The use of laser induced chlorophyll fluorescence (LIF) as a fast and non‑destructive method to investigate water deficit in Arabidopsis

Chlorophyll fluorescence measurements have been widely applied as non-destructive methods to study the photosynthetic efficiency of plants, under control or stress conditions. Compared to most protocols of pulse amplitude modulation (PAM) fluorometry, laser induced chlorophyll fluorescence (LIF) has the advantages of not requiring pulses to be delivered at close range, allowing the remote analysis of a great number of individual plants in a short period of time. Such analyses are extremely useful, for instance, when doing large phenotyping screens of Arabidopsis thaliana mutants or ecotypes. Water deficit is a major abiotic stress compromising plant growth and productivity. Arabidopsis has been adopted as the main model organism in plant sciences and it has been widely used in plant stress studies. However, reports on the applications of LIF techniques to this model plant are scarce. Here we report the use of LIF to investigate changes in chlorophyll a (Chl a) fluorescence signature under progressive drought of potted Arabidopsis plants (Slow Stress) and under fast dehydration of detached leaves (Fast Stress). Results show that the two dehydration methods cause distinct modifications on the red/far-red Chl a fluorescence ratio (F690/F730) and on the wavelengths of Chl a fluorescence maxima. These differences are likely related to distinct changes in water content, photosynthetic pigments, anthocyanins, fatty acid composition and other metabolic adaptations, which are differently regulated in Slow and Fast Stress. Of particular interest are Chl a emission fluorescence changes, which take place under progressive drought, before a substantial decrease in leaf water content. Additionally, no differences were found on LIF emission spectra between fully expanded and young leaves. However, the choice of leaf surface influenced fluorescence emission, with the abaxial surface displaying lower fluorescence and higher F690/F730 ratios. Results suggest that LIF is a fast and non-destructive tool suitable for high-throughput phenotyping of Arabidopsis under water deficit.