Abstract
Fremyella diplosiphon is a freshwater, filamentous cyanobacterium that exhibits light-dependent regulation of photosynthetic pigment accumulation and cellular and filament morphologies in a well-known process known as complementary chromatic adaptation (CCA). One of the techniques used to investigate the molecular bases of distinct aspects of CCA is confocal laser scanning microscopy (CLSM). CLSM capitalizes on the autofluorescent properties of cyanobacterial phycobiliproteins and chlorophyll a. We employed CLSM to perform spectral scanning analyses of F. diplosiphon strains grown under distinct light conditions. We report optimized utilization of CLSM to elucidate the molecular basis of the photoregulation of pigment accumulation and morphological responses in F. diplosiphon.
Highlights
Photosynthetic organisms depend upon light for carbon fixation and production of reductant
To understand the dynamic molecular processes used for adapting to changes in ambient light, we are investigating the functions of biliproteins, light-absorbing pigments centrally involved in both photosynthesis and the regulation of photomorphogenesis in cyanobacteria, algae, and plants
The freshwater filamentous cyanobacterium Fremyella diplosiphon exhibits a well-known light-dependent acclimation process known as complementary chromatic adaptation (CCA)
Summary
Photosynthetic organisms depend upon light for carbon fixation and production of reductant. During CCA, F. diplosiphon exhibits light-dependent changes in the protein composition of the photosynthetic light-harvesting complexes, i.e., phycobilisomes (PBSs), and cell and filament morphologies [1]. The PBSs are attached to the thylakoid membrane and transfer energy to chlorophyll a (Chl a) in the reaction centers of photosynthetic photosystems (reviewed by [5]) By nature of their spectral properties, PBPs are highly autofluorescent proteins. We investigated differential laser excitation and collection of emission data to optimize PBP detection and localization, as well as cellular and filament morphology analysis These studies resulted in the identification of conditions that allow detailed investigations into the molecular bases of the regulation of distinct aspects of CCA through comparative CLSM analyses of wild-type and mutant strains of F. diplosiphon
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