Green synthesis of a dual-functional sulfur nanofertilizer to promote growth and enhance salt stress resilience in faba bean.

BACKGROUND: Salinity is a major abiotic stress, and the use of saline water in the agricultural sector will incur greater demand under the current and future climate changing scenarios. The objective of this study was to develop a dual-functional nanofertilizer capable of releasing a micronutrient that nourishes plant growth while enhancing salt stress resilience in faba bean (Vicia faba L.). RESULTS: Moringa oleifera leaf extract was used to synthesize sulfur nanoparticles (SNPs), which were applied as a foliar spray at different concentrations (0, 25, 50, and 100 mg/l) to mitigate the negative effects of salt stress (150 mM NaCl) on faba bean plants. The SNPs were characterized and found to be spherical in shape with an average size of 10.98 ± 2.91 nm. The results showed that salt stress had detrimental effects on the growth and photosynthetic performance (Fv/Fm) of faba bean compared with control, while foliar spraying with SNPs improved these parameters under salinity stress. SNPs application also increased the levels of osmolytes (soluble sugars, amino acids, proline, and glycine betaine) and nonenzymatic antioxidants, while reducing the levels of oxidative stress biomarkers (MDA and H2O2). Moreover, SNPs treatment under salinity stress stimulated the activity of antioxidant enzymes (ascorbate peroxidase (APX), and peroxidase (POD), polyphenol oxidase (PPO)) and upregulated the expression of stress-responsive genes: chlorophyll a-b binding protein of LHCII type 1-like (Lhcb1), ribulose bisphosphate carboxylase large chain-like (RbcL), cell wall invertase I (CWINV1), ornithine aminotransferase (OAT), and ethylene-responsive transcription factor 1 (ERF1), with the greatest upregulation observed at 50 mg/l SNPs. CONCLUSION: Overall, foliar application of sulfur nanofertilizers in agriculture could improve productivity while minimizing the deleterious effects of salt stress on plants. Therefore, this study provides a strong foundation for future research focused on evaluating the replacement of conventional sulfur-containing fertilizers with their nanoforms to reduce the harmful effects of salinity stress and enhance the productivity of faba beans.

Transcriptional and physiological analyses uncover the mineralization and uptake mechanisms of phytic acid in symbiotically grown Vicia faba plants.

Legume-rhizobia symbiosis requires high phosphorus (P) in the form of ATP to convert atmospheric nitrogen (N) into ammonia. The fixed ammonia is converted to NH4+ by H+-ATPase via protonation. To the best of our knowledge, most of these research works resort to using only inorganic P (Pi) to the neglect of the organic P (Po) counterpart. As it stands, the potential regulating roles of plasma membrane (PM) H+-ATPases during legume-rhizobia symbiosis in response to phytic acid supply and how it alters and modulates the regulation of PM H+-ATPases remain obscure. To contribute to the above hypothesis, we investigate the mechanisms that coordinately facilitate the growth, uptake, and transcript expression of PM H+-ATPase gene isoforms in response to different P sources when hydroponically grown Vicia faba plants were exposed to three P treatments, viz., low- and high-Pi (2.0 and 200 µM KH2PO4; LPi and HPi), and phytic acid (200 µM; Po) and inoculated with Rhizobium leguminosarum bv. viciae 384 for 30 days. The results consistently reveal that the supply of Po improved not only the growth and biomass, but also enhanced photosynthetic parameters, P uptake and phosphatase activities in symbiotically grown Vicia faba relative to Pi. The supply of Po induced higher transcriptional expression of all PM H+-ATPase gene isoforms, with possible interactions between phosphatases and H+-ATPase genes in Vicia faba plants when exclusively reliant on N derived from nodule symbiosis. Overall, preliminary results suggest that Po could be used as an alternative nutrition in symbiotic crops to improve plant growth.

Potential risks of nicotine on the germination, growth, and nutritional properties of broad bean.

