4.7 Article

A chlorophyll fluorescence-based method for the integrated characterization of the photophysiological response to light stress

期刊

JOURNAL OF EXPERIMENTAL BOTANY
卷 68, 期 5, 页码 1123-1135

出版社

OXFORD UNIV PRESS
DOI: 10.1093/jxb/erw492

关键词

Chlorophyll fluorescence; imaging fluorometry; light-response curves; multi-actinic; photoacclimation; photoinactivation; photoinhibition; photoprotection; photorepair

资金

  1. Fundacao para a Ciencia e a Tecnologia (FCT) [SFRH/BD/86788/2012]
  2. CESAM
  3. Fundação para a Ciência e a Tecnologia [SFRH/BD/86788/2012] Funding Source: FCT

向作者/读者索取更多资源

This work introduces a new experimental method for the comprehensive description of the physiological responses to light of photosynthetic organisms. It allows the integration in a single experiment of the main established manipulative chlorophyll fluorescence-based protocols. It enables the integrated characterization of the photophysiology of samples regarding photoacclimation state (generating non-sequential light-response curves of effective PSII quantum yield, electron transport rate or non-photochemical quenching), photoprotection capacity (running light stress-recovery experiments, quantifying non-photochemical quenching components) and the operation of photoinactivation and photorepair processes (measuring rate constants of photoinactivation and repair for different light levels and the relative quantum yield of photoinactivation). The new method is based on a previously introduced technique, combining the illumination of a set of replicated samples with spatially separated actinic light beams of different intensity, and the simultaneous measurement of the fluorescence emitted by all samples using an imaging fluorometer. The main novelty described here is the independent manipulation of light intensity and duration of exposure for each sample, and the control of the cumulative light dose applied. The results demonstrate the proof of concept for the method, by comparing the responses of cultures of Chlorella vulgaris acclimated to low and high light regimes, highlighting the mapping of light stress responses over a wide range of light intensity and exposure conditions, and the rapid generation of paired light-response curves of photoinactivation and repair rate constants. This approach represents a chlorophyll fluorescence 'protocol of everything', contributing towards the high throughput characterization of the photophysiology of photosynthetic organisms.

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