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Article

Regulation of Sulfate Uptake and Assimilation in Barley (Hordeum vulgare) as Affected by Rhizospheric and Atmospheric Sulfur Nutrition

by
Ties Ausma
* and
Luit J. De Kok
Laboratory of Plant Physiology, Groningen Institute for Evolutionary Life Sciences, University of Groningen, 9747 AG Groningen, The Netherlands
*
Author to whom correspondence should be addressed.
Plants 2020, 9(10), 1283; https://doi.org/10.3390/plants9101283
Submission received: 31 August 2020 / Revised: 22 September 2020 / Accepted: 24 September 2020 / Published: 28 September 2020
(This article belongs to the Special Issue Selected Papers from the 28th International Symposium of the Scientific Centre for Fertilizers (CIEC) “Fertilization and Nutrient Use Efficiency in Mediterranean Environments”)
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Abstract

:
To study the regulation of sulfate metabolism in barley (Hordeum vulgare), seedlings were exposed to atmospheric hydrogen sulfide (H2S) in the presence and absence of a sulfate supply. Sulfate deprivation reduced shoot and root biomass production by 60% and 70%, respectively, and it affected the plant’s mineral nutrient composition. It resulted in a 5.7- and 2.9-fold increased shoot and root molybdenum content, respectively, and a decreased content of several other mineral nutrients. Particularly, it decreased shoot and root total sulfur contents by 60% and 70%, respectively. These decreases could be ascribed to decreased sulfate contents. Sulfate deficiency was additionally characterized by significantly lowered cysteine, glutathione and soluble protein levels, enhanced dry matter, nitrate and free amino acid contents, an increased APS reductase (APR) activity and an increased expression and activity of the root sulfate uptake transporters. When sulfate-deprived barley was exposed to 0.6 µL L−1 atmospheric H2S, the decrease in biomass production and the development of other sulfur deficiency symptoms were alleviated. Clearly, barley could use H2S, absorbed by the foliage, as a sulfur source for growth. H2S fumigation of both sulfate-deprived and sulfate-sufficient plants downregulated APR activity as well as the expression and activity of the sulfate uptake transporters. Evidently, barley switched from rhizospheric sulfate to atmospheric H2S as sulfur source. Though this indicates that sulfate utilization in barley is controlled by signals originating in the shoot, the signal transduction pathway involved in the shoot-to-root regulation must be further elucidated.

1. Introduction

Barley (Hordeum vulgare) is cultivated globally for the production of alcoholic beverages and animal fodder [1,2]. Since the crop is frequently grown on light textured soils, it is prone to sulfur (S) deficiency [1]. S is an indispensable macronutrient for plants that is needed for the synthesis of proteins and other organic compounds [3]. An insufficient S supply may limit endosperm modification during malting by lowering the grain’s content of, e.g., B- and D-hordein proteins [1,2]. Moreover, it may lower kernel S-methylmethionine (SMM) and dimethylsulfoxide (DMSO) levels, which are the precursors for dimethylsulfide (DMS) synthesis [1]. DMS decisively affects the aroma, taste and flavor of beverages [1,2].
Plants generally acquire S as sulfate via the roots [3]. In shoot and root plastids, sulfate is activated to adenosine 5’-phosphosulfate, which is next reduced to sulfite by APS reductase (APR) [3]. Sulfite is subsequently reduced to sulfide, which is incorporated in cysteine by the cysteine synthase complex [3]. This incorporation couples S assimilation to nitrogen (N) assimilation [3]. From cysteine, numerous S-containing organic compounds can be synthesized [3].
The rate of sulfate metabolism into cysteine, which is tuned to the plant’s S demand for growth, is controlled by the activity of the root sulfate uptake transporters and APR [3]. Thus, sulfate deprivation generally enhances the expression of the root sulfate transporters, the root’s sulfate uptake capacity and the expression and activity of APR [3].
Atmospheric hydrogen sulfide (H2S) is potentially phytotoxic, though species differ considerably in their H2S susceptibility [4]. Whereas prolonged exposure to 0.03 μL L−1 H2S (a realistic level for industrially- and agriculturally polluted areas) inhibited the biomass production of sensitive dicot species, monocot species, including barley, are rather H2S insusceptible [4]. These species can tolerate up to 1.5 μL L−1 H2S, without negative effects on plant biomass production [4,5]. In monocots, the meristem is sheltered by leaves. Therefore, H2S can hardly penetrate the meristem, which may explain the H2S tolerance of these species [4,5].
Plants can use H2S, absorbed by the foliage, as a S source for growth [4,6,7]. Plants may even grow with atmospheric H2S as the sole S source (viz. when no sulfate is supplied to the root) [4,6]. H2S fumigation of sulfate-deprived plants may fully alleviate the development of S deficiency symptoms. In the many tested plant species, the rate of foliar H2S uptake followed Michaelis–Menten kinetics with respect to the atmospheric H2S level [8]. The kinetics are controlled by the rate of H2S incorporation into cysteine and subsequently other molecules [4,8]. Typically, shoot cysteine content and that of the tripeptide glutathione increase significantly upon H2S fumigation, indicating that absorbed H2S is metabolized with high affinity in these thiols [4].
The fumigation of plants with H2S is a powerful way to obtain insights into the regulation of sulfate uptake and assimilation [4]. Atmospheric H2S is directly incorporated in cysteine in shoots and, consequently, studying the impact of H2S fumigation on the sulfate transporters and APR, in the presence and absence of a sulfate supply, may reveal if these enzymes are regulated by signals originating in the shoot or root environment [4].
Thus, in the current research, the interaction between atmospheric H2S and rhizospheric sulfate nutrition was analyzed in barley. Plants were H2S fumigated in the presence and absence of a sulfate supply. The mineral nutrient composition, S and N metabolite content, APR activity, and the expression and activity of the root sulfate transporters were analyzed.

