Structural and functional disorder in the photosynthetic apparatus of radish plants under magnesium deficiency.

Magnesium (Mg) is one of the significant macronutrients which is involved in the structural stabilisation of plant tissues and many enzymes such as PSII. The latter efficiency and performance were analysed, using chlorophyll (Chl) a fluorescence induction kinetics and microscopic images, to detect the changes in structure and function of photosynthetic apparatus of radish plants grown under Mg deficiency (Mgdef). Plants grown under Mgdef showed less PSII connectivity and fewer active primary electron acceptors (QA) oxidizing reaction centres than control plants. Confocal and electron microscopy analyses showed an increased amount of starch in chloroplasts, and 3,3'-diaminobenzidine (DAB)-uptake method revealed higher H2O2 accumulation under Mgdef. Prominent changes in the Chl a fluorescence parameters such as dissipated energy flux per reaction centre (DIo/RC), relative variable fluorescence at 150µs (Vl), and the sum of the partial driving forces for the events involved in OJIP fluorescence rise (DFabs) were observed under Mg deficiency. The latter also significantly affected some other parameters such as dissipated energy fluxes per cross-section (DIo/CSo), performance index for energy conservation from photons absorbed by PSII antenna until the reduction of PSI acceptors (PItotal), and relative variable fluorescence at 300µs (Vk). This work emphasises the use of chlorophyll fluorescence in combination with microscopic and statistical analyses to diagnose the effects of nutrients deficiency stress on plants at an early stage of its development as demonstrated for the example of Mgdef. Due to the short growth period and simple cultivation conditions of radish plant we recommend it as a new standard (model) plant to study nutrients deficiency and changes in plant photosynthetic efficiency under stress conditions.

Chlorophyll fluorescence as a tool for nutrient status identification in rapeseed plants.

In natural conditions, plants growth and development depends on environmental conditions, including the availability of micro- and macroelements in the soil. Nutrient status should thus be examined not by establishing the effects of single nutrient deficiencies on the physiological state of the plant but by combinations of them. Differences in the nutrient content significantly affect the photochemical process of photosynthesis therefore playing a crucial role in plants growth and development. In this work, an attempt was made to find a connection between element content in (i) different soils, (ii) plant leaves, grown on these soils and (iii) changes in selected chlorophyll a fluorescence parameters, in order to find a method for early detection of plant stress resulting from the combination of nutrient status in natural conditions. To achieve this goal, a mathematical procedure was used which combines principal component analysis (a tool for the reduction of data complexity), hierarchical k-means (a classification method) and a machine-learning method-super-organising maps. Differences in the mineral content of soil and plant leaves resulted in functional changes in the photosynthetic machinery that can be measured by chlorophyll a fluorescent signals. Five groups of patterns in the chlorophyll fluorescent parameters were established: the 'no deficiency', Fe-specific deficiency, slight, moderate and strong deficiency. Unfavourable development in groups with nutrient deficiency of any kind was reflected by a strong increase in F o and ΔV/Δt 0 and decline in φ Po, φ Eo δ Ro and φ Ro. The strong deficiency group showed the suboptimal development of the photosynthetic machinery, which affects both PSII and PSI. The nutrient-deficient groups also differed in antenna complex organisation. Thus, our work suggests that the chlorophyll fluorescent method combined with machine-learning methods can be highly informative and in some cases, it can replace much more expensive and time-consuming procedures such as chemometric analyses.

Frequently asked questions about chlorophyll fluorescence, the sequel.

Using chlorophyll (Chl) a fluorescence many aspects of the photosynthetic apparatus can be studied, both in vitro and, noninvasively, in vivo. Complementary techniques can help to interpret changes in the Chl a fluorescence kinetics. Kalaji et al. (Photosynth Res 122:121-158, 2014a) addressed several questions about instruments, methods and applications based on Chl a fluorescence. Here, additional Chl a fluorescence-related topics are discussed again in a question and answer format. Examples are the effect of connectivity on photochemical quenching, the correction of F V /F M values for PSI fluorescence, the energy partitioning concept, the interpretation of the complementary area, probing the donor side of PSII, the assignment of bands of 77 K fluorescence emission spectra to fluorescence emitters, the relationship between prompt and delayed fluorescence, potential problems when sampling tree canopies, the use of fluorescence parameters in QTL studies, the use of Chl a fluorescence in biosensor applications and the application of neural network approaches for the analysis of fluorescence measurements. The answers draw on knowledge from different Chl a fluorescence analysis domains, yielding in several cases new insights.

Identification of nutrient deficiency in maize and tomato plants by in vivo chlorophyll a fluorescence measurements.

The impact of some macro (Ca, S, Mg, K, N, P) and micro (Fe) nutrients deficiency on the functioning of the photosynthetic machinery in tomato (Solanum lycopersicum L.) and maize (Zea mays L.) plants grown in hydroponic cultures were investigated. Plants grown on a complete nutrient solution (control) were compared with those grown in a medium, which lacked one of macro- or microelements. The physiological state of the photosynthetic machinery in vivo was analysed after 14-days of deficient condition by the parameters of JIP-test based on fast chlorophyll a fluorescence records. In most of the nutrient-deficient samples, the decrease of photochemical efficiency, increase in non-photochemical dissipation and decrease of the number of active photosystem II (PSII) reaction centres were observed. However, lack of individual nutrients also had nutrient-specific effects on the photochemical processes. In Mg and Ca-deficient plants, the most severe decrease in electron donation by oxygen evolving complex (OEC) was indicated. Sulphur deficiency caused limitation of electron transport beyond PSI, probably due to decrease in the PSI content or activity of PSI electron acceptors; in contrary, Ca deficiency had an opposite effect, where the PSII activity was affected much more than PSI. Despite the fact that clear differences in nutrient deficiency responses between tomato and maize plants were observed, our results indicate that some of presented fluorescence parameters could be used as fluorescence phenotype markers. The principal component analysis of selected JIP-test parameters was presented as a possible species-specific approach to identify/predict the nutrient deficiency using the fast chlorophyll fluorescence records.