This study evaluated the allelopathy, uptake and accumulation, and potential agricultural and food safety risks of nicotine in broad bean (Vicia faba L.) during seed germination and seedling growth. Nicotine stress has an allelopathic inhibitory effect on seeds and a hormesis effect on germinated seeds and seedlings, which has an enhancement effect (<50 mg kg-1) and an inhibition effect (>100 mg kg-1) on the germinated seeds and an enhancement effect (<100 mg kg-1) and an inhibition effect (>200 mg kg-1) on the seedlings. Exogenous nicotine can be absorbed by broad bean roots from nicotine-contaminated soil and accumulated in the main organs of the seedlings, especially the leaves, which exceeded the maximum residue level (0.03 mg kg-1 DW) at 50 mg kg-1. Moreover, nicotine resulted in a bitter taste in the edible broad bean leaves, disrupting the balance of basic nutritional properties, decreasing sucrose, and increasing bitter substances such as choline and procyanidin. These results demonstrated that residual nicotine in the soil not only poses potential risks to sustainable agricultural development but also a food safety risk for consumers. The present study provides insight into the potential risks of nicotine in agroecosystems.

Investigating the underlying mechanism of cadmium-induced plant adaptive response to genotoxic stress.

Plants as sessile organisms have developed some unique strategies to withstand environmental stress and adaptive response (AR) is one of them. In the present study Cadmium (Cd)-induced AR was evaluated to ameliorate the genotoxicity of a known chemical mutagen ethyl methanesulphonate (EMS) based on cytotoxicity, genotoxicity and oxidative stress in two model plant systems Allium cepa L. and Vicia faba L. Priming the plants with cadmium chloride (CdCl2, 25 and 50 µM) reduced the genotoxicity of EMS (0.25 mM). Cd-induced AR was evident by the magnitude of adaptive response (MAR) values calculated for cytotoxicity, genotoxicity and biochemical parameters. In addition the involvement of some major metabolic pathways and epigenetic modifications in AR was investigated. Metabolic blockers of protein kinase cascades, DNA repair, oxidative stress and de novo translation interfered with the adaptive response implying their role in AR whereas, inhibitors involved in post-replication repair and autophagy were ineffective implicating that they probably have no role in the AR studied. Moreover to find the role of DNA methylation in AR, methylation-sensitive comet assay was carried out. Simultaneously 5-methyl- 2'-deoxycytidine (5mdC) levels were quantified by HPLC (high performance liquid chromatography). AR was eliminated in cells treated with a demethylating agent, 5-aza- 2'deoxycytidine (AZA). Results implied a contribution of DNA hypermethylation. To the best of our knowledge this is a first report correlating DNA methylation to Cd-induced adaptive response in plants undergoing genotoxic stress.

The transgenerational effects of solar short-UV radiation differed in two accessions of Vicia faba L. from contrasting UV environments.

BACKGROUND AND AIMS: UVB radiation can rapidly induce gene regulation leading to cumulative changes for plant physiology and morphology. We hypothesized that a transgenerational effect of chronic exposure to solar short UV modulates the offspring's responses to UVB and blue light, and that the transgenerational effect is genotype dependent. METHODS: We established a factorial experiment combining two Vicia faba L. accessions, two parental UV treatments (full sunlight and exclusion of short UV, 290-350 nm), and four offspring light treatments from the factorial combination of UVB and blue light. The accessions were Aurora from southern Sweden, and ILB938 from Andean region of Colombia and Ecuador. KEY RESULTS: The transgenerational effect influenced morphological responses to blue light differently in the two accessions. In Aurora, when UVB was absent, blue light increased shoot dry mass only in plants whose parents were protected from short UV. In ILB938, blue light increased leaf area and shoot dry mass more in plants whose parents were exposed to short UV than those that were not. Moreover, when the offspring was exposed to UVB, the transgenerational effect decreased in ILB938 and disappeared in Aurora. For flavonoids, the transgenerational effect was detected only in Aurora: parental exposure to short UV was associated with a greater induction of total quercetin in response to UVB. Transcript abundance was higher in Aurora than in ILB938 for both CHALCONE SYNTHASE (99-fold) and DON-GLUCOSYLTRANSFERASE 1 (19-fold). CONCLUSIONS: The results supported both hypotheses. Solar short UV had transgenerational effects on progeny responses to blue and UVB radiation, and they differed between the accessions. These transgenerational effects could be adaptive by acclimation of slow and cumulative morphological change, and by early build-up of UV protection through flavonoid accumulation on UVB exposure. The differences between the two accessions aligned with their adaptation to contrasting UV environments.

