In the lycophyte Selaginella martensii is the "extra-qT" related to energy spillover? Insights into photoprotection in ancestral vascular plants

Lycophytes are early diverging vascular plants, representing a minor group as compared to the dominating euphyllophytes, mostly angiosperms. Having maximally developed in a CO2-rich atmosphere, extant lycophytes are characterized by a low carbon fixing capacity, which is compensated by a marked ability to induce the non-photochemical quenching of chlorophyll fluorescence (NPQ). Different kinetic components contribute to NPQ, in particular the fast relaxing high-energy quenching qE, the middle relaxing qT, and the slowly relaxing qI. Unlike angiosperms, lycophytes enhance the qT component under high light, originating from an "extra-qT". In this research, we analyze whether in Selaginella martensii the extra-qT can reflect a photosystem (PS) I-based quenching mechanism activated upon saturation of qE capacity. From comparative analyses of fluorescence quenching parameters, carbon fixation, in vivo low- and room-temperature fluorescence spectroscopy, and thylakoid protein phosphorylation, it is proposed that the extra-qT is not mechanistically separate from the ordinary qT. The results suggest a relationship between qT and photoprotective energy spillover to PSI, which is activated upon sensing the excitation energy pressure inside PSII and is possibly facilitated by phosphorylation of Lhcb6, a minor antenna protein of PSII. Energy spillover emphasizes 77K fluorescence emission from PSI core (F714) and becomes more relevant at irradiance levels corresponding to the CO2-limited, potentially photoinhibiting phase of photosynthesis. At the highest irradiances, when Lhcb6 phosphorylation potential has been saturated, the major LHCII increases in turn its phosphorylation level, probably leading to the full exploitation of PSI as a safe excitation sink. It is suggested that the low photosynthetic capacity of lycophytes could allow an easier experimental access to the use of PSI as a safe excitation quencher for PSII, a debated, emerging issue about thylakoid photoprotection in angiosperms.