Pitfalls, artefacts and open questions in chlorophyll thermoluminescence of leaves or algal cells.

Thermoluminescence of intact photosynthetic organisms, leaves or algal cells, raises specific problems. The constitutive S2/3Q B (-) B bands constitute major probes of the state of photosystem II in vivo. The presence of a dark-stable acidic lumen causes a temperature downshift of B bands, specially the S3 B band, providing a lumen pH indicator. This is accompanied by a broadening of the S3 B band that becomes an envelope of elementary B bands. The occasional AT, Q and C bands are briefly examined in an in vivo context. It is emphasized that freezing below the nucleation temperature is not necessary for physiological studies, but a source of artefacts, hence should be avoided. In intact photosynthetic structures, a dark-electron transfer from stroma reductants to the quinonic acceptors of photosystem II via the cyclic/chlororespiratory pathways, strongly stimulated by moderate warming, gives rise to the afterglow (AG) luminescence emission that reflects chloroplast energy status. The decomposition of complex TL signals into elementary bands is necessary to determine the maximum temperature T m and the area of each of them. A comparison of TL signals after 1 flash and 2 flashes prevents from confusing the three main bands observed in vivo, i.e. the S2 and S3 B bands and the AG band. Finally, the thermoluminescence bands arising sometimes above 50 °C are mentioned. The basic principles of (thermo)luminescence established on isolated thylakoids should not be applied directly without a careful examination of in vivo conditions.