北方粳稻光合速率、气孔导度对光强和CO2浓度的响应

 以东北地区主栽的粳稻（Oryza sativa var. japonica）品种为对象,用美国LI-cor公司生产的Li 6400光合作用测定仪控制光强、CO2浓度和温度等环境条件,阐述了光合作用和气孔导度对光和CO2浓度的响应特征及其耦合关系。结果表明,光合速率随光强或CO2浓度的提高而增大,均遵循米氏响应;在不同CO2浓度下,表观量子效率随CO2浓度的提高而增大,但CO2浓度达到800 μmol•mol－1以上时,表观量子效率有所减小;在不同光强下,表观羧化效率也随光的增强而增大,但光强达到1 600 μmol•m-2•s-1以上时,表观羧化效率也有所减小;在光强和CO2浓度协同作用下,光合速率的响应遵循双底物的米氏方程,在光强和CO2浓度均趋于饱和时,北方粳稻（品种：辽粳294）剑叶的潜在最大光合速率为71.737 8 μmol•m-2•s-1,表观量子效率为0.056 0 μmolCO2•μmol-1 photons,表观羧化效率为0.103 1 μmol•m-2•s-1/μmol•mol－1。气孔导度也随光的增强而增大,对光强的响应规律也可以用Michaelis-Menten曲线模拟,而叶面CO2浓度的提高会使气孔导度减小,气孔导度（Gs）对叶面CO2浓度（Cs）的响应可以用Gs=Gmax,c/(1+Cs/Cs0)的双曲线方程模拟。在光强(PFD)和CO2浓度协同作用下,气孔导度可以用式Gs=Gmax(PFD/PFDc)/［(1+PFD/PFDc)(1+Cs/Cs0)］+Gct估算,当CO2浓度趋于0而光强趋于饱和时,北方粳稻的潜在最大气孔导度（Gmax）为0.670 9 mol•m-2•s-1。在光强和CO2浓度协同作用下,Ball-Berry模型及其修正形式依然能很好地表达气孔导度-光合速率的耦合关系,并且用叶面饱和水汽压差（Ds）修正耦合关系中的相对湿度可以提高模拟精度。     

Leaf factors affecting the relationship between chlorophyll fluorescence and the rate of photosynthetic electron transport as determined from CO2 uptake

CO2 uptake and chlorophyll fluorescence were measured under non-photorespiratory conditions in leaves from 14 plant species. The rate of CO2-dependent electron transport (JCO2) was calculated as four times rate of gross photosynthesis. The quantum yield of electron transport in photosystem II was estimated from the ratio delta F/Fm', where delta F is the difference between steady-state and maximal fluorescence in the light. As photon flux density (PFD) increased, JCO2 increased linearly first, and then reached saturation. The product (delta F/Fm')PFD, which is a function of electron transport rate, showed a similar response. Therefore, the relationship between (delta F/Fm') PFD versus JCO2 was proportional. However, under high light, a linear correlation was not always maintained. Factors affecting the linear correlation were analyzed by measuring CO2 uptake and chlorophyll fluorescence under illumination from either the upper (adaxial) or lower (abaxial) leaf surface, and by using plants with anatomically symmetric leaves having palisade tissues on both sides. Consequently, it was shown that the parameter delta F/Fm' is based on chlorophyll fluorescence emitted from chloroplasts present near the illuminated surface. Further, it was suggested that this restriction of the origin of fluorescence actually measured is significant in a leaf with high chlorophyll content, resulting in the deviation from linearity in the relationship between JCO2 and (delta F/Fm')PFD.     

