A new principle photosynthesis capacity biosensor based on quantitative measurement of delayed fluorescence in vivo

Delayed fluorescence (DF) is an excellent marker for evaluating plant photosynthesis. Compared with common methods for measuring the photosynthesis rate based on consumption of CO₂, DF technique can quantify the plant photosynthesis capacity more accurately and faster under its physiological status with less interference from the environment. We previously reported a method for measuring photosynthesis using DF of chloroplast [Wang, C.L., Xing, D., Chen, Q., 2004. Biosens. Bioelectron. 20, 454–459]. In the study, a novel fast and portable photosynthesis capacity biosensor system was developed, which was composed of light-emitting diode lattice as excitation light source, Channel Photomultiplier DC-Module to achieve DF, single-chip microcomputer as control center, hermetic dark sample chamber, battery power supply and CO₂, humidity and temperature controller. Compared with our previous work, the system was portable and can directly measure plant photosynthesis capacity in vivo in less than 10 s. A database in the software to carry out data acquisition and processing was developed to translate maximal DF intensity to net photosynthesis rate (Pn). In addition, local-control and remote-control mode can be chosen in the system. To demonstrate the utility of the system, it was applied to evaluate maximum Pn of four different plant species samples (Queen Rape Myrtle (var. rubra), soybean (Lu Hei No. 1), maize (Jin Dan No. 39) and rice (Jing Dao No. 21)) in field. The results were compared with that using commercial photosynthesis system LI-6400 and the uncertainty was less than ±5%. The new principle of photosynthesis measurement is a challenge and breakthrough to conventional method of gas exchange and may be a potential technique of next generation photosynthesis measurement.     

A novel method for measuring photosynthesis using delayed fluorescence of chloroplast

Photosynthesis is the most important chemical reaction in the world. The measurement of plant photosynthesis rate plays an important role in agriculture. Light-induced delayed fluorescence (DF) in plants is an intrinsic label of the efficiency of charge separation at P680 in photosystem II (PS II). In this paper, we have developed a biosensor that can accurately measure the plant photosynthesis ability by means of DF. Compared with common methods for measuring the photosynthesis rate based on consumption of CO2, the proposed technique can quantify the plant photosynthesis ability with less influence of the environment. The biosensor is an all-weather measuring instrument, it has its own illumination power and utilizes intrinsic DF as the measurement marker. The current investigation has revealed that, there is a good correspondence between the results measured by the biosensor and that by commercially available portable photosynthesis system under controlled conditions. We thus conclude that DF is an excellent marker for evaluating plant photosynthesis ability under its biological status with less interferences of the environment.     

窄叶短柱茶光合特性研究

用LI-6400型便携式光合作用测定系统对窄叶短柱茶Camellia fluviatilis的光合特性进行研究。结果表明：窄叶短柱茶净光合速率日变化呈单峰型曲线,峰值出现在13:00左右,为18.65 μmol·m^-2·s^-1;通过光响应曲线拟合得知,其光补偿点为12 μmol·m^-2·s^-1,光饱和点为500 μmol·m^-2·s^-1。相关性分析表明,窄叶短柱茶净光合速率与光强、大气温度具有极显著正相关性（P≤0.01）,相关系数分别为0.715和0.939;与胞间CO₂浓度存在极显著负相关（P≤0.01）, 相关系数为-0.752;与大气CO₂浓度存在显著负相关性（P≤0.05),相关系数为-0.605。     

热研8号坚尼草的光合特性研究

2006年9月上旬,在晴天用LI-6400便携式光合仪测定热研8号坚尼草的光合特性。结果显示,热研8号坚尼草的光合日变化呈"单峰型",无"午休"现象,13:00时出现峰值(Pn为26.9 μmol/m²·s),变化趋势与蒸腾速率(Tr)、气孔导度(Gs)等因子相同;叶片光饱和点(LSP)约为1 700 μmol/m²·s,光补偿点(LCP)约为46 μmol/m²·s,表观量子效率(AQY)为0.0535,CO₂饱和点(CSP)为600 μmol/mol,CO₂补偿点(CCP)为3.49 μmol/mol,羧化效率(CE)为0.1168。表明热研8号坚尼草是一种耐荫的C₄型阳性植物。     

