二氧化碳和臭氧浓度升高对春小麦生长及次生代谢的影响

通过开顶式气室(OTCs)研究了OTC对照（自然CO₂浓度约342 μmol·mol^-1,O₃浓度约30 nmol·mol^-1）、高浓度CO₂（550 μmol·mol^-1）、高浓度O₃（浓度为80 nmol·mol^-1）及其交互作用（CO₂ 550 μmol ·mol^-1,O₃ 80 nmol·mol^-1）对春小麦不同发育时期生物量、总酚量、黄酮含量及成熟期产量性状的影响.结果表明：CO₂浓度增加条件下,春小麦生物量和产量性状都显著高于OTC对照（P<0.05）;而O₃浓度升高条件下,小麦生物量降低,株高、穗长、穗粒质量及千粒重也显著低于对照;CO₂和O₃交互作用下各项指标处于二者之间.说明CO₂可以缓解O₃对小麦的负效应,而O₃对CO₂的正效应具有削弱作用,但二者的作用并非简单的叠加.CO₂、O₃浓度增加及其交互作用显著增加了春小麦叶片中的总酚含量,其中两者交互作用的效应更大,但在小麦生长后期,总酚含量增加量比对照有所降低.在小麦生长前期,各处理总黄酮含量均低于对照;而在成熟期,各处理都显著高于对照.     

FACE水稻茎蘖动态模型

借助农田开放式空气CO₂浓度增高（FACE）技术平台,以武香粳14为供试水稻品种,设置不同施N量处理,研究大气CO₂浓度为570 μmol·mol^-1（比对照高200 μmol·mol^-1）的FACE处理对水稻茎蘖动态的影响,并建立了相应的模拟模型：T_t=A₁(1+e^a1-b1t₎-A₂(1+e^a2-b2t)+C×[B₁(1+e^a3-b3t)-B₂(1+e^a4-b4t)]+D.模型以时间为驱动因子,描述了水稻茎蘖数随移栽天数的动态变化过程,对常规及CO₂浓度增加条件下水稻茎蘖的变化均有很好的拟合性.通过不同年份试验数据对模型的检验,预测根均方差（RMSE）最大为44.27个·m^-2,最小为13.96个·m^-2,且相关系数均达到了极显著水平.表明模型的预测程度较高,具有很好的适用性.     

不同氮源下生长的柚树叶片光合参数对高浓度CO2驯化作用的比较

比较研究了在不同形式氮源下生长柚树叶片光合对高浓度 CO2 驯化过程中有关参数变化。植株生长在人工混成土壤中 ,分别浇灌含有 2 mmol L- 1N的 NO- 3 - N,NH+ 4 - N和 NH4 NO3- N溶液。空气 CO2 增高处理时向生长植株的开顶透明罩中通入 74.4Pa CO2 ,以空气 CO2 生长的植株为对照。利用 CI- 30 1 ( CID,Inc) CO2 气体交换系统测定叶片光合速率和通过光合作用相关响应曲线计算光合参数。结果表明 ,在 CO2分压倍增下 ,NO- 3 - N生长植株光饱和光合速率较大气 CO2 分压下的高。而生长在 NH+ 4 - N和 NH4 NO3- N的植株光合速率与大气 CO2 分压下的相近 ,表现对高 CO2 的驯化。在空气 CO2 倍增下无论供给何种形式氮源并不影响Γ* ,但可增高 Rd( P<0 .0 5 )。 CO2 分压倍增下供给 NO- 3 - N植株的 Vcmax和 Jmax较大气分压相应的植株高 ,而 NH+ 4 - N和 NH4 NO3- N植株则与大气 CO2分压的相应植株相似 ( P>0 .0 5 )。无论供给何种形式氮源 ,生长在空气 CO2 分压倍增下不改变叶片单位面积干重 ,叶绿素含量和叶片中氮在 Rubisco、生物能学组分和捕光色素复合体组分的分配系数 ;但能改变叶片中氮含量。植物对高 CO2 的驯化可能受到不同形式氮利用性的影响 ,在对高 CO2 驯化过程亦反映叶片中氮在不同光合功能组分     