Chloroplastic Carbon Dioxide Concentration in Norway Spruce (Picea Abies [L.] Karst.) needles relates to the position within the crown

Differences between sun (E) and shaded (S) foliage were studied in a Norway spruce (Picea abies [L.] Karst.) stand. Response curves describing the dependence of the CO2 assimilation rate (PN) on the CO2 concentration at the catalytic site of ribulose-1,5-bisphosphate carboxylase/oxygenase, RuBPCO (PN-Cc) were estimated using the simultaneous measurements of chlorophyll fluorescence and leaf gas exchange. Higher PN, higher electron transport (Ja), higher carboxylation capacity (Vc), and higher RuBPCO activity () for sun acclimated needles was found. The S-needles had higher portion of internal limitation and higher CO2 compensation concentration () than the E-needles. Because higher degree of limitation of photosynthesis by carboxylation was ascertained, it can be assumed that photosynthesis in shade foliage is limited mainly by lower carboxylation capacity and by low chloroplastic CO2 concentration     

Aluminium modulation of the photosynthetic carbon reduction cycle in Zea mays

Two-weeks-old maize (Zea mays L. cv. XL-72.3) plants were submitted to Al concentrations of 0-81 g m-3 for 20 d, after which the A1 concentration-dependent effects on CO2 uptake by the mesophyll tissue and subsequent CO2 assimilation in the photosynthetic carbon reduction cycle of bundle sheath cells were investigated. The net photosynthetic rate (PN) and stomatal conductance (gs) increased continuously up to 27 g m-3 Al, whereas the intercellular CO2 concentration showed minimum values with the 27 g m-3 Al treatment. Moreover, the starch and saccharide concentrations, and fructose-1,6-bisphosphatase did not change significantly with increasing Al concentrations. The photosynthetic electron transport rates along with photosystems 2 and 1 started falling from 9 g m-3 Al onwards, while thylakoid acyl lipid composition did not show a clear pattern. With the Al concentration at 81 g m-3, NADP-malate dehydrogenase activity decreased to minimum values, whereas the opposite occurred with those of pyruvate dikinase, NADP-malic enzyme, and phosphoenolpyruvate carboxylase. Thus in vivo Al concentrations modulate the photosynthetic reduction cycle, possibly by interacting with the carbon flow rate exported to the cytosol. Although the inhibition of NADP-malate dehydrogenase activity might limit pyruvate dikinase, NADP-malic enzyme, and phosphoenolpyruvate carboxylase activities, in vivo the balance between phosphoenolpyruvate production and its carboxylation remains unaffected.     

Suboptimal nitrogen status sensitizes the photosynthetic apparatus in willow leaves to long term but not short term water stress

The response to drought was compared for willow plants of optimal leaf nitrogen content (100 N) and those of 86% of this content (86 N). Gas exchange measurements revealed that the carboxylation efficiency (CE) of photosynthesis was more sensitive to drought than the photosynthetic capacity in both N regimes. Since the leaf content of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) was found to be much more resistant it is suggested that a decreased specific activity of Rubisco underlies the decreased CE. Although the rate of water consumption was the same for 86 N and 100 N plants the photosynthetic apparatus responded much more rapidly in the 86 N leaves. This increased sensitivity of 86 N leaves was not due to accelerated senescence as judged by comparison with parallel plants subjected to discontinued fertilization; the two categories of treatments resulted in the same loss of leaf nitrogen and Rubisco but drought induced a much more rapid photosynthetic depression. In contrast to the drought situation, 86 N and 100 N plants behaved similarly when compared under short term water stress. First, when single attached leaves were exposed to a sudden drop in air humidity the capacity of CO₂ uptake in both N regimes decreased about 20% over 10 min while the leaf water potential remained high. Second, in freely transpiring leaf discs cut from 86 N and 100 N leaves the same relationship between capacity of O₂ evolution and extent of dehydration was observed. The possible mechanisms underlying the increased susceptibility of 86 N leaves to drought is discussed; the water status of the roots not the leaves is suggested to be the determining factor.Abbreviations CE carboxylation efficiency - 100 N optimal nitrogen regime - 86 N suboptimal nitrogen regime with 86% of the optimal leaf nitrogen content, Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase     

