Targeting redox metabolism of the maize-Azospirillum brasilense interaction exposed to arsenic-affected groundwater.

Arsenic in groundwater constitutes an agronomic problem due to its potential accumulation in the food chain. Among the agro-sustainable tools to reduce metal(oid)s toxicity, the use of plant growth-promoting bacteria (PGPB) becomes important. For that, and based on previous results in which significant differences of As translocation were observed when inoculating maize plants with Az39 or CD Azospirillum strains, we decided to decipher the redox metabolism changes and the antioxidant system response of maize plants inoculated when exposed to a realistic arsenate (AsV ) dose. Results showed that AsV caused morphological changes in the root exodermis. Photosynthetic pigments decreased only in CD inoculated plants, while oxidative stress evidence was detected throughout the plant, regardless of the assayed strain. The antioxidant response was strain-differential since only CD inoculated plants showed an increase in superoxide dismutase, glutathione S-transferase (GST), and glutathione reductase (GR) activities while other enzymes showed the same behavior irrespective of the inoculated strain. Gene expression assays reported that only GST23 transcript level was upregulated by arsenate, regardless of the inoculated strain. AsV diminished the glutathione (GSH) content of roots inoculated with the Az39 strain, and CD inoculated plants showed a decrease of oxidized GSH (GSSG) levels. We suggest a model in which the antioxidant response of the maize-diazotrophs system is modulated by the strain and that GSH plays a central role acting mainly as a substrate for GST. These findings generate knowledge for a suitable PGPB selection, and its scaling to an effective bioinoculant formulation for maize crops exposed to adverse environmental conditions.

Azospirillum brasilense ameliorates the response of Arabidopsis thaliana to drought mainly via enhancement of ABA levels.

Production of phytohormones is one of the main mechanisms to explain the beneficial effects of plant growth-promoting rhizobacteria (PGPR) such as Azospirillum sp. The PGPRs induce plant growth and development, and reduce stress susceptibility. However, little is known regarding the stress-related phytohormone abscisic acid (ABA) produced by bacteria. We investigated the effects of Azospirillum brasilense Sp 245 strain on Arabidopsis thaliana Col-0 and aba2-1 mutant plants, evaluating the morphophysiological and biochemical responses when watered and in drought. We used an in vitro-grown system to study changes in the root volume and architecture after inoculation with Azospirillum in Arabidopsis wild-type Col-0 and on the mutant aba2-1, during early growth. To examine Arabidopsis development and reproductive success as affected by the bacteria, ABA and drought, a pot experiment using Arabidopsis Col-0 plants was also carried out. Azospirillum brasilense augmented plant biomass, altered root architecture by increasing lateral roots number, stimulated photosynthetic and photoprotective pigments and retarded water loss in correlation with incremented ABA levels. As well, inoculation improved plants seed yield, plants survival, proline levels and relative leaf water content; it also decreased stomatal conductance, malondialdehyde and relative soil water content in plants submitted to drought. Arabidopsis inoculation with A. brasilense improved plants performance, especially in drought.

Anatomical, morphological, and phytochemical effects of inoculation with plant growth- promoting rhizobacteria on peppermint (Mentha piperita).

Plant growth-promoting rhizobacteria (PGPR) generally exert their effects through enhancement of plant nutrient status and/or phytohormone production. The effects of PGPR on aromatic plant species are poorly known. We measured plant growth parameters, chlorophyll content, trichome density, stomatal density, and levels of secondary metabolites in peppermint (Mentha piperita) seedlings inoculated with PGPR strains Bacillus subtilis GB03, Pseudomonas fluorescens WCS417r, P. putida SJ04, or a combination of WCS417r + SJ04. The treated plants, in comparison with controls, showed increases in shoot biomass, root biomass, leaf area, node number, trichome density, and stomatal density, and marked qualitative and quantitative changes in monoterpene content. Improved knowledge of the factors that control or affect biosynthesis of secondary metabolites and monoterpene accumulation will lead to strategies for improved cultivation and productivity of aromatic plants and other agricultural crops without the use of chemical fertilizers or pesticides.

Expression of the chloroplast thioredoxins f and m is linked to short-term changes in the sugar and thiol status in leaves of Pisum sativum.

