Stefan Falk

Photosynthesis, photoinhibition and low temperature acclimation in cold tolerant plants.

Cold acclimation requires adjustment to a combination of light and low temperature, conditions which are potentially photoinhibitory. The photosynthetic response of plants to low temperature is dependent upon time of exposure and the developmental history of the leaves. Exposure of fully expanded leaves of winter cereals to short-term, low temperature shiftsinhibits whereas low temperature growthstimulates electron transport capacity and carbon assimilation. However, the photosynthetic response to low temperature is clearly species and cultivar dependent. Winter annuals and algae which actively grow and develop at low temperature and moderate irradiance acquire a resistance to irradiance 5- to 6-fold higher than their growth irradiance. Resistance to short-term photoinhibition (hours) in winter cereals is a reflection of the increased capacity to keep QA oxidized under high light conditions and low temperature. This is due to an increased capacity for photosynthesis. These characteristics reflect photosynthetic acclimation to low growth temperature and can be used to predict the freezing tolerance of cereals. It is proposed that the enhanced photosynthetic capacity reflects an increased flux of fixed carbon through to sucrose in source tissue as a consequence of the combined effects of increased storage of carbohydrate as fructans in the vacuole of leaf mesophyll cells and an enhanced export to the crown due to its increased sink activity. Long-term exposure (months) of cereals to low temperature photoinhibition indicates that this reduction of photochemical efficiency of PS II represents a stable, long-term down regulation of PS II to match the energy requirements for CO2 fixation. Thus, photoinhibition in vivo should be viewed as the capacity of plants to adjust photosynthetically to the prevailing environmental conditions rather than a process which necessarily results in damage or injury to plants. Not all cold tolerant, herbaceous annuals use the same mechanism to acquire resistance to photoinhibition. In contrast to annuals and algae, overwintering evergreens become dormant during the cold hardening period and generally remain susceptible to photoinhibition. It is concluded that the photosynthetic response to low temperatures and susceptibility to photoinhibition are consequences of the overwintering strategy of the plant species.

Growth at Low Temperature Mimics High-Light Acclimation in Chlorella vulgaris'

Structural and functional alterations to the photosynthetic apparatus after growth at low temperature (5[deg]C) were investigated in the green alga Chlorella vulgaris Beijer. Cells grown at 5[deg]C had a 2-fold higher ratio of chlorophyll a/b, 5-fold lower chlorophyll content, and an increased xanthophyll content compared to cells grown at 27[deg]C even though growth irradiance was kept constant at 150 [mu]mol m-2 s-1. Concomitant with the increase in the chlorophyll a/b ratio was a lower abundance of light-harvesting polypeptides in 5[deg]C-grown cells as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and confirmed by western blotting.The differences in pigment composition were found to be alleviated within 12 h of transferring 5[deg]C-grown cells to 27[deg]C. Furthermore, exposure of 5[deg]C-grown cells to a 30-fold lower growth irradiance (5 [mu]mol m-2 s-1) resulted in pigment content and composition similar to that in cells grown at 27[deg]C and 150 [mu]mol m-2 s-1. Although both cell types exhibited similar measuring-temperature effects on CO2-saturated O2 evolution, 5[deg]C-grown cells exhibited light-saturated rates of O2 evolution that were 2.8-and 3.9-fold higher than 27[deg]C-grown cells measured at 27[deg]C and 5[deg]C, respectively. Steady-state chlorophyll a fluorescence indicated that the yield of photosystem II electron transport of 5[deg]C-grown cells was less temperature sensitive than that of 27[deg]C-grown cells. This appears to be due to an increased capacity to keep the primary, stable quinone electron acceptor of photosystem II (QA) oxidized at low temperature in 5[deg]C- compared with 27[deg]C-grown cells regardless of irradiance. We conclude that Chlorella acclimated to low temperature adjusts its photosynthetic apparatus in response to the excitation pressure on photosystem II and not to the absolute external irradiance. We suggest that the redox state of QA may act as a signal for this photosynthetic acclimation to low temperature in Chlorella.

Photosystem II Excitation Pressure and Development of Resistance to Photoinhibition (I. Light-Harvesting Complex II Abundance and Zeaxanthin Content in Chlorella vulgaris)

The basis of the increased resistance to photoinhibition upon growth at low temperature was investigated. Photosystem II (PSII) excitation pressure was estimated in vivo as 1 - qp (photochemical quenching). We established that Chlorella vulgaris exposed to either 5[deg]C/150 [mu]mol m-2 s-1 or 27[deg]C/2200 [mu]mol m-2 s-1 experienced a high PSII excitation pressure of 0.70 to 0.75. In contrast, Chlorella exposed to either 27[deg]C/150 [mu]mol m-2 s-1 or 5[deg]C/20 [mu]mol m-2 s-1 experienced a low PSII excitation pressure of 0.10 to 0.20. Chlorella grown under either regime at high PSII excitation pressure exhibited: (a) 3-fold higher light-saturated rates of O2 evolution; (b) the complete conversion of PSII[alpha] centers to PSII[beta] centers; (c) a 3-fold lower epoxidation state of the xanthophyll cycle intermediates; (d) a 2.4-fold higher ratio of chlorophyll a/b; and (e) a lower abundance of light-harvesting polypeptides than Chlorella grown at either regime at low PSII excitation pressure. In addition, cells grown at 5[deg]C/150 [mu]mol m-2 s-1 exhibited resistance to photoinhibition comparable to that of cells grown at 27[deg]C/2200 [mu]mol m-2 s-1 and were 3- to 4-fold more resistant to photoinhibition than cells grown at either regime at low excitation pressure. We conclude that increased resistance to photoinhibition upon growth at low temperature reflects photosynthetic adjustment to high excitation pressure, which results in an increased capacity for nonradiative dissipation of excess light through zeaxanthin coupled with a lower probability of light absorption due to reduced chlorophyll per cell and decreased abundance of light-harvesting polypeptides.

