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. 2000 Nov;124(3):1275-84.
doi: 10.1104/pp.124.3.1275.

Ultraviolet-B radiation impacts light-mediated turnover of the photosystem II reaction center heterodimer in Arabidopsis mutants altered in phenolic metabolism

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Ultraviolet-B radiation impacts light-mediated turnover of the photosystem II reaction center heterodimer in Arabidopsis mutants altered in phenolic metabolism

I S Booij-James et al. Plant Physiol. 2000 Nov.

Abstract

Ultraviolet-B (UV-B) radiation can have a negative impact on the growth and development of plants. Plants tolerant to UV-B alleviate these effects using UV-screening pigments that reduce the penetration of UV-B into mesophyll tissue. Little is known about the relative contribution of specific phenolic compounds to the screening capacity of leaves. The D1 and D2 proteins constituting the photosystem (PS) II reaction center heterodimer are targets of UV-B radiation and can be used as an in situ sensor for UV penetration into photosynthetic tissue. Degradation of these proteins occurs under very low fluences of UV-B, and is strongly accelerated in the presence of visible light. Using the D1-D2 degradation assay, we characterized UV-B sensitivity of Arabidopsis mutants (tt4, tt5, and fah1) that are genetically altered in their composition of phenolic compounds. We found that changes in phenol metabolism result in altered rates of PSII reaction center heterodimer degradation under mixtures of photosynthetically active radiation and UV-B. A comparison of D2 degradation kinetics showed increased UV sensitivity of the Landsberg (Landsberg erecta) tt5 mutant relative to the Landsberg tt4 mutant and the Landsberg wild type. Despite a lack of flavonoid accumulation, the tt4 mutant is not particularly UV sensitive. However, the tolerance of this mutant to UV-B may reflect the increased accumulation of sinapate esters that strongly absorb in the UV range, and may thus protect the plant against environmentally relevant UV-B radiation. This sinapate-mediated protection is less obvious for the tt4 mutant of Columbia ecotype, indicating that the relative contribution of particular phenolics to the total screening capacity varies with the genetic background. The role of sinapate esters in UV screening is further substantiated by the results with the fah1 mutant where absence of most of the sinapate esters results in a significantly accelerated degradation of D2 under mixed light conditions. Because the latter mutant is not expected to be deficient in flavonoids, the relative contribution of flavonoids as protectants of PSII reaction center heterodimer against UV-B damage in Arabidopsis needs to be re-evaluated vis-a-vis screening by simple phenolics like sinapate esters.

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Figures

Figure 1
Figure 1
The biosynthetic pathway of plant phenolics: the intermediates and enzymes involved. Also shown are the blocked reactions in the Arabidopsis mutants used in the present study. See also Table I.
Figure 2
Figure 2
D1 protein degradation in Arabidopsis Columbia and Landsberg ecotypes, and in mutants fah1, tt4–2YY6, tt4-W85, and tt5. Leaves were pulse-labeled with [35S] Met for 3 h under 50 μmol m−2 s−1 PAR, rinsed, and chased for various periods of time at the PAR fluence indicated, in the presence (0.62 μmol m−2 s−1) or absence of a background of UV-B radiation. Following the exposure to radiation, plants were homogenized and the membrane proteins were isolated and fractionated by SDS/PAGE. Radiolabeled bands were detected by autoradiography and their degradation kinetics were determined as described (Greenberg et al., 1987). Values represent averages of data from several independent experiments (n = 4–8). se of the mean are shown.
Figure 3
Figure 3
D2 protein degradation in Arabidopsis Columbia and Landsberg ecotypes, and in mutants fah1, tt4–2YY6, tt4-W85, and tt5. Leaves were pulse-labeled and chased as described in the legend to Figure 1. Leaves were exposed to the PAR fluence indicated, in the presence (0.62 μmol m−2 s−1) or absence of a background of UV-B radiation. Values represent averages of data from several independent experiments (n = 4–8). se of the mean are shown.

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