Foliar selenium fertilization alters the content of dietary phytochemicals in two rocket species.

Biofortification is the process that aims to enrich crops in micronutrients and valuable compounds. Selenium (Se) biofortification has particularly attracted increasing interest in recent times due to the growing number of individuals suffering from Se deficiency. Selenate and selenite are the Se forms most frequently administered to crops. In this study, Se was applied foliarly as selenate at 2.5, 5, or 10 mg per plant to two rocket species, Diplotaxis tenuifolia and Eruca sativa, grown in soil and the effects in terms of Se enrichment and content of primary and secondary metabolites were comparatively analyzed. We also compared our results with those obtained previously when selenate was supplied to the same species in hydroponics by addition to the nutrient solution. In most cases, the results were the opposite. In E. sativa, foliar Se treatment was more effective in promoting Se accumulation, sulfur (S), cysteine, and glucosinolates. No significant effect of Se was evident on total phenolic content, but there were individual phenols. Among amino acids, the content of proline was increased by Se, perhaps to counteract osmotic stress due to high Se accumulation. In D. tenuifolia, the content of S and cysteine decreased under Se treatment, but the amount of glutathione was steady, suggesting a preferred assimilation of cysteine toward the synthesis of this antioxidant. Consistent, the content of methionine and glucosinolates was reduced. The content of total phenolics was enhanced only by the low Se dosage. In both species, selenocysteine (SeCys) was identified, the content of which was higher compared to plants grown hydroponically. Concluding, most metabolic differences between rocket species were observed at high Se supplementation. Low Se foliar fertilization was effective in an enriching rocket in Se without affecting other phytochemicals. However, the Se dosages sufficient for biofortification could be even lower, as the Se concentration in rocket treated with 2.5 mg Se per plant was still very high and the edible part should not be eaten undiluted. Also, a single method of Se supplementation does not appear to be optimal for all plant species or the same species, as the metabolic responses could be very different.

Hyperaccumulator Stanleya pinnata: In Situ Fitness in Relation to Tissue Selenium Concentration.

Earlier studies have shown that Stanleya pinnata benefits from selenium hyperaccumulation through ecological benefits and enhanced growth. However, no investigation has assayed the effects of Se hyperaccumulation on plant fitness in the field. This research aimed to analyze how variation in Se accumulation affects S. pinnata fitness, judged from physiological and biochemical performance parameters and herbivory while growing naturally on two seleniferous sites. Natural variation in Se concentration in vegetative and reproductive tissues was determined, and correlations were explored between Se levels with fitness parameters, herbivory damage, and plant defense compounds. Leaf Se concentration varied between 13- and 55-fold in the two populations, averaging 868 and 2482 mg kg−1 dry weight (DW). Furthermore, 83% and 31% of plants from the two populations showed Se hyperaccumulator levels in leaves (>1000 mg kg−1 DW). In seeds, the Se levels varied 3−4-fold and averaged 3372 and 2267 mg kg−1 DW, well above the hyperaccumulator threshold. Plant size and reproductive parameters were not correlated with Se concentration. There was significant herbivory pressure even on the highest-Se plants, likely from Se-resistant herbivores. We conclude that the variation in Se hyperaccumulation did not appear to enhance or compromise S. pinnata fitness in seleniferous habitats within the observed Se range.

Selenium transport and metabolism in plants: Phytoremediation and biofortification implications.

