Andrew N. Webber

Acclimation of photosynthetic proteins to rising atmospheric CO2.

In this review we discuss how the photosynthetic apparatus, particularly Rubisco, acclimates to rising atmospheric CO2 concentrations (ca). Elevated ca alters the control exerted by different enzymes of the Calvin cycle on the overall rate of photosynthetic CO2 assimilation, so altering the requirement for different functional proteins. A decreased flux of carbon through the photorespiratory pathway will decrease requirements for these enzymes. From modeling of the response of CO2 uptake (A) to intracellular CO2 concentration (ci) it is shown that the requirement for Rubisco is decreased at elevated ca, whilst that for proteins limiting ribulose 1,5 bisphosphate regeneration may be increased. This balance may be altered by other interactions, in particular plasticity of sinks for photoassimilate and nitrogen supply; hypotheses on these interactions are presented. It is speculated that increased accumulation of carbohydrate in leaves developed at elevated ca may signal the ‘down regulation’ of Rubisco. The molecular basis of this ‘down regulation’ is discussed in terms of the repression of photosynthetic gene expression by the elevated carbohydrate concentrations. This molecular model is then used to predict patterns of acclimation of perennials to long term growth in elevated ca.

Increased Accumulation of Carbohydrates and Decreased Photosynthetic Gene Transcript Levels in Wheat Grown at an Elevated CO2 Concentration in the Field

Repression of photosynthetic genes by increased soluble carbohydrate concentrations may explain acclimation of photosynthesis to elevated CO2 concentration. This hypothesis was examined in a field crop of spring wheat (Triticum aestivum L.) grown at both ambient (approximately 360 [mu]mol mol-1) and elevated (550 [mu]mol mol-1) atmospheric CO2 concentrations using free-air CO2 enrichment at Maricopa, Arizona. The correspondence of steady-state levels of mRNA transcripts (coding for the 83-kD photosystem I apoprotein, sedoheptulose-1,7-bisphosphatase, phosphoribulokinase, phosphoglycerokinase, and the large and small subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase) with leaf carbohydrate concentrations (glucose-6-phosphate, glucose, fructose, sucrose, fructans, and starch) was examined at different stages of crop and leaf development and through the diurnal cycle. Overall only a weak correspondence between increased soluble carbohydrate concentrations and decreased levels for nuclear gene transcripts was found. The difference in soluble carbohydrate concentration between leaves grown at elevated and current ambient CO2 concentrations diminished with crop development, whereas the difference in transcript levels increased. In the flag leaf, soluble carbohydrate concentrations declined markedly with the onset of grain filling; yet transcript levels also declined. The results suggest that, whereas the hypothesis may hold well in model laboratory systems, many other factors modified its significance in this field wheat crop.

A fifth chloroplast-encoded polypeptide is present in the photosystem II reaction centre complex

The small polypeptides present in the pea photosystem II reaction centre complex have been separated by high‐resolution SDS‐polyacrylamide gel electrophoresis and characterised by N‐terminal amino acid sequencing. Two polypeptides are identified as the α‐ and β‐subunits of cytochrome b559 which are the products of the pdbE and psbF genes. A third polypeptide, of 4.5 kDA, is identified as the product of a small open reading frame in chloroplast DNA. The gene for this polypeptide , psbI, is located just downstream of the psbK gene chloroplast DNA.

Acclimation response of spring wheat in a free-air CO2 enrichment (FACE) atmosphere with variable soil nitrogen regimes. 2. Net assimilation and stomatal conductance of leaves.

