Foliar Application of Sulfur-Containing Compounds-Pros and Cons.

Sulfate is taken up from the soil solution by the root system; and inside the plant, it is assimilated to hydrogen sulfide, which in turn is converted to cysteine. Sulfate is also taken up by the leaves, when foliage is sprayed with solutions containing sulfate fertilizers. Moreover, several other sulfur (S)-containing compounds are provided through foliar application, including the S metabolites hydrogen sulfide, glutathione, cysteine, methionine, S-methylmethionine, and lipoic acid. However, S compounds that are not metabolites, such as thiourea and lignosulfonates, along with dimethyl sulfoxide and S-containing adjuvants, are provided by foliar application-these are the S-containing agrochemicals. In this review, we elaborate on the fate of these compounds after spraying foliage and on the rationale and the efficiency of such foliar applications. The foliar application of S-compounds in various combinations is an emerging area of agricultural usefulness. In the agricultural practice, the S-containing compounds are not applied alone in spray solutions and the need for proper combinations is of prime importance.

Floret Biofortification of Broccoli Using Amino Acids Coupled with Selenium under Different Surfactants: A Case Study of Cultivating Functional Foods.

Broccoli serves as a functional food because it can accumulate selenium (Se), well-known bioactive amino-acid-derived secondary metabolites, and polyphenols. The chemical and physical properties of Se are very similar to those of sulfur (S), and competition between sulfate and selenate for uptake and assimilation has been demonstrated. Towards an efficient agronomic fortification of broccoli florets, the working questions were whether we could overcome this competition by exogenously applying the S-containing amino acids cysteine (Cys) or/and methionine (Met), or/and the precursors of Glucosinolate (GSL) types along with Se application. Broccoli plants were cultivated in a greenhouse and at the beginning of floret growth, we exogenously applied sodium selenate in the concentration gradient of 0, 0.2, 1.5, and 3.0 mM to study the impact of increased Se concentration on the organic S (Sorg) content of the floret. The Se concentration of 0.2 mM (Se0.2) was coupled with the application of Cys, Met, their combination, or a mixture of phenylalanine, tryptophane, and Met. The application took place through fertigation or foliar application (FA) by adding isodecyl alcohol ethoxylate (IAE) or a silicon ethoxylate (SiE) surfactant. Fresh biomass, dry mass, and Se accumulation in florets were evaluated, along with their contents of Sorg, chlorophylls (Chl), carotenoids (Car), glucoraphanin (GlRa), glucobrassicin (GlBra), glucoiberin (GlIb), and polyphenols (PPs), for the biofortification efficiency of the three application modes. From the studied selenium concentration gradient, the foliar application of 0.2 mM Se using silicon ethoxylate (SiE) as a surfactant provided the lowest commercially acceptable Se content in florets (239 µg or 0.3 µmol g-1 DM); it reduced Sorg (-45%), GlIb (-31%), and GlBr (-27%); and it increased Car (21%) and GlRa (27%). Coupled with amino acids, 0.2 mM Se provided commercially acceptable Se contents per floret only via foliar application. From the studied combinations, that of Met,Se0.2/FA,IAE provided the lowest Se content per floret (183 µg or 0.2 µmol g-1 DM) and increased Sorg (35%), Car (45%), and total Chl (27%), with no effect on PPs or GSLs. Cys,Met,Se0.2/FA,IAE and amino acid mix,Se0.2/FA,IAE increased Sorg content, too, by 36% and 16%, respectively. Thus, the foliar application with the IAE surfactant was able to increase Sorg, and methionine was the amino acid in common in these treatments, with varying positive effects on carotenoids and chlorophylls. Only the Cys,Met,Se0.2 combination presented positive effects on GSLs, especially GlRa, but it reduced the fresh mass of the floret. The foliar application with SiE as a surfactant failed to positively affect the organic S content. However, in all studied combinations of Se 0.2 mM with amino acids, the Se content per floret was commercially acceptable, the yield was not affected, the content of GSLs was increased (especially that of GlRa and GlIb), and PPs were not affected. The content of GlBr decreased except for the treatment with methionine (Met,Se0.2/FA,SiE) where GlBr remained unaffected. Hence, the combination of Se with the used amino acids and surfactants can provide enhanced biofortification efficiency in broccoli by providing florets as functional foods with enhanced functional properties.