The aim of this review is to synthesize current knowledge of selenium (Se) transport and metabolism in plants, with a focus on implications for biofortification and phytoremediation. Selenium is a necessary human micronutrient, and around a billion people worldwide may be Se deficient. This can be ameliorated by Se biofortification of staple crops. Selenium is also a potential toxin at higher concentrations, and multiple environmental disasters over the past 50 years have been caused by Se pollution from agricultural and industrial sources. Phytoremediation by plants able to take up large amounts of Se is an important tool to combat pollution issues. Both biofortification and phytoremediation applications require a thorough understanding of how Se is taken up and metabolized by plants. Selenium uptake and translocation in plants are largely accomplished via sulfur (S) transport proteins. Current understanding of these transporters is reviewed here, and transporters that may be manipulated to improve Se uptake are discussed. Plant Se metabolism also largely follows the S metabolic pathway. This pathway is reviewed here, with special focus on genes that have been, or may be manipulated to reduce the accumulation of toxic metabolites or enhance the accumulation of nontoxic metabolites. Finally, unique aspects of Se transport and metabolism in Se hyperaccumulators are reviewed. Hyperaccumulators, which can accumulate Se at up to 1000 times higher concentrations than normal plants, present interesting specialized systems of Se transport and metabolism. Selenium hyperaccumulation mechanisms and potential applications of these mechanisms to biofortification and phytoremediation are presented.

Effects of selenium hyperaccumulators on soil selenium distribution and vegetation properties.

PREMISE: The ecological implications of hyperaccumulation have been investigated at the organismal level, but are poorly understood at the plant community level. Questions addressed here were: Does the presence of selenium (Se) hyperaccumulators affect Se distribution and concentration in their native soil, and do hyperaccumulators affect overall vegetation properties and species composition? METHODS: Plant survey and soil Se mapping were performed at three seleniferous sites in Colorado. In season one, plots with and without hyperaccumulators were compared for (1) bare ground, canopy cover, and species richness; (2) relative species abundance; (3) soil Se distribution and concentration. In season two, a smaller-scale design was implemented, focusing on areas 3 m in diameter around hyperaccumulators versus nonhyperaccumulators in 44 paired plots on one site. RESULTS: Plots with hyperaccumulators generally showed more bare ground, less canopy cover, higher species richness, and 2-3-fold higher soil Se levels. These patterns were not consistently significant across all sites; the effects of hyperaccumulators may have been diluted by their low abundance and the relatively large area of survey. In the smaller-scale study, highly significant results were obtained, showing more bare ground, less canopy cover, and higher species richness in plots with hyperaccumulators; soil Se concentration was also higher in plots with hyperaccumulators. CONCLUSIONS: Hyperaccumulators may significantly affect local soil Se concentration and vegetation over at least a 3 m diameter area, or 4× their canopy. These differences may result from the combined positive and negative allelopathic effects observed earlier at the organismal level.

Selenium Metabolism in Hemp (Cannabis sativa L.)-Potential for Phytoremediation and Biofortification.

Selenium (Se) deficiency and toxicity affect over a billion people worldwide. Plants can mitigate both problems, via Se biofortification and phytoremediation. Here we explore the potential of hemp (Cannabis sativa L.) for these phytotechnologies. Field surveys in naturally seleniferous agricultural areas in Colorado, United States, found 15-25 µg of Se/g in seed and 5-10 µg of Se/g dry weight (DW) in flowers and leaves. Thus, 4 g of this hemp seed provides the U.S. recommended daily allowance of 55-75 µg of Se. In controlled greenhouse experiments, hemp seedlings grown in Turface supplied with 40-320 µM selenate showed complete tolerance up to 160 µM and accumulated up to 1300 mg of Se/kg shoot dry weight. Mature hemp grown in Turface supplied with 5-80 µM selenate was completely tolerant up to 40 µM selenate and accumulated up to 200 mg of Se/kg DW in leaves, flowers, and seeds. Synchrotron X-ray fluorescence and X-ray absorption spectroscopies of selenate-supplied hemp showed Se to accumulate mainly in the leaf vasculature and in the seed embryos, with predominant Se speciation in C-Se-C forms (57-75% in leaf and more than 86% in seeds). Aqueous seed extracts were found by liquid chromatography mass spectrometry to contain selenomethionine and methyl-selenocysteine (1:1-3 ratio), both excellent dietary Se sources. Floral concentrations of medicinal cannabidiol (CBD) and terpenoids were not affected by Se. We conclude that hemp has good potential for Se phytoremediation while producing Se-biofortified dietary products.

Distribution and chemical form of selenium in Neptunia amplexicaulis from Central Queensland, Australia.

