The recent discovery of the Entner-Doudoroff (ED) pathway as a third glycolytic route beside Embden-Meyerhof-Parnas (EMP) and oxidative pentose phosphate (OPP) pathway in oxygenic photoautotrophs requires a revision of their central carbohydrate metabolism. In this study, unexpectedly, we observed that deletion of the ED pathway alone, and even more pronounced in combination with other glycolytic routes, diminished photoautotrophic growth in continuous light in the cyanobacterium Synechocystis sp. PCC 6803. Furthermore, we found that the ED pathway is required for optimal glycogen catabolism in parallel to an operating Calvin-Benson-Bassham (CBB) cycle. It is counter-intuitive that glycolytic routes, which are a reverse to the CBB cycle and do not provide any additional biosynthetic intermediates, are important under photoautotrophic conditions. However, observations on the ability to reactivate an arrested CBB cycle revealed that they form glycolytic shunts that tap the cellular carbohydrate reservoir to replenish the cycle. Taken together, our results suggest that the classical view of the CBB cycle as an autocatalytic, completely autonomous cycle that exclusively relies on its own enzymes and CO2 fixation to regenerate ribulose-1,5-bisphosphate for Rubisco is an oversimplification. We propose that in common with other known autocatalytic cycles, the CBB cycle likewise relies on anaplerotic reactions to compensate for the depletion of intermediates, particularly in transition states and under fluctuating light conditions that are common in nature.

Summary Flavonoid synthesis is predominantly regulated at the transcriptional level through the MYB−basic helix‐loop‐helix (bHLH)−WD40 (MBW) (MYB: transcription factor of the myeloblastosis protein family, WD40: tanscription factor with a short structural motif of 40 amino acids which terminates in an aspartic acid–tryptophan dipeptide) complex, and responds to both environmental and developmental stimuli. Although the developmental regulation of flavonoid accumulation in Arabidopsis thaliana has been examined in great detail, the response of the flavonoid synthesis pathway to abiotic stress (particularly low temperature) remains unclear. A screen of a Dissociation element (Ds) transposon‐induced mutation collection identified two lines which exhibited an altered profile of phenylpropanoid accumulation following exposure to low‐temperature stress. One of the mutated genes (BRASSINOSTEROID ENHANCED EXPRESSION1 (BEE1)) encoded a brassinosteroid enhanced expression transcription factor, while the other (G2‐LIKE FLAVONOID REGULATOR (GFR)) encoded a G2‐like flavonoid regulator. Phenylpropanoid‐targeted analysis was performed using high‐performance LC‐MS, and gene expression analysis using quantitative reverse transcription−PCR. In both mutants, the accumulation of quercetins and scopolin was reduced under low‐temperature growing conditions, whereas that of anthocyanin was increased. BEE1 and GFR were both shown to negatively regulate anthocyanin accumulation by inhibiting anthocyanin synthesis genes via the suppression of the bHLH (TRANSPARENT TESTA8 (TT8) and GLABROUS3 (GL3)) and/or the MYB (PRODUCTION OF ANTHOCYANIN PIGMENTS2 (PAP2)) components of the MBW complex. Our results provide new insight into the regulatory control of phenylpropanoid metabolism at low temperatures, and reveal that BEE1 and GFR act as important components of the signal transduction chain.

