Abstract:Differences in the pigment and thylakoid membranes levels among the mutant barley 1832C, Chlorina-f2 was compared with a normal one by means of their absorption spectra and the 4th derivative spectra. Results showed that there was no absorption peaks for Chl b at 651 nm in red and 470 nm in blue regions in the absorption and the 4th derivative spectra in the mutant thylakoid membranes. At the same time there was no absorption peaks of Chl b at 645 nm and 455 nm in the absorption and the 4th derivative spectra in its acetone (80%) extract. Therefore, mutant barley 1832C was proved to be a new type of chlorophyll b-less mutant that could survive independently in nature.