Field-collected, young plants of Ni hyperaccumulator Stackhousia tryonii, grown in a glasshouse for 20 weeks, were exposed to low- (available Ni concentration in the native serpentine soil, i.e. 60 microg g(-1) dry soil) and high- (external application of 1000 ppm) Ni concentrations in the substrate. Nickel concentration in the freeze-dried leaf tissues increased from 3700 microg g(-1) to 13 700 microg g(-1) with soil Ni supplementation, of which >60% was extracted with dilute acid (0.025 M HCl). Nickel supplementation also elicited a 575%, 211%, and 37% increase in the final concentrations of oxalic, citric, and malic acids, respectively, in leaf tissues. Malic acid was the dominant organic acid, followed by citric and oxalic acids. The molar ratio of Ni to malic acid was 1.0, consistent with a role for malate as a ligand for Ni in hyperaccumulating plants, supporting detoxification/transport and storage of this heavy metal in S. tryonii. The total amino acid concentrations in the xylem sap did not change with Ni supplementation (21.7+/-3.7 mM and 17.9+/-5 mM, respectively, for low- and high-nickel-treated plants). Glutamine was the major amino acid in both the low- and high-Ni-treated plants. The concentration of glutamine decreased by >60%, with a corresponding increase in alanine, aspartic acid, and glutamic acid, on exposure to high Ni. A role of amino acids in Ni complexation and transport in S. tryonii is not immediately apparent.

Stackhousia tryonii Bailey is one of the three nickel hyperaccumulators reported from Australia. It is a rare, herbaceous plant that accumulates (Ni) both in leaf and stem tissues. Localisation of Ni in leaf and stem tissues of S. tryonii was studied using two micro-analytical techniques, energy dispersive X-ray spectrometry (EDXS) and micro-proton-induced X-ray emission spectrometry (micro-PIXE). Dimethylglyoxime complexation of Ni was also visualised by bright- and dark-field microscopy, but this technique was considered to create artefacts in the distribution of Ni. Energy dispersive X-ray spectrometric analysis indicated that guard cells possessed a lower Ni concentration than epidermal cells, and that epidermal cells and vascular tissue contained higher levels of Ni than mesophyll, as reported for other Ni hyperaccumulators. The highest Ni concentration was recorded (PIXE quantitative point analysis) in the epidermal cells and vascular tissue (5400 μg g–1 DW), approximately double that recorded in palisade cells (2500 μg g–1 DW). However, concentrations were variable within these tissues, explaining, in part, the similarity between average Ni concentrations of these tissues (as estimated by region selection mode). Stem tissues showed a similar distribution pattern as leaves, with relatively low Ni concentration in the pith (central) region. The majority of Ni (73–85% for leaves; 80–92% for stem) was extracted from freeze-dried sections by water extraction, suggesting that this metal is present in a highly soluble and mobile form in the leaf and stem tissues of S. tryonii.

Summary• Metal concentrations within reproductive tissues of metallophytes are rarely reported. Here, the spatial distribution of nickel (Ni) within the fruits (seeds) of the Ni hyperaccumulator Stackhousia tryonii was investigated.• Two microanalytical techniques, energy dispersive x-ray spectrometry (EDXS) and nuclear microprobe (micro-proton-induced x-ray emission spectrometry; micro-PIXE) were employed for qualitative and quantitative assessment, respectively, of localized Ni, within the fruits of S. tryonii . The results were compared with quantitative analysis made using inductively coupled plasma-optical emission spectrometry (ICP-OES).• Nickel analysis made using micro-PIXE was consistent with bulk (ICP-OES) analysis (at 1800 µg g − 1 d. wt), however, a beam resolution of approx. 2 × 2 µm 2 allowed tissue localization. Nickel was partitioned to the fruit wall (pericarp) (4433 µg g − 1 ), while endospermic and cotyledonary tissues possessed little Ni (309 and 182 µg g − 1 d. wt, respectively).• This distribution is consistent with the interpretation that principal pathway of Ni movement within the fruit is symplastic rather than apoplastic (as the filial generation lacks symplastic connection with the parent).

The hypothesis that hyperaccumulation of certain metals in plants may play a role in osmotic adjustment under water stress (drought) was tested in the context of nickel hyperaccumulator Stackhousia tryonii. Field-collected mature plants of S. tryonii, grown in native ultramafic soil, were pruned to soil level and the re-growth exposed to five levels of water stress (20, 40, 60, 80 and 100% field capacity; FC) for 20 weeks. Water stress had significant (P<0.05) influence on growth (biomass), water potential and shoot Ni concentrations, with progressively more impact as water stress was increased from 80 to 40% FC. Shoot Ni concentration increased significantly from 3,400 microg g(-1) dry weight (at 100% FC) to 9,400 microg g(-1) dry weight (at 20% FC). Assuming that Ni is uniformly distributed through the shoot tissue, the Ni concentration could account for 100% at the 80 and 60% FC conditions, and 50% at the 40 and 20% FC conditions of plant osmotic regulation. The results are consistent with a role of Ni in osmotic adjustment and protection of S. tryonii plants against drought.