Comparative studies of nickel,cobalt, and copper uptake by some nickel hyperaccumulators of the genus Alyssum

The uptake of Ni, Co, and Cu by the nickel hyperaccumulator Alyssum troodii Boiss and the non-accumulator Aurinia saxatilis (L.) Desv. were studied in pot trials using artificial rooting media with varying concentrations of the metals added as soluble salts, singly and in combination. The ability of five other Ni hyperaccumulating species of Alyssum to hyperaccumulate Co was also investigated.Leaves and stems of A. troodii accumulated Ni to almost the same extent (8000–10 000 g g^-1). In roots, the highest Ni concentration was 2000 g g^-1. In leaves of Au. saxatilis, the maximum Ni concentration was only 380 g g^-1 and the level in roots was even lower.In media containing Co, the maximum concentration of this element in A. troodii (2325 g g^-1) was ten times higher than in the non-accumulator species. Slightly less Co was found in stems and roots of both species. Among the other Ni hyperaccumulators, the maximum concentration of Co in leaves ranged from about 1000–8000 g g^-1.Copper concentrations were the same in all organs of both species when they were grown in copper-rich media and were in the range 40–80 g g^-1, showing that neither plant was capable of taking up Cu at levels comparable to those of Ni and Co.When both plants were grown in media containing equal amounts of both Co and Ni, the Co concentrations in plant organs were the same as for specimens grown in media containing Co only. However, the Ni levels were lower in both species. Uptake of Co therefore appeared to suppress Ni uptake.Pot trials showed that the order of tolerance was Ni>Cu>Co for A. troodii and Ni>CoCu for Au. saxatilis, whereas the seedling tests showed the order to be Co>Ni>Cu. At metal concentrations 10 000 g g^-1, the overall tolerance of A. troodii was greater than that of Au. saxatilis which exhibited equally low tolerance to Ni and Cu.We conclude that in A. troodii, A. corsicum Duby, A. heldreichii Hausskn., A. murale Waldstein & Kitaibel, A. pintodasilvae T.R. Dudley, and A. tenium Hálácsy, Ni tolerance and hyperaccumulation conveys the same character towards Co. This behaviour should be investigated in other hyperaccumulators of Ni and/or Co.     

Localisation of nickel and mineral nutrients Ca,K, Fe,Mg by Scanning Electron Microscopy microanalysis in tissues of the nickel-hyperaccumulator Alyssum bertolonii Desv. and the non-accumulator Alyssum montanum L.

Plants of the nickel-hyperaccumulator Alyssum bertolonii Desv. and of the non-accumulator A. montanum L. growing on a serpentine site in Tuscany, Italy, and plants of A. montanum from a nearby non-serpentine site were analysed for metal concentration and localisation. The leaves of A. bertolonii contained 160 times more nickel than those of A. montanum from the same site, thus demonstrating its hyperaccumulation capacity towards this metal. On the other hand, both species showed an inversion of the Ca/Mg ratio in their organs relative to the soil. Nickel localisation in plant tissues was examined by Scanning Electron Microanalysis (SEM/EDX). In A. bertolonii, a specific pattern of nickel distribution was detected, with the highest concentrations present in parenchyma and sclerenchyma cells for the roots; in the shoots, the highest amounts of nickel were found in the stem epidermis, the leaf epidermal surface, and the leaf trichome base. This particular nickel tissue distribution pattern was not found in the non-accumulator A. montanum growing on serpentine soil. Other mineral nutrients, namely Mg, Ca, K, Fe, instead, had a similar distribution in the two species. The A. montanum plants from the non-serpentine site had very low nickel levels in their tissues, and these were of the same magnitude as those found in A. bertolonii plants grown in a greenhouse on commercial horticultural soil with low nickel concentration. In A. bertolonii plants, the tissue-specific allocation patterns appeared to depend on the degree of nickel hyperaccumulation, which is, in turn, directly linked to the soil characteristics.     

