Climate and the U.S. distribution of C4 grass subfamilies and decarboxylation variants of C4 photosynthesis

I compared the C(4) grass flora and climatic records for 32 sites in the United States. Consistent with previous studies, I found that the proportion of the grass flora that uses the NADP malic enzyme (NADP-ME) variant of C(4) photosynthesis greatly increases with increasing annual precipitation, while the proportion using the NAD malic enzyme (NAD-ME) variant (and also the less common phosphoenolpyruvate carboxykinase [PCK] variant) decreases. However the association of grass subfamilies with annual precipitation was even stronger than for the C(4) decarboxylation variants. Analysis of the patterns of distribution by partial correlation analysis showed that the correlations between the frequency of various C(4) types and rainfall were solely due to the association of the C(4) types with particular grass subfamilies. In contrast, there was a strong correlation of the frequency of the different subfamilies with annual precipitation that was independent of the influence of the different C(4) variants. It therefore appears that other, as yet unidentified, characteristics that differ among grass subfamilies may be responsible for their differences in distribution across natural precipitation gradients.     

The activities of PEP carboxylase and the C4 acid decarboxylases are little changed by drought stress in three C4 grasses of different subtypes

The C(4) photosynthetic pathway involves the assimilation of CO(2) by phosphoenolpyruvate carboxylase (PEPC) and the subsequent decarboxylation of C(4) acids. The enzymes of the CO(2) concentrating mechanism could be affected under water deficit and limit C(4) photosynthesis. Three different C(4) grasses were submitted to gradually induced drought stress conditions: Paspalum dilatatum (NADP-malic enzyme, NADP-ME), Cynodon dactylon (NAD-malic enzyme, NAD-ME) and Zoysia japonica (PEP carboxykinase, PEPCK). Moderate leaf dehydration affected the activity and regulation of PEPC in a similar manner in the three grasses but had species-specific effects on the C(4) acid decarboxylases, NADP-ME, NAD-ME and PEPCK, although changes in the C(4) enzyme activities were small. In all three species, the PEPC phosphorylation state, judged by the inhibitory effect of L: -malate on PEPC activity, increased with water deficit and could promote increased assimilation of CO(2) by the enzyme under stress conditions. Appreciable activity of PEPCK was observed in all three species suggesting that this enzyme may act as a supplementary decarboxylase to NADP-ME and NAD-ME in addition to its role in other metabolic pathways.     

Oxygen and electron flow in C4 photosynthesis: Mehler reaction, photorespiration and CO2 concentration in the bundle sheath

The photosynthetic linear electron transport rate in excess of that used for CO₂ reduction was evaluated in Sorghum bicolor Moench. [NADP-malic enzyme (ME)-type C₄ plant], Amaranthus cruentus L. (NAD-ME-type C₄ plant) and Helianthus annuus L. (C₃ plant) leaves at different CO₂ and O₂ concentrations. The electron transport rate (J _F) was calculated from fluorescence using the light partitioning factor (relative PSII cross-section) determined under conditions where excess electron transport was assumed to be negligible: low light intensities, 500 μmol CO₂ · mol⁻¹ and 2% O₂. Under high light intensities there was a large excess of J _F/4 at 10–100% O₂ in the C₃ plant due to photorespiration, but very little in sorghum and somewhat more in amaranth, showing that photorespiration is suppressed, more in the NADP-ME- and less in the NAD-ME-type species. It is concluded that when C₄ photosynthesis is limited by supply of atmospheric CO₂ to the C₄ cycle, the C₃ cycle becomes limited by regeneration of ribulose 1,5-bisphosphate (RuBP) which in turn limits RuBP oxygenase activity and photorespiration. The rate of excess electron transport over that consumed for CO₂ fixation in C₄ plants was very sensitive to the presence of O₂ in the gas phase, rapidly increasing between 0.01 and 0.1% O₂, and at 2% O₂ it was about two-thirds of that at 21% O₂. This shows the importance of the Mehler O₂ reduction as an electron sink, compared with photorespiration in C₄ plants. However, the rate of the Mehler reaction is still too low to fully account for the extra ATP which is needed in C₄ photosynthesis. Received: 8 November 1997 / Accepted: 26 December 1997     