The Role of Sulfur in Agronomic Biofortification with Essential Micronutrients.

Sulfur (S) is an essential macronutrient for plants, being necessary for their growth and metabolism and exhibiting diverse roles throughout their life cycles. Inside the plant body, S is present either in one of its inorganic forms or incorporated in an organic compound. Moreover, organic S compounds may contain S in its reduced or oxidized form. Among others, S plays roles in maintaining the homeostasis of essential micronutrients, e.g., iron (Fe), copper (Cu), zinc (Zn), and manganese (Mn). One of the most well-known connections is homeostasis between S and Fe, mainly in terms of the role of S in uptake, transportation, and distribution of Fe, as well as the functional interactions of S with Fe in the Fe-S clusters. This review reports the available information describing the connections between the homeostasis of S and Fe, Cu, Zn, and Mn in plants. The roles of S- or sulfur-derived organic ligands in metal uptake and translocation within the plant are highlighted. Moreover, the roles of these micronutrients in S homeostasis are also discussed.

Phenotypic Acclimation of Maize Plants Grown under S Deprivation and Implications to Sulfur and Iron Allocation Dynamics.

The aim of this work was to study maize root phenotype under sulfur deficiency stress towards revealing potential correlations between the altered phenotypic traits and the corresponding dry mass, sulfur, and iron allocation within plants at the whole-plant level. The dynamics of root morphological and anatomical traits were monitored. These traits were then correlated with plant foliage traits along with dry mass and sulfur and iron allocation dynamics in the shoot versus root. Plants grown under sulfate deprivation did not seem to invest in new root axes. Crown roots presented anatomical differences in all parameters studied; e.g., more and larger xylem vessels in order to maximize water and nutrient transport in the xylem sap. In the root system of S-deficient plants, a reduced concentration of sulfur was observed, whilst organic sulfur predominated over sulfates. A reduction in total iron concentration was monitored, and differences in its subcellular localization were observed. As expected, S-deprivation negatively affected the total sulfur concentration in the aerial plant part, as well as greatly impacted iron allocation in the foliage. Phenotypic adaptation to sulfur deprivation in maize presented alterations mainly in the root anatomy; towards competent handling of the initial sulfur and the induced iron deficiencies.

Multi-Trait Wheat Rhizobacteria from Calcareous Soil with Biocontrol Activity Promote Plant Growth and Mitigate Salinity Stress.

Plant growth promoting rhizobacteria (PGPR) can be functional microbial fertilizers and/or biological control agents, contributing to an eco-spirit and safe solution for chemical replacement. Therefore, we have isolated rhizospheric arylsulfatase (ARS)-producing bacteria, belonging to Pseudomonas and Bacillus genus, from durum wheat crop grown on calcareous soil. These isolates harbouring plant growth promoting (PGP) traits were further evaluated in vitro for additional PGP traits, including indole compounds production and biocontrol activity against phytopathogens, limiting the group of multi-trait strains to eight. The selected bacterial strains were further evaluated for PGP attributes associated with biofilm formation, compatibility, salt tolerance ability and effect on plant growth. In vitro studies demonstrated that the multi-trait isolates, Bacillus (1.SG.7, 5.SG.3) and Pseudomonas (2.SG.20, 2.C.19) strains, enhanced the lateral roots abundance and shoots biomass, mitigated salinity stress, suggesting the utility of beneficial ARS-producing bacteria as potential microbial fertilizers. Furthermore, in vitro studies demonstrated that compatible combinations of multi-trait isolates, Bacillus sp. 1.SG.7 in a mixture coupled with 5.SG.3, and 2.C.19 with 5.SG.3 belonging to Bacillus and Pseudomonas, respectively, may enhance plant growth as compared to single inoculants.

Regulation of Sulfur Homeostasis in Mycorrhizal Maize Plants Grown in a Fe-Limited Environment.

