Near-infrared in vivo measurements of photosystem I and its lumenal electron donors with a recently developed spectrophotometer.

In photosynthesis research, non-invasive in vivo spectroscopic analyses have been used as a practical tool for studying photosynthetic electron transport. Klas-NIR spectrophotometer has been recently developed by Klughammer and Schreiber (Photosynth Res 128:195-214, 2016) for in vivo measurements of redox changes of P700, plastocyanin (Pcy) and ferredoxin (Fd). Here we show examples using the Klas-NIR spectrophotometer for the evaluation of the redox states and quantities of these components in plant leaves and cyanobacterial suspensions. The redox poise under light of the electron transport components is different in leaves from higher plants compared with cyanobacteria. During a short illumination with an actinic light, P700, Pcy, and Fd are kept reduced in barley leaves but are oxidized in cyanobacteria. During far-red light illumination, P700 and Pcy are mostly oxidized in the leaves but are partially kept reduced in cyanobacteria. In the cyanobacterium, Thermosynechococcus elongatus, which has no Pcy but uses cytochrome c6 (cyt c6) as the electron donor to photosystem I, a cyt c6 signal was detected in vivo. To show the potential of Klas-NIR spectrophotometer for studying different developmental stages of a leaf, we performed measurements on fully mature and early senescing barley leaves. Pcy content in leaves decreased during senescence at an early stage. The Pcy loss was quantitatively analyzed using Klas-NIR spectrophotometer, giving absolute ratios of Pcy to PSI of 2.5 and 1.6 in younger and older leaves, respectively. For quantification of the signals in vivo, in vitro data (Sétif et al. in Photosynth Res142:307-319, 2019) obtained with Klas-NIR spectrophotometer were used.