An active Mehler-peroxidase reaction sequence can prevent cyclic PS I electron transport in the presence of dioxygen in intact spinach chloroplasts

Simultaneous measurements of 9-aminoacridine (9-AA) fluorescence quenching, O₂-uptake and chlorophyll fluorescence of intact spinach chloroplasts were carried out to assess the relationship between the transthylakoidal pH and linear electron flux passing through Photosystem II. Three different types of O₂-dependent electron flow were investigated: (1) Catalysed by methyl viologen; (2) in the absence of a catalyst and presence of an active ascorbate peroxidase (Mehler-peroxidase reaction); (3) in the absence of a catalyst and with the ascorbate peroxidase being inhibited by KCN (Mehler reaction). The aim of this study was to assess the relative contribution of pH-formation which is not associated with electron flow through Photosystem II and, which should reflect Photosystem I cyclic flow under the different conditions. The relationship between the extent of 9-AA fluorescence quenching and O₂-uptake rate was found to be almost linear when methyl viologen was present. In the absence of methyl viologen (Mehler reaction) an increase of 9-AA fluorescence quenching to a value of 20% at low light intensities was associated with considerably less O₂-uptake than in the presence of methyl viologen, indicating the involvement of cyclic flow. These findings are in agreement with a preceding study of Kobayashi and Heber (1994). However, when no KCN was added, such that the complete Mehler-peroxidase reaction sequence was operative, the relationship between 9-AA fluorescence quenching and the flux through PS II, as measured via the chlorophyll fluorescence parameter F/Fm × PAR, was identical to that observed in the presence of methyl viologen. Under the assumption that methyl viologen prevents cyclic flow, it is concluded that there is no significant contribution of cyclic electron flow to pH-generation in intact spinach chloroplasts.     

Factors Limiting Photosynthetic Recovery in Sweet Pepper Leaves After Short-Term Chilling Stress Under Low Irradiance

The effects of chilling treatment (4 °C) under low irradiance, LI (100 mol m^–2 s^–1) and in the dark on subsequent recovery of photosynthesis in chilling-sensitive sweet pepper leaves were investigated by comparing the ratio of quantum yields of photosystem (PS) 2 and CO₂ assimilation, _PS2/_CO2, measured in normal air (21 % O₂, NA) and low O₂-air (2% O₂, LOA), and by analyzing chlorophyll (Chl) a fluorescence parameters. Chilling treatment in the dark had little effect on F_v/F_m and _PS2/_CO2, but it caused the decrease of net photosynthetic rate (P _N) under saturating irradiance after 6-h chilling treatment, indicating that short-term chilling alone did not induce PS2 photoinhibition. Furthermore, photorespiration and Mehler reaction also did not obviously change during subsequent recovery after chilling stress in the dark. During chilling treatment under LI, there were obvious changes in F_v/F_m and _PS2/_CO2, determined in NA or LOA. F_v/F_m could recover fully in 4 h at 25 °C, and _PS2/_CO2 increased at the end of the treatment, as determined in both NA and LOA. During subsequent recovery, _PS2/_CO2 in LOA decreased faster than in NA. Thus the Mehler reaction might play an important role during chilling treatment under LI, and photorespiration was an important process during the subsequent recovery. The recovery of PN under saturating irradiance determined in NA and LOA took about 50 h, implying that there were some factors besides CO₂ assimilation limiting the recovery of photosynthesis. From the progress of reduced P700 and the increase of the Mehler reaction during chilling under LI we propose that active oxygen species were the factors inducing PS1 photoinhibition, which prevented the recovery of photosynthesis in optimal conditions because of the slow recovery of the oxidizable P700.     

