The Morphological Parameters and Cytosolic pH of Cells of Root Zones in Tobacco Plants (Nicotiana tabacum L.): Nonlinear Effects of NaCl Concentrations.

Salinity impacts important processes in plants, reducing their yield. The effect of salinity on the cytosolic pH (pHcyt) has been little studied. In this research, we employed transgenic tobacco plants expressing the pH sensor Pt-GFP to investigate the alterations in pHcyt in cells across various root zones. Furthermore, we examined a wide spectrum of NaCl concentrations (ranging from 0 to 150 mM) and assessed morphological parameters and plant development. Our findings revealed a pattern of cytosolic acidification in cells across all root zones at lower NaCl concentrations (50, 100 mM). Interestingly, at 150 mM NaCl, pHcyt levels either increased or returned to normal, indicating a nonlinear effect of salinity on pHcyt. Most studied parameters related to development and morphology exhibited an inhibitory influence in response to NaCl. Notably, a nonlinear relationship was observed in the cell length within the elongation and differentiation zones. While cell elongation occurred at 50 and 100 mM NaCl, it was not evident at 150 mM NaCl. This suggests a complex interplay between stimulating and inhibitory effects of salinity, contributing to the nonlinear relationship observed between pHcyt, cell length, and NaCl concentration.

Comparison of the Efficiency of Hyperspectral and Pulse Amplitude Modulation Imaging Methods in Pre-Symptomatic Virus Detection in Tobacco Plants.

Early detection of pathogens can significantly reduce yield losses and improve the quality of agricultural products. This study compares the efficiency of hyperspectral (HS) imaging and pulse amplitude modulation (PAM) fluorometry to detect pathogens in plants. Reflectance spectra, normalized indices, and fluorescence parameters were studied in healthy and infected areas of leaves. Potato virus X with GFP fluorescent protein was used to assess the spread of infection throughout the plant. The study found that infection increased the reflectance of leaves in certain wavelength ranges. Analysis of the normalized reflectance indices (NRIs) revealed indices that were sensitive and insensitive to infection. NRI700/850 was optimal for virus detection; significant differences were detected on the 4th day after the virus arrived in the leaf. Maximum (Fv/Fm) and effective quantum yields of photosystem II (ΦPSII) and non-photochemical fluorescence quenching (NPQ) were almost unchanged at the early stage of infection. ΦPSII and NPQ in the transition state (a short time after actinic light was switched on) showed high sensitivity to infection. The higher sensitivity of PAM compared to HS imaging may be due to the possibility of assessing the physiological changes earlier than changes in leaf structure.

Effect of low-dose ionizing radiation on spatiotemporal parameters of functional responses induced by electrical signals in tobacco plants.

Plants growing under an increased radiation background may be exposed to additional stressors. Plant acclimatization is formed with the participation of stress signals that cause systemic responses-a change in the activity of physiological processes. In this work, we studied the mechanisms of the effect of ionizing radiation (IR) on the systemic functional responses induced by electrical signals. Chronic ß-irradiation (31.3 µGy/h) have a positive effect on the morphometric parameters and photosynthetic activity of tobacco plants (Nicotiana tabacum L.) at rest. An additional stressor causes an electrical signal, which, when propagated, causes a temporary change in chlorophyll fluorescence parameters, reflecting a decrease in photosynthesis activity. Irradiation did not significantly affect the electrical signals. At the same time, more pronounced photosynthesis responses are observed in irradiated plants: both the amplitude and the leaf area covered by the reaction increase. The formation of such responses is associated with changes in pH and stomatal conductance, the role of which was analyzed under IR. Using tobacco plants expressing the fluorescent pH-sensitive protein Pt-GFP, it was shown that IR enhances signal-induced cytoplasmic acidification. It was noted that irradiation also disrupts the correlation between the amplitudes of the electrical signal, pH shifts, changes in chlorophyll fluorescence parameters. Also stronger inhibition of stomatal conductance by the signal was shown in irradiated plants. It was concluded that the effect of IR on the systemic response induced by the electrical signal is mainly due to its effect on the stage of signal transformation into the response.

Cell-Type-Specific Length and Cytosolic pH Response of Superficial Cells of Arabidopsis Root to Chronic Salinity.