Sodium nitroprusside (SNP) improves tolerance to arsenic (As) toxicity in Vicia faba through the modifications of biochemical attributes, antioxidants, ascorbate-glutathione cycle and glyoxalase cycle.

The present study was conducted to evaluate the effect of arsenic (As) toxicity and the mitigating role of nitric oxide (NO) donor sodium nitroprusside (SNP) on Vicia faba. Arsenics stress decreased the growth and biomass yield, and photosynthetic pigments, but it enhanced As accumulation. Supplementation of NO enhanced the afore-mentioned parameters except As accumulation which decreased in both shoot and root. Supplementation of NO enhanced the shoot tolerance index (Shoot TI%), root tolerance index (Root TI%) but it declined the As translocation factor (TF). Application of NO alleviated the As-induced decline in net assimilation rate, stomatal conductance, transpiration and leaf relative water content. The levels of proline and glycine betaine (GB) further increased due to NO application, whereas malondialdehyde (MDA), hydrogen peroxide (H2O2), electrolyte leakage (EL) and methylglyoxal (MG) declined considerably. Activities of enzymatic antioxidants such as superoxide dismutase (SOD) and catalase (CAT) increased under As stress. Supplementation of NO up-regulated the enzymes involved in Asc-Glu cycle and glyoxalase cycle under As toxicity. Another experiment was setup to authenticate whether NO was certainly able to alleviate As toxicity. For this purpose, the NO scavenger [2-(4-carboxy-2 phenyl)-4,4,5,5-tertamethylimidazoline-1-oxyl-3-oxide (cPTIO)] was added to As and NO supplemented plants. Addition of cPTIO to NO supplemented As-treated plants showed the same effect when As alone was supplied to plants. In conclusion, addition of NO to the growth medium maintained the plant performance under As toxicity through modulation of physio-biochemical attributes, antioxidant enzymes, and the Asc-Glu and glyoxalase systems.

Genotoxicity response of Vicia faba seedlings to cadmium in soils as characterized by direct soil exposure and micronucleus test.

To overcome the drawbacks of the Vicia faba root tip micronucleus test in soil using the solution extract method, we conducted a potting experiment by direct soil exposure. Cadmium was spiked into 3 typical soils (brown soil, red soil, and black soil) to simulate environmental concentrations (0.625, 1.25, 2.5, 5, and 10 mg kg-1). Multiple Vicia faba tissues (primary root tips, secondary root tips, and leaf tips) were sampled, and mitotic index (MI), chromosome aberration frequency (CA), and micronucleus frequency (MN) were used as endpoints after a seedling period of 5 days. The results showed a response between Cd concentrations and multiple sampling tissues of Vicia faba, and the secondary root tips responded to Cd stress the most, followed by primary root tips and leaf tips. Soil physicochemical properties (e.g., pH, total phosphorus, total organic carbon, etc.) influenced the genotoxicity of Cd, and pH was the dominant factor, which resulted in the genetic toxicity response of Cd in soils in the order: red soil > brown soil > black soil. The lowest observable effect concentration (LOEC) of Cd was 1.25 mg kg-1 for both brown soil and red soil and 2.5 mg kg-1 for black soil. In view of this, we suggested that soil properties should be considered in evaluating genotoxicity risk of Cd in soil, especially with soil pH range, and the secondary root tips should be taken as suitable test tissues in the MN test due to its more sensible response feature to Cd stress in soil.