大气CO2浓度升高对香蕉光合作用及光合碳循环过程中叶氮分配的影响

生长在高CO₂浓度(700±5μl·L^-1)1周的香蕉叶片,其光合速率(Pn,μmol·m^-2·s^-1)为5.14±0.32,较生长在大气CO₂浓度(356±301μl·L^-1)的高22.1%,而生长在较高CO₂浓度下8周,叶片Pn较生长在大气CO₂浓度的低18.1%,表现香蕉叶片对较长期高CO₂浓度的驯化和光合作用抑制.生长在高CO₂浓度的香蕉叶片有较低光下呼吸速率(R_d),而不包括光下呼吸的CO₂补偿点则变幅较小.最大羧化速率(Vcmax)和电子传递速率(J)分别较生长在大气CO₂浓度的低30.5%和14.8%,根据气体交换速率计算的表观量子产率(α,mol CO₂·mol^-1光量子),生长在较高CO₂浓度下8周的叶片为0.014±0.01,而生长在大气CO₂浓度下的为0.025±0.005.较高CO₂浓度下叶片的表观量子产率降低44%.光能转换效率electrons·quanta^-1)亦从0.203降低至0.136.生长在较高CO₂浓度下香蕉叶片的叶氮在Rubicos分配系数(PR)、叶氮在生物力能学组分分配系数(P_B)和叶氮在光捕组分的分配系数(P_L)均较生长在大气CO₂浓度低,表明在高CO₂浓度下较长期生长(8周)的香蕉叶片多个光合过程受抑制,光合活性明显降低.     

大田威廉斯香蕉叶片光合特性研究

用Li-6400型便携式光合作用测定系统研究大田威廉斯香蕉叶片光合日变化、光响应曲线和CO₂响应曲线,运用非直角双曲线模型并结合SPSS软件的非线性回归估算其光合作用参数。结果表明,香蕉叶片光合日进程呈单峰曲线,没有明显的午休现象,最大净光合速率出现在10:20左右;光饱和点与光补偿点分别为778.19和61.09μmol/m²·s,表观量子效率为0.0414;CO₂饱和点与补偿点分别为608.97和72.13μmol/mol,羧化效率为0.094。此外,对影响香蕉光合速率的主要环境因子分析表明,在土壤供水充分的条件下,限制光合速率的因子是光合有效辐射而非气孔导度。     

N availability does not modify plant-mediated responses of Trichoplusia ni to elevated CO2

Aims Elevated CO₂ and increased N availability can alter a variety of plant physiological processes leading to changes in the nutritional quality of leaf tissue for herbivores. Numerous experiments have examined the responses of herbivores to environmental change; however the potential effects of simultaneous change in multiple factors on leaf-chewing insect herbivores are less well understood. The plant-mediated effects of elevated CO₂ and high N on the performance of a generalist leaf-chewing insect herbivore, Trichoplusia ni, were investigated.Methods Newly hatched T. ni larvae were introduced to Amaranthus viridis and Polygonum persicaria plants grown under ambient and elevated CO₂ and low and high N conditions. Insect performance was assessed by measuring larvae weight after ten days of feeding. Plant photosynthesis, biomass, leaf area and specific leaf weight were measured to determine the effects of elevated CO₂, N and insect feeding on plant performance.Important findings Elevated CO₂ did not have strong effects on plant or insect performance, only affecting a few responses under low or high N conditions, but not both. Growth under high nitrogen improved almost all measures of plant performance. Trichoplusia ni performed significantly better on Amaranthus viridis (C 4) compared to Polygonum persicaria (C 3), despite similar leaf C:N ratios in both species. The performance of T. ni caterpillars was only improved by the high nitrogen treatment when they were feeding on P. persicaria, the host they performed poorly on. The only interactions between N and CO₂ affecting plant performance were seen for leaf photosynthesis of P. persicaria and leaf area of A. viridis. Contrary to the predictions, there were no significant CO₂ by N interactions affecting T. ni performance.     