水分胁迫下苹果叶片的H_2O_2代谢

轻度水分胁迫下苹果叶片Pr迅速升高 ,CAT活性变化不大 ,NaHSO3 处理能显著降低叶内H2 O2 含量 ,表明光呼吸的加强促进了H2 O2 产生可能是叶内H2 O2大量积累的主要原因 ;中度水分胁迫下叶片AsA含量的下降和Mehler反应的增强都非常明显 ,DDTC和AsA处理都能有效降低叶内H2 O2 积累 ,但MV处理的作用不大 ,说明叶片H2 O2 主要来源于Mehler反应 ,AsA降解造成叶片对H2 O2 清除能力的下降是其积累的根本原因 ;严重水分胁迫时 ,NaHSO3 和DDTC都不能有效地减轻叶内H2 O2 积累 ,光呼吸和Mehler反应都可能不是H2 O2 的主要来源     

Responses of wild C4 and C3 grass (Poaceae) species to elevated atmospheric CO2 concentration: a meta-analytic test of current theories and perceptions

C4 plants contribute ≈ 20% of global gross primary productivity, and uncertainties regarding their responses to rising atmospheric CO₂ concentrations may limit predictions of future global change impacts on C4-dominated ecosystems. These uncertainties have not yet been considered rigorously due to expectations of C4 low responsiveness based on photosynthetic theory and early experiments. We carried out a literature review (1980–97) and meta-analysis in order to identify emerging patterns of C4 grass responses to elevated CO₂, as compared with those of C3 grasses. The focus was on nondomesticated Poaceae alone, to the exclusion of C4 dicotyledonous and C4 crop species. This provides a clear test, controlled for genotypic variability at family level, of differences between the CO₂-responsiveness of these functional types. Eleven responses were considered, ranging from physiological behaviour at the leaf level to carbon allocation patterns at the whole plant level. Results were also assessed in the context of environmental stress conditions (light, temperature, water and nutrient stress), and experimental growing conditions (pot size, experimental duration and fumigation method). Both C4 and C3 species increased total biomass significantly in elevated CO₂, by 33% and 44%, respectively. Differing tendencies between types in shoot structural response were revealed: C3 species showed a greater increase in tillering, whereas C4 species showed a greater increase in leaf area in elevated CO₂. At the leaf level, significant stomatal closure and increased leaf water use efficiency were confirmed in both types, and higher carbon assimilation rates were found in both C3 and C4 species (33% and 25%, respectively). Environmental stress did not alter the C4 CO₂-response, except for the loss of a significant positive CO₂-response for above-ground biomass and leaf area under water stress. In C3 species, stimulation of carbon assimilation rate was reduced by stress (overall), and nutrient stress tended to reduce the mean biomass response to elevated CO₂. Leaf carbohydrate status increased and leaf nitrogen concentration decreased significantly in elevated CO₂ only in C3 species. We conclude that the relative responses of the C4 and C3 photosynthetic types to elevated CO₂ concur only to some extent with expectations based on photosynthetic theory. The significant positive responses of C4 grass species at both the leaf and the whole plant level demand a re-evaluation of the assumption of low responsiveness in C4 plants at both levels, and not only with regard to water relations. The combined shoot structural and water use efficiency responses of these functional types will have consequential implications for the water balance of important catchments and range-lands throughout the world, especially in semiarid subtropical and temperate regions. It may be premature to predict that C4 grass species will lose their competitive advantage over C3 grass species in elevated CO₂.     

