S-deprived Plants Research Articles

Silicon Supply Improves Nodulation and Dinitrogen Fixation and Promotes Growth in Trifolium incarnatum Subjected to a Long-Term Sulfur Deprivation.

In many crops species, sulfur (S) deprivation negatively affects growth, seed yield quality and plant health. Furthermore, silicon (Si) is known to alleviate many nutritional stresses but the effects of Si supply on plants subjected to S deficiency remain unclear and poorly documented. The objective of this study was to evaluate whether Si supply would alleviate the negative effects of S deprivation on root nodulation and atmospheric dinitrogen (N2) fixation capacity in Trifolium incarnatum subjected (or not) to long-term S deficiency. For this, plants were grown for 63 days in hydroponic conditions with (500 µM) or without S and supplied (1.7 mM) or not with Si. The effects of Si on growth, root nodulation and N2 fixation and nitrogenase abundance in nodules have been measured. The most important beneficial effect of Si was observed after 63 days. Indeed, at this harvest time, a Si supply increased growth, the nitrogenase abundance in nodules and N2 fixation in S-fed and S-deprived plants while a beneficial effect on the number and total biomass of nodules was only observed in S-deprived plants. This study shows clearly for the first time that a Si supply alleviates negative effects of S deprivation in Trifolium incarnatum.

Assessing the Effect of Silicon Supply on Root Sulfur Uptake in S-Fed and S-Deprived Brassica napus L.

Silicon (Si) is known to alleviate many nutritional stresses. However, in Brassica napus, which is a highly S-demanding species, the Si effect on S deficiency remains undocumented. The aim of this study was to assess whether Si alleviates the negative effects of S deficiency on Brassica napus and modulates root sulfate uptake capacity and S accumulation. For this, Brassica napus plants were cultivated with or without S and supplied or not supplied with Si. The effects of Si on S content, growth, expression of sulfate transporter genes (BnaSultr1.1; BnaSultr1.2) and sulfate transporters activity in roots were monitored. Si supply did not mitigate growth or S status alterations due to S deprivation but moderated the expression of BnaSultr1.1 in S-deprived plants without affecting the activity of root sulfate transporters. The effects of Si on the amount of S taken-up and on S transporter gene expression were also evaluated after 72 h of S resupply. In S-deprived plants, S re-feeding led to a strong decrease in the expression of both S transporter genes as expected, except in Si-treated plants where BnaSultr1.1 expression was maintained over time. This result is discussed in relation to the similar amount of S accumulated regardless of the Si treatment.

Sulfur Deprivation Modulates Salicylic Acid Responses via Nonexpressor of Pathogenesis-Related Gene 1 in Arabidopsis thaliana.

Mineral nutrients are essential for plant growth and reproduction, yet only a few studies connect the nutritional status to plant innate immunity. The backbone of plant defense response is mainly controlled by two major hormones: salicylic acid (SA) and jasmonic acid (JA). This study investigated changes in the macronutrient concentration (deficiency/excess of nitrogen, phosphorus, potassium, magnesium, and sulfur) on the expression of PR1, a well-characterized marker in the SA-pathway, and PDF1.2 and LOX2 for the JA-pathway, analyzing plants carrying the promoter of each gene fused to GUS as a reporter. After histochemical GUS assays, we determined that PR1 gene was strongly activated in response to sulfur (S) deficiency. Using RT-PCR, we observed that the induction of PR1 depended on the function of Non-expressor of Pathogenesis-Related gene 1 (NPR1) and SA accumulation, as PR1 was not expressed in npr1-1 mutant and NahG plants under S-deprived conditions. Plants treated with different S-concentrations showed that total S-deprivation was required to induce SA-mediated defense responses. Additionally, bioassays revealed that S-deprived plants, induced resistance to the hemibiotrophic pathogen Pseudomonas syringae pv. DC3000 and increase susceptibility to the necrotrophic Botrytis cinerea. In conclusion, we observed a relationship between S and SA/JA-dependent defense mechanisms in Arabidopsis.