Soil salinity negatively affects the growth, development and yield of plants. Acidification of the cytosol in cells of glycophytes was reported under salinity, while various types of plant cells can have a specific reaction under the same conditions. Transgenic Arabidopsis plants expressing the pH sensor Pt-GFP in the cytosol were used in this work for determination of morphometric changes and cytosolic pH changes in the superficial cells of Arabidopsis roots under chronic salinity in vitro. We did not find changes in the length of the root cap cells, while there was a decrease in the length of the differentiation zone under 50, 75 mM NaCl and the size of the epidermal cells of the differentiation zone under 75 mM NaCl. The most significant changes of cytosolic pH to chronic salinity was noted in columella (decrease by 1 pH unit at 75 mM NaCl) and epidermal cells of the differentiation zone (decrease by 0.6 and 0.4 pH units at 50 and 75 mM NaCl, respectively). In developed lateral root cap cells, acidification of cytosol by 0.4 units occurred only under 75 mM NaCl in the medium. In poorly differentiated lateral cells of the root cap, there were no changes in pH under chronic salinity.

The Role of Phialocephala fortinii in Improving Plants' Phosphorus Nutrition: New Puzzle Pieces.

Plants' mineral nutrition in acidic soils can be facilitated by phosphate solubilizing fungi inhabiting the root systems of these plants. We attempt to find dark septate endophyte (DSE) isolates in the roots of wild-heather plants, which are capable of improving plants' phosphorus nutrition levels. Bright-field and confocal laser scanning microscopy were used for the visualization of endophytes. A model system of co-cultivation with Vaccinium macrocarpon Ait. was used to study a fungal isolate's ability to supply plants with phosphorus. Fungal phytase activity and phosphorus content in plants were estimated spectrophotometrically. In V. vitis-idaea L. roots, we obtained a Phialocephala fortinii Wang, Wilcox DSE2 isolate with acid phytase activity (maximum 6.91 ± 0.17 U on 21st day of cultivation on potato-dextrose broth medium) and the ability to accumulate polyphosphates in hyphae cells. The ability of the isolate to increase both phosphorus accumulation and biomass in V. macrocarpon is also shown. The data obtained for the same isolate, as puzzle pieces put together, indicate the possible mediation of P. fortinii DSE2 isolate in the process of phosphorus intake from inorganic soil reserves to plants.

Salt-Induced Changes in Cytosolic pH and Photosynthesis in Tobacco and Potato Leaves.

Salinity is one of the most common factors limiting the productivity of crops. The damaging effect of salt stress on many vital plant processes is mediated, on the one hand, by the osmotic stress caused by large concentrations of Na+ and Cl- outside the root and, on the other hand, by the toxic effect of these ions loaded in the cell. In our work, the influence of salinity on the changes in photosynthesis, transpiration, water content and cytosolic pH in the leaves of two important crops of the Solanaceae family-tobacco and potato-was investigated. Salinity caused a decrease in photosynthesis activity, which manifested as a decrease in the quantum yield of photosystem II and an increase in non-photochemical quenching. Along with photosynthesis limitation, there was a slight reduction in the relative water content in the leaves and a decrease in transpiration, determined by the crop water stress index. Furthermore, a decrease in cytosolic pH was detected in tobacco and potato plants transformed by the gene of pH-sensitive protein Pt-GFP. The potential mechanisms of the salinity influence on the activity of photosynthesis were analyzed with the comparison of the parameters' dynamics, as well as the salt content in the leaves.

Pre-Symptomatic Detection of Viral Infection in Tobacco Leaves Using PAM Fluorometry.

Chlorophyll fluorescence imaging was used to study potato virus X (PVX) infection of Nicotiana benthamiana. Infection-induced changes in chlorophyll fluorescence parameters (quantum yield of photosystem II photochemistry (ΦPSII) and non-photochemical fluorescence quenching (NPQ)) in the non-inoculated leaf were recorded and compared with the spatial distribution of the virus detected by the fluorescence of GFP associated with the virus. We determined infection-related changes at different points of the light-induced chlorophyll fluorescence kinetics and at different days after inoculation. A slight change in the light-adapted steady-state values of ΦPSII and NPQ was observed in the infected area of the non-inoculated leaf. In contrast to the steady-state parameters, the dynamics of ΦPSII and NPQ caused by the dark-light transition in healthy and infected areas differed significantly starting from the second day after the detection of the virus in a non-inoculated leaf. The coefficients of correlation between chlorophyll fluorescence parameters and virus localization were 0.67 for ΦPSII and 0.76 for NPQ. In general, the results demonstrate the possibility of reliable pre-symptomatic detection of the spread of a viral infection using chlorophyll fluorescence imaging.