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High throughput procedure utilising chlorophyll fluorescence imaging to phenotype dynamic photosynthesis and photoprotection in leaves under controlled gaseous conditions

McAusland, Lorna; Murchie, Erik; Atkinson, Jonathan

High throughput procedure utilising chlorophyll fluorescence imaging to phenotype dynamic photosynthesis and photoprotection in leaves under controlled gaseous conditions Thumbnail


Authors

Dr ERIK MURCHIE erik.murchie@nottingham.ac.uk
Professor of Applied Plant Physiology



Abstract

© 2019 The Author(s). Background: As yields of major crops such as wheat (T. aestivum) have begun to plateau in recent years, there is growing pressure to efficiently phenotype large populations for traits associated with genetic advancement in yield. Photosynthesis encompasses a range of steady state and dynamic traits that are key targets for raising Radiation Use Efficiency (RUE), biomass production and grain yield in crops. Traditional methodologies to assess the full range of responses of photosynthesis, such a leaf gas exchange, are slow and limited to one leaf (or part of a leaf) per instrument. Due to constraints imposed by time, equipment and plant size, photosynthetic data is often collected at one or two phenological stages and in response to limited environmental conditions. Results: Here we describe a high throughput procedure utilising chlorophyll fluorescence imaging to phenotype dynamic photosynthesis and photoprotection in excised leaves under controlled gaseous conditions. When measured throughout the day, no significant differences (P > 0.081) were observed between the responses of excised and intact leaves. Using excised leaves, the response of three cultivars of T. aestivum to a user - defined dynamic lighting regime was examined. Cultivar specific differences were observed for maximum PSII efficiency (F v′/F m′ - P < 0.01) and PSII operating efficiency (F q′/F m′ - P = 0.04) under both low and high light. In addition, the rate of induction and relaxation of non-photochemical quenching (NPQ) was also cultivar specific. A specialised imaging chamber was designed and built in-house to maintain gaseous conditions around excised leaf sections. The purpose of this is to manipulate electron sinks such as photorespiration. The stability of carbon dioxide (CO2) and oxygen (O2) was monitored inside the chambers and found to be within ± 4.5% and ± 1% of the mean respectively. To test the chamber, T. aestivum 'Pavon76' leaf sections were measured under at 20 and 200 mmol mol-1 O2 and ambient [CO2] during a light response curve. The F v′/F m′was significantly higher (P < 0.05) under low [O2] for the majority of light intensities while values of NPQ and the proportion of open PSII reaction centers (qP) were significantly lower under > 130 μmol m-2 s-1 photosynthetic photon flux density (PPFD). Conclusions: Here we demonstrate the development of a high-throughput (> 500 samples day-1) method for phenotyping photosynthetic and photo-protective parameters in a dynamic light environment. The technique exploits chlorophyll fluorescence imaging in a specifically designed chamber, enabling controlled gaseous environment around leaf sections. In addition, we have demonstrated that leaf sections do not different from intact plant material even > 3 h after sampling, thus enabling transportation of material of interest from the field to this laboratory based platform. The methodologies described here allow rapid, custom screening of field material for variation in photosynthetic processes.

Citation

McAusland, L., Murchie, E., & Atkinson, J. (2019). High throughput procedure utilising chlorophyll fluorescence imaging to phenotype dynamic photosynthesis and photoprotection in leaves under controlled gaseous conditions. Plant Methods, 15, https://doi.org/10.1186/s13007-019-0485-x

Journal Article Type Article
Acceptance Date Aug 14, 2019
Online Publication Date Sep 18, 2019
Publication Date Sep 18, 2019
Deposit Date Aug 19, 2019
Publicly Available Date Mar 29, 2024
Journal Plant Methods
Electronic ISSN 1746-4811
Publisher Springer Verlag
Peer Reviewed Peer Reviewed
Volume 15
Article Number 109
DOI https://doi.org/10.1186/s13007-019-0485-x
Keywords Biotechnology; Plant Science; Genetics
Public URL https://nottingham-repository.worktribe.com/output/2448005
Publisher URL https://plantmethods.biomedcentral.com/articles/10.1186/s13007-019-0485-x
Related Public URLs https://plantmethods.biomedcentral.com/
Additional Information Received: 24 May 2019; Accepted: 14 August 2019; First Online: 18 September 2019; : Not applicable.; : Not applicable.; : The authors declare that they have no competing interests.

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