Thioredoxins (TRXs) f and m are key components in the light regulation of photosynthetic metabolism via thiol-dithiol modulation in chloroplasts of leaves; however, little is known about the factors modulating the expression of these proteins. To investigate the effect of sugars as photosynthetic products on the expression of PsTRX f and m1 genes, sucrose and glucose were externally supplied to pea plants during the day. There was an increase in the mRNA levels of PsTRX f and m1 genes in response mainly to glucose. When leaf discs were incubated for up to 4h in the dark, glucose also led to an increase in both mRNA and protein levels of TRXs f and m, while sucrose had no substantial effect. Expression of PsDOF7, a carbon metabolism-related transcription factor gene, was also induced by glucose. Protein-DNA interaction showed that PsDOF7 binds specifically to the DOF core located in PsTRX f and m1 gene promoters. Transient expression in agroinfiltrated pea leaves demonstrated that PsDOF7 activated transcription of both promoters. The incubation of leaf discs in dithiotreitol (DTT) to increase the redox status led to a marked increase in the mRNA and protein levels of both TRXs within 4h. The increase in TRX protein levels occurred after 1h DTT feeding, implying a rapid effect of the thiol status on TRX f and m1 protein turnover rates, while transcriptional regulation took 3h to proceed. These results show that the protein levels of both TRXs are under short-term control of the sugar and thiol status in plants.

Vicia villosa Roth: a cover crop to phytoremediate arsenic polluted environments.

Vicia villosa Roth is a legume species with a growing application in Argentina as a cover crop (CC), a practice that favors the sustainable development of agricultural systems. However, several areas where the use of this CC provides numerous advantages are affected by high concentrations of arsenic (As). Thus, in the present work we studied hairy vetch ability to cope with arsenate [As(V)], arsenite [As(III)], and the mixture of both along with oxidative stress indexes [chlorophyll content, malondialdehyde (MDA) equivalents] as well as anatomical and histological changes in the root structure. The results obtained suggested a different behavior of hairy vetch depending on its growth stage and on metal(oid) concentration. The roots treated with the contaminant showed less turgidity, thickening of the epidermal and subepidermal parenchymal outer layers, and the presence of dark deposits. The morpho-anatomic parameters (cortex length, vascular cylinder diameter, total diameter, and vascular cylinder area) were altered in plants treated with As(V) and As(V)/As(III) whereas the roots of plants treated with As(III) did not show significant differences respect to the control. Moreover V. villosa could tolerate and remove As from soil, thus the use of this legume species seems an attractive approach to remediate As while protecting contaminated soils.

Unraveling the impact of arsenic on the redox response of peanut plants inoculated with two different Bradyrhizobium sp. strains.

Arsenic (As) can be present naturally in groundwater from peanut fields, constituting a serious problem, as roots can accumulate and mobilize the metalloid to their edible parts. Understanding the redox changes in the legume exposed to As may help to detect potential risks to human health and recognize tolerance mechanisms. Thirty-days old peanut plants inoculated with Bradyrhizobium sp. strains (SEMIA6144 or C-145) were exposed to a realistic arsenate concentration, in order to unravel the redox response and characterize the oxidative stress indexes. Thus, root anatomy, reactive oxygen species detection by fluorescence microscopy and, ROS histochemical staining along with the NADPH oxidase activity were analyzed. Besides, photosynthetic pigments and damage to lipids and proteins were determined as oxidative stress indicators. Results showed that at 3 µM AsV, the cross-section areas of peanut roots were augmented; NADPH oxidase activity was significantly increased and O2˙¯and H2O2 accumulated in leaves and roots. Likewise, an increase in the lipid peroxidation and protein carbonyls was also observed throughout the plant regardless the inoculated strain, while chlorophylls and carotenes were increased only in those inoculated with Bradyrhizobium sp. C-145. Interestingly, the oxidative burst, mainly induced by the NADPH oxidase activity, and the consequent oxidative stress was strain-dependent and organ-differential. Additionally, As modifies the root anatomy, acting as a possibly first defense mechanism against the metalloid entry. All these findings allowed us to conclude that the redox response of peanut is conditioned by the rhizobial strain, which contributes to the importance of effectively formulating bioinoculants for this crop.

Arsenic stress effects on root water absorption in soybean plants: Physiological and morphological aspects.

Soybean (Glycine max L.) is often cultivated in areas contaminated with arsenic (As), which negatively affects plant growth and reduces crop yield. The deleterious effects may be due, at least in part, to disturbances in the water status, as was reported for some plants exposed to heavy metals. However, to our knowledge, these mechanisms have not been studied in depth in soybean plants exposed to As. The aim of the present work was to analyze possible changes in water relations and the responses developed in soybean plants under arsenate (AsV) and arsenite (AsIII) stress. We discuss physiological and morphological aspects of the As stress response, such as root absorption rate, water content, stomatal conductance, water and osmotic potential, accumulation of compatible solutes, leaf conducting tissues and stomata characteristics. AsV and AsIII caused a significant decrease in root absorption rate, which could reduce metalloid uptake. On the other hand, water content decreased at the beginning of the treatment but was re-established after 4 and 8 d. This was correlated with a decrease in stomatal conductance and a reduction in leaf water and osmotic potential due to the accumulation of proline and soluble sugars. Besides, smaller leaf xylem vessels and abnormal stomata were observed in plants under As treatment. These mechanisms increased the plant's ability to retain water and therefore to avoid dehydration. Thus, the results of the present work contribute to the understanding of how soybean responds to As, by describing key tolerance strategies to the metalloid.