热带季节雨林冠层树种绒毛番龙眼的光合生理生态特性

采用Li-6400便携式光合作用测定仪,对西双版纳热带季节雨林冠层树种绒毛番龙眼成树树冠上、中、下3层叶片进行了测定,分析西双版纳热带季节雨林冠层树木的光合作用.结果表明,绒毛番龙眼成树具有喜光的光合特性,光饱和点较高(1 000～1 500 μmol·m^-2·s^-1),而光补偿点较低(7.7～15.3 μmol·m^-2·s^-1),对光环境有较强的适应和调节能力,光合有效辐射是影响绒毛番龙眼光合日进程的关键因子;12月,叶片处于成熟期,生长良好,光合能力较强,树冠上层净光合速率（P_n）日变化为单峰型,最大净光合速率（A_max）约为8.9 μmol CO₂·m^-2·s^-1;4月处于新老树叶更替期,光合能力下降,树冠上层P_n日变化为双峰型,中午出现“午休”现象,树冠上层A_max约为4.3 μmol CO₂·m^-2·s^-1;7月上、中层叶片P_n为单峰型,下层出现“午休”.如人为使CO₂浓度在短期内迅速升高,则绒毛番龙眼的P_n会增加,而气孔导度和蒸腾速率降低;CO₂浓度从400 μmol·mol^-1升高到800 μmol·mol^-1时,干季水分利用效率（WUE）提高约50%～100％,雨季WUE较低.     

空气CO2增高条件下荔枝叶片光合作用和超氧自由基产率

研究结果表明,生长在77±5PaCO2分压下30d的荔枝幼树,其光合速率较大气CO2分压（39.3Pa）下的低23％,光下线粒体呼吸速率和不包含光下呼吸的CO2补偿点亦略有降低。空气CO2增高使叶片最大羧化速率（Vcmax）和最大电子传递速率（Jmax）降低,表明大气增高CO2分压下叶片的光I(PSI)能量水平较低,呈片超氧自由基产率亦降低39％,叶片感染荔枝霜疫霉病率则从生长在大气CO2分压下的1.8％增至9.5％,可能较低光合和呼吸代谢诱致较低的超氧自由基产率,而使叶片易受病害侵染。叶片受病害侵染后表现为超氧自由基的激增。在全球大气CO2分压增高趋势下须加强对荔枝霜疫霉病的控制。     

内蒙古锡林河流域羊草草原净初级生产力及其对全球气候变化的响应

利用CENTURY模型对内蒙古锡林河流域羊草草原在未来气候变化以及大气CO₂浓度增高条件下的年地上净初级生产力(annual aboveground net primary productivity,ANPP)动态进行了模拟研究.结果表明：CENTURY模型可以较好地预测ANPP的变化.进一步的情景模拟发现,虽然全球气候变化所引起的温度和降水改变、以及大气CO₂浓度升高都会影响ANPP,但降水是关键的影响因子.多个全球气候模型(GCM) 预测该地区未来降水量会减少,故可能导致其ANPP降低,但在以下气候变化情景下研究区ANPP可能会升高：1)CO₂浓度倍增,温度升高2 ℃,降水保持不变或增加10%～20%;2)CO₂浓度保持不变,温度升高2 ℃,降水增加20%.气候变化将对内蒙古锡林河流域羊草草原产生显著影响.     

不同光强下焕镛木和观光木的光合参数变化

 生长在全日光强下的焕镛木(Woonyoungia septentrionalis)和观光木(Tsoongiodendron lotungensis)幼树叶片的最大光合速率、表观量子产率和光能转换效率均较生长在40%和20%日光强的高。当生长光强从全日光强降低至40%日光强时,焕镛木的表观量子产率和光能转换效率分别降低13.1%和6.3%,而观光木则相应分别降低23.8%和33.4%。生长光强降低至40%日光强时,焕镛木的Rubisco最大羧化速率(Vcmax)未见变化;而最大电子传递速率（Jmax）则降低14.1%,表明Jmax对光强降低的响应较Vcmax敏感。当生长光强从全日光强降低到40%和20%日光强时,观光木的Vcmax分别降低7.7%和31.7%,而Jmax则分别降低9.7%和42%。光强从全日光强降低至40%日光强,焕镛木叶氮在Rubisco和捕光叶绿素蛋白复合体中的分配系数没有明显改变,而叶氮在生物力能学组分中的分配系数降低则较为明显（20.4%）,表明生长光强降低对叶氮在光合电子传递链组分分配的影响较在Rubisco的大。结果表明,焕镛木表现阳生树种特性,在迁地保育中宜选择向阳小生境种植,而观光木较耐荫,可种植在较遮荫的环境。     