Melatonin alleviates low-sulfur stress by promoting sulfur homeostasis in tomato plants

Despite involvement of melatonin (MT) in plant growth and stress tolerance, its role in sulfur (S) acquisition and assimilation remains unclear. Here we report that low-S conditions cause serious growth inhibition by reducing chlorophyll content, photosynthesis and biomass accumulation. S deficiency evoked oxidative stress leading to the cell structural alterations and DNA damage. In contrast, MT supplementation to the S-deprived plants resulted in a significant diminution in reactive oxygen species (ROS) accumulation, thereby mitigating S deficiency-induced damages to cellular macromolecules and ultrastructures. Moreover, MT promoted S uptake and assimilation by regulating the expression of genes encoding enzymes involved in S transport and metabolism. MT also protected cells from ROS-induced damage by regulating 2-cysteine peroxiredoxin and biosynthesis of S-compounds. These results provide strong evidence that MT can enhance plant tolerance to low-S-induced stress by improving S uptake, metabolism and redox homeostasis, and thus advocating beneficial effects of MT on increasing the sulfur utilization efficiency.

Effect of sulphur deprivation on osmotic potential components and nitrogen metabolism in oilseed rape leaves: identification of a new early indicator.

Identification of early sulphur (S) deficiency indicators is important for species such as Brassica napus, an S-demanding crop in which yield and the nutritional quality of seeds are negatively affected by S deficiency. Because S is mostly stored as SO4 (2-) in leaf cell vacuoles and can be mobilized during S deficiency, this study investigated the impact of S deprivation on leaf osmotic potential in order to identify compensation processes. Plants were exposed for 28 days to S or to chlorine deprivation in order to differentiate osmotic and metabolic responses. While chlorine deprivation had no significant effects on growth, osmotic potential and nitrogen metabolism, Brassica napus revealed two response periods to S deprivation. The first one occurred during the first 13 days during which plant growth was maintained as a result of vacuolar SO4 (2-) mobilization. In the meantime, leaf osmotic potential of S-deprived plants remained similar to control plants despite a reduction in the SO4 (2-) osmotic contribution, which was fully compensated by an increase in NO3 (-), PO4 (3-) and Cl(-) accumulation. The second response occurred after 13 days of S deprivation with a significant reduction in growth, leaf osmotic potential, NO3 (-) uptake and NO3 (-) reductase activity, whereas amino acids and NO3 (-) were accumulated. This kinetic analysis of S deprivation suggested that a ([Cl(-)]+[NO3 (-)]+[PO4 (3-)]):[SO4 (2-)] ratio could provide a relevant indicator of S deficiency, modified nearly as early as the over-expression of genes encoding SO4 (2-) tonoplastic or plasmalemmal transporters, with the added advantage that it can be easily quantified under field conditions.

Sulfate resupply accentuates protein synthesis in coordination with nitrogen metabolism in sulfur deprived Brassica napus

To investigate the regulatory interactions between S assimilation and N metabolism in Brassica napus, de novo synthesis of amino acids and proteins was quantified by 15N and 34S tracing, and the responses of transporter genes, assimilatory enzymes and metabolites pool involving in nitrate and sulfate metabolism were assessed under continuous sulfur supply, sulfur deprivation and sulfate resupply after 3 days of sulfur (S) deprivation. S-deprived plants were characterized by a strong induction of sulfate transporter genes, ATP sulfurylase (ATPS) and adenosine 5′-phosphosulfate reductase (APR), and by a repressed activity of nitrate reductase (NR) and glutamine synthetase (GS). Sulfate resupply to the S-deprived plants strongly increased cysteine, amino acids and proteins concentration. The increase in sulfate and cysteine concentration caused by sulfate resupply was not matched with the expression of sulfate transporters and the activity of ATPS and APR which were rapidly decreased by sulfate resupply. A strong induction of O-acetylserine(thiol)lyase (OASTL), NR and GS upon sulfate resupply was accompanied with the increase in cysteine, amino acids and proteins pool. Sulfate resupply resulted in a strong increase in de novo synthesis of amino acids and proteins, as evidenced by the increases in N and S incorporation into amino acids (1.8- and 2.4-fold increase) and proteins (2.2-and 6.3-fold increase) when compared to S-deprived plants. The results thus indicate that sulfate resupply followed by S-deprivation accelerates nitrate assimilation for protein synthesis.

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.