Polyphyletic Alyssum cuneifolium (Brassicaceae) revisited: Morphological and genome size differentiation of recently recognized allopatric taxa

Alyssum cuneifolium has been recognized as a perennial alpine species growing in five isolated European mountain ranges: the Pyrenees, Western Alps, Apennines, Pirin Mts and Mt Smolikas. Recent molecular systematic studies revealed that the disjunct populations from distant mountains are not closely related and belong to five independent species: A. cacuminum (Spain, Pyrenees), A. cuneifolium (Italy, Apennines), A. flexicaule (France, Western Alps), A. pirinicum (Bulgaria, Pirin Mts), and A. spruneri (Greece, Mt Smolikas). The present study brings the thorough morphometric analysis of the segregated taxa. We found minor morphological differences between them. Whereas A. pirinicum can be clearly distinguished, the other taxa are recognizable only at the level of population means of investigated characters. The morphological similarity of these distantly related species is obviously the result of adaptation to similar high‐alpine scree habitats. It is not clear, however, whether this adaptation is environmentally controlled or whether it is also genetically fixed and whether it reflects parallel evolution towards similar morphotypes. The observed morphological patterns and their assumed correlation with environmental factors are discussed using examples from other Alyssum taxa. Three different ploidy levels have been reported for the species under study. In the present article, we examine variation in relative nuclear genome size. The Alpine and Pyrenean species have larger relative monoploid genome sizes than the Apennine and Balkan ones, probably reflecting the evolutionary history of the group. A nomenclatural account of the study species is presented, and lectotypes of A. cuneifolium and of two other names are selected.     

Hyperaccumulator Alyssum murale relies on a different metal storage mechanism for cobalt than for nickel

The nickel (Ni) hyperaccumulator Alyssum murale has been developed as a commercial crop for phytoremediation/phytomining Ni from metal-enriched soils. Here, metal co-tolerance, accumulation and localization were investigated for A. murale exposed to metal co-contaminants. A. murale was irrigated with Ni-enriched nutrient solutions containing basal or elevated concentrations of cobalt (Co) or zinc (Zn). Metal localization and elemental associations were investigated in situ with synchrotron X-ray microfluorescence (SXRF) and computed-microtomography (CMT). A. murale hyperaccumulated Ni and Co (> 1000 microg g(-1) dry weight) from mixed-metal systems. Zinc was not hyperaccumulated. Elevated Co or Zn concentrations did not alter Ni accumulation or localization. SXRF images showed uniform Ni distribution in leaves and preferential localization of Co near leaf tips/margins. CMT images revealed that leaf epidermal tissue was enriched with Ni but devoid of Co, that Co was localized in the apoplasm of leaf ground tissue and that Co was sequestered on leaf surfaces near the tips/margins. Cobalt-rich mineral precipitate(s) form on leaves of Co-treated A. murale. Specialized biochemical processes linked with Ni (hyper)tolerance in A. murale do not confer (hyper)tolerance to Co. A. murale relies on a different metal storage mechanism for Co (exocellular sequestration) than for Ni (vacuolar sequestration).     

Hyperaccumulation of nickel by two Alyssum species from the serpentine soils of Iran

Serpentine soils, which contain relatively high concentrations of nickel and some other metals, are the preferred substrate for some plants, especially those that accumulate Ni in their tissues. In temperate regions more Ni-hyperaccumulator plants are found in Alyssum than in any other genus. In this study, serpentine soils of two areas (Marivan and Dizaj) in the west/northwest of Iran and also perennial Alyssum plants growing on these soils were analyzed for Ni and some other metals. The highest concentrations of total metals in the soils of these areas for Ni, Cr, Co and Mn were 1,350, 265, 94 and 1,150 μg g⁻¹, respectively, while concentrations of Fe, Mg and Ca reached 3.55%, 16.8% and 0.585% respectively. The concentration of exchangeable Ni in these soils is up to 4.5 μg g⁻¹. In this study two Alyssum species, A. inflatum and A. longistylum, have been collected from Marivan and Dizaj, respectively. Analysis of leaf dry matter shows that they can contain up to 3,700 and 8,100 μg Ni g⁻¹, respectively. This is the first time that such high Ni concentrations have been found in these species. The concentrations of other metals determined in these species were in the normal range for serpentine plants, except for Ca, which was higher, up to 5.3% and 3.5%, respectively     

Degradation of Alyssum murale biomass in soil

The Ni-hyperaccumulating plant Alyssum murale accumulates exceptionally high concentrations of nickel in its aboveground biomass. The reasons for hyperaccumulation remain unproven; however, it has been proposed that elemental alelopathy might be important. High-Ni leaves shed by the plant may create a "toxic zone" around the plant where germination or growth of competing plants is inhibited. The efficacy of this argument will partially depend upon the rate at which leaves degrade in soil and free metals are released, and the subsequent rate at which metals are bound to soil constituents. To test the degradation of biomass of hyperaccumulators, A. murale was grown on both high- and low-Ni soils to achieve high- (12.0 g Ni/kg) and low- (0.445 g Ni/kg) Ni biomass. Shredded leaf and stem biomass were added to a serpentine soil from Oregon that was originally used to grow high-Ni biomass and a low-Ni control soil from Maryland. Biomass Ni was readily soluble and extractable, suggesting near immediate release as biomass was added to soil Extractable nickel in soil amended with biomass declined rapidly over time due to Ni binding in soil These results suggest that Ni released from biomass of Ni hyperaccumulators may significantly affect their immediate niche only for short periods of time soon after leaf fall, but repeated application may create high Ni levels under and around hyperaccumulators.     