Selenium (Se), a trace element essential for human and animal biological processes, is deficient in many agricultural soils. Some extremely rare plants can naturally accumulate extraordinarily high concentrations of Se. The native legume Neptunia amplexicaulis, endemic to a small area near Richmond and Hughenden in Central Queensland, Australia, is one of the strongest Se hyperaccumulators known on Earth, with foliar concentrations in excess of 4000 µg Se g-1 previously recorded. Here, we report on the Se distribution at a whole plant level using laboratory micro X-ray Fluorescence Microscopy (µXRF) and scanning electron microscopy (SEM-EDS), as well as on chemical forms of Se in various tissues using liquid chromatography-mass spectrometry (LC-MS) and synchrotron X-ray absorption spectroscopy (XAS). The results show that Se occurs in the forms of methyl-selenocysteine and seleno-methionine in the foliar tissues, with up to 13 600 µg Se g-1 total in young leaves. Selenium was found to accumulate primarily in the young leaves, flowers, pods and taproot, with lower concentrations present in the fine-roots and stem and the lowest present in the oldest leaves. Trichomes were not found to accumulate Se. We postulate that Se is (re)distributed in this plant via the phloem from older leaves to newer leaves, using the taproot as the main storage organ. High concentrations of Se in the nodes (pulvini) indicate this structure may play an important a role in Se (re)distribution. The overall pattern of Se distribution was similar in a non-Se tolerant closely related species (Neptunia gracilis), although the prevailing Se concentrations were substantially lower than in N. amplexicaulis.

Identification and physiological comparison of plant species that show positive or negative co-occurrence with selenium hyperaccumulators.

In these studies we identified and compared the properties of plant species that showed positive or negative co-occurrence with selenium (Se) hyperaccumulators in their natural habitat. The main questions addressed were: which species are most abundant directly adjacent to hyperaccumulators, and which are absent? How do Se accumulation and tolerance compare in species found to positively or negatively co-occur with hyperaccumulators? Approaches included field surveys, X-ray microprobe analysis of field samples, and a lab Se tolerance and accumulation study. When 54 hyperaccumulators across two naturally seleniferous sites were surveyed for their five nearest neighboring species, and the relative abundance of these species around hyperaccumulators compared to that in the overall vegetation, some species were identified to positively or negatively co-occur with hyperaccumulators. Several positively co-occurring species showed high Se accumulation capability (up to 900 mg Se per kg dry weight), which may reflect Se tolerance. Leaf X-ray microprobe analysis found relatively more organic forms of Se in two positively co-occurring species than in a negatively co-occurring one. There were elevated soil Se levels around Se hyperaccumulators, and neighbors of Se hyperaccumulators had a higher tissue Se concentration as compared to when the same species grew elsewhere in the area. The elevated soil Se levels around Se hyperaccumulators - likely resulting from litter deposition- may significantly affect the local plant community, facilitating Se-tolerant plant community members but lowering the fitness of Se-sensitive members.

Selenium tolerance, accumulation, localization and speciation in a Cardamine hyperaccumulator and a non-hyperaccumulator.

Cardamine violifolia (family Brassicaceae) is the first discovered selenium hyperaccumulator from the genus Cardamine with unique properties in terms of selenium accumulation, i.e., high abundance of selenolanthionine. In our study, a fully comprehensive experiment was conducted with the comparison of a non-hyperaccumulator Cardamine species, Cardamine pratensis, covering growth characteristics, chlorophyll fluorescence, spatial selenium/sulfur distribution patterns through elemental analyses (synchrotron-based X-Ray Fluorescence and ICP-OES) and speciation data through selenium K-edge micro X-ray absorption near-edge structure analysis (µXANES) and strong cation exchange (SCX)-ICP-MS. The results revealed remarkable differences in contrast to other selenium hyperaccumulators as neither Cardamine species showed evidence of growth stimulation by selenium. Also, selenite uptake was not inhibited by phosphate for either of the Cardamine species. Sulfate inhibited selenate uptake, but the two Cardamine species did not show any difference in this respect. However, µXRF derived speciation maps and selenium/sulfur uptake characteristics provided results that are similar to other formerly reported hyperaccumulator and non-hyperaccumulator Brassicaceae species. µXANES showed organic selenium, "C-Se-C", in seedlings of both species and also in mature C. violifolia plants. In contrast, selenate-supplied mature C. pratensis contained approximately half "C-Se-C" and half selenate. SCX-ICP-MS data showed evidence of the lack of selenocystine in any of the Cardamine plant extracts. Thus, C. violifolia shows clear selenium-related physiological and biochemical differences compared to C. pratensis and other selenium hyperaccumulators.