Redox signaling mediates the expression of a sulfate‐deprivation‐inducible microRNA395 in Arabidopsis

MicroRNA395 (miR395) is a conserved miRNA that targets a low-affinity sulfate transporter (AST68) and three ATP sulfurylases (APS1, APS3 and APS4) in higher plants. In this study, At2g28780 was confirmed as another target of miR395 in Arabidopsis. Interestingly, several dicots contained genes homologous to At2g28780 and a cognate miR395 complementary site but possess a gradient of mismatches at the target site. It is well established that miR395 is induced during S deprivation in Arabidopsis; however, the signaling pathways that mediate this regulation are unknown. Several findings in the present study demonstrate that redox signaling plays an important role in induction of miR395 during S deprivation. These include the following results: (i) glutathione (GSH) supplementation suppressed miR395 induction in S-deprived plants (ii) miR395 is induced in Arabidopsis seedlings exposed to Arsenate or Cu(2+) , which induces oxidative stress (iii), S deprivation-induced oxidative stress, and (iv) compromised induction of miR395 during S deprivation in cad2 mutant (deficient in GSH biosynthesis) that is defective in glutaredoxin-dependent redox signaling and ntra/ntrb (defective in thioredoxin reductases a and b) double mutants that are defective in thioredoxin-dependent redox signaling. Collectively, these findings strongly support the involvement of redox signaling in inducing the expression of miR395 during S deprivation in Arabidopsis.

Supply of sulphur to S-deficient young barley seedlings restores their capability to cope with iron shortage.

The effect of the S nutritional status on a plant's capability to cope with Fe shortage was studied in solution cultivation experiments in barley (Hordeum vulgare L. cv. Europa). Barley is a Strategy II plant and responds to Fe deficiency by secretion of chelating compounds, phytosiderophores (PS). All PS are derived from nicotianamine whose precursor is methionine. This suggests that a long-term supply of an inadequate amount of S could reduce a plant's capability to respond to Fe deficiency by limiting the rate of PS biosynthesis. The responses of barley (Hordeum vulgare L. cv. Europa) plants grown for 12 d on Fe-free nutrient solutions (NS) containing 0 or 1.2 mM SO42−, was examined after 24 h or 48 h from transfer to NS containing 1.2 mM SO42−. After the supply of S was restored to S-deprived plants, an increase in PS release in root exudates was evident after 24 h of growth in S-sufficient NS and the increment reached values up to 4-fold higher than the control 48 h after S resupply. When S was supplied to S-deficient plants, leaf ATPS (EC 2.7.7.4) and OASTL (EC 4.2.99.8) activities exhibited a progressive recovery. Furthermore, root HvST1 transcript abundance remained high for 48 h following S resupply and a significant increase in the level of root HvYS1 transcripts was also found after only 24 h of S resupply. Data support the idea that the extent to which the plant is able to cope with Fe starvation is strongly associated with its S nutritional status. In particular, our results are indicative that barley plants fully recover their capability to cope with Fe shortage after the supply of S is restored to S-deficient plants.

Chromate Differentially Affects the Expression of a High‐Affinity Sulfate Transporter and Isoforms of Components of the Sulfate Assimilatory Pathway in Zea mays (L.)

In this study the chromate accumulation and tolerance were investigated in ZEA MAYS L. in relation to sulfur availability since sulfate may interact with chromate for transport into the cells. Chromate inhibited sulfate uptake when supplied to plants for a short-term period, whereas phosphate uptake remained unchanged. Sulfate absorption was also reduced in S-starved (-S) and S-supplied (+S) plants treated for 2 d with 0.2 mM chromate and the concomitant repression of the root high-affinity sulfate root transporter ZMST1;1 transcript accumulation was observed. Conversely, the plasma membrane H (+)-ATPase MHA2 was unaffected by chromate in +S plants, allowing to exclude a general effect of chromate on the active membrane transport. As observed for sulfate uptake, chromate uptake was enhanced in -S condition and decreased in both +S and -S plants after 2 d of Cr treatment. Chromate reduced the concentration of sulfur and sulfate in +S plants to the basal level of -S plants, and maximum chromium accumulation was recorded in S-deprived plants. Analysis of transcript abundance of genes involved in sulfate assimilation revealed differential regulation by chromate, which was only partly related to sulfur availability and to the levels of thiols. This work shows for the first time that chromate specifically represses sulfate uptake, and such repression occurs without the implication of the candidate regulatory metabolites of the sulfate transport system in plants.