Nickel and zinc hyperaccumulation by Alyssum murale and Thlaspi caerulescens (Brassicaceae) do not enhance survival and whole-plant growth under drought stress

Nickel and Zn hyperaccumulation by Alyssum murale and Thlaspi caerulescens bear substantial energetic costs and should confer benefits to the plant. This research determined whether metal hyperaccumulation can increase osmotic adjustment and resistance to water stress (drought). Alyssum murale and Thlaspi caerulescens treated with low or high concentrations of Ni or Zn were exposed to moderate (?0·4 MPa) and severe (?1·0 MPa) water stresses using aqueous polyethylene glycol. In the absence of metals both water deficits inhibited shoot growth. Nickel and Zn hyperaccumulation did not ameliorate growth inhibition by either level of water stress. The water stress did not induce major changes in shoot metal concentrations of these constitutive hyperaccumulators. Moreover, metal hyperaccumulation had minimal effects on the osmolality of leaf‐sap extracts, relative water content of the shoots, or rate of evapotranspiration. It is concluded that Ni or Zn hyperaccumulation does not augment whole‐plant capacity for drought resistance in A. murale and T. caerulescens.     

Identification of Glucosinolates in Seeds of Three Brassicaceae Species Known to Hyperaccumulate Heavy Metals

Plants from the Brassicaceae family are known to contain secondary metabolites called glucosinolates. Our goal was to establish by LC/MS the glucosinolate profile of seeds of three Brassicaceae species known to hyperaccumulate heavy metals. We investigated Alyssum fallacinum auct. non Hausskn ., Iberis intermedia Guers ., and Noccaea caerulescens (J. Presl & C. Presl ) F. K. Mey . Our results indicate that A. fallacinum seeds contain glucoiberin and glucoibervirin, which had not been previously identified in this plant. Furthermore, we report for the first time the presence of glucoiberin, glucoibervirin, glucotropaeolin, and sinigrin in I. intermedia. We have detected for the first time glucoconringiin in N. caerulescens. In addition, glucosinalbin, 4‐hydroxyglucobrassicin, and glucomoringin were also detected.     

Selection and Combustion of Ni-Hyperaccumulators for the Phytomining Process

Ni recovery from serpentine soils by phytomining has proved feasible. Phytomining involves the crop of hyperaccumulating plants with high Ni contents and the valorization of Ni by pyro or hydrometallurgical process. In order to evaluate the Ni content of different plants, we analyzed the organs of 14 hyperaccumulators from three genera: Alyssum, Leptoplax and Bornmuellera. The highest concentration was recorded in the leaves of Leptoplax (34.3 ± 0.7 mg g^?1 _DM). Additionally, we investigated biomass combustion which is the first step of the process we designed to obtain a nickel salt. We showed that temperature and duration were important parameters to ensure a good quality of ashes. At the bench scale, the best conditions were 550°C and 3 h. In this way, we obtained ashes in which Ni could reach 20 wt%. Biomass ashes can be considered as a bio-ore for recovering metal value.     

Improving the growth of Ni-hyperaccumulating plants in serpentine quarry tailings

Phytomining techniques based on metal-hyperaccumulating plants can be implemented in serpentine quarry wastes for Ni recovery. However, strategies must be developed to overcome the unfavourable plant growth conditions that these substrates present and to optimize Ni yields. In this study, the Ni hyperaccumulators Alyssum serpyllifolium, Alyssum inflatum, and Alyssum bracteatum were evaluated for their Ni phytoextraction efficiency from quarry tailings. Effects of two organic amendments, composted municipal sewage sludge and cow manure, on plant growth and physiological status and Ni removal were determined. Organic amendments were incorporated at two addition rates (5% and 20% w/w). The best-performing hyperaccumulators were A. inflatum and A. serpyllifolium. Organic amendments improved plant biomass production, photosynthetic efficiency and nutrition, but reduced shoot Ni concentrations. However, the stimulation in biomass resulted in significantly enhanced Ni yields. The most promising results were found using low addition rates and after manure incorporation.