Frequently asked questions about in vivo chlorophyll fluorescence: practical issues.

The aim of this educational review is to provide practical information on the hardware, methodology, and the hands on application of chlorophyll (Chl) a fluorescence technology. We present the paper in a question and answer format like frequently asked questions. Although nearly all information on the application of Chl a fluorescence can be found in the literature, it is not always easily accessible. This paper is primarily aimed at scientists who have some experience with the application of Chl a fluorescence but are still in the process of discovering what it all means and how it can be used. Topics discussed are (among other things) the kind of information that can be obtained using different fluorescence techniques, the interpretation of Chl a fluorescence signals, specific applications of these techniques, and practical advice on different subjects, such as on the length of dark adaptation before measurement of the Chl a fluorescence transient. The paper also provides the physiological background for some of the applied procedures. It also serves as a source of reference for experienced scientists.

Experimental in vivo measurements of light emission in plants: a perspective dedicated to David Walker.

This review is dedicated to David Walker (1928-2012), a pioneer in the field of photosynthesis and chlorophyll fluorescence. We begin this review by presenting the history of light emission studies, from the ancient times. Light emission from plants is of several kinds: prompt fluorescence (PF), delayed fluorescence (DF), thermoluminescence, and phosphorescence. In this article, we focus on PF and DF. Chlorophyll a fluorescence measurements have been used for more than 80 years to study photosynthesis, particularly photosystem II (PSII) since 1961. This technique has become a regular trusted probe in agricultural and biological research. Many measured and calculated parameters are good biomarkers or indicators of plant tolerance to different abiotic and biotic stressors. This would never have been possible without the rapid development of new fluorometers. To date, most of these instruments are based mainly on two different operational principles for measuring variable chlorophyll a fluorescence: (1) a PF signal produced following a pulse-amplitude-modulated excitation and (2) a PF signal emitted during a strong continuous actinic excitation. In addition to fluorometers, other instruments have been developed to measure additional signals, such as DF, originating from PSII, and light-induced absorbance changes due to the photooxidation of P700, from PSI, measured as the absorption decrease (photobleaching) at about 705 nm, or increase at 820 nm. In this review, the technical and theoretical basis of newly developed instruments, allowing for simultaneous measurement of the PF and the DF as well as other parameters is discussed. Special emphasis has been given to a description of comparative measurements on PF and DF. However, DF has been discussed in greater details, since it is much less used and less known than PF, but has a great potential to provide useful qualitative new information on the back reactions of PSII electron transfer. A review concerning the history of fluorometers is also presented.

Fluorescence parameters as early indicators of light stress in barley.

Photosynthetic efficiency of two Syrian barley landraces Arabi Aswad and Arabi Abiad grown under different light intensities were studied by the application of qualitative and quantitative analysis of chlorophyll a fluorescence. Different values of fluorescence parameters, quantum efficiencies, specific and phenomenological energy fluxes were obtained for each cultivar. Both low and high light stresses decreased photosystem II (PSII) activity in barley seedlings depending on the stress type and its duration. Cultivar Arabi Aswad was more tolerant to high light while Arabi Abiad was more tolerant to low light stress. The results allowed us to select chlorophyll a fluorescence parameters related to energy flux within PSII which were specifically affected under low or high light stress. We found that the performance index parameter is a sensitive indicator to explore the effect of light changes on PSII activity immediately after stress application, while maximal quantum yield of PSII and phenomenological parameters were only modified after a long period of stress application indicating PSII damage. Thus, we recommend the former parameter for early detection of light stress.

Chlorophyll a fluorescence--A useful tool for the early detection of temperature stress in spring barley (Hordeum vulgare L.).

The photosynthetic activity of two Syrian barley (Hordeum vulgare L.) landraces, Arabi Abiad (A. Abiad) and Arabi Aswad (A. Aswad), grown under low- and high- temperature stresses, were studied by analyzing the measured chlorophyll fluorescence signals. Both the applied stresses influenced photosystem II (PSII) activity. However, the effects depend on the stress type and the duration of its application. Phenomenological parameters were shifted shortly after the application of both stresses, whereas fluorescence ratios and yield values were altered most significantly after 7 days of stress imposition. The earliest changes in PSII activity of both cultivars were observed in the case of high temperature treatment. The maximal quantum efficiency of the photosynthetic apparatus (F(v)/F(M)) did not alter after stress application. Therefore, we could not recommend this parameter for early detection of such stress. In contrast, the results from the present investigation strongly indicate that the most significantly changed chlorophyll a fluorescence parameters could be used as an efficient tool for the early diagnosis of temperature stress in barley.