Spermine and lutein quench chlorophyll fluorescence in isolated PSII antenna complexes.

Non photochemical quenching is a spontaneous mechanism that protects plants and algae from photodamage. In the last two decades, carotenoids through the xanthophylls cycle have been proposed to play a key role in quenching of chlorophyll. More recently, the involvement of endogenous polyamines in energy-dependent component of non photochemical quenching has been suggested by several research groups. In the present contribution, the combined effect of spermine and the xanthophylls, zeaxanthin and lutein on the fluorescence of antenna complexes of photosystem II was tested in vitro. Lutein caused significant quenching on trimeric and monomeric antenna complexes, whereas zeaxanthin under our experimental conditions had negligible effect. Spermine has been shown to allow fluorescence quenching to be induced in isolated antenna in the absence of ΔpH and to accelerate quenching formation. The simultaneous treatment of spermine and lutein maximizes quenching even at relatively low concentrations.

Spermine is a potent modulator of proton transport through LHCII.

The effect of spermine on proton transport across large unilamellar liposomes containing incorporated complexes of the PSII antenna has been studied with the application of a pH-sensitive dye entrapped inside the vesicles. Both monomeric LHCbs and trimeric LHCII increased the permeability of proteoliposomes to protons when in a partly aggregated state within the lipid membrane. We have previously shown that a spermine-induced conformational change in LHCII results in its aggregation and ultimately in the enhancement of excitation energy as heat (qE). In this paper, spermine-induced aggregation of LHCII was found to facilitate proton transport across the proteoliposomes, indicating that a second protective mechanism (other than qE) might exist and might be regulated in vivo by polyamines when photosynthesis is saturated in excess light.

Polyamines induce aggregation of LHC II and quenching of fluorescence in vitro.

Dissipation of excess excitation energy within the light-harvesting complex of Photosystem II (LHC II) is a main process in plants, which is measured as the non-photochemical quenching of chlorophyll fluorescence or qE. We showed in previous works that polyamines stimulate qE in higher plants in vivo and in eukaryotic algae in vitro. In the present contribution we have tested whether polyamines can stimulate quenching in trimeric LHC II and monomeric light-harvesting complex b proteins from higher plants. The tetramine spermine was the most potent quencher and induced aggregation of LHC II trimers, due to its highly cationic character. Two transients are evident at 100 µM and 350 µM for the fluorescence and absorbance signals of LHC II respectively. On the basis of observations within this work, some links between polyamines and the activation of qE in vivo is discussed.

Chemical bonding of chlorophylls and plant aminic axial ligands impact harvesting of visible light and quenching of fluorescence.

In the present work, we tested the mode of interaction of all three polyamines (putrescine, spermidine and spermine) with chlorophyll a and b, as well as pheophytin a and b. The results showed that all three polyamines bind to the Mg ion of chlorophyll ring as probed by Raman spectroscopy. The coordination of spermine with Chl b has the most interesting features from all pigments tested. Spermine induces reversible increases and decreases of the fluorescence yield of Chl b at about 661 nm. Interestingly, equilibrium between a high-fluorescence yield conformation and a low yield is feasible by the interaction of chlorophyll b and aminic ligands. Furthermore, absorption data for the diagnostic regions of 518 and 535 nm are provided for all combinations of pigments and ligands. The significance and consistence of these results with respect to photochemical and bioenergetic principles are discussed.