Arsenic toxicity in soybean seedlings and their attenuation mechanisms.

Even though vast areas contaminated with arsenic (As) are under soybean (Glycine max) cultivation, little is known about the growth and intrinsic antioxidant metabolism of soybean in response to As exposure. Thus, an evaluation was carried out of plant growth, root anatomy, antioxidant system and photosynthetic pigment content under arsenate (As(V)) and arsenite (As(III)) treatment. Soybean seedling growth was significantly affected at 25 µM or higher concentrations of As(V) or As(III), and the toxic effect on root growth was associated with cell death of root tips. Microscopic analysis of cross-sections of As-treated root showed a reduction in the cortex area, dark deposits in cortex cells and broken cells in the outer layer. Similarly, in the vascular cylinder, dark deposits within xylem vessel elements and phloem cell walls were observed. In all the analyzed parameters, the deleterious effect was more evident under As(III) than As(V) treatment. Arsenic-treated soybean seedlings showed increased activity of antioxidant enzymes [total peroxidases (Px) and superoxide dismutase (SOD)] in root and shoot harvested after 2 and 5 d of treatment. However, a reduction in chlorophyll content and an increase in membrane lipids peroxidation were observed. It is suggested that root structural alterations induced by As, such as the particular pattern of dark depositions in the vascular system, could be associated with an adaptation or detoxification mechanism to prevent As translocation to the aboveground tissues.

Arsenic stress induces changes in lipid signalling and evokes the stomata closure in soybean.

Soybean (Glycine max) is often exposed to high arsenic (As) level in soils or through irrigation with groundwater. In previous studies on As-treated soybean seedlings we showed deleterious effect on growth, structural alterations mainly in root vascular system and induction of antioxidant enzymes. However, there are not reports concerning signal transduction pathways triggered by the metalloid in order to develop adaptive mechanisms. Phosphatidic acid (PA), a key messenger in plants, can be generated via phospholipase D (PLD) or via phospholipase C (PLC) coupled to diacylglycerol kinase (DGK). Thus, changes in PA and in an enzyme involved in its metabolism (PLD) were analysed in soybean seedlings treated with 25 µM AsV or AsIII. The present study demonstrated that As triggers the PA signal by PLD and also via PLC/DGK mainly after 48 h of As treatment. DGPP, other lipid messenger produced by phosphorylation of PA by PAK increased in As treated roots. Arsenic also induced rapid and significant stomatal closure after 1.5 h of treatment, mainly with AsIII, probably as an adaptive response to the metalloid to reduce water loss by transpiration. This report constitute the first evidence that shows the effects of As on lipid signalling events in soybean seedlings which would be crucial in adaptation and survival of soybean seedlings under As stress.

Morphophysiological plasticity in a wheat variety in response to NaCl stress and its alleviation by exogenous abscisic acid.

Nowadays, soil salinity is the most unfavourable abiotic factors for plant growth, causing important yield loss of many crops. A partial solution to this situation is to establish crop varieties in these areas affected which are tolerant to stress. The aim of this study was to evaluate in a wheat variety, the morphophysiological plasticity to sodium chloride (NaCl) stress and the effect of exogenous Abscisic Acid (ABA) on physiological variables. This was carried out by using the BI3000 wheat variety, for regional adaptability experiments. The germination percentage, coleoptile and radicle growth and root anatomic were evaluated, both seedling irrigated with water or saline solution. On the other hand, ABA sprays were applied to wheat plants and their biomass, pigment, stomatal behaviour and cellular membrane injuries were determined after salt treatments. In this study, it was possible to determine that the BI3000 wheat variety can grow in high electrical conductivity, with good germination and seedling growth. This variety showed less radical anatomic variations under salinity, what allows a faster plasticity to adapt. ABA applications suggest a protective role in plants under salinity, due to an increase in chlorophyll and carotene content, stability of cell membranes and stomatal behavior. This study is a contribution to a better understanding of the morphophysiological responses of glycophytic plants to salt stress. This have been pointed out as a useful approach to show more tolerance to salt stress crops in the future and it suggests that ABA could help improve agriculture production in areas affected by this stress.