A comparison of measured and calculated net community CO2 exchange: Scaling from leaves to communities

Leaf net photosynthesis is crucial for detecting the mechanism of photosynthesis, whereas community net photosynthesis is useful for understanding the photosynthetic capacity of communities and its relationship with environmental factors. In particular, we need to scale up eco-physiological models from leaf scale to canopy level to study carbon cycling at regional or global scale. We hypothesized that accumulated leaf net photosynthetic rate (P_c) at community scale, i.e., calculated based on leaf net photosynthetic rate (P_n) and leaf area index (LAI), equals to measured net community CO₂ exchange (NCE). The purpose of this study is to verify this hypothesis. Our field study was carried out in Duolun, Nei Mongol, China, where we constructed single-species communities by sowing Medicago sativa ‘Aohan’ seeds in three plots (3 m × 5 m) on May 30, 2012. On August 16, 2014, P_n of five healthy leaves of M. sativa ‘Aohan’ in each plot were measured with a LI-6400 portable photosynthesis system at 10:00, and net ecosystem CO₂ exchange (NEE) in each plot was measured simultaneously with a LI-8100 system connected with a assimilation chamber (0.5 m × 0.5 m × 0.5 m). P_c was calculated based on P_n, number of leaves (n), LAI percentage of healthy leaves (r) and percentage of received effective light by leaves (m). NCE was derived from NEE and ecosystem respiration rate (R_eco). P_c was 3.52 μmol CO₂·m^-2·s^-1, and very close to NCE (3.56 μmol CO₂·m^-2·s^-1), suggesting that leaf-scale photosynthesis may accurately predict community-scale photosynthesis. However, our method could not separate community respiration from soil respiration, and future studies, should be designed to counteract this effect. Scaling up from leaf photosynthesis to community photosynthesis should also consider vertical structure of communities and nonlinear responses of leaf photosynthesis to changes in light quantum.     

梭梭和多枝柽柳的枝干光合及其主要影响因子

荒漠植物长期演化过程中保留了枝干光合(Pg)的特性,枝干光合有效减少局部碳损失,且在维持植物正常生理代谢方面发挥重要作用.本研究以古尔班通古特沙漠南缘荒漠植物梭梭和多枝柽柳为对象,利用Li-Cor 6400便携式光合仪与特制叶室相结合监测枝干与叶片光合速率,同时辅以枝干/叶片功能性状(叶绿素含量、含水量、叶/枝面积、碳...     

叶片和群落尺度净光合速率关系的探讨

叶片净光合速率(P_n)是研究光合作用机理的基本尺度; 而群落净光合速率(P_c)是研究群落光合能力及其与外部环境因子间关系的更好尺度, 特别是区域乃至全球尺度碳循环的研究, 需要将叶片尺度的生理生态模型扩展到冠层尺度。理论上, 群落内所有叶片的累积P_n与实测群落净气体交换速率(NCE)是相等的, 但在野外实际观测中, 两者之间的相互关系目前尚未见报道。该文选取敖汉苜蓿(Medicago sativa ‘Aohan’)人工草地, 采用美国LI-COR公司生产的便携式光合测定系统LI-6400测定P_n, 结合叶面积指数等参数推算P_c, 利用LI-8100连接同化箱测定生态系统净气体交换速率(NEE), 加上土壤呼吸速率, 得到NCE。结果表明: P_c为3.52 μmol CO₂·m^-2·s^-1, 与实测NCE (3.56 μmol CO₂·m^-2·s^-1)基本相等。这表明: 可利用P_n, 结合叶面积指数、群落叶片数目、健康叶片比例和群落可接收有效光照的平均比例等4个关键参数, 准确地换算P_c。然而, 利用同化箱式法测定群落呼吸速率时, 不可避免地会包含土壤呼吸, 所以在观测NCE时, 需要同时测定土壤呼吸。此外, 在冠层模型中, 群落垂直结构和光量子的非线性响应不可忽视。     

CO2加富下水氮耦合对黄瓜叶片光合作用和超微结构的影响

以‘津优35号’黄瓜为试材,采用裂区-再裂区设计,研究了CO₂加富下水氮耦合对黄瓜叶片光合作用和超微结构的影响.主区设大气CO₂浓度(400 μmol·mol^-1,A)和加富CO₂浓度(800±20 μmol·mol^-1,E)2个CO₂浓度处理,裂区设无干旱胁迫(田间持水量的95%,W)和干旱胁迫(田间持水量的75%,D)2个水分处理,再裂区设施氮量450 kg·hm^-2(低氮,N₁)和900 kg·hm^-2(高氮,N₂)2个氮素处理.结果表明: 在干旱和高氮条件下,CO₂加富提高了黄瓜的株高,且使高氮下的叶面积显著增加.正常灌溉条件下,高氮处理的光合速率、气孔导度和蒸腾速率高于低氮处理,而干旱条件下则相反;CO₂加富提高了黄瓜叶片的水分利用效率,并且随着施氮量的增加,其水分利用效率提高.干旱胁迫下,黄瓜近轴面气孔密度增加,而CO₂加富和高氮却显著降低了气孔密度.高氮处理增加了黄瓜叶片叶绿体数量而减少了淀粉粒数量,干旱胁迫使叶绿体数量减少,但使淀粉粒数量呈上升趋势.干旱胁迫增加了叶绿体长度和宽度,显著增加了淀粉粒的大小,而高氮降低了叶绿体和淀粉粒的长度和宽度.CO₂加富和高氮均使基粒厚度和片层数增加(ADN₂除外),并且EDN₂处理的片层数显著高于ADN₂.综上所述,CO₂加富和适宜的水、氮条件能促进黄瓜叶片叶绿体类囊体膜系的发育,显著增加基粒厚度和基粒片层数,有效改善黄瓜的叶绿体结构,增强光合性能,提高黄瓜植株对CO₂和水、氮的吸收利用能力.     