Rapid disruption of nitrogen metabolism and nitrate transport in spinach plants deprived of sulphate

Hydroponically grown spinach plants were deprived of an external source of sulphate after an initial period when the S-supply was sufficient. The time-course of events following this treatment was monitored. The first responses were found in the uptake and translocation of NO(3)(-) and the uptake of SO(4)(2-). The former declined by approximately 50%, the effect being most significant at higher [NO(3)(-)](ext.) while the latter increased 6-fold over a 4 d period. Growth in the absence of external SO(4)(2-) resulted in exhaustion of internal SO(4)(2-) pools, the effect being seen first in roots, then in young leaves and, after a marked delay, in mature leaves. In young leaves, there were dramatic increases in the [NO(3)(-)] and the content of arginine in the first 2 d of S-deprivation. The concentration of glutamine, the most abundant amino acid in S-sufficient conditions, also more than doubled in S-deficient young leaves. The changes in arginine levels were also found in older leaves, but the change in glutamine level was not seen. Assays of nitrate reductase activity (NRA) and nitrate reductase (NR) mRNA from young leaves of S-replete and S-deprived plants revealed a divergence in activity and content only late in the experiments (between days 4 and 8) when results were expressed on a unit leaf basis. However, there were also time-dependent changes in the protein content that kept the specific activities (NRA:protein and RNA:protein) more or less unchanged. The results imply that the impact of S-deficiency on N-utilization are more sensitively monitored by simple measurements of the chemical composition of young leaves than by measurements of NRA or NR transcript abundance. They also suggest that protein synthesis in young leaves is strongly dependent on a continuous supply of SO(4)(2-) from outside the plant.

SO42- Deprivation Has an Early Effect on the Content of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase and Photosynthesis in Young Leaves of Wheat.

Wheat (Triticum aestivum cv Chinese Spring) supplied with 0.45 mM SO42- for 14 d with relative growth rates (RGR) of 0.22 to 0.24 d-1 was deprived of S for 7 to 8 d. There was no significant effect on RGR or leaf development (leaf 2 length was constant; leaf 3 expanded for 2-4 d; leaf 4 emerged and elongated throughout the experiment) during the S deprivation. In controls the net assimilation rate (A) closely reflected leaf ontogeny. S deprivation affected A in all leaves, particularly leaf 4, in which A remained at 8 to 10 [mu]mol CO2 m-2 s-1, whereas in controls A rose steadily to >20 [mu]mol CO2 m-2 s-1. In leaf 2, with a fully assembled photosynthetic system, A decreased in S-deprived plants relative to controls only at the end of the experiment. Effects on A were not due to altered stomatal conductance or leaf internal [CO2] ([C]i); decreases in the initial slope of A/[C]i curves indicated an effect of S deprivation on the carboxylase efficiency. Measurement of Rubisco activity and large subunit protein abundance paralleled effects on A and A/[C]i in S-deprived leaves. Negative effects on photosynthesis in S-deprived plants are discussed in relation to mobilization of S reserves, including Rubisco, emphasizing the need for continuous S supply during vegetative growth.

Sulfur Deprivation and Nitrogen Metabolism in Maize Seedlings

The objective of this experiment was to elucidate the manner in which N metabolism is influenced by S nutrition. Maize (Zea mays L.) seedlings supplied with Hoagland solution minus SO(4) (2-) exhibited S deficiency symptoms 12 days after emergence. Prior to development of these symptoms, a decline in leaf blade nitrate reductase (NR, EC 1.6.6.1) activity was observed in S-deprived seedlings compared to normal seedlings. Twelve days after emergence, in vitro NR activity was diminished 50% compared to normal seedlings. Glutamine synthetase (EC 6.3.1.2) and NAD-glutamate dehydrogenase (EC 1.4.1.2) activities were less severely affected (19 and 13%, respectively, at day 12). NADP-glutamate dehydrogenase (EC 1.4.1.4) activity and leaf blade fresh weight were not altered by S deprivation. Concentrations of soluble protein and chlorophyll (a and b) in leaf blades were reduced 18 and 25%, respectively, at day 12. A significantly higher concentration of NO(3) (-)-N was observed for leaf blade and stem (culms, leaf sheaths, and unfurled leaves) fractions (46 and 31%, respectively) in S-deprived plants. In contrast to the other parameters measured, NR activity in S-deprived seedlings could be readily restored to the normal level by addition of SO(4) (2-). The apparent preferential effect of S deprivation on NR activity could be causally related to the observed changes in NO(3) (-)-N and soluble protein concentration.