Zinc and Silicon Nano-Fertilizers Influence Ionomic and Metabolite Profiles in Maize to Overcome Salt Stress.

Salinity stress is a major factor affecting the nutritional and metabolic profiles of crops, thus hindering optimal yield and productivity. Recent advances in nanotechnology propose an avenue for the use of nano-fertilizers as a potential solution for better nutrient management and stress mitigation. This study aimed to evaluate the benefits of conventional and nano-fertilizers (nano-Zn/nano-Si) on maize and subcellular level changes in its ionomic and metabolic profiles under salt stress conditions. Zinc and silicon were applied both in conventional and nano-fertilizer-using farms under stress (100 mM NaCl) and normal conditions. Different ions, sugars, and organic acids (OAs) were determined using ion chromatography and inductively coupled plasma mass spectroscopy (ICP-MS). The results revealed significant improvements in different ions, sugars, OAs, and other metabolic profiles of maize. Nanoparticles boosted sugar metabolism, as evidenced by increased glucose, fructose, and sucrose concentrations, and improved nutrient uptake, indicated by higher nitrate, sulfate, and phosphate levels. Particularly, nano-fertilizers effectively limited Na accumulation under saline conditions and enhanced maize's salt stress tolerance. Furthermore, nano-treatments optimized the potassium-to-sodium ratio, a critical factor in maintaining ionic homeostasis under stress conditions. With the growing threat of salinity stress on global food security, these findings highlight the urgent need for further development and implementation of effective solutions like the application of nano-fertilizers in mitigating the negative impact of salinity on plant growth and productivity. However, this controlled environment limits the direct applicability to field conditions and needs future research, particularly long-term field trials, to confirm such results of nano-fertilizers against salinity stress and their economic viability towards sustainable agriculture.

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.

Effectiveness of three nitrification inhibitors on mitigating trace gas emissions from different soil textures under surface and subsurface drip irrigation.

Nitrification inhibitors (NIs) and drip irrigation are recommended to mitigate trace gas emissions from agricultural soils. However, studies comparing the effect of different NIs on the release of trace gases from soils with contrasting textures under subsurface (SBD) and surface (SD) drip irrigation are lacking. Therefore, this study aimed to assess the effectiveness of three NIs in mitigating nitrous oxide (N2O), carbon dioxide (CO2), and methane (CH4) emissions from two soils with different textures under SBD, with pipe buried in 10 cm depth, and SD. Two greenhouse experiments were carried out with silt loam and loamy sand soil textures cultivated with wheat under SBD and SD to assess the effectiveness of the NIs Dicyandiamide (DCD), 3,4-Dimethylpyrazole phosphate (DMPP), and 3-Methylpyrazol combined with Triazol (MP + TZ). Ammonium sulfate was applied at a rate of 0.18 g N kg soil-1. The measured variables were daily and cumulative N2O-N, CO2-C, and CH4-C emissions, as well as soil NH4+-N and NO3--N concentrations. The NIs and SBD had additive effects on reducing N2O-N emissions in the silt loam, but not in the loamy sand soil texture. Under SBD, total N2O-N emissions were 44% and 52% lower than under SD in the silt loam and loamy sand soil textures, respectively. Moreover, DMPP kept the highest NH4+-N concentrations and promoted the lowest N2O-N release. CO2-C and CH4-C total emissions were not affected by the treatments. Our findings supported the hypothesis that SBD decreases N2O-N emissions relative to SD. Among the investigated NIs, DMPP has the highest effectiveness in retarding nitrification and mitigating N2O-N release under the studied treatments. Finally, in coarse-textured soils, the use of NIs could be sufficient to significantly abate N2O-N emissions.

Uptake, subcellular distribution, and translocation of foliar-applied phosphorus: Short-term effects on ion relations in deficient young maize plants.

One crucial aspect for successful foliar application is the uptake of the nutrient into the symplast for metabolization by the plant. Our aim was to determine the subcellular distribution of foliar-applied P in leaves, the translocation of this element within the whole plant, and its impact on the ion status of P-deficient maize plants within the first 48 h of treatment. Maize plants with P deficiency were sprayed with 200 mM KH2PO4. After 6, 24, and 48 h, the 5th leaf of each plant was harvested for the isolation of apoplastic washing fluid, cell sap, and vascular bundle sap and for the examination of transporter gene expression. The remaining tissues were divided into 4th leaf, older and younger shoots, and root for total P determination. No accumulation of foliar-applied P was measured in the apoplast. P was mostly taken up into the cytosol within the first 6 h and was associated with increased mRNA levels of PHT1 transporters. A strong tendency towards rapid translocation into the younger shoot and an increase in NO3- uptake or a decrease in organic acid translocation were observed. The apoplast seems to exert no effect on the uptake of foliar-applied P into the epidermal and mesophyll cells of intact leaves. Instead, the plant responds with the rapid translocation of P and changes in ion status to generate further growth. The effect of the absorbed foliar-applied P is assumed to be a rapid process with no transient storage in the leaf apoplast.