Aluminium modulation of the photosynthetic carbon reduction cycle in Zea mays

Two-weeks-old maize (Zea mays L. cv. XL-72.3) plants were submitted to Al concentrations of 0-81 g m-3 for 20 d, after which the A1 concentration-dependent effects on CO2 uptake by the mesophyll tissue and subsequent CO2 assimilation in the photosynthetic carbon reduction cycle of bundle sheath cells were investigated. The net photosynthetic rate (PN) and stomatal conductance (gs) increased continuously up to 27 g m-3 Al, whereas the intercellular CO2 concentration showed minimum values with the 27 g m-3 Al treatment. Moreover, the starch and saccharide concentrations, and fructose-1,6-bisphosphatase did not change significantly with increasing Al concentrations. The photosynthetic electron transport rates along with photosystems 2 and 1 started falling from 9 g m-3 Al onwards, while thylakoid acyl lipid composition did not show a clear pattern. With the Al concentration at 81 g m-3, NADP-malate dehydrogenase activity decreased to minimum values, whereas the opposite occurred with those of pyruvate dikinase, NADP-malic enzyme, and phosphoenolpyruvate carboxylase. Thus in vivo Al concentrations modulate the photosynthetic reduction cycle, possibly by interacting with the carbon flow rate exported to the cytosol. Although the inhibition of NADP-malate dehydrogenase activity might limit pyruvate dikinase, NADP-malic enzyme, and phosphoenolpyruvate carboxylase activities, in vivo the balance between phosphoenolpyruvate production and its carboxylation remains unaffected.     

Light microhabitats,growth and photosynthesis of an epiphytic bromeliad in a tropical dry forest

In the tropical dry forest of Dzibilchaltún, Yucatan, Mexico, light microhabitats, tissue acidity changes, chlorophyll fluorescence and growth were investigated for the epiphytic bromeliad Tillandsia brachycaulos Schltdl. Most individuals occurred in sites between 20 and 60% of the height of the tree, and directly on the main trunk or branches closed to the main trunk. During the dry season, individuals received about nine times more photon flux density (PFD) than during the rainy season. Individuals that occurred under 30–59% of the daily ambient PFD showed greater leaf length increase and monthly leaf production, and produced more flowers than individuals in the brighter (>60% of ambient PFD) and the darker (11–29% of ambient PFD) light microhabitats. Well-watered plants acclimated to deep shade (6% of ambient PFD) showed values of tissue acidity similar to plants under higher light treatments. Well-watered plants under 60% of daily ambient PFD showed low values of maximum quantum efficiency at midday during most of the year but showed similar values of tissue acidity than plants under 6 and 30% of ambient PFD. Leaf temperatures of plants in low sites within the canopy and on the forest floor were significantly higher than the air during about 1 h, indicating that those plants could not dissipate the excess of heat. Individuals located in the most shaded microhabitats could also receive less amount of water from rainfall and dew. Therefore, it would be less damaging for this epiphytic bromeliad to occur in the most exposed microhabitats than in the most shaded microhabitats in this tropical dry forest.     

Effects of Growth Temperature on the Responses of Ribulose-1,5-Biphosphate Carboxylase,Electron Transport Components,and Sucrose Synthesis Enzymes to Leaf Nitrogen in Rice,and Their Relationships to Photosynthesis

