Formation of Chlorophyll b, and the Fluorescence Properties and Photochemical Activities of Isolated Plastids from Greening Pea Seedlings

Chlorophyll b was first detectable after 10 minutes of illumination of etiolated pea seedlings (Pisum sativum L. var Greenfeast) with continuous white light. The chlorophyll a/b ratio decreased from 300 at 10 minutes to 15 after 1 hour. There was little change in the chlorophyll a/b ratio between 1 and 2 hours, and it declined to 3 between 2 and 5 hours of illumination. In red light, the time courses of total chlorophyll synthesis and chlorophyll a/b ratio were similar to those in white light for the first 5 hours of illumination. But with increasing time of illumination with red light, there was an increase in the chlorophyll a/b ratio to 7 after 30 hours. Illumination with white light of very low intensity also gave high chlorophyll a/b ratios. Seedlings which had been illuminated for varying periods and then returned to darkness always showed an increase in chlorophyll a/b ratio during the dark period. It is concluded that the synthesis of chlorophyll b is controlled by light.The onset of the transfer of excitation energy from carotenoids to chlorophyll a correlated with the appearance of chlorophyll b. The detection of chlorophyll b and the onset of energy transfer from carotenoids to chlorophyll a preceded by several hours the appearance of Hill activity in the isolated pea plastids. The detection of the large fluorescence emission band at 735 nm at 77 K also preceded the observation of Hill activity, but it correlated with the phase of rapid chlorophyll synthesis. There was a good correlation between Hill activity and the formation of grana. The fluorescence kinetics which are characteristic of mature chloroplasts appeared considerably later than grana or the observation of Hill activity.It is concluded that the formation of photosynthetic membranes and the assembly of the photosystems is not a single step process, at least in the initial stages of chloroplast development from the etioplast.