Functional analysis of the iron-stress induced CP 43' polypeptide of PS II in the cyanobacterium Synechococcus sp. PCC 7942.

Under conditions of iron-stress, the Photosystem II associated chlorophyll a protein complex designated CP 43′, which is encoded by the isiA gene, becomes the major pigment-protein complex in Synechococcus sp. PCC 7942. The isiB gene, which is located immediately downstream of isiA, encodes the protein flavodoxin, which can functionally replace ferredoxin under conditions of iron stress. We have constructed two cyanobacterial insertion mutants which are lacking (i) the CP 43′ apoprotein (designated isiA−) and (ii) flavodoxin (designated isiB−). The function of CP 43′ was studied by comparing the cell characteristics, PS II functional absorption cross-sections and Chl a fluorescence parameters from the wild-type, isiA− and isiB− strains grown under iron-stressed conditions. In all strains grown under iron deprivation, the cell number doubling time was maintained despite marked changes in pigment composition and other cell characteristics. This indicates that iron-starved cells remained viable and that their altered phenotype suggests an adequate acclimation to low iron even in absence of CP 43′ and/or flavodoxin. Under both iron conditions, no differences were detected between the three strains in the functional absorption crossection of PS II determined from single turnover flash saturation curves of Chl a fluorescence. This demonstrates that CP 43′ is not part of the functional light-harvesting antenna for PS II. In the wild-type and the isiB− strain grown under iron-deficient conditions, CP 43′ was present in the thylakoid membrane as an uncoupled Chl-protein complex. This was indicated by (1) an increase of the yield of prompt Chl a fluorescence (Fo) and (2) the persistence after PS II trap closure of a fast fluorescence decay component showing a maximum at 685 nm.

Photosynthetic Adjustment to Temperature

The description of a general mechanism for photosynthetic adjustment to temperature that encompasses all autotrophic species is not possible for three principal reasons: (i) inherent genetic diversity, (ii) differential strategies in growth and development, and (iii) organisms respond to temperature changes rather than to absolute temperature. Thus, ‘high’ and ‘low’ temperature are relative terms and will differ for pyschrophilic, mesophilic and thermophilic organisms. However, given this complexity, some consensus regarding photosynthetic adjustment to temperature is emerging. At low temperature (0–10 °C), photosynthesis is constrained thermodynamically. This may be manifested by chloroplast phosphate limitation due to reduced rates of sucrose synthesis and/or source-sink limitations. In this case, rates of CO2 uptake and O2 evolution are regulated directly through metabolite accumulation (feedback inhibition) and photosynthetic control. Alternatively, feedback inhibition may be regulated indirectly through catabolite repression of photosynthetic genes. Although light may exacerbate susceptibility to photoinhibition at low temperature in many species, cold grown, chilling-tolerant plants exhibit increased capacity for carbohydrate synthesis at low temperature which alleviates phosphate limitation, supplies a cryoprotectant and results in higher photosynthetic capacity than warm-grown plants. However, photosynthetic adjustment in cold-grown higher plants and algae does not reflect adjustment to low temperature per se, but rather, changes in excitation pressure on PS II. In contrast, photosynthesis in chilling-sensitive plants is not only constrained thermodynamically by low temperature but is also severely inhibited developmentally.

Photosynthetic performance and fluorescence in relation to antenna size and absorption cross-sections in rye and barley grown under normal and intermittent light conditions

The size of the Photosystem II light harvesting antenna and the absorption cross-sections of PS I (σPSI) and PS II (σPSII) were examined in relation to photosynthetic performance fluorescence. Wild-type (WT) rye (Secale cereale) and barley (Hordeurn vulgare) as well as the barley chlorophyllb-less chlorina F2 mutant were grown under control and intermittent light (IML) conditions. (σPSII) in control barley F2 was similar to IML grown WT rye and barley, which, in turn was 2.5 to 3.5 times smaller than for control WT plants. In contrast, σPSI was similar for all control plants. This was 2.5 to 4 times larger than for IML-grown WT plants. IML-grown barley mutant plants had the smallest absorption cross-sections. Photosynthetic light response curves revealed that the barley chlorina F2-mutant had rates of oxygen evolution on a per leaf area basis that were only slightly lower than control WT rye and barley while IML-grown plants had strongly reduced photosynthetic performance. Convexity (Θ) for control barley chlorina F2-mutants was equal to the WT controls (0.6–0.7), while all IML-grown plants had a Θ of 0. This indicates that, in contrast to control barley mutants, IML-plants were limited by PS II turn-over rates at all irradiances. However, on a per leaf Chl-basis the IML-grown plants exhibited the highest photosynthetic rates. Thus, the comparatively poor photosynthetic rates for IML-grown plants on a per leaf area basis were not due to less efficient photosynthetic reaction centers, but may rather be due to an increased limitation from PS II turn-over and a reduction in the number of reaction centers per leaf area.