2. Results and Discussion

2.1. Impact of Sulfate Deprivation and H2S Fumigation on Biomass Production and Mineral Nutrient Content

A 7-day sulfate deprivation reduced shoot and root biomass production by 60% and 70%, respectively (Figure 1). Since shoot and root biomass production were similarly reduced, sulfate deprivation hardly affected the shoot-to-root ratio. Shoot and root dry matter contents were enhanced by sulfate deprivation by 1.2-fold and 1.5-fold, respectively, which may be due to an increased level of non-structural carbohydrates (viz. sugars and starch; Figure 1) [3].
Exposure of sulfate-sufficient plants to 0.6 µL L−1 atmospheric H2S hardly affected biomass production and dry matter content (Figure 1). However, upon sulfate deprivation, H2S fumigation alleviated the establishment of S-deficiency signs. The biomass production and dry matter content of sulfate-deprived H2S-fumigated plants were comparable to those of sulfate-sufficient plants (Figure 1). Obviously, in accordance with previous observations [6], barley could use H2S, absorbed by the foliage, as a S source for the synthesis of organic compounds.
Sulfate deprivation resulted in a strong (60%) decrease in the shoot’s total S content, whereas the shoot’s content of most other mineral nutrients was hardly affected (Table 1). In the shoots of sulfate-deprived plants, N content was slightly decreased (by 10%), whereas P and Mo contents were increased by 1.3- and 5.7-fold, respectively. By contrast, sulfate deprivation significantly affected the root’s mineral nutrient composition (Table 1). It decreased S, P, K, Mn, Cu and Zn contents by 70%, 30%, 20%, 50%, 40% and 40%, respectively, and it increased Mo content by 2.9-fold.
H2S fumigation of sulfate-sufficient plants did, apart from a 1.6-fold increased shoot S level, not result in changes in the mineral nutrient composition (Table 1). H2S fumigation of sulfate-deprived plants alleviated the impacts of sulfate deprivation on shoot S, N and P contents and on shoot and root Mo contents (Table 1). By contrast, H2S fumigation hardly alleviated the other impacts of sulfate deprivation on the plant’s mineral nutrient composition (Table 1). Clearly, similar to observations in other plants [7,9,10], in barley there is no strict coupling between the metabolism of S and the majority of other mineral nutrients.

2.2. Impact of Sulfate Deprivation and H2S Fumigation on Sulfur and Nitrogen Metabolite Content

The observed changes in total S content upon sulfate deprivation and H2S fumigation could, at least partly, be attributed to changes in sulfate content. Sulfate deprivation resulted in a 70 and 90% decrease in the shoot and root sulfate content, respectively (Figure 2). H2S fumigation alleviated the decrease in shoot sulfate content (Figure 2). Moreover, H2S fumigation of sulfate-sufficient plants enhanced shoot sulfate levels by 2.2-fold (Figure 2).
The (total water-soluble non-protein) thiol pool represented a minor proportion of total S in the plant (approximately 3%; Figure 2). Sulfate deprivation decreased shoot and root thiol levels by 40% and 50%, respectively (Figure 2). These changes could mainly be attributed to changes in glutathione content, since cysteine accounted for only 5% and 10% of the thiol pool in sulfate-sufficient shoots and roots, respectively (Figure 2). Yet, sulfate deprivation decreased shoot and root cysteine levels by almost 100% (Figure 2). H2S fumigation alleviated the decreases in thiol levels and it even increased shoot cysteine levels by 3.9-fold (Figure 2). Moreover, H2S fumigation of sulfate-sufficient plants increased the total water-soluble non-protein thiol and cysteine level of the shoot by 1.7- and 5.0-fold, respectively (Figure 2). Evidently, H2S is with high affinity assimilated into cysteine and subsequently glutathione.
Sulfate deprivation enhanced shoot nitrate levels by 1.2-fold and shoot and root free amino acid levels by 3.8-fold and 4.7-fold, respectively (Figure 3). Shoot soluble protein levels were 35% lower in sulfate-deprived plants compared to sulfate-sufficient plants (Figure 3).
Sulfate is required for cysteine synthesis and, therefore, S deficiency hampers protein synthesis [3]. This hampering may not only lower protein contents, but it may also result in an accumulation of nitrate and (non-S-containing) free amino acids, since these are precursors for protein synthesis [3]. The impacts of sulfate deprivation on nitrate, free amino acid and soluble protein content were alleviated by H2S fumigation, indicating again that barley used atmospheric H2S for cysteine and subsequently protein synthesis (Figure 3). H2S fumigation of sulfate-sufficient plants did not affect nitrate, free amino acid and soluble protein levels (Figure 3).