Phytotoxicity of silver nanoparticles on Vicia faba: Evaluation of particle size effects on photosynthetic performance and leaf gas exchange.

Nanotechnology is an emerging field in science and engineering, which presents significant impacts on the economy, society and the environment. The nanomaterials' (NMs) production, use, and disposal is inevitably leading to their release into the environment where there are uncertainties about its fate, behaviour, and toxicity. Recent works have demonstrated that NMs can penetrate, translocate, and accumulate in plants. However, studies about the effects of the NMs on plants are still limited because most investigations are carried out in the initial stage of plant development. The present study aimed to evaluate and characterize the photochemical efficiency of photosystem II (PSII) of broad bean (Vicia faba) leaves when subjected to silver nanoparticles (AgNPs) with diameters of 20, 51, and 73 nm as well as to micrometer-size Ag particles (AgBulk). The AgNPs were characterized by transmission electron microscopy and dynamic light scattering. The analyses were performed by injecting the leaves with 100 mg L-1 aqueous solution of Ag and measuring the chlorophyll fluorescence imaging, gas exchange, thermal imaging, and reactive oxygen species (ROS) production. In addition, silver ion (Ag+) release from Ag particles was determined by dialysis. The results revealed that AgNPs induce a decrease in the photochemical efficiency of photosystem II (PSII) and an increase in the non-photochemical quenching. The data also revealed that AgNPs affected the stomatal conductance (gs) and CO2 assimilation. Further, AgNPs induced an overproduction of ROS in Vicia faba leaves. Finally, all observed effects were particle diameter-dependent, increasing with the reduction of AgNPs diameter and revealing that AgBulk caused only a small or no changes on plants. In summary, the results point out that AgNPs may negatively affect the photosynthesis process when accumulated in the leaves, and that the NPs themselves were mainly responsible since negligible Ag+ release was detected.

Physiological and biochemical responses involved in water deficit tolerance of nitrogen-fixing Vicia faba.

Climate change is increasingly impacting the water deficit over the world. Because of drought and the high pressure of the rising human population, water is becoming a scarce and expensive commodity, especially in developing countries. The identification of crops presenting a higher acclimation to drought stress is thus an important objective in agriculture. The present investigation aimed to assess the adaptation of three Vicia faba genotypes, Aguadulce (AD), Luz d'Otonio (LO) and Reina Mora (RM) to water deficit. Multiple physiological and biochemical parameters were used to analyse the response of the three genotypes to two soil water contents (80% and 40% of field capacity). A significant lower decrease in shoot, root and nodule dry weight was observed for AD compared to LO and RM. The better growth performance of AD was correlated to higher carbon and nitrogen content than in LO and RM under water deficit. Leaf parameters such as relative water content, mass area, efficiency of photosystem II and chlorophyll and carotenoid content were significantly less affected in AD than in LO and RM. Significantly higher accumulation of proline was correlated to the higher performance of AD compared to LO and RM. Additionally, the better growth of AD genotype was related to an important mobilisation of antioxidant enzyme activities such as ascorbate peroxidase and catalase. Taken together, these results allow us to suggest that AD is a water deficit tolerant genotype compared to LO and RM. Our multiple physiological and biochemical analyses show that nitrogen content, leaf proline accumulation, reduced leaf hydrogen peroxide accumulation and leaf antioxidant enzymatic activities (ascorbate peroxidase, guaiacol peroxidase, catalase and polyphenol oxidase) are potential biological markers useful to screen for water deficit resistant Vicia faba genotypes.

Herbivore-Triggered Electrophysiological Reactions: Candidates for Systemic Signals in Higher Plants and the Challenge of Their Identification.