柚树叶片CO2驯化的光合参数变化

柚树(Citrus grandis)幼树生长在砂和磋石的生长介质,每周供给0.05mmol P(正常P,P)和0．1mmol P(高磷,2P)的营养液．植株分别生长在空气CO2分压(约39Pa)和倍增CO2分压(81±5Pa)下45d,利用CI-301PS(CID,Inc)光合作用测定系统在较高光强(1150μmol·m^-2·s^-1)下测定叶片光合速率并得出的Pn-Pi关系曲线和在较高CO2分压(PCO2,56Pa)下得出Pn-PAR关系曲线计算有关光合参数。结果表明,大气CO2分压下2P植株最大光合速率较P植株高13．3％,倍增CO2分压下,无论P或2P植株最大光合速率较大气CO2分压下相应植株低,但在倍增CO2分压下2P植株较P植株高,且2P植株有较P植株高的表观量子产率和光能利用效率(P<0．05),但并不改变г^*、Rd和Rubisco羧化速率(Vc)和氧速率的比率(P>0．05)在大气CO2分压下2P植株的Vcmax和Jmax较P植株分别高83％和12．5％,在倍增CO2分压下2P植株的Vcmax和Jmax均较P植株高,柚树在高CO2驯化中改变叶N在Rubisco和捕光组分分配系数,但不改变叶N在光合电子传递链的分配系数,结果表明,增加P供给可以促进高CO2分压下光合碳循环中P的周转,提高倍增CO2分压下植株的光合速率,调节柚树叶片的CO2驯化的光合参数。     

冬小麦光合作用对开放式空气CO2 浓度增高(FACE)的非气孔适应

在同样CO2浓度下测定时,开放式空气CO2浓度增高(FACE,580 μmol CO2 /mol)条件下生长的冬小麦叶片的净光合速率、气孔导度和羧化效率都显著低于普通空气(380 μmol CO2 /mol)中生长的对照叶片.与此相一致,FACE叶片的可溶性蛋白、二磷酸核酮糖羧化酶/加氧酶(Rubisco)和Rubisco活化酶含量也都显著低于对照叶片.这些结果表明,在根系生长不受限制的田间条件下,冬小麦叶片的光合作用对高浓度CO2产生了适应现象,其主要原因可能是碳同化的关键酶Rubisco等含量的降低.     

北京市六种针叶树叶面附着颗粒物的理化特征

对北京市6种针叶树叶面颗粒物附着密度、叶表面微形态、颗粒物矿物与元素组成的研究结果表明：同一树种叶面颗粒物附着密度随大气颗粒物浓度增加而增大;同一地点不同树种叶面颗粒物附着密度存在很大差异,圆柏、侧柏颗粒物附着密度最高,其次为雪松、白皮松,油松、云杉最低;受地面扬尘影响,低矮叶片较高处叶片颗粒物附着密度大;受降雨和新生叶片稀释影响,夏季颗粒物附着密度小于冬季.叶表面粗糙程度越大,颗粒物附着密度越高.SiO₂、CaCO₃、CaMg(CO₃)₂、NaCl、2CaSO₄·H₂O、CaSO₄·2H₂O、Fe₂O₃ 7种主要矿物占叶面颗粒物总质量的10%～30%,其中,SiO₂含量最高,其次为CaMg(CO₃)₂ 、CaSO₄·2H₂O和CaCO₃.此外,还含有蒙脱石、伊利石、高岭石等粘土矿物及长石.21种测定元素占叶面颗粒物总质量的16%～37%,其中Ca、Al、Fe、Mg、K、Na、S 7种元素占测定元素总质量的97%以上,其它痕量污染元素含量很少,并且受采样地点和树种影响较小.     

Responses of Rubisco and Rubisco activase in cucumber seedlings to low temperature and weak light