2.3. Impact of Sulfate Deprivation and H2S Fumigation on Sulfate Uptake and Reduction

APR activity was approximately 10-fold higher in the shoot than root under all treatments, which indicated that in barley, sulfate reduction primarily takes place in shoot chloroplasts (Figure 4). Sulfate deprivation enhanced shoot and root APR activity by 2.6-fold (Figure 4). The specific APR activity of the shoot was even 3.0-fold enhanced, since sulfate deprivation decreased shoot soluble protein content (Figure 3 and Figure 4). In line with previous observations [11,12], sulfate deprivation also resulted in a higher expression and activity of the root sulfate transporters. The expression of HvST1, a high-affinity root sulfate uptake transporter (Km 6.9 µM sulfate) [11], and the root sulfate uptake capacity were, respectively, 24- and 7-fold higher in sulfate-deprived plants than in sulfate-sufficient plants (Figure 5).
H2S fumigation of sulfate-sufficient plants resulted in a 50% decreased shoot and root APR activity (Figure 4). Moreover, it resulted in an 80% and 40% decrease in the root’s HvST1 expression and the sulfate uptake capacity, respectively (Figure 5). Apparently, barley switched from rhizospheric sulfate to atmospheric H2S as S source for biomass production. However, since H2S fumigation resulted in an enhanced shoot sulfate content (Figure 2), root sulfate uptake may, upon H2S fumigation, not be strictly regulated in barley (viz. strictly tuned to the crop’s S demand for growth) [4,13].
H2S fumigation of sulfate-deprived plants decreased shoot and root APR activity by approximately 35% (Figure 4). Moreover, it largely alleviated the impacts of sulfate deprivation on the expression and activity of the root’s sulfate transporters (Figure 5). The sulfate uptake capacity of sulfate-deprived fumigated plants was only slightly (1.4-fold) higher than that of sulfate-sufficient non-fumigated plants (Figure 5).
It deserves mentioning here that the impact of S supply on the sulfate uptake capacity mimicked its impact on shoot and root Mo contents (Table 1; Figure 5). Sulfate deprivation resulted in an enhanced sulfate uptake capacity and Mo content, which were alleviated by H2S fumigation (Table 1; Figure 5). Molybdate is a structural analogue of sulfate and thus the sulfate transporters in barley may be involved in molybdate uptake [9].
Since atmospheric H2S is directly incorporated into cysteine in shoots and since H2S fumigation strongly affected the activity of APR and the expression and activity of the sulfate transporters in both sulfate-sufficient and sulfate-deprived plants, our findings indicate that these enzymes are controlled by signals originating in the shoot. Moreover, our observations indicate that the in-situ sulfate concentration in the rhizosphere hardly controls these enzymes.
The nature of the shoot-derived regulatory signals remains ambiguous. Although Vidmar and co-workers [12] suggested glutathione to regulate the sulfate transporters in barley, in the current study glutathione levels did not correlate with the HvST1 expression, sulfate uptake capacity, or APR activity (Figure 2, Figure 4 and Figure 5). This discrepancy may be understood by presuming that variation in glutathione content is accompanied with variation in the content of other metabolites that actually regulate the sulfate transporters.
Sulfate and metabolites from nitrate assimilation (e.g., nitrate and free amino acids) have also been assumed to regulate the sulfate transporters and APR [3]. N and S are needed for cysteine synthesis and thus the rates of nitrate and sulfate assimilation are tuned to each other [3]. However, sulfate, nitrate and free amino acid contents did not clearly correlate with the HvST1 expression, sulfate uptake capacity, or APR activity (Figure 2, Figure 3, Figure 4 and Figure 5). The absence of such correlations makes it tempting to speculate that the activity and expression of the sulfate transporters and APR are instead merely regulated by the sink strength of the shoot for organic sulfur (viz. source-sink dynamics) [4].