Selenium Accumulation, Speciation and Localization in Brazil Nuts (Bertholletia excelsa H.B.K.).

More than a billion people worldwide may be selenium (Se) deficient, and supplementation with Se-rich Brazil nuts may be a good strategy to prevent deficiency. Since different forms of Se have different nutritional value, and Se is toxic at elevated levels, careful seed characterization is important. Variation in Se concentration and correlations of this element with other nutrients were found in two batches of commercially available nuts. Selenium tissue localization and speciation were further determined. Mean Se levels were between 28 and 49 mg kg-1, with up to 8-fold seed-to-seed variation (n = 13) within batches. Brazil nut Se was mainly in organic form. While present throughout the seed, Se was most concentrated in a ring 1 to 2 mm below the surface. While healthy, Brazil nuts should be consumed in moderation. Consumption of one seed (5 g) from a high-Se area meets its recommended daily allowance; the recommended serving size of 30 g may exceed the allowable daily intake (400 µg) or even its toxicity threshold (1200 µg). Based on these findings, the recommended serving size may be re-evaluated, consumers should be warned not to exceed the serving size and the seed may be sold as part of mixed nuts, to avoid excess Se intake.

On the Ecology of Selenium Accumulation in Plants.

Plants accumulate and tolerate Se to varying degrees, up to 15,000 mg Se/kg dry weight for Se hyperaccumulators. Plant Se accumulation may exert positive or negative effects on other species in the community. The movement of plant Se into ecological partners may benefit them at low concentrations, but cause toxicity at high concentrations. Thus, Se accumulation can protect plants against Se-sensitive herbivores and pathogens (elemental defense) and reduce surrounding vegetation cover via high-Se litter deposition (elemental allelopathy). While hyperaccumulators negatively impact Se-sensitive ecological partners, they offer a niche for Se-tolerant partners, including beneficial microbial and pollinator symbionts as well as detrimental herbivores, pathogens, and competing plant species. These ecological effects of plant Se accumulation may facilitate the evolution of Se resistance in symbionts. Conversely, Se hyperaccumulation may evolve driven by increasing Se resistance in herbivores, pathogens, or plant neighbors; Se resistance also evolves in mutualist symbionts, minimizing the plant's ecological cost. Interesting topics to address in future research are whether the ecological impacts of plant Se accumulation may affect species composition across trophic levels (favoring Se resistant taxa), and to what extent Se hyperaccumulators form a portal for Se into the local food chain and are important for Se cycling in the local ecosystem.

Selenium Biofortification Differentially Affects Sulfur Metabolism and Accumulation of Phytochemicals in Two Rocket Species (Eruca Sativa Mill. and Diplotaxis Tenuifolia) Grown in Hydroponics.

Biofortification can be exploited to enrich plants in selenium (Se), an essential micronutrient for humans. Selenium as selenate was supplied to two rocket species, Eruca sativa Mill. (salad rocket) and Diplotaxis tenuifolia (wild rocket), at 0⁻40 µM in hydroponics and its effects on the content and profile of sulphur (S)-compounds and other phytochemicals was evaluated. D. tenuifolia accumulated more total Se and selenocysteine than E. sativa, concentrating up to ~300 mg Se kg-1 dry weight from 10⁻40 µM Se. To ensure a safe and adequate Se intake, 30 and 4 g fresh leaf material from E. sativa grown with 5 and 10⁻20 µM Se, respectively or 4 g from D. tenuifolia supplied with 5 µM Se was estimated to be optimal for consumption. Selenium supplementation at or above 10 µM differentially affected S metabolism in the two species in terms of the transcription of genes involved in S assimilation and S-compound accumulation. Also, amino acid content decreased with Se in E. sativa but increased in D. tenuifolia and the amount of phenolics was more reduced in D. tenuifolia. In conclusion, selenate application in hydroponics allowed Se enrichment of rocket. Furthermore, Se at low concentration (5 µM) did not significantly affect accumulation of phytochemicals and plant defence S-metabolites.