Using 'Jinyou 3' cucumber seedlings as test materials, this paper studied their photosynthetic rate (P(n)), Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and Rubisco activase (RCA) activities, and gene expression of Rubisco and RCA under optimal temperature and weak light (WL: 25 degrees C/18 degrees C, 100 micromol x m(-2) x s(-1)), suboptimal temperature and weak light (ST+WL: 18 degrees C/12 degrees C, 100 micromol x m(-2) x s(-1)), and low temperature and weak light (LT+WL: 10 degress C/5 degrees C, 100 micromol x m(-2) x s(-1)). Comparing with the control (25 degrees C/18 degrees C, 400 micromol x m(-2) x s(-1)), treatments WL, ST+WL, and LT+WL all led to a remarkable decrease in leaf area and dry matter mass. At initial stage, the P(n), Rubisco activity, rbcL and rbcS expression, RCA activity, and CsRCA expression in the three treatments declined by a big margin; 5-7 days later, these parameters tended to be less changed in treatment WL, ascended slowly in treatment ST+WL, and decreased continuously in treatment LT+WL. These results suggested that the photosynthetic apparatus of test cucumber seedlings could gradually adapt to weak light or suboptimal temperature and weak light. The Rubisco and RCA activities and the gene expression of Rubisco and RCA showed the similar responses to low temperature and weak light as the P(n), suggesting that the decline in Rubisco and RCA activities and gene expression in cucumber seedlings under low temperature and weak light could be the important reason leading to the decrease of P(n).     

The effects of Rubisco activase on C4 photosynthesis and metabolism at high temperature

The activation of Rubisco in vivo requires the presence of the regulatory protein Rubisco activase. This enzyme facilitates the release of sugar phosphate inhibitors from Rubisco catalytic sites thereby influencing carbamylation. T(1) progeny of transgenic Flaveria bidentis (a C(4) dicot) containing genetically reduced levels of Rubisco activase were used to explore the role of the enzyme in C(4) photosynthesis at high temperature. A range of T(1) progeny was screened at 25 degrees C and 40 degrees C for Rubisco activase content, photosynthetic rate, Rubisco carbamylation, and photosynthetic metabolite pools. The small isoform of F. bidentis activase was expressed and purified from E. coli and used to quantify leaf activase content. In wild-type F. bidentis, the activase monomer content was 10.6+/-0.8 micromol m(-2) (447+/-36 mg m(-2)) compared to a Rubisco site content of 14.2+/-0.8 micromol m(-2). CO(2) assimilation rates and Rubisco carbamylation declined at both 25 degrees C and 40 degrees C when the Rubisco activase content dropped below 3 mumol m(-2) (125 mg m(-2)), with the status of Rubisco carbamylation at an activase content greater than this threshold value being 44+/-5% at 40 degrees C compared to 81+/-2% at 25 degrees C. When the CO(2) assimilation rate was reduced, ribulose-1,5-bisphosphate and aspartate pools increased whereas 3-phosphoglycerate and phosphoenol pyruvate levels decreased, demonstrating an interconnectivity of the C(3) and C(4) metabolites pools. It is concluded that during short-term treatment at 40 degrees C, Rubisco activase content is not the only factor modulating Rubisco carbamylation during C(4) photosynthesis.     

Temperature response of photosynthesis and internal conductance to CO2: results from two independent approaches

The internal conductance to CO(2) transfer from intercellular spaces to chloroplasts poses a major limitation to photosynthesis, but few studies have investigated its temperature response. The aim of this study was to determine the temperature response of photosynthesis and internal conductance between 10 degrees C and 35 degrees C in seedlings of a deciduous forest tree species, Quercus canariensis. Internal conductance was estimated via simultaneous measurements of gas exchange and chlorophyll fluorescence ("variable J method"). Two of the required parameters, the intercellular photocompensation point (C(i)*) and rate of mitochondrial respiration in the light (R(d)), were estimated by the Laisk method. These were used to calculate the chloroplastic photocompensation point (Gamma*) in a simultaneous equation with g(i). An independent estimate of internal conductance was obtained by a novel curve-fitting method based on the curvature of the initial Rubisco-limited portion of an A/C(i) curve. The temperature responses of the rate of Rubisco carboxylation (V(cmax)) and the RuBP limited rate of electron transport (J(max)) were determined from chloroplastic CO(2) concentrations. The rate of net photosynthesis peaked at 24 degrees C. C(i)* was similar to reports for other species with a C(i)* of 39 micromol mol(-1) at 25 degrees C and an activation energy of 34 kJ mol(-1). Gamma* was very similar to the published temperature response for Spinacia oleracea from 20 degrees C to 35 degrees C, but was slightly greater at 10 degrees C and 15 degrees C. J(max) peaked at 30 degrees C, whereas V(cmax) did not reach a maximum between 10 degrees C and 35 degrees C. Activation energies were 49 kJ mol(-1) for V(cmax) and 100 kJ mol(-1) for J(max). Both methods showed that internal conductance doubled from 10 degrees C to 20 degrees C, and then was nearly temperature-independent from 20 degrees C to 35 degrees C. Hence, the temperature response of internal conductance could not be fitted to an Arrhenius function. The best fit to estimated g(i) was obtained with a three-parameter log normal function (R(2)=0.98), with a maximum g(i) of 0.19 mol m(-2) s(-1) at 29 degrees C.     