Photoinhibition and Active Oxygen Species Production in Detached Apple Leaves During Dehydration

In the course of dehydration, the gas exchange and chlorophyll (Chl) fluorescence were measured under irradiance of 800 mol m^–2 s^–1 in detached apple leaves, and the production of active oxygen species (AOS), hydrogen peroxide (H₂O₂), superoxide (O₂ ^–), hydroxyl radical (–OH), and singlet oxygen (¹O₂), were determined. Leaf net photosynthetic rate (P _N) was limited by stomatal and non-stomatal factors at slight (2–3 h dehydration) and moderate (4–5 h dehydration) water deficiency, respectively. Photoinhibition occurred after 3-h dehydration, which was defined by the decrease of photosystem 2 (PS2) non-cyclic electron transport (P-rate). After 2-h dehydration, an obvious rise in H₂O₂ production was found as a result of photorespiration rise. If photorespiration was inhibited by sodium bisulfite (NaHSO₃), the rate of post-irradiation transient increase in Chl fluorescence (Rfp) was enhanced in parallel with a slight decline in P-rate and with an increase in Mehler reaction. At 3-h dehydration, leaf P-rate decrease could be blocked by glycine (Gly) or methyl viologen (MV) pre-treatment, and MV was more effective than Gly at moderate drought time. AOS (H₂O₂ and O₂ ^–), prior to photoinhibition produced from photorespiration and Mehler reaction in detached apple leaves at slight water deficiency, were important in dissipating photon energy which was excess to the demand of CO₂ assimilation. So photoinhibition could be effectively prevented by the way of AOS production.     

Intact chloroplasts display pH 5 optimum of O2-reduction in the absence of methyl viologen: Indirect evidence for a regulatory role of superoxide protonation

The pH-dependence of light-driven O₂-reduction in intact spinach chloroplasts is studied by means of chlorophyll fluorescence quenching analysis and polarographic O₂-uptake measurements. Most experiments are carried out in presence of KCN, which blocks activities of Calvin cycle, ascorbate peroxidase and superoxide dismutase. pH is varied by equilibration with external buffers in presence of nigericin. Vastly different pH-optima for O₂-dependent electron flow are observed in the presence and absence of the redox catalyst methyl viologen. Both fluorescence quenching analysis and O₂-uptake reveal a distinct pH 5 optimum of O₂-reduction in the absence of methyl viologen. In the presence of this catalyst, O₂-reduction is favoured in the alkaline region, with an optimum around pH 8, similar to other types of Hill reaction. It is suggested that in the absence of methyl viologen the extent of irreversibility of O₂-reduction is determined by the rate of superoxide protonation. This implies that O₂-reduction takes place within the aprotic phase of the thylakoid membrane and that superoxide-reoxidation via oxidized PS I donors competes with protonation. Superoxide protonation is proposed to occur at the internal surface of the thylakoid membrane. There is no competition between superoxide reoxidation and protonation when in the presence of methyl viologen the site of O₂-reduction is shifted into the protic stroma phase. In confirmation of this interpretation, fluorescence measurements in the absence of KCN reveal, that non-catalysed O₂-dependent electron flow is unique in beingstimulated by the transthylakoidal pH-gradient. On the basis of these findings a major regulatory role of O₂-dependent electron flow under excess light conditions is postulated.     

Photosystem II energy use,non-photochemical quenching and the xanthophyll cycle in Sorghumbicolor grown under drought and free-air CO2 enrichment(FACE) conditions

The present study was carried out to test the hypothesis thatelevated atmospheric CO₂ (Ca) will alleviate over‐excitationof the C₄ photosynthetic apparatus and decrease non‐photochemicalquenching (NPQ) during periods of limited water availability. Chlorophyll a fluorescencewas monitored in Sorghum bicolor plants grown under a free‐aircarbon‐dioxide enrichment (FACE) by water‐stress (Dry) experiment.Under Dry conditions elevated Ca increased the quantum yield ofphotosystem II (φPSII) throughout the day throughincreases in both photochemical quenching coefficient (q_p)and the efficiency with which absorbed quanta are transferred toopen PSII reaction centres (F_v′/F_m′).However, in the well‐watered plants (Wets) FACE enhanced φPSIIonly at midday and was entirely attributed to changes in F_v′/F_m′_. Underfield conditions, decreases in φPSII under Dry treatmentsand ambient Ca corresponded to increases in NPQ but the de‐epoxidation stateof the xanthophyll pool (DPS) showed no effects. Water‐stress didnot lead to long‐term damage to the photosynthetic apparatus asindicated by φPSII and carbon assimilation measuredafter removal of stress conditions. We conclude that elevated Caenhances photochemical light energy usage in C₄ photosynthesisduring drought and/or midday conditions. Additionally,NPQ protects against photo‐inhibition and photodamage. However,NPQ and the xanthophyll cycle were affected differently by elevatedCa and water‐stress.     

