The delayed luminescence (DL) of photosystem 2 (PS2) after infiltration of 7-d-old etiolated barley leaves with chlorophyllides (Chlide) a or b followed by 2.5 h dark incubation was studied. Chlide a caused a very weak DL of PS2 just at the beginning of irradiation and the intensity of this DL was not higher when the infiltration medium contained 2 mM of NADPH. Chlide b was a somewhat more efficient inducer of PS2 formation in the dark and NADPH enhanced this efficiency 4.5 times though it did not affect the amount of esterified Chlides. The photoconversion of endogenous Pchlide led to a much higher intensity of the DL in comparison with the infiltration of Chlides, while the total amount of chlorophyll (Chl) formed was almost unchanged. The use of Chlide b together with the acetone extract from green leaves, devoid of pigments, resulted in the DL intensity comparable with that observed after Pchlide photoconversion followed by 2.5 h incubation in the dark. Dark formation of active PS2 in etiolated leaves was shown for the first time. Thus the dark formation of active PS2 may require Chl b, NADPH, and some unidentified water-soluble factor(s), synthesized in the dark after a short irradiation of etiolated leaves and inherent in green leaves. and V. P. Domanskiï, L.I. Fradkin.