Sulfur is an essential macronutrient for growth of higher plants. The entry of the sulfate anion into the plant, its importation into the plastids for assimilation, its long-distance transport through the vasculature, and its storage in the vacuoles require specific sulfate transporter proteins. In this study, mycorrhizal and non-mycorrhizal maize plants were grown for 60 days in an S-deprived substrate, whilst iron was provided to the plants in the sparingly soluble form of FePO4. On day 60, sulfate was provided to the plants. The gene expression patterns of a number of sulfate transporters as well as sulfate assimilation enzymes were studied in leaves and roots of maize plants, both before as well as after sulfate supply. Prolonged sulfur deprivation resulted in a more or less uniform response of the genes' expressions in the roots of non-mycorrhizal and mycorrhizal plants. This was not the case neither in the roots and leaves after the supply of sulfur, nor in the leaves of the plants during the S-deprived period of time. It is concluded that mycorrhizal symbiosis modified plant demands for reduced sulfur, regulating accordingly the uptake, distribution, and assimilation of the sulfate anion.

Impact of Elemental Sulfur on the Rhizospheric Bacteria of Durum Wheat Crop Cultivated on a Calcareous Soil.

Previous experiments have shown that the application of fertilizer granules containing elemental sulfur (S0) as an ingredient (FBS0) in durum wheat crops produced a higher yield than that produced by conventional ones (F), provided that the soils of the experimental fields (F vs. FBS0) were of comparable quality and with the Olsen P content of the field's soil above 8 mg kg-1. In this experiment the FBS0 treatment took place in soil with Olsen P at 7.8 mg kg-1, compared with the F treatment's soil with Olsen P of 16.8 mg kg-1, aiming at reducing the imbalance in soil quality. To assess and evaluate the effect of FBS0 on the dynamics of the rhizospheric bacteria in relation to F, rhizospheric soil at various developmental stages of the crops was collected. The agronomic profile of the rhizospheric cultivable bacteria was characterized and monitored, in connection with the dynamics of phosphorus, iron, organic sulfur, and organic nitrogen, in both the rhizosoil and the aerial part of the plant during development. Both crops were characterized by a comparable dry mass accumulation per plant throughout development, while the yield of the FBS0 crop was 3.4% less compared to the F crop's one. The FBS0 crop's aerial part showed a transient higher P and Fe concentration, while its organic N and S concentrations followed the pattern of the F crop. The incorporation of S0 into the conventional fertilizer increased the percentage of arylsulfatase (ARS)-producing bacteria in the total bacterial population, suggesting an enhanced release of sulfate from the soil's organic S pool, which the plant could readily utilize. The proportion of identified ARS-producing bacteria possessing these traits exhibited a maximum value before and after topdressing. Phylogenetic analysis of the 68 isolated ARS-producing bacterial strains revealed that the majority of the isolates belonged to the Pseudomonas genus. A large fraction also possessed phosphate solubilization, and/or siderophore production, and/or ureolytic traits, thus improving the crop's P, Fe, S, and N balance. The aforementioned findings imply that the used FBS0 substantially improved the quality of the rhizosoil at the available phosphorus limiting level by modulating the abundance of the bacterial communities in the rhizosphere and effectively enhancing the microbially mediated nutrient mobilization towards improved plant nutritional dynamics.

An Exploration of the Roles of Ferric Iron Chelation-Strategy Components in the Leaves and Roots of Maize Plants.

Plants have developed sophisticated mechanisms for acquiring iron from the soil. In the graminaceous species, a chelation strategy is in charge, in order to take up ferric iron from the rhizosphere. The ferric iron chelation-strategy components may also be present in the aerial plant parts. The aim of this work was to search for possible roles of those components in maize leaves. To this end, the expression patterns of ferric iron chelation-strategy components were monitored in the leaves and roots of mycorrhizal and non-mycorrhizal sulfur-deprived maize plants, both before and after sulfate supply. The two levels of sulfur supply were chosen due to the strong impact of this nutrient on iron homeostasis, whilst mycorrhizal symbiosis was chosen as a treatment that forces the plant to optimize its photosynthetic efficiency, in order to feed the fungus. The results, in combination with the findings of our previous works, suggest a role for the aforementioned components in ferric chelation and/or unloading from the xylem vessels to the aerial plant parts. It is proposed that the gene expression of the DMA exporter ZmTOM1 can be used as an early indicator for the establishment of a mycorrhizal symbiotic relationship in maize.

The Effect of Granular Commercial Fertilizers Containing Elemental Sulfur on Wheat Yield under Mediterranean Conditions.