In stressed plants, electrophysiological reactions (elRs) are presumed to contribute to long-distance intercellular communication between distant plant parts. Because of the focus on abiotic stress-induced elRs in recent decades, biotic stress-triggered elRs have been widely ignored. It is likely that the challenge to identify the particular elR types (action potential [AP], variation potential, and system potential [SP]) was responsible for this course of action. Thus, this survey focused on insect larva feeding (Spodoptera littoralis and Manduca sexta) that triggers distant APs, variation potentials, and SPs in monocotyledonous and dicotyledonous plant species (Hordeum vulgare, Vicia faba, and Nicotiana tabacum). APs were detected only after feeding on the stem/culm, whereas SPs were observed systemically following damage to both stem/culm and leaves. This was attributed to the unequal vascular innervation of the plant and a selective electrophysiological connectivity of the plant tissue. However, striking variations in voltage patterns were detected for each elR type. Further analyses (also in Brassica napus and Cucurbita maxima) employing complementary electrophysiological approaches in response to different stimuli revealed various reasons for these voltage pattern variations: an intrinsic plasticity of elRs, a plant-specific signature of elRs, a specific influence of the applied (a)biotic trigger, the impact of the technical approach, and/or the experimental setup. As a consequence, voltage pattern variations, which are not irregular but rather common, need to be included in electrophysiological signaling analysis. Due to their widespread occurrence, systemic propagation, and respective triggers, elRs should be considered as candidates for long-distance communication in higher plants.

Rapid modulation of ultraviolet shielding in plants is influenced by solar ultraviolet radiation and linked to alterations in flavonoids.

The accumulation of ultraviolet (UV)-absorbing compounds (flavonoids and related phenylpropanoids) and the resultant decrease in epidermal UV transmittance (TUV ) are primary protective mechanisms employed by plants against potentially damaging solar UV radiation and are critical components of the overall acclimation response of plants to changing solar UV environments. Whether plants can adjust this UV sunscreen protection in response to rapid changes in UV, as occurs on a diurnal basis, is largely unexplored. Here, we use a combination of approaches to demonstrate that plants can modulate their UV-screening properties within minutes to hours, and these changes are driven, in part, by UV radiation. For the cultivated species Abelmoschus esculentus, large (30-50%) and reversible changes in TUV occurred on a diurnal basis, and these adjustments were associated with changes in the concentrations of whole-leaf UV-absorbing compounds and several quercetin glycosides. Similar results were found for two other species (Vicia faba and Solanum lycopersicum), but no such changes were detected in Zea mays. These findings reveal a much more dynamic UV-protection mechanism than previously recognized, raise important questions concerning the costs and benefits of UV-protection strategies in plants and have practical implications for employing UV to enhance crop vigor and quality in controlled environments.

Glucose- and mannose-induced stomatal closure is mediated by ROS production, Ca(2+) and water channel in Vicia faba.

Sugars act as vital signaling molecules that regulate plant growth, development and stress responses. However, the effects of sugars on stomatal movement have been unclear. In our study, we explored the effects of monosaccharides such as glucose and mannose on stomatal aperture. Here, we demonstrate that glucose and mannose trigger stomatal closure in a dose- and time-dependent manner in epidermal peels of broad bean (Vicia faba). Pharmacological studies revealed that glucose- and mannose-induced stomatal closure was almost completely inhibited by two reactive oxygen species (ROS) scavengers, catalase (CAT) and reduced glutathione (GSH), was significantly abolished by an NADPH oxidase inhibitor, diphenylene iodonium chloride (DPI), whereas they were hardly affected by a peroxidase inhibitor, salicylhydroxamic acid (SHAM). Furthermore, glucose- and mannose-induced stomatal closure was strongly inhibited by a Ca(2+) channel blocker, LaCl3 , a Ca(2+) chelator, ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid (EGTA) and two water channel blockers, HgCl2 and dimethyl sulfoxide (DMSO); whereas the inhibitory effects of the water channel blockers were essentially abolished by the reversing agent ß-mercaptoethanol (ß-ME). These results suggest that ROS production mainly via NADPH oxidases, Ca(2+) and water channels are involved in glucose- and mannose-induced stomatal closure.

