Enhancing plant photochemistry analysis: a novel approach to chlorophyll a fluorescence measurement under environmental stress

Chlorophyll a fluorescence (ChlF) has been a pivotal tool in understanding plant photochemistry, offering insights into the energy transfer processes within chloroplasts and the efficiency of Photosystem II (PSII). Researchers have relied on quantifying ChlF through specific measures such as F0, Fm, and Fv under various lighting conditions to assess photosynthetic activities. Nonetheless, the technique encounters obstacles owing to the intrinsic uncertainties in gauging the absolute magnitude of ChlF and the fluctuation in baseline levels affected by environmental conditions. This complicates the interpretation of ChlF data and its correlation with photosynthetic efficiency.

In February 2024, Plant Phenomics published a research article entitled by “Dynamic Analysis of Chlorophyll a Fluorescence in Response to Time-Variant Excitations during Strong Actinic Illumination and Application in Probing Plant Water Loss”.

In this study, researchers employed a dynamic approach to measuring Chlorophyll a fluorescence (ChlF) in plants by using a pseudo-random binary sequence (PRBS) as a time-variant multiple-frequency illumination excitation under strong actinic light. This method allowed for the observation of ChlF responses not only in the time domain but also, and more importantly, in the frequency domain, emphasizing amplitude gain variations at different frequencies. By analyzing these responses amidst conditions of varying moisture levels, the study revealed significant insights into how nonphotochemical quenching (NPQ) and moisture loss impact ChlF dynamics. The results revealed that NPQ activities introduce nonlinearity and nonstationarity in ChlF responses, particularly affecting low-frequency responses below 0.03 rad/s, which exhibit a linear correlation with NPQ levels. This correlation suggests that low-frequency gain could serve as a compensatory measure for the variabilities encountered in ChlF measurements. Furthermore, the study demonstrated that while the low-frequency amplitude gain provides a direct link to NPQ, the high-frequency gain more closely correlates with plant moisture loss, suggesting that ChlF kinetics are markedly influenced by both NPQ and moisture status in a nonlinear manner.

In summary, this research not only underscores the complexity of ChlF responses under varying environmental conditions but also presents a novel methodological framework that leverages dynamic characteristics of ChlF to enhance the accuracy and utility of ChlF measurements in plant physiological research. The findings advocate for the use of frequency domain analysis to better account for variations in ChlF measurements induced by strong actinic illumination, offering a path toward mitigating these variabilities and improving the reliability of ChlF as a tool for assessing plant health and stress responses.

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References

Authors

Junqing  Chen¹, Ya  Guo², and Jinglu  Tan^1*

Affiliations

¹Department  of  Chemical  and  Biomedical  Engineering,  University  of  Missouri,  Columbia,  MO,  USA. 

²School of IOT, Jiangnan University, Wuxi, Jiangsu, China.

About  Jinglu  Tan

Jinglu Tan a professor in the Department of Chemical and Biomedical Engineering at University  of  Missouri. He serves as Director for the Division of Food, Nutrition & Exercise Sciences, a joint division between the College of Agriculture, Food & Natural Resources and the School of Medicine. His current research includes modeling and sensing of photoelectron transduction in plants and energy metabolism in muscles.