The demand to develop fertilizers with higher sulfur use efficiency has intensified over the last decade, since sulfur deficiency in crops has become more widespread. The aim of this study was to investigate whether fertilizers enriched with 2% elemental sulfur (ES) via a binding material of organic nature improve yield when compared to the corresponding conventional ones. Under the scanning electron microscope, the granules of the ES-containing fertilizer were found to be covered by a layer of crystal-like particles, the width of which was found to be up to 60 µm. Such a layer could not be found on the corresponding conventional fertilizer granules. Several fertilization schemes with or without incorporated ES were tested in various durum wheat varieties, cultivated in commercial fields. The P-Olsen content of each commercial field was found to be correlated with the corresponding relative change in the yields (YF/YFBES) with a strong positive relationship. The content of 8 ppm of available soil phosphorus was a turning point. At higher values the incorporation of ES in the fertilization scheme resulted in higher yield, while at lower values it resulted in lower yield, compared with the conventional one. The experimental field trials that established following a randomized block design, were separated in two groups: One with P-Olsen ranging between 18⁻22 ppm and the other between 12⁻15 ppm, the results of which corroborated the aforementioned finding. The use of ES in all portions of fertilization schemes provided higher relative yields. The coexistence of ES with sulfate in the granule was more efficient in terms of yield, when compared to the granule enriched with ES alone under the same fertilization scheme and agronomic practice. The application of fertilizer mixtures containing the urease inhibitor N-(n-butyl) thiophosphoric triamide (NBPT), ES and ammonium sulfate resulted in even higher relative yields. Yield followed a positive linear relationship with the number of heads per square meter. In this correlation, the P-Olsen content separated the results of the two groups of blocks, where the applied linear trend line in each group presented the same slope.

Arbuscular mycorrhizal symbiosis alters the expression patterns of three key iron homeostasis genes, ZmNAS1, ZmNAS3, and ZmYS1, in S deprived maize plants.

Nicotianamine is an essential molecule for Fe homeostasis in plants, its primary precursor is the S-containing compound methionine, and it is biosynthesized by the enzyme family of nicotianamine synthases (NASs). In maize, a graminaceous plant that follows Strategy II for Fe uptake, ZmNAS genes can be subgrouped into two classes, according to their roles and tissue specific expression profiles. In roots, the genes of class I provide NA for the production of deoxymugineic acid (DMA), which is secreted to the rhizosphere and chelates Fe(III). The Fe(III)-DMA complex is then inserted to the root via a ZmYS1 transporter. The genes of class II provide NA for local translocation and detoxification of Fe in the leaves. Due to the connection between S and Fe homeostasis, S deficiency causes Fe deprivation responses to graminaceous plants and when S is supplied, these responses are inverted. In this study, maize plants were grown in pots with sterile river sand containing FePO4 and were inoculated with the mycorrhizal fungus Rhizophagus irregularis. The plants were grown under S deficient conditions until day 60 from sowing and on that day sulfate was provided to the plants. In order to assess the impact of AM symbiosis on Fe homeostasis, the expression patterns of ZmNAS1, ZmNAS3 (representatives of ZmNAS class I and class II), and ZmYS1 were monitored before and after S supply by means of real time RT-PCR and they were used as indicators of the plant Fe status. In addition, total shoot Fe concentration was determined before and after S supply. AM symbiosis prevented Fe deprivation responses in the S deprived maize plants and iron was possibly provided directly to the mycorrhizal plants through the fungal network. Furthermore, sulfate possibly regulated the expression of all three genes revealing its potential role as signal molecule for Fe homeostasis.

New insights into trophic aerenchyma formation strategy in maize (Zea mays L.) organs during sulfate deprivation.