2.4. Impact of Sulfate Deprivation and H2S Fumigation in Barley versus Other Species

In the present research, the interaction between atmospheric H2S and rhizospheric sulfate nutrition was analyzed in barley. Previously, this interaction has detailly been analyzed in species from the genus Brassica [14,15,16,17,18]. The current data show that this interaction and, therefore, the regulation of sulfate metabolism, partly differs between barley and Brassica. Sulfate deprivation of Brassica increased the expression of the sulfate uptake transporters, which was associated with a strongly enhanced sulfate uptake capacity [15,18]. Sulfate deprivation of Brassica also enhanced the expression and activity of APR in shoots and roots [14,16,17]. Finally, in Brassica it resulted in a lower shoot-to-root ratio [14,18]. Notably, sulfate deprivation of barley hardly affected the shoot-to-root ratio (Figure 1). In barley, the shoot-to-root ratio is approximately 2.5, whereas in Brassica it is approximately 6 (at an ample sulfate supply; Figure 1) [18]. Therefore, barley constitutively has relatively more roots than Brassica and, upon sulfate deprivation, it may increase its sulfate uptake capacity without the need to allocate resources to root synthesis.
H2S fumigation of sulfate-sufficient Brassica downregulated shoot and root APR activity as well as the expression and activity of the root sulfate uptake transporters [4,14,15,17]. Clearly, similar to barley, Brassica switched from rhizospheric sulfate to atmospheric H2S as S source. Thus, in both plants sulfate utilization is controlled by signals originating in the shoot. In line with this, cutting of the shoot of curly kale (Brassica oleracea) rapidly decreased the plants’ sulfate uptake capacity. Similar to barley, in Brassica the nature of the shoot-derived regulatory signals remains ambiguous [4].
Dissimilar to Brassica, in barley H2S fumigation resulted in an enhanced shoot sulfate content (Figure 2) [15]. Notably, in spruce (Picea abies) spinach (Spinacia oleracea), red clover (Trifolium pratense) soybean (Glycine max), common bean (Phaseolus vulgaris) and onion (Allium cepa) fumigation with an atmospheric H2S level that is sufficient to cover the S requirement for growth, also resulted in an enhanced sulfate content [4,13,19,20]. Thus, potentially, upon H2S fumigation in most plants, sulfate uptake is less strictly regulated than in Brassica (viz. less strictly tuned to the plant’s S demand for growth).
Fumigation of sulfate-deprived Brassica with ≥0.06 µL L−1 H2S alleviated the sulfate-deprived decreases in biomass production and in cysteine and glutathione contents [14,15,18]. Additionally, it alleviated the enhanced expression of APR upon sulfate deprivation. However, dissimilar to barley, in Brassica H2S fumigation hardly affected the sulfate-deprived enhancement of the expression and activity of the root sulfate uptake transporters (Figure 5) [15,16,17,18]. Moreover, it hardly affected the sulfate-deprived decrease in the shoot-to-root ratio [15,16,17,18]. This indicates that, dissimilar to barley, in Brassica sulfate uptake is, besides by shoot-derived signals, controlled by the in-situ sulfate concentration in the rhizosphere. It has been assumed that, upon sulfate deprivation, Brassica forms special roots that are typified by a high sulfate transporter expression and activity, which cannot be modified by the plant’s internal sulfur status [17]. Since H2S exposure of sulfate-deprived barley strongly alleviated the increased expression and activity of the sulfate transporters, apparently barley hardly forms such roots upon sulfate deprivation.

3. Materials and Methods

3.1. Plant Material and H2S Fumigation

Barley (cv. KWS Irina; Wiersum Plantbreeding; Winschoten; The Netherlands) was germinated on boxes, containing 15 l aerated tap water, in a climate-controlled room with temperature, humidity, light intensity and light duration settings identical to those described in [7]. After 7 days, the seedlings were transferred to stainless-steel boxes (60 plants per box) holding 13 l aerated 50% Hoagland nutrient solutions. Nutrient solutions either contained 1 mM sulfate (+S; sulfate-sufficient; solution’s composition being 2.5 mM CaCl2, 2.5 mM KCl, 0.5 mM KH2PO4, 1 mM MgSO4, 3.75 mM NH4NO3, 23.4 µM H3BO3, 4.8 µM MnCl2, 0.48 µM ZnSO4, 0.16 µM CuSO4, 0.26 µM Na2MoO4 and 45 µM Fe3+EDTA) or 0 mM sulfate (-S; sulfate-deprived; all sulfate salts replaced by chloride salts). The boxes were placed in 50 L cylindrical stainless-steel cabinets (0.6 m diameter) with a polymethyl methacrylate top with temperature, humidity, light intensity and light duration settings identical to those described in [7]. In two independent (identical) experiments, the plants were exposed to 0 or 0.6 µL L−1 atmospheric H2S (for details on H2S application, see [7]).
After 7 days of treatment, plants were harvested 3 h after the start of the light period. After the root was rinsed in ice-cold de-mineralized water (3 × 20 s), the shoot and root were separately weighted. The shoot and root biomass productions were calculated by subtracting the initial, pre-treatment weight from the weights at harvest. For the measurement of the dry matter content and the mineral nutrient composition, shoots and roots were dried overnight at 80 °C. For the determination of the S and N metabolite content, APR activity and sulfate transporter expression, shoots and roots were frozen at −80 °C.

3.2. Mineral Nutrient Content

The mineral nutrient content of shoots and roots was determined from dried plant material, that was pulverized using a Retsch Mixer-Mill (Retsch type MM2, Haan, Germany). Total N content was measured via the Dumas procedure, using an automated elemental analyzer (model EA 1110; Interscience, New York, NY, USA) with Eager 200 for Windows [21]. The levels of other minerals were analyzed via inductively coupled plasma mass spectrometry (ICP-MS) with an Agilent 7700 ICP-MS (Agilent Technologies, Santa Clara, CA, USA) [22].

3.3. Sulfur and Nitrogen Metabolite Content

Water-soluble non-protein thiols were isolated from freshly harvested plant material and their content was quantified colorimetrically according to [23]. Sulfate and nitrate were isolated from frozen plant material according to [24] and their contents were quantified with ion chromatography (IC) as described in [25]. Free amino acids and soluble proteins were isolated from frozen plant material and quantified as described by [26].

3.4. APR Activity and Sulfate Uptake Capacity

For the determination of APR activity, frozen shoots and roots were ground with liquid N2. The resulting plant powder was homogenized in 1 mL 50 mM K2PO4 buffer (pH 8.0) that contained 30 mM Na2SO4, 500 µM AMP and 10 mM DTE. After centrifugation, APR activity was measured as described in [27]. For the determination of the sulfate uptake capacity, plants were incubated for 1 h on a 25% Hoagland nutrient solution that was labeled with 500 µM 35S-sulfate (2 MBq L−1), after which the sulfate uptake capacity was assessed following [16].