参棚透光率对西洋参叶片光合作用的影响

研究了参棚透光率与西洋参叶片净光合速率之间的关系.结果表明,西洋参叶片光饱和点、净光合速率及其日变型均随参棚透光率的不同,存在一定的差异.4年生西洋参叶片在12%、30%、42%3种透光率下(气温29.0℃左右),光饱和点分别为171.0、323.0和429.0μmol·m^-2·s^-1,净光合速率最大值为6.54mg·dm^-2·h^-1(CO₂),出现在透光率为30%的参棚下;3年生西洋参在透光率不超过25.8%的参棚下,叶片净光合速率日变化呈单峰型,透光率大于25.8%时,呈双峰型,参叶“光合午休”现象明显.单相关分析表明,光量子通量密度是影响西洋参净光合速率的主要因子;回归分析结果表明,各影响因子对参叶净光合速率的综合影响显著.     

土壤水分胁迫和大气CO2浓度对光合分馏及后光合分馏的影响

对植物光合和后光合分馏进行分析,有助于提升对植物生理和水分管理等的认识。本研究通过测定大气、侧柏叶片和枝条韧皮部可溶性化合物的δ¹³C,探讨了光合作用时大气和叶片间碳同位素的分馏(ΔC_a-leaf)和光合作用后叶片到枝条间的碳同位素分馏(ΔC_leaf-phlo)对土壤含水量(SWC)和大气CO₂浓度(C_a)的响应。结果表明: ΔC_a-leaf在SWC为田间持水量(FC)的95%～100%(95%～100%FC)且C_a为400 μmol·mol^-1时达到最大值(13.06‰),在SWC为35%～45%FC且C_a为800 μmol·mol^-1时达到最小值(8.63‰);气孔导度和叶肉细胞导度均与ΔC_a-leaf呈显著线性正相关,相关系数分别为0.43和0.44;而ΔC_leaf-phlo并未受到SWC和C_a的显著影响。本研究不仅可以提高对碳同位素的分馏机制的认识,而且可以为植物对未来气候变化的生存适应性提供理论依据。     

CO2加富下水氮耦合对黄瓜叶片光合作用和超微结构的影响

以‘津优35号’黄瓜为试材,采用裂区-再裂区设计,研究了CO₂加富下水氮耦合对黄瓜叶片光合作用和超微结构的影响.主区设大气CO₂浓度(400 μmol·mol^-1,A)和加富CO₂浓度(800±20 μmol·mol^-1,E)2个CO₂浓度处理,裂区设无干旱胁迫(田间持水量的95%,W)和干旱胁迫(田间持水量的75%,D)2个水分处理,再裂区设施氮量450 kg·hm^-2(低氮,N₁)和900 kg·hm^-2(高氮,N₂)2个氮素处理.结果表明: 在干旱和高氮条件下,CO₂加富提高了黄瓜的株高,且使高氮下的叶面积显著增加.正常灌溉条件下,高氮处理的光合速率、气孔导度和蒸腾速率高于低氮处理,而干旱条件下则相反;CO₂加富提高了黄瓜叶片的水分利用效率,并且随着施氮量的增加,其水分利用效率提高.干旱胁迫下,黄瓜近轴面气孔密度增加,而CO₂加富和高氮却显著降低了气孔密度.高氮处理增加了黄瓜叶片叶绿体数量而减少了淀粉粒数量,干旱胁迫使叶绿体数量减少,但使淀粉粒数量呈上升趋势.干旱胁迫增加了叶绿体长度和宽度,显著增加了淀粉粒的大小,而高氮降低了叶绿体和淀粉粒的长度和宽度.CO₂加富和高氮均使基粒厚度和片层数增加(ADN₂除外),并且EDN₂处理的片层数显著高于ADN₂.综上所述,CO₂加富和适宜的水、氮条件能促进黄瓜叶片叶绿体类囊体膜系的发育,显著增加基粒厚度和基粒片层数,有效改善黄瓜的叶绿体结构,增强光合性能,提高黄瓜植株对CO₂和水、氮的吸收利用能力.     