The photosynthetic apparatus of higher plants acclimates to irradiance. Among the features which are changing is the pool size of the pigments belonging to the violaxanthin cycle, in which zeaxanthin is formed. In high light grown leaves, the violaxanthin cycle pool size is up to five times larger than in low light. The changes are reversible on a time scale of several days. Since it has been published that violaxanthin cycle pigments do not transfer absorbed energy to chlorophyll, we hypothesized that excitation of chlorophyll fluorescence in the blue spectral region may be reduced in high light-acclimated leaves. Fluorescence excitation spectra of leaves of the Arabidopsis thaliana tt3 mutant showed strong differences between high and low light-acclimated plants from 430 to 520 nm. The resulting difference spectrum was similar to carotenoids but shifted by about 20 nm to higher wavelengths. A good correlation was observed between the fluorescence excitation ratio F 470/F 660 and the violaxanthin cycle pool size when leaves were acclimated to a range of irradiances. In parallel to the decline of F 470/F 660 with high light acclimation also the quantum yield of photosynthetic oxygen evolution in blue light decreased. The data confirm that violaxanthin cycle carotenoids do not transfer absorbed light to chlorophyll. It is proposed to use the ratio F 470/F 660 as an indicator for the light acclimation status of the chloroplasts in a leaf.

Up to 40% of incident light was screened in red Berberis leaves in vivo by anthocyanins, resulting also in up to 40% reduction of light-limited photosynthesis. The biological function of anthocyanins in leaves has been strongly discussed, but the hypothesis of a screening function is favored by most authors. For an evaluation of the function as photoprotective pigments, a quantification of their screening of the mesophyll is important. Here, chlorophyll fluorescence excitation of leaves of a red and a green variety of Berberis thunbergii was used to estimate the extent of screening by anthocyanins at 545 nm and over the whole photosynthetically active wavelength range. Growth at high light (430 µmol m s) resulted in 90% screening at 545 nm corresponding to 40-50% screening over the whole wavelength range, depending on the light source. The concomitant reduction of photosynthetic quantum yield was of the same size as the calculated reduction of light reaching the chloroplasts. The induction of anthocyanins in the red variety also enhanced the epoxidation state of the violaxanthin cycle under growth conditions, indicating that red leaves were suffering less from excessive irradiance. Pool sizes of violaxanthin cycle carotenoids indicated a shade acclimation of the light harvesting complexes in red leaves. The observed reduction of internal light in anthocyanic leaves has by necessity a photoprotective effect.

The accumulation of foliar phenolics constitutes one strategy of plants against the potentially harmful effects of ultraviolet-B and A (UV-B, UV-A) radiation. These compounds protect photosensitive tissues by shielding and antioxidative function. It is unknown, however, whether seasonal acclimation to natural conditions may modify the UV-B effect on phenylpropanoid composition and localisation, and thus their screening efficiency. To address this debate, a field experiment with the wildtype of Arabidopsis thaliana accession Landsberg erecta (Ler) was implemented over a whole year with plants exposed to different UV-filter treatments. While seasonal increases of UV-B radiation had a slight negative effect on the amount of hydroxycinnamic acids (HCAs), low temperatures increased foliar HCAs. HCAs, however, did not contribute substantially to seasonal changes of in vivo UV absorbance. Kaempferol and quercetin derivatives increased significantly under ambient UV-B radiation, and low temperature interacted with this effect. A shift of epidermal UV-A shielding from kaempferol to quercetin derivatives was elucidated in UV-B presence. Despite this, a substantial 20-fold increase of quercetin derivatives, during periods with high irradiance and low temperature, did not affect UV absorbance leading to the conclusion that quercetin accumulation was not exclusively in epidermal vacuoles. Using confocal microscopy, the potential occurrence of quercetin in mesophyll cells was demonstrated in plants grown with experimental UV-B radiation at low temperature for the first time in A. thaliana. The presented study discusses the idea that cross-talk of UV-B radiation and temperature might adjust the physiological function of quercetin from an (epidermal) screening to an antioxidant substance. | INTRODUCTIONPlants rely on diverse mechanisms to attain resistance against the potentially deleterious effects of ultraviolet-A and B radiation (UV-A: 315-400 nm, UV-B: 280-315 nm). Among others, the accumulation of phenolic compounds is one important strategy. However, there is an ongoing debate about the major function of foliar phenolic compounds (reviewed in Winkel-Shirley 2002;Agati et al. 2020). Strong arguments support the idea that these aromatic substances have a dual function in leaves: they are antioxidants on the one hand