Effects of growth temperature on the photosynthetic gas-exchange rates and their underlying biochemical properties were examined in young, fully expanded leaves of rice (Oryza sativa L.). The plants were grown hydroponically under day/night temperature regimes of 18/15[deg]C, 23/18[deg]C, and 30/23[deg]C and all photosynthetic measurements were made at a leaf temperature of 25[deg]C and an irradiance of 1800 [mu]mol quanta m-2 s-1. Growth temperature affected the photosynthetic CO2 response curve. The relative ratio of the initial slope to the CO2-saturated photosynthesis increased with rising growth temperature. This was caused mainly by an increase in CO2-limited photosynthesis for a given leaf nitrogen content with rising growth temperature. However, there was no difference in ribulose-1,5-bisphosphate carboxylase (Rubisco) content at any given leaf nitrogen content among temperature treatments. In addition, the activation state and catalytic turnover rate of Rubisco were not affected by growth temperature. The increase in CO2-limited photosynthesis with rising growth temperature was the result of an increase in the CO2 transfer conductance between the intercellular airspaces and the carboxylation sites. The amounts of total chlorophyll and light-harvesting chlorophyll a/b protein II increased for the same leaf nitrogen content with rising growth temperature, but the amounts of cytochrome f and coupling factor 1 and the activities of cytosolic fructose-1,6-bisphosphatase and sucrose-phosphate synthase were the same between plants grown at 23/18[deg]C and those grown at 30/23[deg]C. Similarly, CO2-saturated photosynthesis was not different for the same leaf nitrogen content between these treatments. For the 18/15[deg]C-grown plants, a slight decrease in the amounts of cytochrome f and coupling factor 1 and an increase in the activities of cytosolic fructose-1,6-bisphosphatase and sucrose-phosphate synthase were found, but these were not reflected in CO2-saturated photosynthesis.     

Elevated CO2 differentially affects photosynthetic induction response in two Populus species with different stomatal behavior

To understand dynamic photosynthetic characteristics in response to fluctuating light under a high CO(2) environment, we examined photosynthetic induction in two poplar genotypes from two species, Populus koreana 9 trichocarpa cv. Peace and Populus euramericana cv. I-55, respectively. Stomata of cv. Peace barely respond to changes in photosynthetic photon flux density (PFD), whereas those of cv. I-55 show a normal response to variations in PFD at ambient CO(2). The plants were grown under three CO2 regimes (380, 700, and 1,020 μmol CO(2) mol(-1) in air) for approximately 2 months. CO2 gas exchange was measured in situ in the three CO2 regimes under a sudden PFD increase from 20 to 800 μmol m(-2) s(-1). In both genotypes, plants grown under higher CO(2) conditions had a higher photosynthetic induction state, shorter induction time, and reduced induction limitation to photosynthetic carbon gain. Plants of cv. I-55 showed a much larger increase in induction state and decrease in induction time under high CO(2) regimes than did plants of cv. Peace. These showed that, throughout the whole induction process, genotype cv. I-55 had a much smaller reduction of leaf carbon gain under the two high CO(2) regimes than under the ambient CO(2) regime, while the high CO(2) effect was smaller in genotype cv. Peace. The results suggest that a high CO(2) environment can reduce both biochemical and stomatal limitations of leaf carbon gain during the photosynthetic induction process, and that a rapid stomatal response can further enhance the high CO(2) effect.     

转C4光合酶基因水稻的CO2交换和荧光特性

用转PEPC、PPDK、NADP-ME、PEPC+PPDK酶基因水稻(Oryza sativa L.)及原种为材料 ,研究了光合作用对光照、温度、CO2的响应和光抑制条件下的叶绿素荧光特性,结果如下: 1.转C4光合酶基因水稻的饱和光合速率比原种高,其中转PEPC、PEPC+PPDK双基因水稻的光饱和点比原种高200 μmol*m-2*s-1,饱和光合速率比原种分别高51.6%和 58.5%;转PEPC基因水稻的羧化效率比原种高49.3%,CO2补偿点降低26.2%;在高温(35 ℃)下,转PEPC基因水稻的光合速率比原种高17.5%.2.经光抑制处理8 d后,转PEPC、PEPC +PPDK酶基因水稻的PSⅡ光化学效率(Fv/Fm)和光化学猝灭(qP)下降20%- 30%,非光化学猝灭(qN)增加了约30%;但原种的Fv/Fm和qP下降了5 0%多,qN变化不明显,表明转C4光合基因水稻耐光抑制能力增强.这些结果为用生物技术提高水稻光合效率研究提供了新的依据和途径.     