3.5. Expression of HvST1

To determine the expression of HvST1, total RNA was isolated from frozen plant material, made free of genomic DNA and transcribed into cDNA as described by [18]. The acquired cDNA samples were diluted 1:50 in distilled water. During quantitative PCR (qPCR), ADP-ribosylation factor 1-like protein (ADP) was used as reference gene, since its expression is stable across several environmental conditions [28]. Primers for ADP and HvST1 were retrieved from [28] and [29], respectively. The qPCR reaction mixture contained 2 µL 1:50 diluted cDNA, 12.5 µL 2x Bio-Star qPCR-Mastermix SYBR Blue (GeneON GmbH; Ludwigshafen; Germany), 0.75 µL ROX (GeneON GmbH), 0.75 µL of each primer (10 µM stock) and 8.25 µL deionized water. Reactions were run in triplicate on an Applied Biosystems 7300 Real Time PCR system (Applied Biosystems, Foster City, CA, USA) with an initial denaturation of 5 min at 95 °C, followed by 50 cycles of 15 s denaturation at 95 °C, 15 s annealing at 60 °C and 30 s elongation at 72 °C. The program was finished by denaturation from 65 °C to 95 °C to generate melting curves (to verify a-gene-specificity of the primers). The LinRegPCR software (version 2014.2; Heart Failure Research Centre; Amsterdam, The Netherlands) was used to baseline-correct the qPCR data, after which the initial number of gene transcripts (N0) in a sample was determined with the mean PCR efficiency per primer set (which was between 95% and 100%) [30,31,32]. For the calculation of the relative expression level of HvST1, the N0 value of HvST1 was divided by the N0 value of ADP.

3.6. Statistical Analyses

GraphPad Prism (version 8.4.2; GraphPad Software, San Diego, CA, USA) was used for statistical analyses. Treatment means were compared using a two-way analysis of variance (ANOVA) with a Tukey’s HSD test as post-hoc test at the P ≤ 0.05 level.

4. Conclusions

H2S fumigation alleviates S deficiency in barley, which could grow with H2S as the only S source. Moreover, in barley there was a strong interaction between the metabolism of atmospheric H2S and rhizospheric sulfate. H2S exposure downregulated APR activity and the expression and activity of the root sulfate transporters. Clearly, similar to observations in Brassica, in barley shoot-derived signals regulate sulfate utilization. However, dissimilar to Brassica, in barley the in-situ sulfate concentration in the rhizosphere hardly regulates sulfate utilization. The nature of the shoot regulatory signals needs to be elucidated in further studies.

Author Contributions

T.A. conceived and designed the research. T.A. performed the experiments and analyzed the data. T.A. and L.J.D.K. wrote the manuscript. All authors have read and agreed to the published version of the manuscript.

Funding

T.A. receives funding from The Netherlands Organization for Scientific Research (NWO) via ALW Graduate Program Grant 2017.015.

