Limitations to photosynthesis by proton motive force-induced photosystem II photodamage
- Michigan State University, United States
- Max-Planck-Institut für Molekulare Pflanzenphysiologie, Germany
- Imperial College London, United Kingdom
- Washington State University, United States
Figures
Figure 1
γ-subunit mutations alter photosynthetic proton efflux.
Sequence alignment of Arabidopsis ATPC1 and ATPC2 regulatory region (A). Amino acid differences incorporated into ATPC1 to generate minira are indicated by symbols (♦). Amino acid numbers are based …
Figure 2 with 16 supplements
Dynamic light conditions enhance pmf dependent phenotypes.
Whole plant fluorescent images were captured over three days under the illumination conditions displayed in Panel A and listed in Supplementary file 2. Plants were illuminated over the 16-hr …
Figure 2—figure supplement 1
Whole plant fluorescence imaging phenotyping of minira 3–1 mutant.
Three week old plants were imaged over three consecutive 16-hr photoperiods and fluorescent measurements taken at the end of each light transition for LEF (A–C), qE (D–F), and qI (G–I) for Ws-2 and m…
Figure 2—figure supplement 2
Increased pH-dependent quenching correlates with increased photoinhibitory quenching.
Whole plant fluorescent phenotypes were measured over three consecutive photoperiods. Day one consisted of a single irradiance level (open symbols), day two of sinusoidal irradiance (half filled …
Figure 2—figure supplement 3
Whole plant fluorescence imaging phenotyping of minira 11–1 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 4
Whole plant fluorescence imaging phenotyping of minira 14–1 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 5
Whole plant fluorescence imaging phenotyping of minira 12–2 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 6
Whole plant fluorescence imaging phenotyping of minira 8–1 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 7
Whole plant fluorescence imaging phenotyping of minira 6–2 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 8
Whole plant fluorescence imaging phenotyping of minira 4–2 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 9
Whole plant fluorescence imaging phenotyping of minira 6–1 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 10
Whole plant fluorescence imaging phenotyping of minira 7–1 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 11
Whole plant fluorescence imaging phenotyping of minira 3–2 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 12
Whole plant fluorescence imaging phenotyping of minira 4–1 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 13
Whole plant fluorescence imaging phenotyping of minira 9–1 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 14
Whole plant fluorescence imaging phenotyping of minira 4–3 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 15
Whole plant fluorescence imaging phenotyping of minira 12–3 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 2—figure supplement 16
Whole plant fluorescence imaging phenotyping of minira 2–2 mutant.
For details, please refer to the legend of Figure 2—figure supplement 1.
Figure 3
Elevated pmf leads to PSII photodamage.
Detached leaves were infiltrated with either water (A,C) or a 3 mM solution of lincomycin (B,D) and treated with 1000 μmol photons m−2s−1 red light for the times indicated. Following dark …
Figure 4
The dependence of photoinhibition on the redox state of QA.
The redox state of the primary electron acceptor QA was assayed using the qL fluorescence parameter concurrently with photoinhibitory quenching qI at 100 (solid symbols), 300 (half filled symbols), …
Figure 5 with 3 supplements
Photoinhibition is strongly correlated with Δψ but not ΔpH in minira lines.
Photoinhibition, estimated by the qI fluorescence parameter, is plotted against either the ΔpH or Δψ components of pmf, estimated by the ECSss (A) and ECSinv (B) parameters, as described in …
Figure 5—figure supplement 1
Reduction kinetics of P700+.
Light-dependent P700+ reduction half-times (mean ± s.d., n = 3) of wild type, minira 3–1 and minira 14–1 (A). Quenching of the 810 nm absorbance signal was followed during a brief dark interval and …
Figure 5—figure supplement 2
The electric field component of the pmf dominates under high pmf conditions.
ECS measurements were performed to determine the partitioning of the light-driven pmf between ΔpH and Δψ. Measurements were performed at 100 (solid symbols), 300 (half filled symbols), and 500 μmol …
Figure 5—figure supplement 3
Tobacco ATPC1 antisense knockdown increase Δψ partitioning under high pmf conditions.
The partitioning of the pmf in wild type Samsun (black) and ATPC1 (red) γ-subunit antisense line were determined from the deconvoluted ECS signal at ΔA520 nm (A). Following the light-dark transition …
Figure 6
In vitro manipulation of the pmf Δψ alters PSII S2QA− recombination rates.
Isolated spinach thylakoids in the presence of 5 μM spinach ferredoxin and 10 μm sodium ascorbate were treated with 3–3,4-dichlorophenyl 1,1-dimethylurea (DCMU) to block PSII forward electron …
Figure 7 with 2 supplements
Induction of Δψ and 1O2 production under fluctuating light in wild type plants.