Resistance of Faba Bean and Pea Germplasm to Callosobruchus chinensis (Coleoptera: Bruchidae) and Its Relationship With Quality Components.

In total, 339 faba bean (Vicia faba L.) and 100 pea (Pisum sativum L.) accessions were screened for their ability to resist Callosobruchus chinensis L. in free choice laboratory tests. Four, 15, and 43 faba bean varieties were highly resistant, resistant, and moderately resistant to C. chinensis, respectively. Three immune, three highly resistant, and six resistant accessions were discovered among the pea germplasm. The faba bean and pea varieties presented a hundred-kernel weight reduction varied from 0.18 to 35.36% for faba bean varieties and 0 to 56.53% for pea varieties. Varieties with brown and black seed color had significantly fewer wormholes and higher C. chinensis resistance than varieties with light-color seeds. Resistance to C. chinensis showed a significant, positive correlation with catechin, total polyphenol, and γ-aminobutyric acid contents, but a significant, negative correlation with oligosaccharide content. Correlation coefficients (r) between infestation rate of faba bean and total phenol, catechin, and oligosaccharide contents were -0.9723, -0.8071, and 0.7631, respectively. The values of r for pea resistance and total phenol, catechin, and oligosaccharide content were -0.8846, -0.7666, and 0.8308, respectively. The results suggest that quality components in faba bean and pea have a great role in resistance against C. chinensis.

Single-bilayer graphene oxide sheet impacts and underlying potential mechanism assessment in germinating faba bean (Vicia faba L.).

This study investigates the impact of different single-bilayer graphene oxide sheet (hereafter 'graphene oxide', GO; size: 0.5-5 µm) concentrations (0, 100, 200, 400, 800 and 1,600 mg L(-1)) and underlying potential mechanisms in germinating faba bean (Vicia faba L.) seedlings. The study revealed both positive and negative concentration-dependent GO-effects on V. faba. Significant negative impacts of GO concentrations (ordered by magnitude of effect: 1600>200>100 mg GO L(-1)) were indicated by decreases in growth parameters and the activity of H2O2-decomposing enzymes (ascorbate peroxidase, APX; catalase, CAT), and by increases in the levels of electrolyte leakage (EL), H2O2, and lipid and protein oxidation. The positive impacts of 400 and 800 mg GO L(-1) included significant improvements in V. faba health status indicated by decreased levels of EL, H2O2, and lipid and protein oxidation, and by enhanced H2O2-decomposing APX and CAT activity, and increased proline and seed-relative water content. V. faba seedlings-polypeptide patterns strongly substantiated these GO-concentration effects. Overall, the positive effects of these two GO concentrations (800>400 mg L(-1)) on V. faba seedlings indicate their safe nature and allow to suggest further studies.

Response of bean (Vicia faba L.) plants to low sink demand by measuring the gas exchange rates and chlorophyll a fluorescence kinetics.

BACKGROUND: The decline of photosynthesis in plants under low sink demand is well known. Previous studies focused on the relationship between stomatal conductance (gs) and net photosynthetic rate (Pn). These studies investigated the effect of changes in Photosystem II (PSII) function on the Pn decline under low sink demand. However, little is known about its effects on different limiting steps of electron transport chain in PSII under this condition. METHODOLOGY/PRINCIPAL FINDING: Two-month-old bean plants were processed by removing pods and flowers (low sink demand). On the 1(st) day after low sink demand treatment, a decline of Pn was accompanied by a decrease in gs and internal-to-ambient CO2 concentration ratio (Ci/Ca). From the 3(rd) to 9(th) day, Pn and gs declined continuously while Ci/Ca ratio remained stable in the treatment. Moreover, these values were lower than that of control. Wk (a parameter reflecting the damage to oxygen evolving complex of the donor side of PSII) values in the treatment were significantly higher than their corresponding control values. However, RCQA (a parameter reflecting the number of active RCs per excited cross-section of PSII) values in the treatment were significantly lower than control from the 5(th) day. From the 11(th) to 21(st) day, Pn and gs of the treatment continued to decline and were lower than control. This was accompanied by a decrease of RCQA, and an increase of Wk. Furthermore, the quantum yield parameters φPo, φEo and ψEo in the treatment were lower than in control; however, Ci/Ca values in the treatment gradually increased and were significantly higher than control on the 21(st) day. CONCLUSIONS: Stomatal limitation during the early stage, whereas a combination of stomatal and non-stomatal limitation during the middle stage might be responsible for the reduction of Pn under low sink demand. Non-stomatal limitation during the late stages after the removal of the sink of roots and pods may also cause Pn reduction. The non-stomatal limitation was associated with the inhibition of PSII electron transport chain. Our data suggests that the donor side of PSII was the most sensitive to low sink demand followed by the reaction center of PSII. The acceptor side of PSII may be the least sensitive.