A/C(i) curve analysis across a range of woody plant species: influence of regression analysis parameters and mesophyll conductance

The analysis and interpretation of A/C(i) curves (net CO(2) assimilation rate, A, versus calculated substomatal CO(2) concentration, C(i)) is dependent upon a number of underlying assumptions. The influence of the C(i) value at which the A/C(i) curve switches between the Rubisco- and electron transport-limited portions of the curve was examined on A/C(i) curve parameter estimates, as well as the effect of mesophyll CO(2) conductance (g(m)) values on estimates of the maximum rate of Rubisco-mediated carboxylation (V(cmax)). Based on an analysis using 19 woody species from the Pacific Northwest, significant variation occurred in the C(i) value where the Rubisco- and electron transport-limited portions of the curve intersect (C(i_t)), ranging from 20 Pa to 152 Pa and averaging c. 71 Pa and 37 Pa for conifer and broadleaf species, respectively. Significant effects on estimated A/C(i) parameters (e.g. V(cmax)) may arise when preliminary estimates of C(i_t), necessary for the multiple regression analyses, are set either too high or too low. However, when the appropriate threshold is used, a significant relationship between A/C(i) and chlorophyll fluorescence estimates of carboxylation is achieved. The use of the V(cmax) parameter to describe accurately the Rubisco activity from the A/C(i) curve analysis is also dependent upon the assumption that C(i) is approximately equal to chloroplast CO(2) concentrations (C(c)). If leaf mesophyll conductance is low, C(c) will be much lower than C(i) and will result in an underestimation of V(cmax) from A/C(i) curves. A large range of mesophyll conductance (g(m)) values was observed across the 19 species (0.005+/-0.002 to 0.189+/-0.011 mol m(-2) s(-1) for Tsuga heterophylla and Quercus garryana, respectively) and, on average, g(m) was 1.9 times lower for the conifer species (0.058+/-0.017 mol m(-2) s(-1) for conifers versus 0.112+/-0.020 mol m(-2) s(-1) for broadleaves). When this mesophyll limitation was accounted for in V(cmax) estimates, considerable variation still existed between species, but the difference in V(cmax) between conifer and broadleaf species was reduced from c. 11 micromol m(-2) s(-1) to 4 micromol m(-2) s(-1). For example, A/C(i) curve estimates of V(cmax) were 31.2+/-6.2 and 42.2+/-4.4 micromol m(-2) s(-1), and A/C(c) curve estimates were 41.2+/-7.1 micromol m(-2) s(-1) and 45.0+/-4.8 micromol m(-2) s(-1), for the conifer and broadleaf species, respectively.     

Temperature response of photosynthesis in transgenic rice transformed with 'sense' or 'antisense' rbcS

The responses of chlorophyll fluorescence, gas exchange rate and Rubisco activation state to temperature were examined in transgenic rice plants with 130 and 35% of the wild-type (WT) Rubisco content by transformation with rbcS cDNA in sense and antisense orientations, respectively. Although the optimal temperatures of PSII quantum efficiency and CO(2) assimilation were found to be between 25 and 32 degrees C, the maximal activation state of Rubisco was found to be between 16 and 20 degrees C in all genotypes. The Rubisco flux control coefficient was also the highest between 16 and 20 degrees C in the WT and antisense lines [>0.88 at an intercellular CO(2) pressure (Ci) of 28 Pa]. Gross photosynthesis at Ci = 28 Pa per Rubisco content in the WT between 12 and 20 degrees C was close to that of the antisense lines where high Rubisco control is present. Thus, Rubisco activity most strongly limited photosynthesis at cool temperatures. These results indicated that a selective enhancement of Rubisco content can enhance photosynthesis at cool temperatures, but in the sense line with enhanced Rubisco content Pi regeneration limitation occurred. Above 20 degrees C, the Rubisco flux control coefficient declined. This decline was associated with a decline in Rubisco activation. The activation state of Rubisco measured at each temperature decreased with increasing Rubisco content, and the slope of activation to Rubisco content was independent of temperature. We discuss the possibility that the decline in Rubisco activation at intermediate and high temperatures is part of a regulated response to a limitation in other photosynthetic processes.     