Acknowledgments

We thank Stanislav Kopriva (University of Cologne; Germany) for facilitating the APR, IC and ICP-MS measurements. Moreover, we thank Marten Staal, Nelly D. Eck and Erik J. Bunskoeke (University of Groningen; The Netherlands) for their technical support.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Zhao, F.J.; Fortune, S.; Barbosa, V.L.; McGrath, S.P.; Stobart, R.; Bilsborrow, P.E.; Booth, E.J.; Brown, A.; Robson, P. Effects of sulphur on yield and malting quality of barley. J. Cereal Sci. 2006, 43, 369–377. [Google Scholar] [CrossRef]
  2. Holopainen, U.R.M.; Rajala, A.; Jauhiainen, L.; Wilhelmson, A.; Home, S.; Kaupilla, R.; Peltonen-Sainio, P. Influence of sulphur application on hordein composition and malting quality of barley (Hordeum vulgare L.) in Northern European growing conditions. J. Cereal Sci. 2015, 62, 151–158. [Google Scholar] [CrossRef]
  3. Hawkesford, M.J.; De Kok, L.J. Managing sulphur metabolism in plants. Plant Cell Environ. 2006, 29, 382–395. [Google Scholar] [CrossRef]
  4. Ausma, T.; De Kok, L.J. Atmospheric H2S: Impact on plant functioning. Front. Plant Sci. 2019, 10, 743. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  5. Stulen, I.; Posthumus, F.S.; Amâncio, S.; Masselink-Beltman, I.; Müller, M.; De Kok, L.J. Mechanism of H2S phytotoxicity. In Sulfur Nutrition and Sulfur Assimilation in Higher Plants: Molecular, Biochemical and Physiological Aspects, 1st ed.; Brunold, C., Rennenberg, H., De Kok, L.J., Davidian, J.C., Eds.; Paul Haupt: Bern, Switzerland, 2000; pp. 381–382. [Google Scholar]
  6. Stuiver, C.E.E.; De Kok, L.J. Atmospheric H2S as sulphur source for sulphur deprived Brassica oleracea L. and Hordeum vulgare L. In Sulphur Metabolism in Higher Plants: Molecular, Ecophysiological and Nutritional Aspects, 1st ed.; Cram, W.J., De Kok, L.J., Stulen, I., Brunold, C., Rennenberg, H., Eds.; Backhuys Publishers: Leiden, The Netherlands, 1997; pp. 292–294. [Google Scholar]
  7. Ausma, T.; Parmar, S.; Hawkesford, M.J.; De Kok, L.J. Impact of atmospheric H2S, salinity and anoxia on sulfur metabolism in Zea mays. In Sulfur Metabolism in Higher Plants: Fundamental, Environmental and Agricultural Aspects, 1st ed.; De Kok, L.J., Hawkesford, M.J., Haneklaus, S.H., Schnug, E., Eds.; Springer: Dordrecht, The Netherlands, 2017; pp. 93–101. [Google Scholar]
  8. De Kok, L.J.; Stahl, K.; Rennenberg, H. Fluxes of atmospheric hydrogen sulfide to plant shoots. New Phytol. 1989, 112, 533–542. [Google Scholar] [CrossRef] [PubMed]
  9. Zuidersma, E.I.; Ausma, T.; Stuiver, C.E.E.; Prajapati, D.H.; Hawkesford, M.J.; De Kok, L.J. Molybdate toxicity in Chinese cabbage is not the direct consequence of changes in sulphur metabolism. Plant Biol. 2020, 22, 331–336. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  10. Reich, M.; Shahbaz, M.; Prajapati, D.H.; Parmar, S.; Hawkesford, M.J.; De Kok, L.J. Interactions of sulfate with other nutrients as revealed by H2S fumigation of Chinese cabbage. Front. Plant Sci. 2016, 7, 541. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  11. Smith, F.W.; Hawkesford, M.J.; Ealing, P.M.; Clarkson, D.T.; Van den Berg, P.J.; Belcher, A.R.; Warrilow, A.G.S. Regulation of expression of a cDNA from barley roots encoding a high affinity sulphate transporter. Plant J. 1999, 12, 875–884. [Google Scholar] [CrossRef] [PubMed]
  12. Vidmar, J.J.; Schjoerring, J.K.; Touraine, B.; Glass, A.D.M. Regulation of the hvst1 gene encoding a high-affinity sulfate transporter from Hordeum vulgare. Plant Mol. Biol. 1999, 40, 883–892. [Google Scholar] [CrossRef]
  13. Tausz, M.; Weidner, W.; Wonisch, A.; De Kok, L.J.; Grill, D. Uptake and distribution of 35S-sulfate in needles and roots of spruce seedlings as affected by exposure to SO2 and H2S. J. Exp. Bot. 2003, 50, 211–220. [Google Scholar] [CrossRef]
  14. Westerman, S.; De Kok, L.J.; Stuiver, C.E.E.; Stulen, I. Interaction between metabolism of atmospheric H2S in the shoot and sulfate uptake by the roots of curly kale (Brassica oleracea). Physiol. Plant 2000, 109, 443–449. [Google Scholar] [CrossRef]
  15. Buchner, P.; Stuiver, C.E.E.; Westerman, S.; Wirtz, M.; Hell, R.; Hawkesford, M.J.; De Kok, L.J. Regulation of sulfate uptake and expression of sulfate transporter genes in Brassica oleracea L. as affected by atmospheric H2S and pedospheric sulfate nutrition. Plant Physiol. 2004, 136, 3396–3408. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  16. Koralewska, A.; Posthumus, F.S.; Stuiver, C.E.E.; Buchner, P.; De Kok, L.J. The characteristic high sulfate content in Brassica oleracea is controlled by the expression and activity of sulfate transporters. Plant Biol. 2007, 9, 654–661. [Google Scholar] [CrossRef] [PubMed]
  17. Koralewska, A.; Stuiver, C.E.E.; Posthumus, F.S.; Kopriva, S.; Hawkesford, M.J.; De Kok, L.J. Regulation of sulfate uptake, expression of the sulfate transporters Sultr1;1 and Sultr1;2, and APS reductase in Chinese cabbage (Brassica pekinensis) as affected by atmospheric H2S nutrition and sulfate deprivation. Funct. Plant Biol. 2008, 35, 318–327. [Google Scholar] [CrossRef]
  18. Aghajanzadeh, T.; Hawkesford, M.J.; De Kok, L.J. Atmospheric H2S and SO2 as sulfur sources for Brassica juncea and Brassica rapa: Regulation of sulfur uptake and assimilation. Env. Exp. Bot. 2016, 124, 1–10. [Google Scholar] [CrossRef]
