Browsing by Author "Clark, Gregory Thomas"
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- ItemPhosphate absorption by Arabidopsis thaliana : the effects of phosphorus nutritional status : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Plant Biology and Biotechnology at Massey University, Palmerston North, New Zealand(Massey University, 1997) Clark, Gregory ThomasThe effect of phosphorus nutritional status on phosphate uptake within the concentration range of the high affinity uptake mechanism, and subsequent translocation to the shoot was investigated in the plant species Arabidopsis thaliana. Plants of different nutritional status were generated by exposure to different set phosphate concentrations throughout an aseptic hydroponic growing period. Alternatively phosphorus deficiency was induced by growth at high concentrations of phosphate followed by a period of 5 days in phosphate-free hydroponic solution. In effect these growth conditions resulted in plants of distinguishable phenotypic character with respect to phosphate absorption, phosphate translocation, arsenate sensitivity and root-shoot ratio. To determine absorption kinetics nutrient depletion trials were carried out in which phosphate uptake was measured by monitoring the loss of phosphate from depletion solutions of set initial phosphate concentration to which the root systems of intact plants were exposed. Km and Vmax kinetic parameters were calculated from the depletion trial data using the software package "Igor Pro". Influx and net phosphate uptake was determined by setting the initial phosphate concentration of the depletion trials using either 32 P labelled KH 2 P0 4 or non-labelled KH 2 P0 4 respectively. Radioactivity was measured by counting the Cerenkov radiation in a scintillation counter. Non-labelled phosphate depletion was measured by either spectrophotometric assay or ion chromatography. To asses the effect of the phosphate analogue arsenate on phosphate influx, 32 P labelled phosphate uptake was measured with arsenate (KH 2 AsO 4 ) present in the depletion solution at a concentration of 20 µM. Phosphate translocation was determined by counting the Cerenkov radiation in the roots and shoots separately of plants that had been exposed to the 32 P labelled depletion solutions. Under the conditions of this project, phosphorus deficient plants exhibited alterations in the kinetic parameters Km and Vmax for phosphate uptake that were dependent on how the deficiency was induced. For plants that were grown continuously at low phosphate concentrations Km was decreased without a concomitant change in Vmax. For plants that were grown at high concentrations of phosphate followed by a 5 day period of phosphate starvation, a significant increase in Vmax was recorded without an associated change to Km. Phosphate uptake was found to be severely inhibited by the presence of arsenate in the depletion solution. Greatest inhibition however was found not to occur at the level of absorption into the plant root system but rather appeared to be at a site involved in phosphate loading into the xylem. Inhibition at this site was also found to be greatest in low phosphorus status plants. From these results it is suggested that plants of low phosphorus status possess high affinity phosphate xylem loading mechanisms, induced under conditions of phosphorus deficiency, which have a greater susceptibility to arsenate competitive inhibition and toxicity than equivalent xylem loading mechanisms in high phosphorus status plants.
- ItemTissue-specific responses to water deficit in the New Zealand xerophytic tussock species Festuca novae-zelandiae : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Plant Biology at Massey University, Palmerston North, New Zealand(Massey University, 2005) Clark, Gregory ThomasFestuca novae-zelandiae (Hack.) Cockayne is an endemic New Zealand perennial tussock forming grass of the family Poaceae. Morphologically F. novae-zelandiae exhibits a number of leaf adaptations associated with dehydration postponement as reflected in the climatic distribution of this species and its occurrence as a physiognomic dominant grass in semi arid short-tussock grasslands. Biochemical studies into the drought tolerance of this species have indicated the occurrence of tissue specific responses with respect to abscisic acid (ABA) and proline accumulation and protein turnover suggestive of a preferential protection of the tiller base and associated meristematic zones at the expense of lamina tissues. Further tissue specific biochemical responses to water-deficit stress in F. novae-zelandiae have been investigated. Changes in water-soluble carbohydrates (WSC) were monitoured over a 49-day dry-down period (decline in soil water content from 30% to 4%) in consecutive leaf segments comprising the leaf base (meristem region), elongation zone, the enclosed and exposed lamina, as well as basal sheath segments from the two next oldest leaves. In fully hydrated leaf tissues polymers of fructose (fructans) were the main WSC present and were mainly low molecular weight fructans of the inulin and neokestose series with the average degree of polymerization (DP) of fructan pools from 6 to 9. The highest fructan concentrations were present towards the leaf base. Fructan concentrations decreased over the course of the dry-down, although remained significantly higher in the meristem region of the tiller base with respect to any other tissue, until tissue water content fell below 45%. By day 49 of the dry-down period, the average DP of the fructan pool in tissues was from 3 to 5. Sucrose content increased in each tissue during the course of the dry-down, and was highest at the leaf base, where a concentration of 200 µmol g-1 dry weight was measured after 49 days of dry-down. The negative correlation between fructan and sucrose content, which indicates an inter-conversion dependent on tissue water content, suggests that, in this specises, fructans serve as a carbohydrate storage pool, while sucrose stabilises the meristem during extreme water deficit.