Selenium distribution and speciation in the hyperaccumulator Astragalus bisulcatus and associated ecological partners.

The goal of this study was to investigate how plant selenium (Se) hyperaccumulation may affect ecological interactions and whether associated partners may affect Se hyperaccumulation. The Se hyperaccumulator Astragalus bisulcatus was collected in its natural seleniferous habitat, and x-ray fluorescence mapping and x-ray absorption near-edge structure spectroscopy were used to characterize Se distribution and speciation in all organs as well as in encountered microbial symbionts and herbivores. Se was present at high levels (704-4,661 mg kg(-1) dry weight) in all organs, mainly as organic C-Se-C compounds (i.e. Se bonded to two carbon atoms, e.g. methylselenocysteine). In nodule, root, and stem, up to 34% of Se was found as elemental Se, which was potentially due to microbial activity. In addition to a nitrogen-fixing symbiont, the plants harbored an endophytic fungus that produced elemental Se. Furthermore, two Se-resistant herbivorous moths were discovered on A. bisulcatus, one of which was parasitized by a wasp. Adult moths, larvae, and wasps all accumulated predominantly C-Se-C compounds. In conclusion, hyperaccumulators live in association with a variety of Se-resistant ecological partners. Among these partners, microbial endosymbionts may affect Se speciation in hyperaccumulators. Hyperaccumulators have been shown earlier to negatively affect Se-sensitive ecological partners while apparently offering a niche for Se-resistant partners. Through their positive and negative effects on different ecological partners, hyperaccumulators may influence species composition and Se cycling in seleniferous ecosystems.

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.

Characterization of rhizosphere fungi from selenium hyperaccumulator and nonhyperaccumulator plants along the eastern Rocky Mountain Front Range.

PREMISE OF STUDY: Selenium-hyperaccumulator plants can store over 1% (dry mass) Se in their tissues, despite the toxicity of this element at high concentrations across eukaryotes. These levels of Se can have widespread effects on the plant's ecological partners, including herbivores and pathogens. Still other partners seem to have coevolved Se tolerance. This is the first known study addressing the rhizosphere mycoflora of Se hyperaccumulators and aims to evaluate the rhizospheric fungal diversity and Se tolerance to further the knowledge of how these organisms interact with their host plants and survive in these extreme habitats. METHODS: Rhizosphere fungi were isolated from Se-hyperaccumulator and nonaccumulator plant species collected from five sites in Colorado and Wyoming; four seleniferous sites and one nonseleniferous site. 259 isolates were identified to genus or species and evaluated for Se tolerance. KEY RESULTS: Among the 24 represented genera, 11 comprised 86% of the isolates. The majority of isolates from the seleniferous sites were unaffected by 10 mg·L(-1) Se, irrespective of host plant (hyperaccumulator vs. nonaccumulator), while rhizosphere fungi from a control, nonseleniferous site were highly sensitive to Se at 10 mg·L(-1) and as a group were significantly less (α = 0.05) tolerant than the isolates from the seleniferous sites. CONCLUSIONS: Even though Se is a commonly used antifungal agent, these results suggest that rhizosphere fungi from seleniferous habitats have widespread Se tolerance, likely an adaptive advantage in their Se-rich habitat.

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.

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.

Overexpression of ATP sulfurylase in Indian mustard: effects on tolerance and accumulation of twelve metals.

Indian mustard [Brassica juncea (L.) Czern.] transgenics overexpressing ATP sulfurylase (APS plants) were shown previously to have higher levels of total thiols, S, and Se. The present study explores the effect of ATP sulfurylase overexpression on tolerance and accumulation of other metals, both oxyanions and cations, reasoning that some anions may react directly with ATP sulfurylase, while other ions may be bound by its thiol end products. The APS transgenics were compared with wild-type plants with respect to tolerance and accumulation of As, Cd, Cr, Cu, Hg, Mn, Mo, Ni, Pb, V, W, and Zn, supplied individually in agar medium (seedlings) or in hydroponics (mature plants). At the seedling stage, APS transgenics were more tolerant than wild type to As(III), As(V), Cd, Cu, Hg, and Zn, but less tolerant to Mo and V. The APS seedlings had up to 2.5-fold higher shoot concentrations of As(III), As(V), Hg, Mo, Pb, and V, and somewhat lower Cr levels. Mature APS plants contained up to 2.5-fold higher shoot concentrations of Cd, Cr, Cu, Mo, V, and W than wild type. They also contained 1.5- to 2-fold higher levels of the essential elements Fe, Mo, and S in most of the treatments. Mature APS plants showed no differences in metal tolerance compared with the wild type. Overexpression of ATP sulfurylase may be a promising approach to create plants with enhanced phytoextraction capacity for mixtures of metals.