Characterization of Selenium Accumulation, Localization and Speciation in Buckwheat-Implications for Biofortification.

Buckwheat is an important crop species in areas of selenium (Se) deficiency. To obtain better insight into their Se metabolic properties, common buckwheat (Fagopyrum esculentum) and tartary buckwheat (F. tataricum) were supplied with different concentrations of Se, supplied as selenate, selenite, or Astragalus bisulcatus plant extract (methyl-selenocysteine). Se was supplied at different developmental stages, with different durations, and in the presence or absence of potentially competing ions, sulfate, and phosphate. The plants were analyzed for growth, Se uptake, translocation, accumulation, as well as for Se localization and chemical speciation in the seed. Plants of both buckwheat species were supplied with 20 µM of either of the three forms of Se twice over their growth period. Both species accumulated 15-40 mg Se kg-1 DW in seeds, leaves and stems, from all three selenocompounds. X-ray microprobe analysis showed that the Se in seeds was localized in the embryo, in organic C-Se-C form(s) resembling selenomethionine, methyl-selenocysteine, and γ-glutamyl-methylselenocysteine standards. In short-term (2 and 24 h) Se uptake studies, both buckwheat species showed higher Se uptake rate and shoot Se accumulation when supplied with plant extract (methyl-selenocysteine), compared to selenite or selenate. In long-term (7 days) uptake studies, both species were resistant to selenite up to 50 µM. Tartary buckwheat was also resistant to selenate up to 75 µM Se, but >30 µM selenate inhibited common buckwheat growth. Selenium accumulation was similar in both species. When selenite was supplied, Se levels were 10-20-fold higher in root (up to 900 mg Se kg-1 DW) than shoot, but 4-fold higher in shoot (up to 1,200 mg Se kg-1 DW) than root for selenate-supplied plants. Additionally, sulfate and phosphate supply affected Se uptake, and conversely selenate enhanced S and P accumulation in both species. These findings have relevance for crop Se biofortification applications.

Fungal Endophyte Alternaria tenuissima Can Affect Growth and Selenium Accumulation in Its Hyperaccumulator Host Astragalus bisulcatus.

Endophytes can enhance plant stress tolerance by promoting growth and affecting elemental accumulation, which may be useful in phytoremediation. In earlier studies, up to 35% elemental selenium (Se0) was found in Se hyperaccumulator Astragalus bisulcatus. Since Se0 can be produced by microbes, the plant Se0 was hypothesized to be microbe-derived. Here we characterize a fungal endophyte of A. bisulcatus named A2. It is common in seeds from natural seleniferous habitat containing 1,000-10,000 mg kg-1 Se. We identified A2 as Alternaria tenuissima via 18S rRNA sequence analysis and morphological characterization. X-ray microprobe analysis of A. bisulcatus seeds that did or did not harbor Alternaria, showed that both contained >90% organic seleno-compounds with C-Se-C configuration, likely methylselenocysteine and glutamyl-methylselenocysteine. The seed Se was concentrated in the embryo, not the seed coat. X-ray microprobe analysis of A2 in pure culture showed the fungus produced Se0 when supplied with selenite, but accumulated mainly organic C-Se-C compounds when supplied with selenate. A2 was completely resistant to selenate up to 300 mg L-1, moderately resistant to selenite (50% inhibition at ∼50 mg Se L-1), but relatively sensitive to methylselenocysteine and to Se extracted from A. bisulcatus (50% inhibition at 25 mg Se L-1). Four-week old A. bisulcatus seedlings derived from surface-sterilized seeds containing endophytic Alternaria were up to threefold larger than seeds obtained from seeds not showing evidence of fungal colonization. When supplied with Se, the Alternaria-colonized seedlings had lower shoot Se and sulfur levels than seedlings from uncolonized seeds. In conclusion, A. tenuissima may contribute to the Se0 observed earlier in A. bisulcatus, and affect host growth and Se accumulation. A2 is sensitive to the Se levels found in its host's tissues, but may avoid Se toxicity by occupying low-Se areas (seed coat, apoplast) and converting plant Se to non-toxic Se0. These findings illustrate the potential for hyperaccumulator endophytes to affect plant properties relevant for phytoremediation. Facultative endophytes may also be applicable in bioremediation and biofortification, owing to their capacity to turn toxic inorganic forms of Se into non-toxic or even beneficial, organic forms with anticarcinogenic properties.