苋菜的光合特性

宽菜Amaranthus cruentus cv.生长在调控的温室条件。在光强0至800μmol.m~(-2)S~(-1),光合速率(PN,μmol.CO_2m~(-2)、s~(-1))随光强(PFD,μmol、m~(-2)、s~(-1))增高而增大,其关系为PN=56.82 PFD×10~(-3)—2.13。光补偿点为60μmol.m~(-2)、s~(-1)。叶片在1400 μmol.m~(-2)、s~(-1)达到光合光饱和点。在叶温35℃,叶片/空气水蒸汽压陡度20 m Pa、Pa~(-1)和外界CO_2浓度340μ1、1~(-1),光饱和光合速率为51.63±4.90μ mol.CO_2、m~(-2)、S~(-1)。在光强0至600μmol.m~(-2)、s~(-1),气孔传道率随光强增高而增大。光强高于600μmol.m~(-2)、s~(-1),气孔传道率变化较小。细胞间CO_2浓度为120μ1.1~(-1)由于细胞间CO_2浓度在光合速率——CO_2关系曲线的转折点,可能表明光合作用不受气孔限制。结果表明,苋菜适于高光强环境生长,在干旱条件下具有高的光合速率。     

Small decreases in SBPase cause a linear decline in the apparent RuBP regeneration rate, but do not affect Rubisco carboxylation capacity

The response of net photosynthetic CO(2) uptake (A) to increasing leaf intercellular CO(2) concentration (c(i)) was determined in antisense Nicotiana tabacum plants, derived from six independent transformation lines, displaying a range of sedoheptulose-1, 7-bisphosphatase (SBPase) activities. The maximum in vivo ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylation (V(c,max)) and RuBP regeneration (J(max)) rates were calculated from the steady-state measurements of the A to c(i) response curves. In plants with reductions in SBPase activity of between 9% and 60%, maximum RuBP regeneration capacity declined linearly (r(2)=0.79) and no significant change in apparent in vivo Rubisco activity (V(c,max)) was observed in these plants. No correlation between V(c,max) and a decrease in capacity for RuBP regeneration was observed (r(2)=0.14) in the SBPase antisense plants. These data demonstrate that small decreases in SBPase activity limit photosynthetic carbon assimilation by reducing the capacity for RuBP regeneration.     

Relationships between quantum yield for CO2 assimilation, activity of key enzymes and CO2 leakiness in Amaranthus cruentus, a C4 dicot, grown in high or low light

In C(4) photosynthesis, a part of CO(2) fixed by phosphoenolpyruvate carboxylase (PEPC) leaks from the bundle-sheath cells. Because the CO(2) leak wastes ATP consumed in the C(4) cycle, the leak may decrease the efficiency of CO(2) assimilation. To examine this possibility, we studied the light dependence of CO(2) leakiness (phi), estimated by the concurrent measurements of gas exchange and carbon isotope discrimination, initial activities of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and pyruvate, orthophosphate dikinase (PPDK), the phosphorylation state of PEPC and the CO(2) assimilation rate using leaves of Amaranthus cruentus (NAD-malic enzyme subtype, dicot) plants grown in high light (HL) and low light (LL). phi was constant at photon flux densities (PFDs) >200 micromol m(-2) s(-1) and was around 0.3. At PFDs <150 micromol m(-2) s(-1), phi increased markedly as PFD decreased. At 40 micromol m(-2) s(-1), phi was 0.76 in HL and 0.55 in LL leaves, indicating that the efficiency of CO(2) assimilation at low PFD was greater in LL leaves. The activities of Rubisco and PPDK, and the phosphorylated state of PEPC all decreased as PFD decreased. Theoretical calculations with a mathematical model clearly showed that the increase in phi with decreasing PFD contributed to the decrease in the CO(2) assimilation rate. It was also shown that the 'conventional' quantum yield of photosynthesis obtained by fitting the straight line to the light response curve of the CO(2) assimilation rate at the low PFD region is seriously overestimated. Ecological implications of the increase in phi in LL are discussed.     