The effect of temperature on C(4)-type leaf photosynthesis parameters

C(4)-type photosynthesis is known to vary with growth and measurement temperatures. In an attempt to quantify its variability with measurement temperature, the photosynthetic parameters - the maximum catalytic rate of the enzyme ribulose 1.5-bisphosphate carboxylase/oxygenase (Rubisco) (V(cmax)), the maximum catalytic rate of the enzyme phosphoenolpyruvate carboxylase (PEPC) (V(pmax)) and the maximum electron transport rate (J(max)) - were examined. Maize plants were grown in climatic-controlled phytotrons, and the curves of net photosynthesis (A(n)) versus intercellular air space CO(2) concentrations (C(i)), and A(n) versus photosynthetic photon flux density (PPFD) were determined over a temperature range of 15-40 degrees C. Values of V(cmax), V(pmax) and J(max) were computed by inversion of the von Caemmerer & Furbank photosynthesis model. Values of V(pmax) and J(max) obtained at 25 degrees C conform to values found in the literature. Parameters for an Arrhenius equation that best fits the calculated values of V(cmax), V(pmax) and J(max) are then proposed. These parameters should be further tested with C(4) plants for validation. Other model key parameters such as the mesophyll cell conductance to CO(2) (g(i)), the bundle sheath cells conductance to CO(2) (g(bs)) and Michaelis-Menten constants for CO(2) and O(2) (K(c), K(p) and K(o)) also vary with temperature and should be better parameterized.     

Seasonal change in the balance between capacities of RuBP carboxylation and RuBP regeneration affects CO2 response of photosynthesis in Polygonum cuspidatum

The balance between the capacities of RuBP (ribulose-1,5-bisphosphate) carboxylation (V(cmax)) and RuBP regeneration (expressed as the maximum electron transport rate, J(max)) determines the CO(2) dependence of the photosynthetic rate. As it has been suggested that this balance changes depending on the growth temperature, the hypothesis that the seasonal change in air temperature affects the balance and modulates the CO(2) response of photosynthesis was tested. V(cmax) and J(max) were determined in summer and autumn for young and old leaves of Polygonum cuspidatum grown at two CO(2) concentrations (370 and 700 micromol mol(-1)). Elevated CO(2) concentration tended to reduce both V(cmax) and J(max) without changing the J(max):V(cmax) ratio. The seasonal environment, on the other hand, altered the ratio such that the J(max):V(cmax) ratio was higher in autumn leaves than summer leaves. This alternation made the photosynthetic rate more dependent on CO(2) concentration in autumn. Therefore, when photosynthetic rates were compared at growth CO(2) concentration, the stimulation in photosynthetic rate was higher in young-autumn than in young-summer leaves. In old-autumn leaves, the stimulation of photosynthesis brought by a change in the J(max):V(cmax) ratio was partly offset by accelerated leaf senescence under elevated CO(2). Across the two seasons and the two CO(2) concentrations, V(cmax) was strongly correlated with Rubisco and J(max) with cytochrome f content. These results suggest that seasonal change in climate affects the relative amounts of photosynthetic proteins, which in turn affect the CO(2) response of photosynthesis.     

C4 photosynthesis at low temperature. A study using transgenic plants with reduced amounts of Rubisco

C(4) plants are rare in the cool climates characteristic of high latitudes and elevations, but the reasons for this are unclear. We tested the hypothesis that CO(2) fixation by Rubisco is the rate-limiting step during C(4) photosynthesis at cool temperatures. We measured photosynthesis and chlorophyll fluorescence from 6 degrees C to 40 degrees C, and in vitro Rubisco and phosphoenolpyruvate carboxylase activity from 0 degrees C to 42 degrees C, in Flaveria bidentis modified by an antisense construct (targeted to the nuclear-encoded small subunit of Rubisco, anti-RbcS) to have 49% and 32% of the wild-type Rubisco content. Photosynthesis was reduced at all temperatures in the anti-Rbcs plants, but the thermal optimum for photosynthesis (35 degrees C) did not differ. The in vitro turnover rate (kcat) of fully carbamylated Rubisco was 3.8 mol mol(-)(1) s(-)(1) at 24 degrees C, regardless of genotype. The in vitro kcat (Rubisco Vcmax per catalytic site) and in vivo kcat (gross photosynthesis per Rubisco catalytic site) were the same below 20 degrees C, but at warmer temperatures, the in vitro capacity of the enzyme exceeded the realized rate of photosynthesis. The quantum requirement of CO(2) assimilation increased below 25 degrees C in all genotypes, suggesting greater leakage of CO(2) from the bundle sheath. The Rubisco flux control coefficient was 0.68 at the thermal optimum and increased to 0.99 at 6 degrees C. Our results thus demonstrate that Rubisco capacity is a principle control over the rate of C(4) photosynthesis at low temperatures. On the basis of these results, we propose that the lack of C(4) success in cool climates reflects a constraint imposed by having less Rubisco than their C(3) competitors.     