Saccharomyces cerevisiae-induced stomatal closure mainly mediated by salicylhydroxamic acid-sensitive peroxidases in Vicia faba.

Saccharomyces cerevisiae induced stomatal closure in a dose-dependent manner on Vicia faba L. (cv. Daqingpi). Using pharmacological inhibitors in this study, we found that stomatal closure was completely inhibited by salicylhydroxamic acid (SHAM) and reduced glutathione (GSH), whereas slightly inhibited by diphenyleneiodonium chloride (DPI), suggesting that H2O2 was mostly produced by cell wall peroxidases. The specific NO scavenger (cPTIO), NO synthase (NOS) inhibitor NG-nitro-l-arginine methyl ester (l-NAME) and sodium azide (NaN3; inhibitor of nitrate reductase) prevented yeast-induced stomatal closure, suggesting that NO in guard cells of V. faba is derived from both NOS-like enzyme and nitrate reductase. Results of HgCl2 and ß-mercaptoethanol (ME) treatment (as a functional inhibitor of water channels and its reversing agent, respectively) suggest that water channels are involved in yeast-induced stomatal movements. CoCl2 (the blocker of calcium channel), LaCl3 (Ca(2+) antagonist) and EGTA (Ca(2+) chelator) also impaired yeast-induced stomatal closure. Thus, it is concluded that H2O2, NO, water channels and Ca(2+) are involved in yeast-induced stomatal closure.

Ratiometric monitoring of transient apoplastic alkalinizations in the leaf apoplast of living Vicia faba plants: chloride primes and PM-H+-ATPase shapes NaCl-induced systemic alkalinizations.

Transient apoplastic alkalinization has been discussed as a general stress factor, and is thought to represent a root-to-shoot signal that transmits information regarding an ongoing NaCl stress event from the site of the trigger to the distant plant tissue. Surprisingly, despite this importance, a number of gaps exist in our knowledge of NaCl-induced apoplastic pH alkalinization. This study was designed in order to shed light onto the mechanisms responsible for the initiation and transiency of leaf apoplastic alkalinization under conditions of NaCl stress as supplied to roots. An H(+)-sensitive fluorescence probe, in combination with ratiometric microscopy imaging, was used for in planta live recording of leaf apoplastic pH. The use of a nonionic solute demonstrated that the alkalinization is induced in response to ionic, and not osmotic, components of NaCl stress. Tests with Cl(-)- or Na(+)-accompanying counter-ions strengthened the idea that the stress factor itself, namely Cl(-), is transferred from root to shoot and elicits the pH alterations. Investigations with a plasma membrane ATPase inhibitor suggest that ATPase activity influences the course of the alkalinization by having a shaping re-acidifying effect on the alkalinization.

Antioxidant and prooxidant effects of lanthanum ions on Vicia faba L. seedlings under cadmium stress, suggesting ecological risk.