Atmospheric CO2 concentration continues to rise. It is important, therefore, to determine what acclimatory changes will occur within the photosynthetic apparatus of wheat (Triticum aestivum L. cv. Yecora Rojo) grown in a future high-CO2 world at ample and limited soil N contents. Wheat was grown in an open field exposed to the CO2 concentration of ambient air [370 μmol (CO2) mol−1; Control] and air enriched to ∼200 μmol (CO2) mol−1 above ambient using a Free-Air CO2 Enrichment (FACE) apparatus (main plot). A High (35 g m−2) or Low (7 and 1.5 g m−2 for 1996 and 1997, respectfully) level of N was applied to each half of the main CO2 treatment plots (split-plot). Under High-N, FACE reduced stomatal conductance (gs) by 30% at mid-morning (2 h prior to solar noon), 36% at midday (solar noon) and 27% at mid-afternoon (2.5 h after solar noon), whereas under Low-N, gs was reduced by as much as 31% at mid-morning, 44% at midday and 28% at mid-afternoon compared with Control. But, no significant CO2 × N interaction effects occurred. Across seasons and growth stages, daily accumulation of carbon (A′) was 27% greater in FACE than Control. High-N increased A′ by 18% compared with Low-N. In contrast to results for gs, however, significant CO2 × N interaction effects occurred because FACE increased A′ by 30% at High-N, but by only 23% at Low-N. FACE enhanced the seasonal accumulation of carbon (A′′) by 29% during 1996 (moderate N-stress), but by only 21% during 1997 (severe N-stress). These results support the premise that in a future high-CO2 world an acclimatory (down-regulation) response in the photosynthetic apparatus of field-grown wheat is anticipated. They also demonstrate, however, that the stimulatory effect of a rise in atmospheric CO2 on carbon gain in wheat can be maintained if nutrients such as nitrogen are in ample supply.

Detection of calcium binding by photosystem II polypeptides immobilised onto nitrocellulose membrane

Photosystem II calcium‐binding polypeptides have been detected by their ability to selectively bind 45Ca when immobilised onto nitrocellulose membrane following SDS‐polyacrylamide gel electrophoresis. Two calcium‐binding polypeptides of 26 kDa and 24 kDa are shown to be components of LHCII. The 24 kDa polypeptide was further characterised by N‐terminal amino acid sequence analysis and shown to be the product of a type II cab gene. A third polypeptide of 33 kDa bound calcium more weakly and was not positively identified.

The effect of ionic stress on photosynthesis in Dunaliella tertiolecta : Chlorophyll fluorescence kinetics and spectral characteristics.

A comparison of the effects of ionic stress and an uncoupler on long-term fluorescence transients (the ‘Kautsky effect’) in the green alga Dunaliella tertiolecta indicated that the large quenching induced by ionic stress was caused by a pH gradient across the thylakoid membrane. This possiblity was given support by the increase in the slow phase of 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea-induced fluorescence relaxation in algae subjected to ionic stress. Low-temperature fluorescence emission spectra indicated that salt stress enhanced photosystem-I emission in the dark, and a comparison of simultaneous emissions at 695 and 720 nm at room temperature indicated a further increase in photosystem-I emission during the fluorescence transients. Taken together with the decrease in the fast phase of 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea-induced fluorescence relaxation in stressed algae, our results indicate that ionic stress stimulates cyclic electron flow, and that non-cyclic flow is inhibited. The effect of sucrose-induced osmotic stress was similar to, but less marked than, the effects of NaCl and KCl; the effect of decreasing the external salinity was small.

Acclimation response of spring wheat in a free-air CO2 enrichment (FACE) atmosphere with variable soil nitrogen regimes. 3. Canopy architecture and gas exchange.

The response of whole-canopy net CO2 exchange rate (CER) and canopy architecture to CO2 enrichment and N stress during 1996 and 1997 for open-field-grown wheat ecosystem (Triticum aestivum L. cv. Yecora Rojo) are described. Every Control (C) and FACE (F) CO2 treatment (defined as ambient and ambient +200 μmol mol−1, respectively) contained a Low- and High-N treatment. Low-N treatments constituted initial soil content amended with supplemental nitrogen applied at a rate of 70 kg N ha−1 (1996) and 15 kg N ha−1 (1997), whereas High-N treatments were supplemented with 350 kg N ha−1 (1996 and 1997). Elevated CO2 enhanced season-long carbon accumulation by 8% and 16% under Low-N and High-N, respectively. N-stress reduced season-long carbon accumulation 14% under ambient CO2, but by as much as 22% under CO2 enrichment. Averaging both years, green plant area index (GPAI) peaked approximately 76 days after planting at 7.13 for FH, 6.00 for CH, 3.89 for FL, and 3.89 for CL treatments. Leaf tip angle distribution (LTA) indicated that Low-N canopies were more erectophile than those of High-N canopies: 48° for FH, 52° for CH, and 58° for both FL and CL treatments. Temporal trends in canopy greenness indicated a decrease in leaf chlorophyll content from the flag to flag-2 leaves of 25% for FH, 28% for CH, 17% for CL, and 33% for FL during 1997. These results indicate that significant modifications of canopy architecture occurs in response to both CO2 and N-stress. Optimization of canopy architecture may serve as a mechanism to diminish CO2 and N-stress effects on CER.

