During senescence of flowering plants, only breakdown products derived from
chlorophyll a were detected although b disappears, too (Matile et al., 199
6, Plant Physiol 112: 1403-1409). We investigated the possibility of chloro
phyll b reduction during dark-induced senescence of barley (Hordeum vulgare
L.) leaves. Plastids isolated From senescing leaves were lysed and incubat
ed with NADPH. We found 7(1)-hydroxy-chlorophyll a, 7(1)-hydroxy-chlorophyl
lide ct, and, after incubation with Zn-pheophorbide b, also Zn-7(1)-hydroxy
-pheophorbide a, indicating activity of chlorophyll(ide) b reductase. The h
ighest activity was found at day 2 of senescence when chlorophyll breakdown
reached its highest rate. Chlorophyllase reached its highest activity unde
r the same conditions only at days 4-6 of senescence. Based on the chloroph
yll b reductase activity of plastids at day 2.5 of senescence (=100%), the
bulk of activity (83%) was found in the thylakoids and only traces (5%) in
the envelope fraction. Chlorophyll b reduction is considered to be an early
and obligatory step of chlorophyll b breakdown.