Distribution of excitation energy in photosynthesis: a study of heat loss, photo chemical activity and fluorescence emission in intact calls of cyanobacterium.

Photosynthetic state transitions were investigated in the cyanobacterium Synechococcus sp. PCC 6301 by studying fluorescence emission, heat loss, and PS I activity in intact cells brought to state 1 and state 2. 77K fluorescence emission spectra were modelled with a sum of 6 components corresponding to PBS, PS II, and PS I emissions. The modelled data showed a large decrease in PS II fluorescence accompanied with a small increase in the PS I fluorescence upon transition to state 2 for excitation wavelengths absorbed by both PBS and ChI ll.. The fluorescence changes seen with ChI .a. excitations do not support the predictions of the mobile PBS model of state transition in PBS-containing organisms. Measurements of heat loss from intact cells in the two states were similar for both ChI it. and PBS excitations over three orders of magnitude of laser flash intensity. This suggests that the PBS does not become decoupled from PS II in state 2 as proposed by the PBS detachment model of state transition in PBS-containing organisms. PS I activity measurements done on intact cells showed no difference in the two states, in contrast with the predictions of all of the existing models of state transitions. Based on these results a model for state transition In PBScontaining organisms is proposed, with a PS II photoprotectory function.

Synechococcus sp. PCC 7002 CpcB lyase null mutations alter phycocyanin chromophore function and, consequently, affect the redistribution of excitation energy via the light state transition /

Phycobilisomes are the major light harvesting complexes for cyanobacteria and phycocyanin is the primary phycobiliprotein of the phycobilisome rod. The phycocyanobilin lyases responsible for chromophorylating the phycocyanin p subunit (CpcB) have been recently identified in the cyanobacterium Synechococcus sp. PCC 7002. Surprisingly, mutants missing the CpcB lyases were nevertheless capable of producing pigmented phycocyanin. 10K absorbance measurements revealed that the energy states of the p phycocyanin chromophores were only subtly shifted; however, 77K steady state fluorescence emission spectroscopy showed excitation energy transfer involving the targeted chromophores to be highly disrupted. Such evidence suggests that phycobilin orientation within the binding domain is specifically modified. We hypothesized that alternate, less specific lyases are able to act on the p binding sites. A phycocyanin linker-polypeptide deficient mutant was similarly characterized. The light state transition, a short term adaptation of the photosynthetic light harvesting apparatus resulting in the redistribution of excitation energy among the photo systems, was shown to be dominated by the reallocation of phycocyanin-absorbed excitation energy. Treatment with a high M phosphate buffer effectively prevented the redistribution of both chlorophyll a- and phycobilisome- absorbed excitation energy, suggesting that the two effects are not strictly independent. The mutant strains required a larger redistribution of excitation energy between light states, perhaps to compensate for their loss in phycobilisome antenna function.

A relationship between photosynthesis and translocation in plants stressed by ionizing radiation

Since previous investigations have shown that low levels of ionizing radiation can induce a reduction in the rates of apparent photosynthesis and in the magnitude of photoassimilated l4C exported out of a leaf, the present studies were designed and conducted to determine the relationship, if any, between the radiation effects on these two physiological processes. The experiments were particularly designed to determine if the radiation-induced reduction in export is the result of the reduction in photosynthesis and hence availability of materials for translocation or the result of a reduction in the amount of energy available for the vein loading process. This study has shown that the radiation-induced reduction in l4C export out of a leaf is likely related to a loss of energy available for the vein loading process rather than a reduction in the supply of materials available for export due to reduced C02 uptake. The process of photophosphorylation was shown to be reduced by exposure to radiation to an extent similar to the reduction in the export of l4C which was also observed. Both of these processes returned to their pre-irradiation rates 120 minutes following radiatruon exposure. The rate of photosynthetic C02 uptake was also reduced by radiation exposur~ howeve~ this process did not return to the control level nor was the extent of reduction as large as observed for photophosphorylation and photoassimilate export. The observed relationship between the reductions of export and photoph~sphorylation pointed to the utilization of photosynthetically produced ATP in the vein loading process. The radiation-induced reduction in the export of l4C was observed at the highest light intensity used in this study which would also imply the involvement of the photophosphorylation process as an energy seurce for vein loading. The lack of radiation-induced reduction in export at low light intensities was interpreted as being due to the utilization of respiratory derived ATP, a process known to be insensitive to radiation at the levels used in this study, as the energy source for the vein loading process. Studies using plants not stressed by radiation showed that there was an increase in export of 14C with higher light intensities. In summary, the data has been interpreted as showing that at high light intensities the ATP, produced by photophosphorylation, is available for use in the vein loading process. The site of ATP utilization could not be determined from the data obtained in this study but possible sites have been indicated from the work done by other physiologists and are discussed in the thesis.