The present study combined chemical analyses and biological measurements to investigate biphasic effects of La on Cd stress in leaves of Vicia faba seedlings, which were hydroponically cultivated for 15 d in the combination of 6 µM CdCl(2) and 2 to 480 µM La(NO(3))(3), respectively. The results showed that contents of Cd first elevated above and then declined below the 6 µM single Cd treatment when 2 to 30 µM extraneous La were combined. Contents of mineral nutrients altered differentially and became imbalanced. No distinct band was observed in catalase (CAT), guaiacol peroxidase (GPX), or ascorbate peroxidase (APX) patterns, but in superoxide dismutase (SOD) isozymes by the supplementation with 8 to 480 µM of extraneous La. Superoxide dismutase and APX activities changed as a U-shaped curve; however, CAT and GPX changed as an inverted U-shaped curve along with increasing La. Moreover, heat shock protein 70 (HSP 70) production was reduced below the single treatment of Cd at 2 to 8 µM of extraneous La and enhanced thereafter. Thus, La at lower concentrations promoted antioxidation against Cd stress; La at higher concentrations turned to prooxidant effects, implicating potential ecological risk. Heat shock protein 70, combined with the antioxidant enzymes, constitutes an integrative defense system, which can be used to estimate the degree of antioxidation or prooxidation of extraneous La to Cd-induced oxidative stress in the seedlings.

Phosphatidic acid inhibits blue light-induced stomatal opening via inhibition of protein phosphatase 1 [corrected].

Stomata open in response to blue light under a background of red light. The plant hormone abscisic acid (ABA) inhibits blue light-dependent stomatal opening, an effect essential for promoting stomatal closure in the daytime to prevent water loss. However, the mechanisms and molecular targets of this inhibition in the blue light signaling pathway remain unknown. Here, we report that phosphatidic acid (PA), a phospholipid second messenger produced by ABA in guard cells, inhibits protein phosphatase 1 (PP1), a positive regulator of blue light signaling, and PA plays a role in stimulating stomatal closure in Vicia faba. Biochemical analysis revealed that PA directly inhibited the phosphatase activity of the catalytic subunit of V. faba PP1 (PP1c) in vitro. PA inhibited blue light-dependent stomatal opening but did not affect red light- or fusicoccin-induced stomatal opening. PA also inhibited blue light-dependent H(+) pumping and phosphorylation of the plasma membrane H(+)-ATPase. However, PA did not inhibit the autophosphorylation of phototropins, blue light receptors for stomatal opening. Furthermore, 1-butanol, a selective inhibitor of phospholipase D, which produces PA via hydrolysis of phospholipids, diminished the ABA-induced inhibition of blue light-dependent stomatal opening and H(+) pumping. We also show that hydrogen peroxide and nitric oxide, which are intermediates in ABA signaling, inhibited the blue light responses of stomata and that 1-butanol diminished these inhibitions. From these results, we conclude that PA inhibits blue light signaling in guard cells by PP1c inhibition, accelerating stomatal closure, and that PP1 is a cross talk point between blue light and ABA signaling pathways in guard cells.

Alleviation effect of alginate-derived oligosaccharides on Vicia faba root tip cells damaged by cadmium.

Cadmium has been shown to prevent Vicia faba growth by inhibiting cell mitosis. In this study we investigated the role of Alginate-derived Oligosaccharides (ADO) in alleviating Vicia faba root tip cells damaged by 6 and 8 mg L(-1) CdCl2. Micronucleus assay and chromosomal aberration assay were used to determine mitotic index, micronucleus frequency and chromosomal aberration frequency. The results showed that micronucleus frequency of Vicia faba root tip cells was inhibited under all the ADO concentrations. Especially, the inhibition ratio of 0.125% ADO highly reached 66.11 and 67.17% in 6 and 8 mg L(-1) CdCl2, respectively. Furthermore, the mitotic index increased (p < 0.05) and chromosomal aberration frequency decreased (p < 0.05) under all the ADO concentrations. This indicated that ADO had a significant alleviation effect on Vicia faba root tip cells damaged by cadmium.