Mechanisms regulating the thermal acclimation of dark respiration in snow tussock and ryegrass
Degree GrantorUniversity of Canterbury
Degree NameMaster of Science
The aim of this research was to identify the mechanisms that underpin changes in respiratory capacity during acclimation to temperature. Dark respiration, enzyme activities and leaf ultrastructure were measured from ryegrass (Lolium perenne) in controlled environmental chambers and two species of native grass (Chionochloa rubra & C. pallens) growing at different altitudinal ranges on Mount Hutt, Canterbury, New Zealand. The overall hypothesis was that the changes in both mitochondrial numbers and enzyme activity underpin the greater respiratory capacity observed in response to decreasing temperatures. Gas exchange measurements were carried out to measure rates of dark respiration (Rd) in leaves of both ryegrass and tussocks. Respiratory homeostasis (full acclimation) was achieved in ryegrass leaves but only partial acclimation in both species of tussock plants. Dark respiration rates for warm-grown ryegrass were greatly reduced compared to cool-grown grasses. Rd was lower for C. rubra growing at the base of the mountain (450m) compared to plants at a higher altitude (1060m). The dark respiration rates were also lower for C. pallens growing at 1070m than at 1600m. When comparing Rd between high and low altitude plants, it was significantly lower in low altitude plants at 450m than at 1600m. Oxygen consumption was measured in intact leaves and roots, crude mitochondria and isolated mitochondria from ryegrass to investigate whether a change in respiratory capacity was involved with changes in Rd. Mitochondrial respiratory capacity was slightly reduced in warm leaves and roots (not significantly). The respiratory capacity results from isolated mitochondria for C. rubra (at 450m and 1060m) and C. pallens (at 1070m and 1600m) were consistent with the hypothesis that plants from warm sites have lower respiratory capacity in comparison to plants from cool sites. Based on these results and those of previous studies, it was concluded that respiratory flux for any given temperature is not simply determined by maximal capacities of the respiratory apparatus but rather a combination of the availability of substrate supply, the demand for respiratory products (i.e. ATP) and/or the maximal capacity of respiratory enzymes. Utilizing transmission electron micrographs, it was found that mitochondria were significantly less abundant in warm-grown than cool-grown ryegrass mesophyll cells. Mitochondria dimensions increased slightly between the cool and warm treatment. At lower altitudes (C. rubra), there was a significant decrease in mitochondria numbers with decreasing elevation. At higher altitudes (C. pallens), there was no noticeable change in mitochondria numbers between 1070m and 1600m. It was concluded that mitochondrial abundance for the controlled and field experiments, and mitochondrial sizes in the field, were associated with changes in Rd. The maximal activities of fumarase and succinate dehydrogenase (SDH) in isolated mitochondria from leaves of ryegrass and tussock were measured spectrophotometrically. The results in the controlled experiment indicate that enzymes other than fumarase and SDH could be responsible for the increased respiratory capacity observed in cold acclimated leaves of ryegrass. However, fumarase maximal activity was significantly reduced in C. rubra at low altitude compared with C. pallens growing at high altitude - this suggests that it may be involved in the differences in respiratory capacity and Rd between the two sites. Succinate dehydrogenase did not differ significantly in response to altitude. The large difference between the two field sites for fumarase activity is comparable to the large difference in Rd and reduction in mitochondrial abundance and dimensions seen between the two sites. This supports the overall hypothesis that cool-grown plants keep up with energy demands at low temperatures by increasing enzyme concentrations/capacity. The results of this study are supportive of the hypothesis that growth in low altitudes and warm conditions will result in the reduction of Rd as a consequence of: (1) temperature sensitivity of the respiratory apparatus, resulting in the reduction of the respiratory capacities of mitochondria; (2) a reduction in mitochondria size and numbers; and as a consequence of this (3) a reduction in the activities of mitochondrial enzymes. However, these responses are species specific and vary according to the range of temperatures experienced by plants in the field and controlled environments.