Function of 3' non-coding sequences and stop codon usage in expression of the chloroplast psaB gene in Chlamydomonas reinhardtii.

The rate of mRNA decay is an important step in the control of gene expression in prokaryotes, eukaryotes and cellular organelles. Factors that determine the rate of mRNA decay in chloroplasts are not well understood. Chloroplast mRNAs typically contain an inverted repeat sequence within the 3′ untranslated region that can potentially fold into a stem-loop structure. These stem-loop structures have been suggested to stabilize the mRNA by preventing degradation by exonuclease activity, although such a function in vivo has not been clearly established. Secondary structures within the translation reading frame may also determine the inherent stability of an mRNA. To test the function of the inverted repeat structures in chloroplast mRNA stability mutants were constructed in the psaB gene that eliminated the 3′ flanking sequences of psaB or extended the open reading frame into the 3′ inverted repeat. The mutant psaB genes were introduced into the chloroplast genome of Chlamydomonas reinhardtii. Mutants lacking the 3′ stem-loop exhibited a 75% reduction in the level of psaB mRNA. The accumulation of photosystem I complexes was also decreased by a corresponding amount indicating that the mRNA level is limiting to PsaB protein synthesis. Pulse-chase labeling of the mRNA showed that the decay rate of the psaB mRNA was significantly increased demonstrating that the stem-loop structure is required for psaB mRNA stability. When the translation reading frame was extended into the 3′ inverted repeat the mRNA level was reduced to only 2% of wild-type indicating that ribosome interaction with stem-loop structures destabilizes chloroplast mRNAs. The non-photosynthetic phenotype of the mutant with an extended reading frame allowed us to test whether infrequently used stop codons (UAG and UGA) can terminate translation in vivo. Both UAG and UGA are able to effectively terminate PsaB synthesis although UGA is never used in any of the Chlamydomonas chloroplast genes that have been sequenced.

Acclimation response of spring wheat in a free-air CO2 enrichment (FACE) atmosphere with variable soil nitrogen regimes. 1. Leaf position and phenology determine acclimation response

We have examined the photosynthetic acclimation of wheat leaves grown at an elevated CO2 concentration, and ample and limiting N supplies, within a field experiment using free-air CO2 enrichment (FACE). To understand how leaf age and developmental stage affected any acclimation response, measurements were made on a vertical profile of leaves every week from tillering until maturity. The response of assimilation (A) to internal CO2 concentration (Ci) was used to estimate the in vivo carboxylation capacity (Vcmax) and maximum rate of ribulose-1,5-bisphosphate limited photosynthesis (Asat). The total activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), and leaf content of Rubisco and the Light Harvesting Chlorophyll a/b protein associated with Photosystem II (LHC II), were determined. Elevated CO2 did not alter Vcmax in the flag leaf at either low or high N. In the older shaded leaves lower in the canopy, acclimatory decline in Vcmax and Asat was observed, and was found to correlate with reduced Rubisco activity and content. The dependency of acclimation on N supply was different at each developmental stage. With adequate N supply, acclimation to elevated CO2 was also accompanied by an increased LHC II/Rubisco ratio. At low N supply, contents of Rubisco and LHC II were reduced in all leaves, although an increased LHC II/Rubisco ratio under elevated CO2 was still observed. These results underscore the importance of leaf position, leaf age and crop developmental stage in understanding the acclimation of photosynthesis to elevated CO2 and nutrient stress.

Photosystem I reaction-centre proteins contain leucine zipper motifs: A proposed role in dimer formation

The photosystem I (PS I) reaction‐centre polypeptides, encoded by the psaA and psaB genes, are shown to contain several highly conserved leucine repeats, consisting of a leucine residue every seventh amino acid, similar to the leucine zipper motifs known to mediate DNA‐binding polypeptide dimerisation. In each of the PSI reaction‐centre subunits the leucine zipper motif precedes highly conserved cysteine residues which have been proposed to ligate the interpolypeptide [4Fe‐4S] centre, Fx. We propose that PS I reaction‐centre dimerisation and [4Fe‐4S] centre formation are mediated through the leucine zipper.