(A) Illumination conditions and measurement points used in the experiments. Fluctuating light conditions (replicating Figure 2 day three) are shown as connected points, with open squares …
Figure 7—figure supplement 1
Uncoupling Δψ decreases SOSG fluorescence in minira 3–1.
Minira 3–1 leaves were vacuum infiltrated with either SOSG (solid triangles) or SOSG and 50 μM valinomycin (crossed triangles) to decrease the photosynthetic Δψ. Leaves were illuminated at a …
Figure 7—figure supplement 2
Fluctuations in light intensity result in transient ECS spikes.
Wild type plants were measured under fluctuating light (A) and the ECS measurements taken 10 s after each intensity fluctuation from lower to higher light (A, open squares). The resulting …
Figure 8
Schemes for the trans-thylakoid Δψ-induced acceleration of recombination reactions in PSII and subsequent production of 1O2.
(A) The relative positions of PSII electron transfer cofactors with respect to the electric field (double-headed arrow) imposed across the thylakoid membrane (dotted lines). The red and blue arrows …
Videos
Video 1
(phi2_movie_labeled_day1): Whole plant PSII quantum efficiency (ΦII) during constant illumination.
False-colored chlorophyll fluorescence images of whole plants obtained during constant, 100 μmol photons m−2 s−1 actinic illumination over a 16 hr photoperiod. Measurements and calculations were …
Video 2
(phi2_movie_labeled_day2): Whole plant PSII quantum efficiency (ΦII) during sinusoidal illumination.
False-colored chlorophyll fluorescence images of whole plants obtained during sinusoidal actinic illumination over a 16 hr photoperiod. Measurements and calculations were performed as described in …
Video 3
(phi2_movie_labeled_day3): Whole plant PSII quantum efficiency (ΦII) during fluctuating sinusoidal illumination.
False-colored chlorophyll fluorescence images of whole plants obtained during fluctuating sinusoidal actinic illumination over a 16 hr photoperiod. Measurements and calculations were performed as …
Video 4
qE_movie_labeled_day1): Whole plant pH-dependent quenching (qE) during constant illumination.
False-colored chlorophyll fluorescence images of whole plants obtained during constant, 100 μmol photons m−2 s−1 actinic illumination over a 16 hr photoperiod. Measurements and calculations were …
Video 5
(qE_movie_labeled_day2): Whole plant pH-dependent quenching (qE) during sinusoidal illumination.
False-colored chlorophyll fluorescence images of whole plants obtained during sinusoidal actinic actinic illumination over a 16 hr photoperiod. Measurements and calculations were performed as …
Video 6
(qE_movie_labeled_day3): Whole plant pH-dependent quenching (qE) during fluctuating sinusoidal illumination.
False-colored chlorophyll fluorescence images of whole plants obtained during fluctuating sinusoidal actinic illumination over a 16 hr photoperiod. Measurements and calculations were performed as …
Video 7
(qI_movie_labeled_day1): Whole plant photoinhibitory quenching (qI) during constant illumination.
False-colored chlorophyll fluorescence images of whole plants obtained during constant, 100 μmol photons m−2 s−1 actinic illumination over a 16 hr photoperiod. Measurements and calculations were …
Video 8
(qI_movie_labeled_day2): Whole plant photoinhibitory quenching (qI) during sinusoidal illumination.
False-colored chlorophyll fluorescence images of whole plants obtained during sinusoidal actinic actinic illumination over a 16 hr photoperiod. Measurements and calculations were performed as …
Video 9
(qI_movie_labeled_day3): Whole plant photoinhibitory quenching (qI) during fluctuating sinusoidal illumination.
False-colored chlorophyll fluorescence images of whole plants obtained during fluctuating sinusoidal actinic illumination over a 16 hr photoperiod. Measurements and calculations were performed as …
Additional files
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Supplementary file 1
Oligonucleotides used for site directed mutagenesis of atpc1.
(a) Oligonucleotide sequences utilized for adapter ligation mutagenesis. (b) Oligonucleotide sequences utilized for adapter ligation mutagenesis to introduce secondary mutations. (c) Oligonucleotide sequences utilized for splicing by overlap extension PCR. (d) Synthetic gene constructs incorporating multiple ATPC2 mutations into ATPC1.
- https://doi.org/10.7554/eLife.16921.040
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Supplementary file 2
Timing and light intensity profiles used for chlorophyll fluorescence imaging.
(a) Timing and light profile of imaging day one. (b) Timing and light profile of imaging day two. (c) Timing and light profile of imaging day three.
- https://doi.org/10.7554/eLife.16921.041
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Supplementary file 3
Chlorophyll content of wild-type (Ws-2) and minira leaves.
All measurements were performed on three-week old leaves following in vivo spectroscopic measurements as described in Materials and methods. Data represent the mean ± s.d. for n ≥ 3 leaves. Statistically significant differences (*p<0.05) from wild-type were determined using a t-test.
- https://doi.org/10.7554/eLife.16921.042