Plant selenium hyperaccumulation- Ecological effects and potential implications for selenium cycling and community structure.

BACKGROUND: Selenium (Se) hyperaccumulation occurs in ~50 plant taxa native to seleniferous soils in Western USA. Hyperaccumulator tissue Se levels, 1000-15,000 mg/kg dry weight, are typically 100 times higher than surrounding vegetation. Relative to other species, hyperaccumulators also transform Se more into organic forms. SCOPE OF REVIEW: We review abiotic and biotic factors influencing soil Se distribution and bioavailability, soil being the source of the Se in hyperaccumulators. Next, we summarize the fate of Se in plants, particularly hyperaccumulators. We then extensively review the impact of plant Se accumulation on ecological interactions. Finally, we discuss the potential impact of Se hyperaccumulators on local community composition and Se cycling. MAJOR CONCLUSIONS: Selenium (hyper)accumulation offers ecological advantages: protection from herbivores and pathogens and competitive advantage over other plants. The extreme Se levels in and around hyperaccumulators create a toxic environment for Se-sensitive ecological partners, while offering a niche for Se-resistant partners. Through these dual effects, hyperaccumulators may influence species composition in their local environment, as well as Se cycling. GENERAL SIGNIFICANCE: The implied effects of Se hyperaccumulation on community assembly and local Se cycling warrant further investigations into the contribution of hyperaccumulators and general terrestrial vegetation to global Se cycling and may serve as a case study for how trace elements influence ecological processes. Furthermore, understanding ecological implications of plant Se accumulation are vital for safe implementation of biofortification and phytoremediation, technologies increasingly implemented to battle Se deficiency and toxicity.

Mechanisms of selenium hyperaccumulation in plants: A survey of molecular, biochemical and ecological cues.

BACKGROUND: Selenium (Se) is a micronutrient required for many life forms, but toxic at higher concentration. Plants do not have a Se requirement, but can benefit from Se via enhanced antioxidant activity. Some plant species can accumulate Se to concentrations above 0.1% of dry weight and seem to possess mechanisms that distinguish Se from its analog sulfur (S). Research on these so-called Se hyperaccumulators aims to identify key genes for this remarkable trait and to understand ecological implications. SCOPE OF REVIEW: This review gives a broad overview of the current knowledge about Se uptake and metabolism in plants, with a special emphasis on hypothesized mechanisms of Se hyperaccumulation. The role of Se in plant defense responses and the associated ecological implications are discussed. MAJOR CONCLUSIONS: Hyperaccumulators have enhanced expression of S transport and assimilation genes, and may possess transporters with higher specificity for selenate over sulfate. Genes involved in antioxidant reactions and biotic stress resistance are also upregulated. Key regulators in these processes appear to be the growth regulators jasmonic acid, salicylic acid and ethylene. Hyperaccumulation may have evolved owing to associated ecological benefits, particularly protection against pathogens and herbivores, and as a form of elemental allelopathy. GENERAL SIGNIFICANCE: Understanding plant Se uptake and metabolism in hyperaccumulators has broad relevance for the environment, agriculture and human and animal nutrition and may help generate crops with selenate-specific uptake and high capacity to convert selenate to less toxic, anticarcinogenic, organic Se compounds.

