Exploring the importance of sulfate transporters and ATP sulphurylases for selenium hyperaccumulation\u2014a comparison of Stanleya pinnata and Brassica juncea (Brassicaceae)

Selenium (Se) hyperaccumulation, the capacity of some species to concentrate Se to levels upwards of 0.1% of dry weight, is an intriguing phenomenon that is only partially understood. Questions that remain to be answered are: do hyperaccumulators have one or more Se-specific transporters? How are these regulated by Se and sulfur (S)? In this study, hyperaccumulator Stanleya pinnata was compared with related non-hyperaccumulator Brassica juncea with respect to S-dependent selenate uptake and translocation, as well as for the expression levels of three sulfate/selenate transporters (Sultr) and three ATP sulphurylases (APS). Selenium accumulation went down ~10-fold with increasing sulfate supply in B. juncea, while S. pinnata only had a 2\u20133-fold difference in Se uptake between the highest (5 mM) and lowest sulfate (0 mM) treatments. The Se/S ratio was generally higher in the hyperaccumulator than the non-hyperaccumulator, and while tissue Se/S ratio in B. juncea largely reflected the ratio in the growth medium, S. pinnata enriched itself up to 5-fold with Se relative to S. The transcript levels of Sultr1;2 and 2;1 and APS1, 2, and 4 were generally much higher in S. pinnata than B. juncea, and the species showed differential transcript responses to S and Se supply. These results indicate that S. pinnata has at least one transporter with significant selenate specificity over sulfate. Also, the hyperaccumulator has elevated expression levels of several sulfate/selenate transporters and APS enzymes, which likely contribute to the Se hyperaccumulation and hypertolerance phenotype

Effects of selenium biofortification on crop nutritional quality

Selenium (Se) at very low doses has crucial functions in humans and animals. Since plants represent the main dietary source of this element, Se-containing crops may be used as a means to deliver Se to consumers (biofortification). Several strategies have been exploited to increase plant Se content. Selenium assimilation in plants affects both sulphur (S) and nitrogen (N) metabolic pathways, which is why recent research has also focused on the effect of Se fertilization on the production of S- and N- secondary metabolites with putative health benefits. In this review we discuss the function of Se in plant and human nutrition and the progress in the genetic engineering of Se metabolism to increase the levels and bioavailability of this element in food crops. Particular attention is paid to Se biofortification and the synthesis of compounds with beneficial effects on health

Effects of selenium accumulation on reproductive functions in Brassica juncea and Stanleya pinnata

Selenium (Se) is an essential micronutrient for many organisms, but is also a toxin and environmental pollutant at elevated levels. Due to its chemical similarity to sulphur, most plants readily take up and assimilate Se. Se accumulators such as Brassica juncea can accumulate Se between 0.01% and 0.1% of dry weight (DW), and Se hyperaccumulators such as Stanleya pinnata (Brassicaeae) contain between 0.1% and 1.5% DW of Se. While Se accumulation offers the plant a variety of ecological benefits, particularly protection from herbivory, its potential costs are still unexplored. This study examines the effects of plant Se levels on reproductive functions. In B. juncea, Se concentrations >0.05–0.1% caused decreases in biomass, pollen germination, individual seed and total seed weight, number of seeds produced, and seed germination. In S. pinnata there was no negative effect of increased Se concentration on pollen germination. In cross-pollination of B. juncea plants with different Se levels, both the maternal and paternal Se level affected reproduction, but the maternal Se concentration had the most pronounced effect. Interestingly, high-Se maternal plants were most efficiently pollinated by Se-treated paternal plants. These data provide novel insights into the potential reproductive costs of Se accumulation, interactive effects of Se in pollen grains and in the pistil, and the apparent evolution of physiological tolerance mechanisms in hyperaccumulators to avoid reproductive repercussions

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

Summary 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

Getting to the Root of Selenium Hyperaccumulation—Localization and Speciation of Root Selenium and Its Effects on Nematodes

Elemental hyperaccumulation protects plants from many aboveground herbivores. Little is known about effects of hyperaccumulation on belowground herbivores or their ecological interactions. To examine effects of plant selenium (Se) hyperaccumulation on nematode root herbivory, we investigated spatial distribution and speciation of Se in hyperaccumulator roots using X-ray microprobe analysis, and effects of root Se concentration on root-associated nematode communities. Perennial hyperaccumulators Stanleya pinnata and Astragalus bisulcatus, collected from a natural seleniferous grassland contained 100–1500 mg Se kg−1 root dry weight (DW). Selenium was concentrated in the cortex and epidermis of hyperaccumulator roots, with lower levels in the stele. The accumulated Se consisted of organic (C-Se-C) compounds, indistinguishable from methyl-selenocysteine. The field-collected roots yielded 5–400 nematodes g−1 DW in Baermann funnel extraction, with no correlation between root Se concentration and nematode densities. Even roots containing \u3e 1000 mg Se kg−1 DW yielded herbivorous nematodes. However, greenhouse-grown S. pinnata plants treated with Se had fewer total nematodes than those without Se. Thus, while root Se hyperaccumulation may protect plants from non-specialist herbivorous nematodes, Se-resistant nematode taxa appear to associate with hyperaccumulators in seleniferous habitats, and may utilize high-Se hyperaccumulator roots as food source. These findings give new insight into the ecological implications of plant Se (hyper)accumulation

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