柚树(Citrus grandis)叶片光合作用对补增UV-B辐射的响应

生长在人工光照 4 0 0μmol m- 2 s- 1 下的柚树幼树光合速率的最大值为 1 0 .2± 0 .5μmol m- 2 s- 1 ;而补增UV-B辐射 ( 3.8-4 .2μW cm- 2 ,2 4 5～ 2 97nm,4 5d)的叶片则为 6.4± 0 .8μmol m- 2 s- 1 ,较对照植株降低37.2 %。对照植物的表观量子产率 (固定 mol CO2 mol- 1量子 )为 0 .0 75± 0 .0 1 2 ,而经 UV-B辐射处理植株则为0 .0 4 1± 0 .0 0 8,明显较对照植株低。UV-B辐射处理使植株叶片的光呼吸和不包括光呼吸的 CO2 补偿点增高。对照植株叶片的最大值的 CO2 羧化速率 (μmol m- 2 s- 1 )为 57.1± 1 .5μmol m- 2 s- 1 ,较 UV-B辐射处理的高30 .9% ,而 UV-B辐射处理的植株的光合电子传递速率较对照低 30 %。同时 UV-B辐射植株叶片有较低的光能转化效率 ,其较对照低 39.1 % ,叶片亦含有较低的叶绿素含量。结果表明 ,UV-B辐射明显抑制叶片光合羧化速率和光合电子传递速率 ,UV-B辐射可能抑制包括 Rubisco羧化作用在内的多个光合生理过程 ,降低叶片光合速率。柚树叶片对 UV-B辐射敏感 ,选育抗 UV-B辐射的柚树品种势在必行。     

Partitioning of the Leaf CO2 Exchange into Components Using CO2 Exchange and Fluorescence Measurements

Photorespiration was calculated from chlorophyll fluorescence and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) kinetics and compared with CO2 evolution rate in the light, measured by three gas-exchange methods in mature sunflower (Helianthus annuus L.) leaves. The gas-exchange methods were (a) postillumination CO2 burst at unchanged CO2 concentration, (b) postillumination CO2 burst with simultaneous transfer into CO2-free air, and (c) extrapolation of the CO2 uptake to zero CO2 concentration at Rubisco active sites. The steady-state CO2 compensation point was proportional to O2 concentration, revealing the Rubisco specificity coefficient (Ksp) of 86. Electron transport rate (ETR) was calculated from fluorescence, and photorespiration rate was calculated from ETR using CO2 and O2 concentrations, Ksp, and diffusion resistances. The values of the best-fit mesophyll diffusion resistance for CO2 ranged between 0.3 and 0.8 s cm-1. Comparison of the gas-exchange and fluorescence data showed that only ribulose-1,5-bisphosphate (RuBP) carboxylation and photorespiratory CO2 evolution were present at limiting CO2 concentrations. Carboxylation of a substrate other than RuBP, in addition to RuBP carboxylation, was detected at high CO2 concentrations. A simultaneous decarboxylation process not related to RuBP oxygenation was also detected at high CO2 concentrations in the light. We propose that these processes reflect carboxylation of phosphoenolpyruvate, formed from phosphoglyceric acid and the subsequent decarboxylation of malate.     

Cool-night temperature induces spike emergence and affects photosynthetic efficiency and metabolizable carbohydrate and organic acid pools in <Emphasis Type="Italic">Phalaenopsis aphrodite</Emphasis>

After being acclimated to constant warm (28 degrees C day/28 degrees C night) and cool-night temperature (28 degrees C day/20 degrees C night) regimes in growth chambers for 2 weeks, the two groups of mature Phalaenopsis aphrodite subsp. formosana plants both clearly exhibited a diurnal oscillation of stomatal conductance, net CO(2) uptake rate, malate and starch levels, and the phosphoenolpyruvate carboxylase (EC 4.1.1.31) and NAD(+)-malic enzyme (EC 1.1.1.39) activities. Hence, P. aphrodite is an obligate crassulacean acid metabolism plant. Nevertheless, different night temperature greatly affected both the stomatal conductance and the contribution of ambient and respiratory CO(2) to the nocturnal accumulation of malate. However, the amounts of nocturnal accumulated malate and daily deposited starch appeared to have no significant difference between the two groups. These results demonstrate that P. ahrodite is congruent with the characteristics of CAM plants having great flexibility and plasticity in response to changes in environmental conditions. In addition, the formation of reproductive stem, viz. spike, was noticeably inhibited by a constant warm temperature, but induced by a fluctuating warm day and cool night condition. The relationship between the metabolic pool variation and spike induction of Phalaenopsis is also discussed.     

