Photochemical reactions in dehydrated photosynthetic organisms, leaves, chloroplasts and photosystem II particles: reversible reduction of pheophytin and chlorophyll and oxidation of β-carotene

Abstract

Abstract Photoreactions of dehydrated leaves, isolated broken chloroplasts and PSII membrane fragments of spinach ( Spinacia oleracea ) were studied at different air humidities and compared with photoreactions of dry fronds of a fern, Polypodium vulgare , and a dry lichen, Parmelia sulcata , which in contrast to spinach are insensitive to photoinactivation in the dry state. Even in very dry air, P700 in the reaction center of photosystem I of dry leaves was oxidized, and the primary quinone acceptor Q A in the reaction center of photosystem II was photoreduced by low light. These reactions were only very slowly reversed in the dark and saturated under low light intensity. Light-minus-dark difference absorption spectra of the dry leaves, isolated chloroplasts and PSII membrane fragments measured at higher light intensities revealed absorbance changes of β-carotene at 500 nm (light-dependent bleaching) and 980 nm (light-dependent band formation) and bleaching of chlorophyll at 436 and 680 nm with appearance of bands at 450 and 800 nm. Decrease of chlorophyll fluorescence upon strong illumination indicated photoaccumulation of a quencher. All these changes were kinetically related and readily reversible. They are interpreted to show light-induced oxidation of β-carotene (Car) and reduction of chlorophyll-680 (Chl-680) in the reaction center of photosystem II of the dried leaves, chloroplasts and photosystem II particles. The fluorescence quencher was suggested to be Chl-680 − or Car + in close proximity to P680, the primary electron donor. Appreciable photoaccumulation of reduced pheophytin was only observed in dry leaves after Q A reduction had been lost during heat treatment of hydrated leaves prior to dehydration. The observations are interpreted to show light-dependent cyclic electron flow within the reaction center of photosystem II in which Chl-680 (or Pheo) is reduced by P680 * and Car is oxidized by P680 + with consequent recombination of Car + and Chl-680 − (or Pheo − ). Cyclic electron flow is thought to be moderately photoprotective by reducing the concentration of the strong oxidant P680 + . Nevertheless, prolonged strong illumination caused considerable photodamage in the dry spinach leaves, much less damage in dry fern fronds and no damage in the dry lichen in which distinct photoreactions were not expressed at light intensities which were effective in dry leaves and fern fronds.