Roles of rhizobial symbionts in selenium hyperaccumulation in Astragalus (Fabaceae).

PREMISE OF THE STUDY: Are there dimensions of symbiotic root interactions that are overlooked because plant mineral nutrition is the foundation and, perhaps too often, the sole explanation through which we view these relationships? In this paper we investigate how the root nodule symbiosis in selenium (Se) hyperaccumulator and nonaccumulator Astragalus species influences plant selenium (Se) accumulation. METHODS: In greenhouse studies, Se was added to nodulated and nonnodulated hyperaccumulator and nonaccumulator Astragalus plants, followed by investigation of nitrogen (N)-Se relationships. Selenium speciation was also investigated, using x-ray microprobe analysis and liquid chromatography-mass spectrometry (LC-MS). KEY RESULTS: Nodulation enhanced biomass production and Se to S ratio in both hyperaccumulator and nonaccumulator plants. The hyperaccumulator contained more Se when nodulated, while the nonaccumulator contained less S when nodulated. Shoot [Se] was positively correlated with shoot N in Se-hyperaccumulator species, but not in nonhyperaccumulator species. The x-ray microprobe analysis showed that hyperaccumulators contain significantly higher amounts of organic Se than nonhyperaccumulators. LC-MS of A. bisulcatus leaves revealed that nodulated plants contained more γ-glutamyl-methylselenocysteine (γ-Glu-MeSeCys) than nonnodulated plants, while MeSeCys levels were similar. CONCLUSIONS: Root nodule mutualism positively affects Se hyperaccumulation in Astragalus. The microbial N supply particularly appears to contribute glutamate for the formation of γ-Glu-MeSeCys. Our results provide insight into the significance of symbiotic interactions in plant adaptation to edaphic conditions. Specifically, our findings illustrate that the importance of these relationships are not limited to alleviating macronutrient deficiencies.

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.

Selenium hyperaccumulation by Astragalus (Fabaceae) does not inhibit root nodule symbiosis.

PREMISE OF STUDY: A survey of the root-nodule symbiosis in Astragalus and its interaction with selenium (Se) has not been conducted before. Such studies can provide insight into how edaphic conditions modify symbiotic interactions and influence partner coevolution. In this paper plant-organ Se concentration ([Se]) was investigated to assess potential Se exposure to endophytes. • METHODS: Selenium distribution and molecular speciation of root nodules from Se-hyperaccumulators Astragalus bisulcatus, A. praelongus, and A. racemosus was determined by Se K-edge x-ray absorption spectroscopy. A series of greenhouse experiments were conducted to characterize the response of root-nodule symbiosis in Se-hyperaccumulators and nonhyperaccumulators. • KEY RESULTS: Nodules in three Se-hyperaccumulators (Astragalus crotalariae, A. praelongus, and A. preussii) are reported for the first time. Leaves, flowers, and fruits from Se-hyperaccumulators were routinely above the hyperaccumulator threshold (1,000 µg Se g(-1) DW), but root samples rarely contained that amount, and nodules never exceeded 110 µg Se g(-1) DW. Nodules from A. bisulcatus, A. praelongus, and A. racemosus had Se throughout, with a majority stored in C-Se-C form. Finally, an evaluation of nodulation in Se-hyperaccumulators and nonhyperaccumulators indicated that there was no nodulation inhibition because of plant Se tolerance. Rather, we found that in Se-hyperaccumulators higher levels of Se treatment (up to 100 µM Se) corresponded with higher nodule counts, indicating a potential role for dinitrogen fixation in Se-hyperaccumulation. The effect was not found in nonhyperaccumulators. • CONCLUSIONS: As the evolution of Se hyperaccumulation in Astragalus developed, root-nodule symbiosis may have played an integral role.

Molybdenum accumulation, tolerance and molybdenum-selenium-sulfur interactions in Astragalus selenium hyperaccumulator and nonaccumulator species.

Some species hyperaccumulate selenium (Se) upwards of 0.1% of dry weight. This study addressed whether Se hyperaccumulators also accumulate and tolerate more molybdenum (Mo). A field survey revealed on average 2-fold higher Mo levels in three hyperaccumulator Astragali compared to three nonaccumulator Astragali, which were not significantly different. Next, a controlled study was performed where hyperaccumulators Astragalus racemosus and Astragalus bisulcatus were compared with nonaccumulators Astragalus drummondii and Astragalus convallarius for Mo accumulation and tolerance, alone or in the presence of Se. When grown on agar media with 0, 12, 24 or 48 mg L(-1) molybdate and/or 0, 1.6 or 3.2 mg L(-1) selenate, all species decreased in biomass with increasing Mo supply. Selenium did not impact biomass at the supplied levels. All Astragali accumulated Mo upwards of 0.1% of dry weight. Selenium levels were up to 0.08% in Astragalus racemosus and 0.04% Se in the other species. Overall, there was no correlation between Se hyperaccumulation and Mo accumulation capacity. However, the hyperaccumulators and nonaccumulators differed in some respects. While none of the species had a higher tissue Mo to sulfur (S) ratio than the growth medium, nonaccumulators had a higher Mo/S ratio than hyperaccumulators. Also, while molybdate and selenate reduced S accumulation in nonaccumulators, it did not in hyperaccumulators. Furthermore, A. racemosus had a higher Se/S ratio than its medium, while the other species did not. Additionally, Mo and Se treatment affected S levels in nonaccumulators, but not in hyperaccumulators. In conclusion, there is no evidence of a link between Se and Mo accumulation and tolerance in Astragalus. Sulfate transporters in hyperaccumulating Astragali appear to have higher sulfate specificity over other oxyanions, compared to nonaccumulators, and A. racemosus may have a transporter with enhanced selenate specificity relative to sulfate or molybdate.