Effects of leaf age, nitrogen nutrition and photon flux density on the organization of the photosynthetic apparatus in leaves of a vine (Ipomoea tricolor Cav.) grown horizontally to avoid mutual shading of leaves

Effects of leaf age, nitrogen nutrition and photon flux density (PFD) on the organization of the photosynthetic apparatus in leaves were investigated in a vine, Ipomoea tricolor Cav., which was grown horizontally so as to avoid mutual shading of leaves. The plants were grown hydroponically at two nitrate levels under two growth light treatments. For one group of the plants, leaves were exposed to full sunlight. For another group, respective leaves were artificially shaded in a manner that simulated changes in the light gradient with the development of an erect herbaceous canopy: old leaves were placed under progressively shadier conditions with growth of the plants (canopy-type shading). In all the treatments, chlorophyll (Chl) content gradually decreased with leaf age. Photosystem I (PSI) per Chl was constant, independent of leaf age, nitrogen nutrition and/or PFD. Photosystem II (PSII) and cytochrome / per Chl, and Chl a/b ratio were independent of leaf age and/or nitrogen nutrition but decreased with the decrease in growth PFD. Ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39, RuBPCase) per Chl steeply decreased with decrease in PFD. When leaves grown at the same PFD were compared, RuBPCase/Chl was lower in the plants grown under lower nitrogen availability and also decreased with leaf age in the plants grown without shading. These decreases were attributed to the curvilinear relationship between RuBPCase and Chl in leaves grown at full sunlight, that was independent of nitrogen availability and leaf age. From these results, it is concluded that the composition of the photosynthetic apparatus is independent of leaf age but changes depending on the light environment and total amount of photosynthetic components of the leaf.Abbreviations Chl chlorophyll - cyt f cytochrome f - PFD photon flux density - RuBPCase ribulose-1,5-bisphosphate carboxylase The author thanks Drs. K. Sonoike, Y. Kashino, K. Okada, H. Hatanaka, Y. Suzuki and A. Aoyama for technical advise. The author also thanks Drs. I. Terashima, A. Makino (Tohoku University, Sendai, Japan), Dr. J.R. Evans (Research School of Biological Sciences, Australian National University, Canberra) and Prof. A. Watanabe for valuable suggestions.     

Competing carboxylases: circadian and metabolic regulation of Rubisco in C3 and CAM Mesembryanthemum crystallinum L

The temporal co-ordination of ribulose 1·5-bisphosphate carboxylase/oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPc) activities by Mesembryanthemum crystallinum L. in C(3) and crassulacean acid metabolism (CAM) modes was investigated under conventional light-dark (LD) and continuous light (LL) conditions. When C(3) , net CO(2) assimilation rate increased during each subjective night under LL with maximum carboxylation unrelated to Rubisco activation state. The CAM circadian rhythm of CO(2) uptake was more pronounced, with CO(2) assimilation rate maximal towards the end of each subjective night. In vivo and in vitro techniques were integrated to map carboxylase enzyme regulation to the framework provided by CAM LL gas exchange activity. Rubisco was activated in vitro throughout each subjective dark period and consistently deactivated at each subjective dawn, similar to that observed at true dawn in constitutive CAM species. Instantaneous carbon isotope discrimination showed in vivo carboxylase co-dominance during the CAM subjective night, initially by Rubisco and latterly C(4) (PEPc), despite both enzymes seemingly activated in vitro. The circadian rhythm in titratable acidity accumulation was progressively damped over successive subjective nights, but maintenance of PEPc carboxylation capacity ensures that CAM plants do not become progressively more 'C(3) -like' with time under LL.     

