Comparison of cytology and distribution of nickel in roots of Ni-hyperaccumulating and non-hyperaccumulating genotypes of Senecio coronatus

Abstract

Two genotypes of Senecio coronatus (Thunb.) Harv. (Asteraceae) growing on ultramafic outcrops were identified previously: a Ni hyperaccumulator and a non-hyperaccumulator. The aim of the present study was to investigate the cytology of the roots of both genotypes, their Ni content and tissue distribution, and to ascertain whether there was a cytological basis for the differential uptake of Ni. Light and fluorescence microscopy together with histochemical methods were used to study root cytology. X-ray microanalysis by means of a nuclear microprobe—particle-induced X-ray emission (PIXE) and proton backscattering (BS) techniques—was utilized to determine the concentration and distribution of Ni and other elements. Average concentration of Ni and distribution in roots differed significantly between hyperaccumulating and non-hyperaccumulating genotypes. Ni amount in the hyperaccumulating genotype was ca. 60 times higher in the older part of the root (1,760 μg g−1) and ca. 10 times higher (314 μg g−1) in the younger root hair region in comparison with the equivalent parts of the non-accumulating genotype where Ni amounts were 30 μg g−1. Ni distribution pattern was also different in both cases. Cytological differences were observed in the inner cortical region and exodermis of the roots. Distinct groups of specialized cells with an organelle-rich cytoplasm that produced copious numbers of spherical bodies occurred in the inner cortical region of the hyperaccumulator. Such distinct cell groups were absent from the inner cortex of the non-hyperaccumulator. Cortical cells here had a thin parietal cytoplasmic layer and produced fewer spherical bodies. In both genotypes the spherical structures were extruded from the cytoplasm into air spaces between the cells where they coalesced to form amorphous deposits, significantly larger and more abundant in the hyperaccumulator. Histochemical tests identified these deposits as a mixture of lipids, alkaloids and terpenoids. Specialized cells present in the inner cortex of the hyperaccumulating genotype demonstrated significant relative Ni depletion in comparison with the adjacent inner cortex and phloem. Casparian bands were identified in exodermal cell walls of both genotypes but the bands fluoresced more intensely in the non-accumulator suggesting differences in chemical composition and probably also a more efficient apoplastic barrier. This feature was correlated with the observed Ni distribution pattern. The highest Ni enrichment in the hyperaccumulating genotype occurred in the outer cortex; 20 times more than in the adjacent epidermis/exodermis in older portions of roots and 3 times more than in the epidermis/exodermis in younger root hair regions. In contrast, in the non-hyperaccumulating genotype, a higher concentration of nickel was found in the epidermis/exodermis compared to the outer cortex. The Ni ratio between the outer cortex and epidermis/exodermis was about 0.4 in the non-hyperaccumulator. Different cytological features exhibited by the genotypes may represent adaptive responses to the presence of high concentrations of Ni in the soil and subsequent differential uptake of Ni. Basic characteristics and elemental content of soil collected from the close vicinity of roots of two S. coronatus genotypes are reported.