Aerenchyma attributes plant tissues that contain enlarged spaces exceeding those commonly found as intracellular spaces. It is known that sulfur (S) deficiency leads to formation of aerenchyma in maize adventitious roots by lysis of cortical cells. Seven-day-old maize plants were grown in a hydroponics setup for 19 days under S deprivation against full nutrition. At day 17 and 26 from sowing (d10 and d19 of the deprivation, respectively), a detailed analysis of the total sulfur and sulfate allocation among organs as well as a morphometric characterization were performed. Apart from roots, in S-deprived plants aerenchyma formation was additionally found in the second leaf and in the mesocotyl, too. The lamina (LA) of this leaf showed enlarged gas spaces between the intermediate and small vascular bundles by lysis of mesophyll cells and to a greater extent on the d10 compared to d19. Aerenchymatous spaces were mainly distributed along the middle region of leaf axis. At d10, -S leaves invested less dry mass with more surface area, whilst lesser dry mass was invested per unit surface area in -S LAs. In the mesocotyl, aerenchyma was located near the scutelar node, where mesocotyl roots were developing. In -S roots, more dry mass was invested per unit length. Our data suggest that trying to utilize the available scarce sulfur in an optimal way, the S-deprived plant fine tunes the existing roots with the same length or leaves with more surface area per unit of dry mass. Aerenchyma was not found in the scutelar node and the bases of the attached roots. The sheaths, the LAs' bases and the crown did not form aerenchyma. This trophic aerenchyma is a localized one, presumably to support new developing tissues nearby, by induced cell death and recycling of the released material. Reduced sulfur allocation among organs followed that of dry mass in a proportional fashion.

Comparative spatiotemporal analysis of root aerenchyma formation processes in maize due to sulphate, nitrate or phosphate deprivation.

Nitrate (N), phosphate (P) or sulphate (S) deprivation causes aerenchyma formation in maize (Zea mays L.) nodal roots. The exact mechanisms that trigger the formation of aerenchyma under these circumstances are unclear. We have compared aerenchyma distribution across the nodal roots of first whorl (just emerging in 10-day-old seedlings), which were subject to S, N or P deprivation over a period of 10 days in connection with oxygen consumption, ATP concentration, cellulase and polygalacturonase activity in the whole root. The effect of deprivation on aerenchyma formation was examined using light and electron microscopy, along with in situ detection of calcium and of reactive oxygen species (ROS) by fluorescence microscopy. Aerenchyma was not found in the root base regardless of the deprivation. Programmed cell death (PCD) was observed near the root tip, either within the first two days (-N) or a few days later (-S, -P) of the treatment. Roots at day 6 under all three nutrient-deprived conditions showed signs of PCD 1 cm behind the cap, whereas only N-deprived root cells 0.5 cm behind the cap showed severe ultrastructural alterations, due to advanced PCD. The lower ATP concentration and the higher oxygen consumptions observed at day 2 in N-, P- and S-deprived roots compared to the control indicated that PCD may be triggered by perturbations in energy status of the root. The peaks of cellulase activity located between days 3 (-N) and 6 (-P), along with the respective alterations in polygalacturonase activity, indicated a coordination which preceded aerenchyma formation. ROS and calcium seemed to contribute to PCD initiation, with ROS possessing dual roles as signals and eliminators. All the examined parameters presented both common features and characteristic variations among the deprivations.

Dynamics of Aerenchyma distribution in the cortex of sulfate-deprived adventitious roots of maize.

BACKGROUND AND AIMS: Aerenchyma formation in maize adventitious roots is induced in nutrient solution by the deprivation of sulfate (S) under well-oxygenated conditions. The aim of this research was to examine the extent of aerenchyma formation in the cortex of sulfate-deprived adventitious roots along the root axis, in correlation with the presence of reactive oxygen species (ROS), calcium levels and pH of cortex cells and root lignification. METHODS: The morphometry of the second whorl of adventitious (W2) roots, subject to S-deprivation conditions throughout development, was recorded in terms of root length and lateral root length and distribution. W2 roots divided into sectors according to the mean length of lateral roots, and cross-sections of each were examined for aerenchyma. In-situ detection of alterations in ROS presence, calcium levels and pH were performed by means of fluorescence microscopy using H(2)DCF-DA, fluo-3AM and BCECF, respectively. Lignification was detected using the Wiesner test. KEY RESULTS: S-deprivation reduced shoot growth and enhanced root proliferation. Aerenchyma was found in the cortex of 77 % of the root length, particularly in the region of emerging or developing lateral roots. The basal and apical sectors had no aerenchyma and no aerenchyma connection was found with the shoot. S-deprivation resulted in alterations of ROS, calcium levels and pH in aerenchymatous sectors compared with the basal non-aerenchymatous region. Lignified epidermal layers were located at the basal and the proximal sectors. S-deprivation resulted in shorter lateral roots in the upper sectors and in a limited extension of the lignified layers towards the next lateral root carrying sector. CONCLUSIONS: Lateral root proliferation is accompanied by spatially localized induced cell death in the cortex of developing young maize adventitious roots during S-deprivation.