Comparison of ATP sulfurylase 2 from selenium hyperaccumulator Stanleya pinnata and non-accumulator Stanleya elata reveals differential intracellular localization and enzyme activity levels.

BACKGROUND: The plant Stanleya pinnata hyperaccumulates Se up to 0.5% of its dry weight in organic forms, whereas the closely related Stanleya elata does not hyperaccumulate Se. ATP sulfurylase (ATPS) can catalyze the formation of adenosine 5'-phosphoselenate (APSe) from ATP and selenate. We investigated the S. pinnata ATPS2 isoform (SpATPS2) to assess its possible role in Se hyperaccumulation. METHODS: ATPS expression and activity was compared in the two Stanleya species. The ATPS2 protein sequences were modeled. Sub-cellular locations were analyzed using GFP fusions. Enzyme activity of purified recombinant SpATPS2 was measured. RESULTS: ATPS2 transcript levels were six-fold higher in roots of S. pinnata relative to S. elata. Overall root ATPS enzyme activity was two-fold elevated in S. pinnata. Cloning and sequencing of SpATPS2 and S. elata ATPS2 (SeATPS2) showed the predicted SeATPS2 to be canonical, while SpATPS2, although very similar in its core structure, has unique features, including an interrupted plastid targeting signal due to a stop codon in the 5' region of the coding sequence. Indeed GFP fusions revealed that SpATPS2 had exclusive cytosolic localization, while SeATPS2 showed dual localization in plastids and cytosol. SpATPS2 activity was inhibited by both sulfate and selenate, indicating that the enzyme acts on both substrates. CONCLUSIONS: The ATPS2 from S. pinnata differs from non-accumulator ATPS2 in its elevated expression and sub-cellular localization. It likely acts on both selente and sulfate substrates. GENERAL SIGNIFICANCE: These observations shed new light on the role of ATPS2 in the evolution of Se hyperaccumulation in plants. This article is part of a Special Issue entitled Selenium research in biochemistry and biophysics - 200 year anniversary issue, edited by Dr. Elias Arnér and Dr. Regina Brigelius-Flohe.

Transcriptome-wide comparison of selenium hyperaccumulator and nonaccumulator Stanleya species provides new insight into key processes mediating the hyperaccumulation syndrome.

To obtain better insight into the mechanisms of selenium hyperaccumulation in Stanleya pinnata, transcriptome-wide differences in root and shoot gene expression levels were investigated in S. pinnata and related nonaccumulator Stanleya elata grown with or without 20 µm selenate. Genes predicted to be involved in sulphate/selenate transport and assimilation or in oxidative stress resistance (glutathione-related genes and peroxidases) were among the most differentially expressed between species; many showed constitutively elevated expression in S. pinnata. A number of defence-related genes predicted to mediate synthesis and signalling of defence hormones jasmonic acid (JA, reported to induce sulphur assimilatory and glutathione biosynthesis genes), salicylic acid (SA) and ethylene were also more expressed in S. pinnata than S. elata. Several upstream signalling genes that up-regulate defence hormone synthesis showed higher expression in S. pinnata than S. elata and might trigger these selenium-mediated defence responses. Thus, selenium hyperaccumulation and hypertolerance in S. pinnata may be mediated by constitutive, up-regulated JA, SA and ethylene-mediated defence systems, associated with elevated expression of genes involved in sulphate/selenate uptake and assimilation or in antioxidant activity. Genes pinpointed in this study may be targets of genetic engineering of plants that may be employed in biofortification or phytoremediation.

Influence of sulfate supply on selenium uptake dynamics and expression of sulfate/selenate transporters in selenium hyperaccumulator and nonhyperaccumulator Brassicaceae.