Effects of inorganic carbon accumulation on photosynthetic oxygen reduction and cyclic electron flow in the cyanobacterium Synechococcus PCC7942

This paper examines the effect of inorganic carbon transport and accumulation in Synechococcus PCC7942 on fluorescence quenching, photosynthetic oxygen reduction and both linear and cyclic electron flow. The data presented support the previous findings of Miller et al. (1991) that the accumulation of Ci by the CO₂ concentrating mechanism is able to stimulate oxygen photoreduction, particularly so when CO₂ fixation is inhibited by PCR cycle inhibitors such as glycolaldehyde. This effect is found with both high and low-Ci grown cells, but the potential for oxygen photoreduction is about two-fold higher in low-Ci grown cells. This greater potential for O₂ photoreduction is also correlated with a higher ability of low-Ci cells to photoreduce H₂O₂. Experiments with a mutant which transports Ci but does not accumulate it internally, indicates that the stimulation of O₂ photoreduction appears to be a direct effect of the internal accumulation of Ci rather than from its participation in the transport process. In the absence of Ci, no specific partial reactions of photosynthetic electron transport appear to be inhibited, and the PS 1 acceptors PNDA and MV as well as the PS 2 acceptor DMQ can all run electron transport at levels approaching those during active CO₂ fixation. Measurements of P700⁺ show that when the cells are depleted of Ci during photosynthesis, P700 becomes more oxidised. This indicates that the resupply of electrons from the intersystem chain is relatively more restricted under conditions of Ci limitation than is the availability of PS 1 electron acceptors. It is proposed that the accumulated Ci pool can directly stimulate the ability of O₂ to act as a PS 1 acceptor and that the ability of PS 1 acceptors, such as O₂, to relieve restrictions on intersystem electron transfer is perhaps a result of a reduction in cyclic electron flow and a subsequent increase in the oxidation state of the plastoquinone pool.Abbreviations BTP 1,3-bis[tris(hydroxymethyl)-methylaminopropane] - CA carbonic anhydrase' - Ci inorganic carbon (CO₂+HCO₃ ^–+CO₃ ^2–) - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DMQ 2,6-dimethylbenzoquinone - EZ ethoxyzolamide or 6-ethoxy-2-benzothiazole-sulfonamide - FCCP carbonyl cyanide p-trifluoro methoxyphenyl-hydrazone - F steady-state chlorophyll fluorescence - F_m chlorophyll fluorescence during a saturating light pulse - F_o chlorophyll fluorescence in the dark, prior to illumination by actinic light - MV methyl viologen or 1,1-dimethyl-4,4-bipyridinium dichloride - PCR cycle photosynthetic carbon reduction cycle - PNDA N,N-dimethyl-p-nitrosoaniline - _{PS 1} the quantum yield of Photosystem 1 - _{PS 2} the quantum yield of Photosystem 2     

C3 grasses have higher nutritional quality than C4 grasses under ambient and elevated atmospheric CO2