  19. Maas, F.M.; De Kok, L.J.; Hoffmann, I.; Kuiper, P.J.C. Plant responses to H2S and SO2 fumigation. I. Effects on growth, transpiration and sulfur content of spinach. Physiol. Plant 1987, 70, 713–721. [Google Scholar] [CrossRef]
  20. Maas, F.M.; De Kok, L.J.; Peters, J.L.; Kuiper, P.J.C. A comparative study on the effects of H2S and SO2 fumigation on the growth and accumulation of sulfate and sulfhydryl compounds in Trifolium pratense L., Glycine max Merr and Phaseolus vulgaris L. J. Exp. Bot. 1987, 38, 1459–1469. [Google Scholar] [CrossRef]
  21. Van Klink, R.; Van Laar-Wiersma, J.; Vorst, O.; Smit, C. Rewilding with large herbivores: Positive direct and delayed effects of carrion on plant and arthropod communities. PLoS ONE 2020, 15, e0226946. [Google Scholar] [CrossRef]
  22. Almario, J.; Jeena, G.; Wunder, J.; Langen, G.; Zucarro, A.; Coupland, G.; Buchner, M. Root-associated fungal microbiota of nonmycorrhizal Arabis alpina and its contribution to plant phosphorus nutrition. Proc. Natl. Acad. Sci. USA 2017, 114, 9403–9412. [Google Scholar] [CrossRef] [Green Version]
  23. De Kok, L.J.; Buwalda, F.; Bosma, W. Determination of cysteine and its accumulation in spinach leaf tissue upon exposure to excess sulfur. J. Plant Physiol. 1988, 133, 502–505. [Google Scholar] [CrossRef]
  24. Maas, F.M.; Hoffmann, I.; Van Harmelen, M.J.; De Kok, L.J. Refractometric determination of sulfate and anions in plants separated by high performance liquid chromatography. Plant Soil 1986, 91, 129–132. [Google Scholar] [CrossRef]
  25. Huang, X.Y.; Chao, D.Y.; Koprivova, A.; Danku, J.; Wirtz, M.; Muller, S.; Sandoval, F.J.; Bauwe, H.; Roje, S.; Dilkes, B. Nuclear localized MORE SULPHUR ACCUMULATION1 epigenetically regulates sulphur homeostasis in Arabidopsis thaliana. PLoS Genet. 2016, 12, e1006298. [Google Scholar] [CrossRef]
  26. Stuiver, C.E.E.; De Kok, L.J.; Westerman, S. Sulfur deficiency in Brassica oleracea L.: Development, biochemical characterization, and sulfur/nitrogen interactions. Russ. J. Plant Physiol. 1997, 44, 505–513. [Google Scholar]
  27. Durenkamp, M.; De Kok, L.J.; Kopriva, S. Adenosine 5’-phosphosulphate reductase is regulated differently in Allium cepa L. and Brassica oleracea L. upon exposure to H2S. J. Exp. Bot. 2007, 58, 1571–1579. [Google Scholar] [CrossRef] [PubMed]
  28. Ferdous, J.; Li, Y.; Reid, N.; Langridge, P.; Shi, B.J.; Tricker, P.J. Identification of reference genes for quantitative expression of microRNAs and mRNAs in barley under various stress conditions. PLoS ONE 2015, 10, e0118503. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  29. Reid, R.; Gridley, K.; Kawamata, Y.; Zhu, Y. Arsenite elicits anomalous sulfur starvation responses in barley. Plant Physiol. 2013, 162, 401–409. [Google Scholar] [CrossRef] [Green Version]
  30. Ramakers, C.; Ruijter, J.M.; Deprez, R.H.; Moorman, A.F. Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data. Neurosci. Lett. 2003, 339, 62–66. [Google Scholar] [CrossRef]
  31. Ruijter, J.M.; Ramakers, C.; Hoogaars, W.M.; Karlen, Y.; Bakker, O.; van den Hoff, M.J.; Moorman, A.F. Amplification efficiency: Linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Res. 2009, 37, e45. [Google Scholar] [CrossRef] [Green Version]
  32. Dalla Benetta, E.; Beukeboom, L.W.; van de Zande, L. Adaptive differences in circadian clock gene expression patterns and photoperiodic diapause induction in Nasonia vitripennis. Am. Nat. 2019, 193, 881–896. [Google Scholar] [CrossRef]
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Figure 1. Biomass production and dry matter content (DMC) of barley shoots and roots in the presence and absence of a H2S and sulfate supply. Seven-day old seedlings were grown on a 50% Hoagland nutrient solution at 1 mM (+S) or 0 mM sulfate (−S) and exposed to 0 or 0.6 µL L−1 H2S for 7 days. The initial shoot and root weights were 0.16 ± 0.01 and 0.08 ± 0.01 g FW, respectively. Data on biomass production represent the average of 2 experiments with 12–13 measurements with 3-6 plants per measurement (±SD). Data on DMC represent the average of 3 measurements with 6 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
Figure 1. Biomass production and dry matter content (DMC) of barley shoots and roots in the presence and absence of a H2S and sulfate supply. Seven-day old seedlings were grown on a 50% Hoagland nutrient solution at 1 mM (+S) or 0 mM sulfate (−S) and exposed to 0 or 0.6 µL L−1 H2S for 7 days. The initial shoot and root weights were 0.16 ± 0.01 and 0.08 ± 0.01 g FW, respectively. Data on biomass production represent the average of 2 experiments with 12–13 measurements with 3-6 plants per measurement (±SD). Data on DMC represent the average of 3 measurements with 6 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
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Figure 2. Sulfate, total water-soluble non-protein thiol and cysteine content of barley shoots and roots in the presence and absence of a H2S and sulfate supply. For experimental details, see the legend of Figure 1. Data represent the average of 3 measurements with 3 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
