iReenCAM: automated imaging system for kinetic analysis of photosynthetic pigment biosynthesis at high spatiotemporal resolution during early deetiolation

Frontiers in Plant Science

Plant Science

Ioannis Spyroglou¹  Ingrid Kovacova¹  Jan Skalak¹  Dominique Arnaud¹  Jan Hejatko²  Tereza Dobisova²  Veronika Balakhonova²  Klara Panzarova³  Jaromir Pytela³  Radka Koci³  Martin Trtilek³  Zuzana Benedikty³ 

[1] CEITEC - Central European Institute of Technology, Masaryk University, Brno, Czechia;CEITEC - Central European Institute of Technology, Masaryk University, Brno, Czechia;National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, Czechia;Photon Systems Instruments, Drasov, Czechia;

关键词: de-etiolation;    chlorophyll biosynthesis;    fluorescence;    iReenCAM;    cytokinins;    ethylene;    Arabidopsis;   

DOI  :  10.3389/fpls.2023.1093292

 received in 2022-11-08, accepted in 2023-04-04,  发布年份 2023

来源: Frontiers

【 摘 要 】

Seedling de-etiolation is one of the key stages of the plant life cycle, characterized by a strong rearrangement of the plant development and metabolism. The conversion of dark accumulated protochlorophyllide to chlorophyll in etioplasts of de-etiolating plants is taking place in order of ns to µs after seedlings illumination, leading to detectable increase of chlorophyll levels in order of minutes after de-etiolation initiation. The highly complex chlorophyll biosynthesis integrates number of regulatory events including light and hormonal signaling, thus making de-etiolation an ideal model to study the underlying molecular mechanisms. Here we introduce the iReenCAM, a novel tool designed for non-invasive fluorescence-based quantitation of early stages of chlorophyll biosynthesis during de-etiolation with high spatial and temporal resolution. iReenCAM comprises customized HW configuration and optimized SW packages, allowing synchronized automated measurement and analysis of the acquired fluorescence image data. Using the system and carefully optimized protocol, we show tight correlation between the iReenCAM monitored fluorescence and HPLC measured chlorophyll accumulation during first 4h of seedling de-etiolation in wild type Arabidopsis and mutants with disturbed chlorophyll biosynthesis. Using the approach, we demonstrate negative effect of exogenously applied cytokinins and ethylene on chlorophyll biosynthesis during early de-etiolation. Accordingly, we identify type-B response regulators, the cytokinin-responsive transcriptional activators ARR1 and ARR12 as negative regulators of early chlorophyll biosynthesis, while contrasting response was observed in case of EIN2 and EIN3, the components of canonical ethylene signaling cascade. Knowing that, we propose the use of iReenCAM as a new phenotyping tool, suitable for quantitative and robust characterization of the highly dynamic response of seedling de-etiolation.

【 授权许可】

Unknown   
Copyright © 2023 Balakhonova, Dobisova, Benedikty, Panzarova, Pytela, Koci, Spyroglou, Kovacova, Arnaud, Skalak, Trtilek and Hejatko