Likewise, studies integrating humic effects on root and shoot are also few. In this context, the main goal of this work is to summarize some of the results regarding the effects of humic substances on plant development within a hypothetical holistic framework that will allow us to interconnect these findings and disclose some features of the functional crosstalk between the effects on soil, root and shoot. Furthermore, the significance of all these mechanisms in plants growing in the field is also discussed.

Background: The release of phytosiderephores (PS) to the rhizosphere is the main root response to iron (Fe) deficiency in graminaceous plants. We have investigated the role of the Fe status in the shoot as well as of the signaling pathways controlled by three relevant phytoregulators-indolacetic acid (IAA), ethylene and nitric oxide (NO) - in the regulation of this root response in Fe-starved wheat plants. To this end, the PS accumulation in the nutrient solution and the root expression of the genes encoding the nicotianamine aminotransferase (TaNAAT) and ferritin (TaFER) have been evaluated in plants subjected to different treatments. Results: The application of Fe to leaves of Fe-deficient plants prevented the increase in both PS root release and TaNAAT gene expression thus showing the relevant role of the shoot to root communication in the regulation of PS root release and some steps of PS biosynthesis. Experiments with specific hormone inhibitors showed that while ethylene and NO did not positively regulate Fe deficiency induced PS root release, auxin plays an essential role in the regulation of this process. Moreover, the application of IM to Fe-sufficient plants promoted both PS root release and TaNAAT gene expression thus indicating that auxin might be involved in the shoot to root signaling network regulating Fe-deficiency root responses in wheat Conclusions: These results therefore indicate that PS root release in Fe-deficient wheat plants is directly modulated by the shoot Fe status through signaling pathways involving, among other possible effectors, auxin.

The physiological and metabolic mechanisms behind the humic acid-mediated plant growth enhancement are discussed in detail. Experiments using cucumber (Cucumis sativus) plants show that the shoot growth enhancement caused by a structurally well-characterized humic acid with sedimentary origin is functionally associated with significant increases in abscisic acid (ABA) root concentration and root hydraulic conductivity. Complementary experiments involving a blocking agent of cell wall pores and water root transport (polyethylenglycol) show that increases in root hydraulic conductivity are essential in the shoot growth-promoting action of the model humic acid. Further experiments involving an inhibitor of ABA biosynthesis in root and shoot (fluridone) show that the humic acid-mediated enhancement of both root hydraulic conductivity and shoot growth depended on ABA signaling pathways. These experiments also show that a significant increase in the gene expression of the main root plasma membrane aquaporins is associated with the increase of root hydraulic conductivity caused by the model humic acid. Finally, experimental data suggest that all of these actions of model humic acid on root functionality, which are linked to its beneficial action on plant shoot growth, are likely related to the conformational structure of humic acid in solution and its interaction with the cell wall at the root surface.

Higher plants have to cope with fluctuating mineral resource availability. However, strategies such as stimulation of root growth, increased transporter activities, and nutrient storage and remobilization have been mostly studied for only a few macronutrients. Leaves of cultivated crops (Zea mays, Brassica napus, Pisum sativum, Triticum aestivum, Hordeum vulgare) and tree species (Quercus robur, Populus nigra, Alnus glutinosa) grown under field conditions were harvested regularly during their life span and analyzed to evaluate the net mobilization of 13 nutrients during leaf senescence. While N was remobilized in all plant species with different efficiencies ranging from 40% (maize) to 90% (wheat), other macronutrients (K-P-S-Mg) were mobilized in most species. Ca and Mn, usually considered as having low phloem mobility were remobilized from leaves in wheat and barley. Leaf content of Cu-Mo-Ni-B-Fe-Zn decreased in some species, as a result of remobilization. Overall, wheat, barley and oak appeared to be the most efficient at remobilization while poplar and maize were the least efficient. Further experiments were performed with rapeseed plants subjected to individual nutrient deficiencies. Compared to field conditions, remobilization from leaves was similar (N-S-Cu) or increased by nutrient deficiency (K-P-Mg) while nutrient deficiency had no effect on Mo-Zn-B-Ca-Mn, which seemed to be non-mobile during leaf senescence under field conditions.