Figure 2. Sulfate, total water-soluble non-protein thiol and cysteine content of barley shoots and roots in the presence and absence of a H2S and sulfate supply. For experimental details, see the legend of Figure 1. Data represent the average of 3 measurements with 3 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
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Figure 3. Nitrate, free amino acid and soluble protein content of barley shoots and roots in the presence and absence of a H2S and sulfate supply. For experimental details, see the legend of Figure 1. Data represent the average of 3 measurements with 3 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
Figure 3. Nitrate, free amino acid and soluble protein content of barley shoots and roots in the presence and absence of a H2S and sulfate supply. For experimental details, see the legend of Figure 1. Data represent the average of 3 measurements with 3 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
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Figure 4. APR activity in barley shoots and roots in the presence and absence of a H2S and sulfate supply. For experimental details, see the legend of Figure 1. Data represent the average of 3 measurements with 3 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
Figure 4. APR activity in barley shoots and roots in the presence and absence of a H2S and sulfate supply. For experimental details, see the legend of Figure 1. Data represent the average of 3 measurements with 3 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
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Figure 5. Sulfate uptake capacity and expression of the HvST1 transporter in barley roots in the presence and absence of a H2S and sulfate supply. For experimental details, see the legend of Figure 1. Data represent the average of 7 (sulfate uptake capacity) and 3 (HvST1 expression) measurements with 3 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
Figure 5. Sulfate uptake capacity and expression of the HvST1 transporter in barley roots in the presence and absence of a H2S and sulfate supply. For experimental details, see the legend of Figure 1. Data represent the average of 7 (sulfate uptake capacity) and 3 (HvST1 expression) measurements with 3 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
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Table
Table 1. Mineral nutrient content of barley shoots and roots in the presence and absence of a H2S and sulfate supply. For experimental details, see the legend of Figure 1. Data represent the average of 3 measurements with 6 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
Table 1. Mineral nutrient content of barley shoots and roots in the presence and absence of a H2S and sulfate supply. For experimental details, see the legend of Figure 1. Data represent the average of 3 measurements with 6 plants per measurement (±SD). Significant differences between treatments are indicated with different letters (P ≤ 0.05; two-way ANOVA; Tukey’s HSD test as a post-hoc test).
Mineral Nutrient Content0 µL L−1 H2S0.6 µL L−1 H2S
(µmol g−1 DW)+S−S+S−S
Shoot
Calcium103 ± 10 a84 ± 3 a100 ± 29 a81 ± 8.0 a
Copper0.29 ± 0.03 a0.25 ± 0.01 a0.29 ± 0.05 a0.25 ± 0.03 a
Iron1.92 ± 0.12 a1.65 ± 0.07 a2.06 ± 0.45 a1.72 ± 0.20 a
Magnesium81 ± 7 a76 ± 3 a88 ± 16 a71 ± 8 a
Manganese0.73 ± 0.06 ab0.66 ± 0.03 ab0.83 ± 0.14 a0.59 ± 0.05 b
Molybdenum0.04 ± 0.00 c0.25 ± 0.01 a0.04 ± 0.01 c0.19 ± 0.03 b
Nitrogen3521 ± 150 a3216 ± 61 b3412 ± 71 ab3418 ± 69 ab
Phosphorus192 ± 9 b250 ± 11 a204 ± 34 ab180 ± 22 b
Potassium1758 ± 79 a1637 ± 94 a1852 ± 205 a1570 ± 161 a
Sulfur99 ± 4 b36 ± 2 c163 ± 16 a100 ± 12 b
Zinc0.63 ± 0.07 ab0.77 ± 0.05 a0.67 ± 0.11 ab0.50 ± 0.03 b
Root
Calcium26 ± 6 a38 ± 4 a27 ± 3 a21 ± 1 a
Copper2.05 ± 0.43 a1.13 ± 0.02 b1.83 ± 0.15 a1.45 ± 0.11 ab
Iron2.92 ± 1.14 a3.50 ± 0.46 a2.34 ± 0.05 a2.78 ± 0.12 a
Magnesium28 ± 6 a31 ± 3 a30 ± 6 a25 ± 1 a
Manganese2.21 ± 0.37 a1.17 ± 0.08 b2.36 ± 0.21 a1.34 ± 0.11 b
Molybdenum0.14 ± 0.02 b0.42 ± 0.07 a0.17 ± 0.03 b0.18 ± 0.01 b
Nitrogen3231 ± 133 a2917 ± 114 a3174 ± 182 a3152 ± 121 a
Phosphorus149 ± 21 a100 ± 8 b153 ± 12 a134 ± 16 ab
Potassium842 ± 103 a662 ± 21 b844 ± 42 a719 ± 56 ab
Sulfur82 ± 8 a28 ± 0 b94 ± 9 a44 ± 4 b
Zinc0.59 ± 0.12 a0.40 ± 0.05 b0.57 ± 0.04 ab0.47 ± 0.03 ab

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Ausma, T.; De Kok, L.J. Regulation of Sulfate Uptake and Assimilation in Barley (Hordeum vulgare) as Affected by Rhizospheric and Atmospheric Sulfur Nutrition. Plants 2020, 9, 1283. https://doi.org/10.3390/plants9101283

AMA Style

Ausma T, De Kok LJ. Regulation of Sulfate Uptake and Assimilation in Barley (Hordeum vulgare) as Affected by Rhizospheric and Atmospheric Sulfur Nutrition. Plants. 2020; 9(10):1283. https://doi.org/10.3390/plants9101283

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Ausma, Ties, and Luit J. De Kok. 2020. "Regulation of Sulfate Uptake and Assimilation in Barley (Hordeum vulgare) as Affected by Rhizospheric and Atmospheric Sulfur Nutrition" Plants 9, no. 10: 1283. https://doi.org/10.3390/plants9101283

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