Silicon-mediated growth promotion in maize (Zea mays L.) occurs via a mechanism that does not involve activation of the plasma membrane H+-ATPase.

Silicon (Si)-mediated growth promotion of various grasses is well documented. In the present study, Si-induced changes in maize shoot growth and its underlying mechanisms were studied. Maize plants were grown with various concentrations of Si (0-3 mM) in the nutrient solution. Silicon nutrition improved plant expansion growth. Silicon-supplied maize plants (0.8 and 1.2 mM) showed higher plant height and leaf area compared to no-Si amended plants. It was assumed that Si-induced expansion growth was due to positive Si effects on plasma membrane (PM) H+-ATPase. In this context, western blot analysis revealed an increase in PM H+-ATPase abundance by 77% under Si nutrition. However, in vitro measurements of enzyme activities showed no significant effect on apoplast pH, proton pumping, passive H+ efflux and enzyme kinetics such as Km, Vmax, and activation energy. Further, these results were confirmed by in vivo ratiometric analysis of apoplastic pH, which showed non-significant changes upon Si supply. In contrast, 1 mM Si altered the relative transcripts of specific PM H+-ATPase isoforms. Silicon application resulted in a significant decrease of MHA3, and this decrease in transcription seems to be compensated by an increased concentration of H+-ATPase protein. From these results, it can be concluded that changes in cell wall composition and PM H+-ATPase may be responsible for Si-mediated growth improvement in maize.

Silicon decreases cadmium concentrations by modulating root endodermal suberin development in wheat plants.

Silicon (Si) can alleviate cadmium (Cd) toxicity in many plants, but mechanisms underlying this beneficial effect are still lacking. In this study, the roles of Si in time-dependent apoplastic and symplastic Cd absorption by roots of wheat plants were investigated. Results showed that, during short-term Cd exposure, the symplastic pathway of Cd in roots was not significantly affected by Si. Cell wall properties and cell wall-bound Cd regarding the apoplastic pathway were unaffected by Si either. Nevertheless, Cd concentrations in the apoplastic fluid of roots were decreased by Si. The reason could be that Si delayed endodermal suberization of roots resulting in promoted apoplastic Cd translocation to shoots, thus decreasing Cd in the apoplastic fluid of roots after short-term Cd stress. By contrast, after long-term Cd stress, cell wall properties and the expression of genes related to Cd influx and transport were unaffected. Intriguingly, Si up-regulated the expression of the Cd efflux-related gene TaTM20 and repressed apoplastic Cd translocation to shoots, which might contribute to decrease of Cd after long-term Cd exposure. Taken together, these results indicate that Si-dependent decrease in root Cd concentrations during short-term Cd exposure helps plants to mitigate Cd toxicity in the long-term.

Diferulic acids in the cell wall may contribute to the suppression of shoot growth in the first phase of salt stress in maize.

In the first phase of salt stress the elongation growth of maize shoots is severely affected. The fixation of shape at the end of the elongation phase in Poaceae leaves has frequently been attributed to the formation of phenolic cross-links in the cell wall. In the present work it was investigated whether this process is accelerated under salt stress in different maize hybrids. Plants were grown in nutrient solution in a growth chamber. Reduction of shoot fresh mass was 50% for two hybrids which have recently been developed for improved salt resistance (SR 03, SR 12) and 60% for their parental genotype (Pioneer 3906). For SR 12 and Pioneer 3906, the upper three leaves were divided into elongated and elongating tissue and cell walls were isolated from which phenolic substances and neutral sugars were determined. Furthermore, for the newly developed hybrids the activity of phenolic peroxidase in the cell wall was analysed in apoplastic washing fluids and after sequential extraction of cell-wall material with CaCl2 and LiCl. The concentration of ferulic acid, the predominant phenolic cross-linker in the grass cell wall, was about 5mgg(-1) dry cell wall in elongating and in elongated tissue. The concentration of diferulic acids (DFA) was 2-3mgg(-1) dry cell wall in both tissues. Salt stress increased the concentration of ferulic acid (FA) and DFA in the parental genotype Pioneer 3906, but not in SR 12. Both genotypes showed an increase in arabinose, which is the molecule at which FA and DFA are coupled to interlocking arabinoxylan polymers. In SR 12, the activity of phenolic peroxidase was not influenced by salt stress. However, in SR 03 salt stress clearly increased the phenolic peroxidase activity. Results are consistent with the hypothesis that accelerated oxidative fixation of shape contributes to growth suppression in the first phase of salt stress in a genotype-specific manner.

