Abstract
A simple analytical procedure for the analysis of the chlorophyll protein complex was developed using two step (Trlton X-100 → SDS) electrophoresis instead of chlorophyll isolation by sucrose or Percoll density gradient centrifugation. Triton X-100 (polyoxyethyleneglycol p-t-octylphenyl ether) at a 40% concentration dissolved effectively water insoluble proteins from the thylakoid membrane including the chlorophyll protein cornplex. The chlorophyll protein complex appeared as closely overlapping green bands and the bond between protein and chlorophyll was not destroyed after separation by polyacrylamide gel electrophoresis (PAGE) using 3% Triton X-100 in a 7% Davis buffer system. Stripes in the polypeptide pattern of these bands were cut horizontally from the gel, treated with SDS (sodium dodecyl sulfate) and then again subjected to SDS-PAGE. The polypeptide patterns of the chlorophyll protein complex of petunia, chrysanthemum, 60 varieties of rice (Oryza sativa L.) and 11 Oryza species were compared. In the genus Oryza, all the varieties and species showed a similar pattern except for some Oryza species i.e. O, alta (CD genome), O. latifolia (CD), and O. brachyantha (F). Moreover, there was no difference in the polypeptide patterns obtained with this method when compared to the conventional method.