Although climate scenarios have predicted an increase in [CO2] and temperature conditions, to date few experiments have focused on the interaction of [CO2] and temperature effects in wheat development. Recent evidence suggests that photosynthetic acclimation is linked to the photorespiration and N assimilation inhibition of plants exposed to elevated CO2. The main goal of this study was to analyze the effect of interacting [CO2] and temperature on leaf photorespiration, C/N metabolism and N transport in wheat plants exposed to elevated [CO2] and temperature conditions. For this purpose, wheat plants were exposed to elevated [CO2] (400 vs 700 mu molmol(-1)) and temperature (ambient vs ambient+4 degrees C) in CO2 gradient greenhouses during the entire life cycle. Although at the agronomic level, elevated temperature had no effect on plant biomass, physiological analyses revealed that combined elevated [CO2] and temperature negatively affected photosynthetic performance. The limited energy levels resulting from the reduced respiratory and photorespiration rates of such plants were apparently inadequate to sustain nitrate reductase activity. Inhibited N assimilation was associated with a strong reduction in amino acid content, conditioned leaf soluble protein content and constrained leaf N status. Therefore, the plant response to elevated [CO2] and elevated temperature resulted in photosynthetic acclimation. The reduction in transpiration rates induced limitations in nutrient ...

The importance of zinc (Zn) has been of little concern in human nutrition despite a strong decrease of this element in crops since the rise of high yielding varieties. For better food quality, Zn biofortification can be used, but will be optimal only if mechanisms governing Zn management are better known. Using Zn deficiency, we are able to demonstrate that Zn is not remobilized in Brassica napus (B. napus). Thus, remobilization processes should not be targeted by biofortification strategies. This study also complemented previous work by investigating leaf responses to Zn deficiency, especially from proteomic and ionomic points of view, showing for example, an increase in Manganese (Mn) content and of the Mn-dependent protein, Oxygen Evolving Enhancer.

During the last 40 years, crop breeding has strongly increased yields but has had adverse effects on the content of micronutrients, such as Fe, Mg, Zn and Cu, in edible products despite their sufficient supply in most soils. This suggests that micronutrient remobilization to edible tissues has been negatively selected. As a consequence, the aim of this work was to quantify the remobilization of Cu in leaves of Brassica napus L. during Cu deficiency and to identify the main metabolic processes that were affected so that improvements can be achieved in the future. While Cu deficiency reduced oilseed rape growth by less than 19% compared to control plants, Cu content in old leaves decreased by 61.4%, thus demonstrating a remobilization process between leaves. Cu deficiency also triggered an increase in Cu transporter expression in roots (COPT2) and leaves (HMA1), and more surprisingly, the induction of the MOT1 gene encoding a molybdenum transporter associated with a strong increase in molybdenum (Mo) uptake. Proteomic analysis of leaves revealed 33 proteins differentially regulated by Cu deficiency, among which more than half were located in chloroplasts. Eleven differentially expressed proteins are known to require Cu for their synthesis and/or activity. Enzymes that were located directly upstream or downstream of Cu-dependent enzymes were also differentially expressed.

The role of soil humus in soil fertility and crop production has been well established by many studies and practical farming experience. This role is related to the presence of a family of organic substances, known as humic substances, with the capacity to increase the pool of plant-available nutrients in soil for root uptake. The improvement in plant growth and mineral nutrition is directly linked to the physicochemical features of humic substances, and more specifically, to their capacity to form stable chemical complexes with metals. This chapter reviews the main mechanisms behind the action of humic substances in improving plant mineral nutrition. It also discusses the main signalling pathways that might be involved in the regulation of these humic substances.

The main aim of this communication is to discuss the current knowledge about the potential direct mechanisms that are involved in the beneficial action of humic substances on plant development. To this end, we present and discuss here recent results obtained in our laboratory, along with other findings published by other authors. Finally, we propose a hypothetical whole mechanism for explaining the action of humic substances on plant development. In this schema, we point out those steps that remain unclear.

The aims of this chapter are: (i) First, to present new experimental evidenceshowingthe signal-role of nitrate in the correction and improvement of the deleterious effects of ammonium-and/or urea-based nutritionin different plant species, either dicotyledonous or monocotyledonous. Likewise, we discuss the different and complementary mechanisms, which are expressed at diverse but interconnected molecular and physiological levels, responsible for this specific role of nitrate. This discussion is accompanied by the propositionof different hypothesisthat could explain these experimental results and orientate future research in this important field as well. (ii) Second, we also discuss the different methodologies that might be developed in order to apply this basic researchin the development of new nitrogenfertilizerswith higher metabolic efficiency and lower potential environmental risks. (iii) Finally, we propose a framework for future research oriented to do a synthesis-involving and inter-connecting this basic and applied research -related to both the future development of nitrogen-related fertilizersand cropyield ¿quality improvement.