Stanleya pinnata not only hyperaccumulates selenium (Se) to 0.5% of its dry weight, but also exhibits higher tissue Se-to-sulfur (S) ratios than other species and its surroundings. To investigate the mechanisms underlying this Se enrichment, we compared S. pinnata with the nonhyperaccumulators S. elata and Brassica juncea for selenate uptake in long- (9 d) and short-term (1 h) assays, using different concentrations of selenate and competitor sulfate. Different sulfate pre-treatments (0, 0.5, 5 mM, 3 d) were also tested for effects on selenate uptake and sulfate transporters' expression. Relative to nonhyperaccumulators, S. pinnata showed higher rates of root and shoot Se accumulation and less competitive inhibition by sulfate or by high-S pretreatment. The selenate uptake rate for S. pinnata (1 h) was three- to four-fold higher than for nonhyperaccumulators, and not significantly affected by 100-fold excess sulfate, which reduced selenate uptake by 100% in S. elata and 40% in B. juncea. Real-time reverse transcription PCR indicated constitutive upregulation in S. pinnata of sulfate transporters SULTR1;2 (root influx) and SULTR2;1 (translocation), but reduced SULTR1;1 expression (root influx). In S. pinnata, selenate uptake and translocation rates are constitutively elevated and relatively sulfate-independent. Underlying mechanisms likely include overexpression of SULTR1;2 and SULTR2;1, which may additionally have evolved enhanced specificity for selenate over sulfate.

Selenium Biofortification and Phytoremediation Phytotechnologies: A Review.

The element selenium (Se) is both essential and toxic for most life forms, with a narrow margin between deficiency and toxicity. Phytotechnologies using plants and their associated microbes can address both of these problems. To prevent Se toxicity due to excess environmental Se, plants may be used to phytoremediate Se from soil or water. To alleviate Se deficiency in humans or livestock, crops may be biofortified with Se. These two technologies may also be combined: Se-enriched plant material from phytoremediation could be used as green fertilizer or as fortified food. Plants may also be used to "mine" Se from seleniferous soils. The efficiency of Se phytoremediation and biofortification may be further optimized. Research in the past decades has provided a wealth of knowledge regarding the mechanisms by which plants take up, metabolize, accumulate, and volatilize Se and the role plant-associated microbes play in these processes. Furthermore, ecological studies have revealed important effects of plant Se on interactions with herbivores, detrivores, pollinators, neighboring vegetation, and the plant microbiome. All this knowledge can be exploited in phytotechnology programs to optimize plant Se accumulation, transformation, volatilization, and/or tolerance via plant breeding, genetic engineering, and tailored agronomic practices.

The fascinating facets of plant selenium accumulation - biochemistry, physiology, evolution and ecology.

Contents 1582 I. 1582 II. 1583 III. 1588 IV. 1590 V. 1592 1592 References 1592 SUMMARY: The importance of selenium (Se) for medicine, industry and the environment is increasingly apparent. Se is essential for many species, including humans, but toxic at elevated concentrations. Plant Se accumulation and volatilization may be applied in crop biofortification and phytoremediation. Topics covered here include beneficial and toxic effects of Se on plants, mechanisms of Se accumulation and tolerance in plants and algae, Se hyperaccumulation, and ecological and evolutionary aspects of these processes. Plant species differ in the concentration and forms of Se accumulated, Se partitioning at the whole-plant and tissue levels, and the capacity to distinguish Se from sulfur. Mechanisms of Se hyperaccumulation and its adaptive significance appear to involve constitutive up-regulation of sulfate/selenate uptake and assimilation, associated with elevated concentrations of defense-related hormones. Hyperaccumulation has evolved independently in at least three plant families, probably as an elemental defense mechanism and perhaps mediating elemental allelopathy. Elevated plant Se protects plants from generalist herbivores and pathogens, but also gives rise to the evolution of Se-resistant specialists. Plant Se accumulation affects ecological interactions with herbivores, pollinators, neighboring plants, and microbes. Hyperaccumulation tends to negatively affect Se-sensitive ecological partners while facilitating Se-resistant partners, potentially affecting species composition and Se cycling in seleniferous ecosystems.