Temperature dependence of nitrate reductase in the psychrophilic unicellular alga Koliella antarctica and the mesophilic alga Chlorella sorokiniana

Temperature responses of nitrate reductase (NR) were studied in the psychrophilic unicellular alga, Koliella antarctica, and in the mesophilic species, Chlorella sorokiniana. Enzymes from both species were purified to near homogeneity by Blue Sepharose (Pharmacia, Uppsala, Sweden) affinity chromatography and high-resolution anion-exchange chromatography (MonoQ; Pharmacia; Uppsala, Sweden). Both enzymes have a subunit molecular mass of 100 kDa, and K. antarctica NR has a native molecular mass of 367 kDa. NR from K. antarctica used both NADPH and NADH, whereas NR from C. sorokiniana used NADH only. Both NRs used reduced methyl viologen (MVH) or benzyl viologen (BVH). In crude extracts, maximal NADH and MVH-dependent activities of cryophilic NR were found at 15 and 35 degrees C, respectively, and retained 77 and 62% of maximal activity, respectively, at 10 degrees C. Maximal NADH and MVH-dependent activities of mesophilic NR, however, were found at 25 and 45 degrees C, respectively, with only 33 and 23% of maximal activities being retained at 10 degrees C. In presence of 2 microM flavin adenine dinucleotide (FAD), activities of cryophilic NADH:NR and mesophilic NADH:NR were stable up to 25 and 35 degrees C, respectively. Arrhenius plots constructed with cryophilic and mesophilic MVH:NR rate constants, in both presence or absence of FAD, showed break points at 15 and 25 degrees C, respectively. Essentially, similar results were obtained for purified enzymes and for activities measured in crude extracts. Factors by which the rate increases by raising temperature 10 degrees C (Q10) and apparent activation energy (E(a)) values for NADH and MVH activities measured in enzyme preparations without added FAD differed slightly from those measured with FAD. Overall thermal features of the NADH and MVH activities of the cryophilic NR, including optimal temperatures, heat inactivation (with/without added FAD) and break-point temperature in Arrhenius plots, are all shifted by about 10 degrees C towards lower temperatures than those of the mesophilic enzyme. Transfer of electrons from NADH to nitrate occurs via all three redox centres within NR molecule, whereas transfer from MVH requires Mo-pterin prosthetic group only; therefore, our results strongly suggest that structural modification(s) for cold adaptation affect thermodynamic properties of each of the functional domains within NR holoenzyme in equal measure.     

Adaptation of photosynthesis in marama bean Tylosema esculentum (Burchell A. Schreiber) to a high temperature, high radiation, drought-prone environment.

Marama bean, Tylosema esculentum, is a tuberous legume native to the Kalahari region of Southern Africa where it grows under high temperatures (typical daily max 37 degrees C during growing season) and radiation (frequently in excess of 2000 micromol m(-2) s(-1)) in sandy soils with low rainfall. These conditions might be expected to select for increased water-use efficiency of photosynthesis. However, marama was found to give similar leaf photosynthetic rates to other C3 plants for a given internal leaf CO2 concentration and Rubisco content. Under conditions of increasing drought, no increase in water-use efficiency of photosynthesis was observed, but stomata closed early and preceded any change in leaf water potential. The possibility of subtle adaptations of photosynthetic characteristics to its natural environment were investigated at the level of Rubisco kinetics. The specificity factor of marama Rubisco was slightly lower than that of wheat, but the apparent Km for CO2 in air (Km') was about 20% lower than that of wheat. This is consistent with better adaptation for efficient photosynthesis at high temperatures in marama compared to wheat, although the net benefit is predicted to be very small (<0.5% at 35 degrees C). The sequence of marama rbcL gene shows 27 deduced amino acid residue differences from that for wheat, and the possibility that one or more of these cause the difference in Rubisco Km' is discussed.