Carotenoid importance for membrane organization of NADPH protochlorophyllide oxidoreductase (POR) was studied by comparing interaction of two membrane fluorescent probes with proteins in prolamellar bodies isolated from norflurazon-treated wheat plants (cdPLBs) to those isolated form plants with normal carotenoid amount (oPLBs). The tryptophan fluorescence quenching by 1-anilino-8-naphthalene sulfonate (attached to the surface of membrane lipid phase) and pyrene (situated deep into the fatty acid region of membrane lipids) was used to locate the position of POR molecules toward lipid phase, to analyze their supramolecular organization and the light-induced structural transitions. Our results showed that the pigment-protein complexes of cdPLBs were larger than those of oPLBs. Upon flash irradiation the aggregates of both types of PLB dissociated into smaller units but in cdPLBs this process was accompanied by reorientation of the POR molecules closer to the lipid surface and/or dissociation from the lipids. These results revealed that carotenoid deficiency led to a looser attachment of POR to the lipid phase and its early (in comparison with oPLBs) dissociation from the membranes during the light-induced transformation of cdPLBs. This might be one of the reasons for the inability of carotenoid-deficient plants to form functional plastids.