Proteome analysis of sugar beet (Beta vulgaris L.) elucidates constitutive adaptation during the first phase of salt stress.

Salinity is one of the major stress factors responsible for growth reduction of most of the higher plants. In this study, the effect of salt stress on protein pattern in shoots and roots of sugar beet (Beta vulgaris L.) was examined. Sugar beet plants were grown in hydroponics under control and 125 mM salt treatments. A significant growth reduction of shoots and roots was observed. The changes in protein expression, caused by salinity, were monitored using two-dimensional gel-electrophoresis. Most of the detected proteins in sugar beet showed stability under salt stress. The statistical analysis of detected proteins showed that the expression of only six proteins from shoots and three proteins from roots were significantly altered. At this stage, the significantly changed protein expressions we detected could not be attributed to sugar beet adaptation under salt stress. However, unchanged membrane bound proteins under salt stress did reveal the constitutive adaptation of sugar beet to salt stress at the plasma membrane level.

Hydrolytic and pumping activity of H+-ATPase from leaves of sugar beet (Beta vulgaris L.) as affected by salt stress.

Cell wall extensibility plays an important role in plant growth. According to the acid-growth theory, lower apoplastic pH allows extension growth by affecting cell wall extensibility. A lowered apoplastic pH is presumed to activate wall-loosening enzymes that control plant growth. Plasma membrane (PM) H(+)-ATPases play a major role in the apoplastic acidification by H(+) transport from cytosol to the apoplast. A salt-induced decrease in H(+)-pumping activity of plasma membrane H(+)-ATPases in salt-sensitive maize plants has previously been found. This led us to formulate the hypothesis that salt-resistant plant species such as sugar beet (Beta vulgaris L.) may have a mechanism to eliminate the effect of higher salt concentrations on plasma membrane H(+)-ATPase activity. In the present study, sugar beet plants were grown in 1mM NaCl (control) or 150 mM NaCl in hydroponics. H(+)-ATPase hydrolytic and pumping activities were measured in plasma membrane vesicles isolated from sugar beet shoots. We found that plasma membrane H(+)-ATPase hydrolytic and pumping activities were not affected by application of 150 mM NaCl. Moreover, apoplastic pH was also not affected under salt stress. However, a decrease in plant growth was observed. We assume that growth reduction was not due to a decrease in PM-H(+)-ATPase activity, but that other factors may be responsible for growth inhibition of sugar beet plants under salt stress.

The apoplastic pH and its significance in adaptation to salinity in maize (Zea mays L.): Comparison of fluorescence microscopy and pH-sensitive microelectrodes.

The apoplastic ionic milieu contains essential determinants for cell expansion and plant growth. Since pH is a multifunctional basic component of this extracellular space, the knowledge of its behaviour during stress situations is of major importance. In detached leaves of maize (Zea mays L. cvs. Pioneer 3906 and SR 03) the effect of salinity on apoplastic pH was measured to investigate its adaptive role to salt stress applying two different methods: an optical approach using pH-sensitive fluorescent dyes (fluorescein isothiocyanate-dextran (FITC), fluorescein tetramethylrhodamine-dextran (FTMR) and Oregon Green(®) 488), and an electrophysiological technique, pH-sensitive microelectrodes. Both approaches yielded similar results. In the presence of 100mM NaCl, which was added to the growth medium, apoplastic pH of the salt-sensitive maize genotype Pioneer 3906 leaves increased in maximum by 0.4 units (pH microelectrodes) and by 0.3 units (fluorescent dyes); the salt-resistant SR 03 hardly responded. The same treatment reduced leaf growth by 60% in Pioneer 3906, but only by 40% in SR 03. Since according to acid growth considerations apoplastic pH is an important factor in elongation growth, we suggest that this pH increase is a main cause for reduced leaf growth under salt stress conditions.