Photosynthetic capacity was assessed by measuring electron transport rate (ETR), maximum photosystem II (PSII) quantum efficiency (Fv/Fm) and quantum yield of PSII (ΦPSII) of drought‐treated and control plants using Imaging‐PAM (M‐Series, Maxi version; Heinz Walz GmbH) (Baker, 2008 ). Plants were adapted to dark for 15 min before imaging. ETR was determined on the base of a rapid light curve, which was used for the calculation of fluorescence parameters at a photosynthetic photon flux density of 0, 56, 111, 186, 281, 336 and 396 µmol m−2 s−1 light intensities. Two leaf areas were selected on each plant and 20 plants were measured for each genotype. Experiments were repeated four times.
Imaging pam m series maxi version
Manufactured by Walz
Sourced in Germany
The IMAGING-PAM M-Series Maxi Version is a versatile and powerful tool for non-invasive, pulse-amplitude modulated (PAM) chlorophyll fluorescence imaging. It is designed to measure and analyze the photosynthetic performance of plants and other photosynthetic organisms.
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7 protocols using imaging pam m series maxi version
1
Photosynthetic Capacity Evaluation under Drought
2
Photosynthetic Parameters Determination
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Photosynthesis parameters at PSII were determined fluorometrically according to Schreiber et al. (1986) (link) and Genty et al. (1989) (link) with an Imaging PAM (M-series, Maxi version, Walz, Effeltrich, Germany) Induction kinetics of photosynthesis and light saturation curves have been assessed according to the routines implemented in the Imaging PAM software. The plants were dark-adapted for 30 min prior to induction with actinic light for 5 min. Care was taken that the light saturation curves of photosynthesis were measured immediately after induction. The duration at each PFD was 20 s starting with the lowest PFD. The parameters ΦNPQ (quantum efficiency of directed non-photochemical quenching [mainly ‘heat emission’] at PSII) and ΦNO (quantum efficiency of non-directed ‘other’ non-photochemical events) were derived from theoretical considerations discussed in Kramer et al. (2004) (link), simplified by Hendrickson et al. (2004) (link). From the quantum efficiency of PSII electron transport (ΦPSII) non-cyclic electron transport rates (ETR) can be estimated according to Genty et al. (1989) (link). Since neither leaf absorptance nor antenna cross sections of the photosystems and excitation energy distribution between them were determined, ETR is expressed in relative terms.
3
Photosynthesis Parameters Determination
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Photosynthesis parameters at PSII were determined fluorometrically according to Schreiber et al. (1986) (link) and Genty et al. (1989) (link) with an Imaging PAM (M-series, Maxi version, Walz, Effeltrich, Germany). Induction kinetics of photosynthesis and light saturation curves have been assessed according to the routines implemented in the Imaging PAM software. The plants were dark-adapted for 30 min prior to induction with actinic light for 5 min. Care was taken that the light saturation curves of photosynthesis were measured immediately after induction. The duration at each PFD was 20 s starting with the lowest PFD. From the quantum efficiency of PSII photochemistry (ΦPSII) electron transport rates (ETR) can be estimated according to Genty et al. (1989) (link). Since neither leaf absorptance nor antenna cross sections of the photosystems and excitation energy distribution between them were determined, ETR is expressed in relative terms.
4
Chlorophyll a Fluorescence Analysis of Arabidopsis
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The chlorophyll a fluorescence levels of the Arabidopsis leaves (20 days old) were monitored with IMAGING-PAM M-Series Maxi Version (Heinz Walz GmbH, Effeltrich, Germany). The plants were dark-adapted for at least 1 hr before the measurements. After the maximum and initial fluorescence (Fm and F0, respectively) were determined with a delay of 40 s, the plants were illuminated using the following light intensities: 0, 81, 145, 186, 281, 335, 461, 701 and 926 μmol photon m−2 s−1. The duration of the illumination of each light intensity was 3 min. A saturation pulse (800 ms, 2700 μmol photon m−2 s−1) was applied at the end of the 3-min illumination. For each light intensity, the photosynthesis parameters were calculated using the formulas: Y(II) was calculated as (Fm’ − F) / Fm’; qP was calculated as (Fm’ − F) / (Fm’ − F0’); Non-photochemical quenching (NPQ) was calculated as (Fm − Fm’) / Fm’; electron transport rate (ETR) was calculated as Y(II) × light intensity × 0.5 × absorptivity. Fm’ was the maximal fluorescence of light-illuminated plant. F was the current fluorescence yield. F0’ was estimated by F0 / (Fv / Fm + F0 / Fm’) (Oxborough & Baker, 1997 ).
5
Chlorophyll Fluorescence of Seedlings
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Chlorophyll fluorescence in 10-d-old seedlings was monitored with the IMAGING-PAM M-Series Maxi Version (WALZ). Before the measurement, the treated seedlings were incubated in the dark for 1 h. After the maximum and initial fluorescence (Fm Fo) were determined and a delay of 40 s, the plants were illuminated with a light intensity of 296 µmol⋅m−2⋅s−1 for 3 min. A saturation pulse was applied at the end of the 3-min illumination to measure the chlorophyll fluorescence parameters.
6
Monitoring Chlorophyll Fluorescence in Arabidopsis
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The chlorophyll fluorescence levels of Arabidopsis leaves were monitored with IMAGING-PAM M-Series Maxi Version (WALZ, Effeltrich, Germany). The plants were dark-adapted for at least 1 h before measurement. After the maximum (Fm) and initial (Fo) fluorescence values were determined with a delay of 40 s, the plants were illuminated at the following light intensities: 0, 81, 145, 186, 281, 335, 461, 701, and 926 µmol photon m−2 s−1. The duration of illumination at individual light intensities was 3 min. A saturation pulse was applied at the end of each 3 min illumination.
7
Photosynthetic Efficiency Analysis of Arabidopsis Leaves
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The chlorophyll a fluorescence levels of the Arabidopsis leaves (20 days old) were monitored with IMAGING-PAM M-Series Maxi Version (WALZ, Germany). The plants were darkadapted for at least 1 hour before the measurements. After the maximum and initial fluorescence (Fm and F0, respectively) were determined with a delay of 40 s, the plants were illuminated using the following light intensities: 0, 81, 145, 186, 281, 335, 461, 701, and 926 µmol photon m -2 s -1 . The duration of the illumination of each light intensity was 3 min. A saturation pulse (800 ms, 2700 µmol photon m -2 s -1 ) was applied at the end of the 3-min illumination. For each light intensity, the photosynthesis parameters were calculated using the formulas: Y(II) was calculated as (Fm' -F) /Fm' ; qP is calculated as (Fm' -F) /(Fm' -Fo'); NPQ was calculated as (Fm -Fm') /Fm' ; ETR was calculated as Y(II) x light intensity x 0.5 x absorptivity. Fm' is the maximal fluorescence of light-illuminated plant. F is the current fluorescence yield. Fo' was estimated by Fo /(Fv /Fm + Fo /Fm') (Oxborough and Baker, 1997) .
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