Phosphorus nutrition of arsenate-tolerant and nontolerant phenotypes of velvetgrass

Velvetgrass (Holcus lanatus L.), also known as Yorkshire fog grass, has evolved tolerance to high levels of arsenate, and this adaptation involves reduced accumulation of arsenate through the suppression of the high affinity phosphate-arsenate uptake system. To determine the role of P nutrition in arsenate tolerance, inhibition kinetics of arsenate influx by phosphate were determined. The concentration of inhibitor required to reduce maximum influx (V(max)) by 50%, K₁, of phosphate inhibition of arsenate influx was 0.02 mol m^-3 in both tolerant and nontolerant clones. This was compared with the concentration where influx is 50% of maximum, a K(m), for arsenate influx of 0.6 mol m^-3 for tolerants and 0.025 mol m^-3 for nontolerants and, therefore, phosphate was much more effective at inhibiting arsenate influx in tolerant genotypes. The high affinity phosphate uptake system is inducible under low plant phosphate status, this increasing plant phosphate status should increase tolerance by decreasing arsenate influx. Root extension in arsenate solutions of tolerant and nontolerant tillers grown under differing phosphate nutritional regimes showed that indeed, increased plant P status increased the tolerance to arsenate of both tolerant and nontolerant clones. That plant P status increased tolerance again argues that P nutrition has a critical role in arsenate tolerance. To determine if short term flux and solution culture studies were relevant to As and P accumulation in soils, soil and plant material from a range of As contaminated sites were analyzed. As predicted from the short-term competition studies, P was accumulated preferentially to As in arsenate tolerant clones growing on mine spoil soils even when acid extractable arsenate in the soils was much greater than acid extractable phosphate. Though phosphate was much more efficient at competing with arsenate for uptake, plants growing on arsenate contaminated land still accumulated considerable amounts of As. Plants from the differing habitats showed large variation in plant phosphate status, pasture plants having much higher P levels than plants growing on the most contaminated mine spoil soils. The selectivity of the phosphate-arsenate uptake system for phosphate compared with arsenate, coupled with the suppression of this uptake system enabled tolerant clones of the grass velvetgrass to grow on soils that were highly contaminated with arsenate and deficient in phosphate.

The diversity of extradiol dioxygenase (edo) genes in cresol degrading rhodococci from a creosote-contaminated site that express a wide range of degradative abilities.

Analysis of the bacterial population of soil surface samples from a creosote-contaminated site showed that up to 50% of the culturable micro-organisms detected were able to utilise a mixture of cresols. From fifty different microbial isolates fourteen that could utilise more than one cresol isomer were selected and identified by 16S rRNA analysis. Eight isolates were Rhodococcus strains and six were Pseudomonas strains. In general, the Rhodococcus strains exhibited a broader growth substrate range than the Pseudomonas strains. The distribution of various extradiol dioxygenase (edo) genes, previously associated with aromatic compound degradation in rhodococci, was determined for the Rhodococcus strains by PCR detection and Southern-blot hybridization. One strain, Rhodococcus sp. I1 exhibited the broadest growth substrate range and possessed five different edo genes. Gene disruption experiments indicated that two genes (edoC and edoD) were associated with isopropylbenzene and naphthalene catabolism respectively. The other Rhodococcus strains also possessed some of the edo genes and one (edoB) was present in all of the Rhodococcus strains analysed. None of the rhodococcal edo genes analysed were present in the Pseudomonas strains isolated from the site. It was concluded that individual strains of Rhodococcus possess a wide degradative ability and may be very important in the degradation of complex mixtures of substrates found in creosote.