UV-B induced damage and recovery processes in apple leaves as assessed by LIF and PAM fluorescence techniques

Abstract

The capability of laser-induced chlorophyll fluorescence (LIF) and pulse-amplitude-modulated (PAM) fluorescence technique as well as RED/NIR-light reflection measurements for detection and quantification of UV-B induced damages was evaluated in greenhouse experiments with apple seedlings (Malus domestica Borkh.). Photosynthetic recovery from short-term UV-B stress was assessed during 7 days after UV-B treatment with the PAM fluorometer. The exposure of apple leaves to UV-B doses in the range of 10-26 W m^-2 for 180 minutes (UV-B_BE dose = 5.4-14 kJ m^-2) affected neither chlorophyll content nor leaf reflection. Although UV-B damage was not visually evident 2 hours after irradiation, it could be detected by PAM and LIF fluorescence techniques with equivalent success. The intensity of LIF, estimated as the integral of fluorescence spectrum, was reduced after UV-B irradiation by 19-30%. A stronger decrease in F686 compared to F740 fluorescence resulted in significantly lower F686/F740 values in all UV-B treatments.
Apple leaves displayed a strong and significant reduction in maximum fluorescence (Fm) and a slightly increase in ground fluorescence (Fo) 2 hours after UV-B treatment, as documented by PAM fluorescence measurement.
Negative linear regressions between investigated UV-B doses and selected PAM parameters were found with determination coefficients (R²) of 0.50 for Fv, 0.48 for Fv/Fm, and 0.58 for Fv/Fo. Among the PAM and LIF parameters tested, the Fv/Fo ratio appeared most sensitive for detection of UV-B induced damages displaying greatest changes and strongest correlation with the applied UV-B doses. PAM fluorescence images of apple leaves visualised an enhanced spatial heterogeneity of photosynthetic activity with increasing UV-B dose. The disturbance in photosynthetic functionality was followed by a continuous recovery process as indicated by restoring Fo and Fm parameters. A decline in maximum photochemical efficiency Fv/Fm from 0.80 to 0.72 and 0.43 after exposure to 20 W m^-2 for 240 and 360 minutes (UV-B_BE = 14.4 and 21.6 kJ m^-2), respectively, was followed by recovery at 7 x 10^-4 and 5 x 10^-3 units per hour during the first 48 hours after UV-B treatment. The recovery curves of Fm, Fv, Fv/Fm and Fv/Fo parameters during a week after UV-B irradiation were well fitted with exponential rise to maximum function, such as: y = yo + a (1 - e-bx). However, within 7 days after exposure to UV-B light, apple leaves displayed 14% or 4% lower Fm, and 5% or 1% lower Fv/Fm values compared with control plants, indicating only a partial recovery from photoinhibition and irreversible damages in PSII.