Grasses with the C₃ photosynthetic pathway are commonly considered to be more nutritious host plants than C₄ grasses, but the nutritional quality of C₃ grasses is also more greatly impacted by elevated atmospheric CO₂ than is that of C₄ grasses; C₃ grasses produce greater amounts of nonstructural carbohydrates and have greater declines in their nitrogen content than do C₄ grasses under elevated CO₂. Will C₃ grasses remain nutritionally superior to C₄ grasses under elevated CO₂ levels? We addressed this question by determining whether levels of protein in C₃ grasses decline to similar levels as in C₄ grasses, and whether total carbohydrate : protein ratios become similar in C₃ and C₄ grasses under elevated CO₂. In addition, we tested the hypothesis that, among the nonstructural carbohydrates in C₃ grasses, levels of fructan respond most strongly to elevated CO₂. Five C₃ and five C₄ grass species were grown from seed in outdoor open‐top chambers at ambient (370 ppm) or elevated (740 ppm) CO₂ for 2 months. As expected, a significant increase in sugars, starch and fructan in the C₃ grasses under elevated CO₂ was associated with a significant reduction in their protein levels, while protein levels in most C₄ grasses were little affected by elevated CO₂. However, this differential response of the two types of grasses was insufficient to reduce protein in C₃ grasses to the levels in C₄ grasses. Although levels of fructan in the C₃ grasses tripled under elevated CO₂, the amounts produced remained relatively low, both in absolute terms and as a fraction of the total nonstructural carbohydrates in the C₃ grasses. We conclude that C₃ grasses will generally remain more nutritious than C₄ grasses at elevated CO₂ concentrations, having higher levels of protein, nonstructural carbohydrates, and water, but lower levels of fiber and toughness, and lower total carbohydrate : protein ratios than C₄ grasses.     

Mechanisms of energy dissipation in peanut under water stress

Effect of drought on the mechanisms of energy dissipation was evaluated in two-month-old Arachis hypogaea cvs. 57–422, 73–30, and GC 8–35. Plants were submitted to three treatments: control (C), mild water stress (S1), and severe water stress (S2). Photosynthetic performance was evaluated as the Hill and Mehler reactions. These activities were correlated with the contents of the low and high potential forms of cytochrome (cyt) b ₅₅₉, plastoquinone, cyt b ₅₆₃, and cyt f. Additionally, the patterns of carotenoids and chlorophylls (Chls), as well as the alterations of Chl a fluorescence parameters were studied. Under mild water stress the regulatory mechanism at the antennae level was effective for 57–422 and GC 8–35, while in the cv. 73–30 an overcharge of photosynthetic apparatus occurred. Relative to this cv., under S1 the stability of carotene and the dissipative cycle around photosystem (PS) 2 became an important factor for the effective protection of the PS2 reaction centres. The cyclic electron flow around PS1 was important for energy dissipation under S1 only for the cvs. 57–422 and 73–30.     

Activities of Photosystems I and II in Chlamydomonas segnis Adapted and Adapting to Air and Air-enriched with Carbon Dioxide*

Activities of photosystems I and II were compared at a saturating irradiance in air- and 5% CO₂-adapted and adapting Chlamydomonas segnis at the active phase of photosynthesis during the cell cycle. PSII activity was 200% greater in air- than in 5% CO₂-adapted cells, while PSI activity was similar in both types of cells and matched the level of PSII activity in air-adapted cells. As a result, air- and 5% CO₂-adapted cells were characterized by low and high PSI/PSII ratios, respectively. In air-adapted cells, the greater PSII activity (rate of O₂ evolved) exceeded that of photosynthetic (Ps) O₂ evolution, resulting in a Ps/PSII ratio below unity. This was associated with higher levels of catalase activity, lower l -ascorbate content, and higher dehydro-l -ascorbate content than in 5% CO₂-adapted cells. During adaptation to air or 5% CO₂ for 6 h in light, PSI rather than PSII was sensitive to changes in the concentration of CO₂, and the adapting cells acquired the characteristics of air- and 5% CO₂-adapted cells as indicated by PSI/PSII, Ps/PSII, catalase activity, l -ascorbate and dehydro-l -ascorbate contents. The results are discussed in the light of changes in the molecular organization of the thylakoid membranes and enhanced non-cyclic electron transport coupled with O₂-uptake (Mehler reaction) for the generation of the ATP required for CO₂/HCO^?₃-transport in air-adapted and adapting cells.