Abstract

The responses of photosynthetic organisms to light stress are of interest for both fundamental and applied research. Functional traits related to the photoinhibition, the light-induced loss of photosynthetic efficiency, are particularly interesting as this process is a key limiting factor of photosynthetic productivity in algae and plants. The quantitative characterization of light responses is often time-consuming and calls for cost-effective high throughput approaches that enable the fast screening of multiple samples. Here we present a novel illumination system based on the concept of ‘multi-actinic imaging’ of in vivo chlorophyll fluorescence. The system is based on the combination of an array of individually addressable low power RGBW LEDs and custom-designed well plates, allowing for the independent illumination of 64 samples through the digital manipulation of both exposure duration and light intensity. The illumination system is inexpensive and easily fabricated, based on open source electronics, off-the-shelf components, and 3D-printed parts, and is optimized for imaging of chlorophyll fluorescence. The high-throughput potential of the system is illustrated by assessing the functional diversity in light responses of marine macroalgal species, through the fast and simultaneous determination of kinetic parameters characterizing the response to light stress of multiple samples. Although the presented illumination system was primarily designed for the measurement of phenotypic traits related to photosynthetic activity and photoinhibition, it can be potentially used for a number of alternative applications, including the measurement of chloroplast phototaxis and action spectra, or as the basis for microphotobioreactors.

Highlights

  • The description and understanding of how photosynthetic organisms respond to light has long been a central topic in photobiology and photosynthesis research

  • The illumination system described here is a combination of (i) a set of individually addressable LEDs mounted as an orthogonal array on a flat panel, delivering photosynthetically-relevant intensities of photosynthetically active radiation (PAR), and (ii) custom-made multiwell plates, optimized for independent light exposure of samples and for chlorophyll fluorescence imaging

  • The irradiance emitted by the tested LEDs varied linearly with PWM intensity settings, as defined in the microcontroller code, for all colors, separately and combined (Fig. 3A)

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Summary

Introduction

The description and understanding of how photosynthetic organisms respond to light has long been a central topic in photobiology and photosynthesis research. Light is the primary driver and regulatory factor of photosynthesis, but it is one of the main stressors of the photosynthetic apparatus. The photoinactivation of PSII is in turn counteracted by photoprotective energy-dissipation mechanisms, operating to balance light absorption and damage caused by excess light energy (Müller, Li & Niyogi, 2001; Demmig-Adams & Adams, 2006). The quantitative characterization of the light-dependence of photosynthetic processes is of interest for fundamental research on their underlying mechanisms, their diversity, and the evolution of functional traits (Goss & Lepetit, 2015). The interplay between photoprotection and photoinactivation, and of their regulating factors, is of interest for applied research, as these processes are increasingly recognized as main determinants of plant and algal productivity (Murchie & Niyogi, 2011)

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