Estimation of Bundle Sheath Cell Conductance in C4 Species and O2 Insensitivity of Photosynthesis

Low conductance to CO2 of bundle sheath cells is required in C4 photosynthesis to maintain high [CO2] at the site of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Elevated [CO2] allows high CO2 assimilation rates by this enzyme and prevents Rubisco oxygenase activity and O2 inhibition of carboxylation. Bundle sheath conductance to CO2 was estimated by chemically inhibiting phosphoenolpyruvate carboxylase and calculating the slope of the linear response of leaf CO2 uptake to [CO2]. The inhibitor 3,3-dichloro-2-dihydroxyphosphinoylmethyl-2-propenoate was supplied to detached leaves of Panicum maximum, Panicum miliaceum, and Sorghum bicolor at 4 mM. Uptake of CO2 was measured at 210 mL L-1 O2 over the CO2 concentration range of 0.34 to 28 mL L-1. Without the inhibitor, CO2 uptake increased steeply at low [CO2] and saturated at about 1 mL L-1. After inhibition, CO2 uptake was a linear function of [CO2] over much of the range tested. The slope of this CO2 response, taken as bundle sheath conductance, was 2.35, 1.96, and 1.13 mmol m-2 s-1 for P. maximum, P. miliaceum, and S. bicolor, respectively, on a leaf area basis. Conductance based on bundle sheath area was 0.76, 0.93, and 0.54 mmol m-2 s-1, respectively. Uptake of CO2 by leaves of P. maximum supplied with the inhibitor was not affected by reduction of [O2] from 210 to 20 mL L-1 over the range of [CO2] used. Because [CO2] in bundle sheath cells of inhibited leaves is likely to be much lower than ambient, the lack of O2 sensitivity of CO2 uptake cannot be ascribed to lack of O2 reaction with ribulose bisphosphate and is probably due to the low conductance of bundle sheath cells, especially at low ambient [CO2]. The likely result of reducing [O2] from 210 to 20 mL L-1 is to stimulate carboxylation of ribulose bisphosphate, thus further reducing [CO2] in bundle sheath cells and increasing CO2 diffusion to these cells from the mesophyll. However, the increase in diffusion is greatly limited by low conductance of the bundle sheath cell walls. Calculations based on estimated bundle sheath conductance show that changes in bundle sheath [CO2] of 0.085 to 0.5 mL L-1, which might be associated with reduced [O2], would have a negligible effect on CO2 uptake.     

Photosynthetic characteristics and tolerance to photo-oxidation of transgenic rice expressing C4 photosynthesis enzymes

The photosynthetic characteristics of four transgenic rice lines over-expressing rice NADP-malic enzyme (ME), and maize phosphoenolpyruvate carboxylase (PC), pyruvate,orthophosphate dikinase (PK), and PC+PK (CK) were investigated using outdoor-grown plants. Relative to untransformed wild-type (WT) rice, PC transgenic rice exhibited high PC activity (25-fold increase) and enhanced activity of carbonic anhydrase (more than two-fold increase), while the activity of ribulose-bisphosphate carboxylase/oxygenase (Rubisco) and its kinetic property were not significantly altered. The PC transgenic plants also showed a higher light intensity for saturation of photosynthesis, higher photosynthetic CO₂ uptake rate and carboxylation efficiency, and slightly reduced CO₂ compensation point. In addition, chlorophyll a fluorescence analysis indicates that PC transgenic plants are more tolerant to photo-oxidative stress, due to a higher capacity to quench excess light energy via photochemical and non-photochemical means. Furthermore, PC and CK transgenic rice produced 22–24% more grains than WT plants. Taken together, these results suggest that expression of maize C₄ photosynthesis enzymes in rice, a C₃ plant, can improve its photosynthetic capacity with enhanced tolerance to photo-oxidation. This revised version was published online in June 2006 with corrections to the Cover Date.