The aim of the present work was to investigate the toxic effects of zinc oxide nanoparticles (ZnO NPs, particle size < 50 nm) on the physiological and anatomical indices of spring barley (Hordeum sativum L.). The results show that ZnO NPs inhibited H. sativum growth by affecting the chlorophyll fluorescence emissions and causing deformations of the stomatal and trichome morphology, alterations to the cellular organizations, including irregularities of the chloroplasts, and disruptions to the grana and thylakoid organizations. There was a lower number of chloroplasts per cell observed in the H. sativum leaf cells treated with ZnO NPs as compared to the non-treated plants. Cytomorphometric quantification revealed that ZnO NPs decreased the size of the chloroplast by 1.5 and 4 times in 300 and 2000 mg/L ZnO NP-treated plants, respectively. The elemental analysis showed higher Zn accumulation in the treated leaf tissues (3.8 and 10.18-fold with 300 and 2000 mg/L ZnO NPs, respectively) than the untreated. High contents of Zn were observed in several spots in ZnO NP-treated leaf tissues using X-ray fluorescence. Deviations in the anatomical indices were significantly correlated with physiological observations. The accumulation of Zn content in plant tissues that originated from ZnO NPs was shown to cause damage to the structural organization of the photosynthetic apparatus and reduced the photosynthetic activities.

A simple method of a bicolor (multicolor), fast-Fourier, PAM chlorophyll fluorometry has been developed to obtain fluorescence induction curves. Quantum yields of PSII photochemistry were determined with blue and red simultaneously applied pulsed measuring lights for three subsequent 20-min periods of dark-, light-adaptation under actinic light and dark recovery. Measuring lights were cross-combined with blue and red actinic lights and saturation pulses. Coefficients of chromatic divergence were calculated as a ratio of the quantum yields obtained by red measuring light to that obtained by blue measuring light. Adaptation of Ficus benjamina and Hordeum vulgare leaves under blue (but not red) actinic light resulted in the sufficient increase of chromatic divergence. In addition, fraction of active, non(photo)inhibited, PSII centers was shown to be dependent on the color of measuring light. Thus, color of the light sources should be considered when reporting results of parameters evaluated from fluorescence induction curves.