基于稳定碳同位素技术研究大气CO2浓度升高对植物-土壤系统碳循环的影响

大气CO₂浓度升高影响植物光合作用过程和生物量积累,改变植物地上和地下生物量的动态分配.土壤有机质的形成和周转依赖于植物组分的输入,因此,CO₂浓度升高所造成的植物生理和代谢的变化对土壤碳库收支平衡具有重要影响.采用稳定碳同位素(¹³C)技术研究土壤-植物系统的碳循环可阐明大气CO₂浓度升高条件下光合碳在植物各器官的分配特征和时间动态,明确光合碳在土壤中的积累、分解与迁移转化过程以及对土壤有机碳库周转的影响.本文综述了基于¹³C自然丰度法或¹³C示踪技术研究大气CO₂浓度升高对土壤-植物系统碳循环的影响,主要包括:1)对植物光合作用的同位素分馏的影响;2)对植物光合碳(新碳)分配动态的影响;3)对土壤有机碳新老碳库动态以及微生物转化过程的影响.明确上述过程及其调控机制可为预测CO₂浓度升高对陆地生态系统碳循环及源汇效应的长期影响奠定基础.     

Rate limiting factors in leaf photosynthesis. II. Electron transport

Increased scattering of a weak 535 nm measuring beam which indicates the light-dependent formation of a transthylakoid proton gradient in leaves was used to examine the role of the electron-transport chain in limiting photosynthetic carbon assimilation. The proton gradient is supported by electron flux and indicates thylakoid energization. In CO₂-free air, half saturation of thylakoid energization was observed at intensities of red light ranging from 2 to 50 W·m⁻² in different plant species. The differences were attributed to different carbohydrate availability for energy-consuming photorespiratory processes when external CO₂ was absent. Thylakoid energization of shade leaves (Asarum, Fagus) was saturated at lower light intensities than that of sun leaves (Phaseolus, Fagus). When photorespiratory carbohydrate oxidation was suppressed by decreasing the O₂ concentration from 21 to 2% in the absence of CO₂, thylakoid energization saturated at lower light intensities than in CO₂-free air. CO₂ decreased thylakoid energization particularly at low light intensities. Under high intensity illumination, however, thylakoid energization was remarkably high even in the presence of saturating CO₂. Apparently, electron transport was capable of maintaining the energy status of the photosynthetic apparatus at a high level even when photosynthetic carbon fluxes were maximal. This suggests that electron transport is less important in limiting photosynthesis than previously thought.     

基于稳定碳同位素技术研究大气CO2浓度升高对植物-土壤系统碳循环的影响

大气CO₂浓度升高影响植物光合作用过程和生物量积累,改变植物地上和地下生物量的动态分配.土壤有机质的形成和周转依赖于植物组分的输入,因此,CO₂浓度升高所造成的植物生理和代谢的变化对土壤碳库收支平衡具有重要影响.采用稳定碳同位素(¹³C)技术研究土壤-植物系统的碳循环可阐明大气CO₂浓度升高条件下光合碳在植物各器官的分配特征和时间动态,明确光合碳在土壤中的积累、分解与迁移转化过程以及对土壤有机碳库周转的影响.本文综述了基于¹³C自然丰度法或¹³C示踪技术研究大气CO₂浓度升高对土壤-植物系统碳循环的影响,主要包括:1)对植物光合作用的同位素分馏的影响;2)对植物光合碳(新碳)分配动态的影响;3)对土壤有机碳新老碳库动态以及微生物转化过程的影响.明确上述过程及其调控机制可为预测CO₂浓度升高对陆地生态系统碳循环及源汇效应的长期影响奠定基础.     

植物叶肉导度的测定及计算方法综述

叶肉导度(g_m)指叶肉细胞内部的CO₂扩散能力,它是叶肉细胞阻力的倒数.光合作用研究的早期,研究者们多将叶肉细胞对CO₂的扩散阻力视为零,即假定g_m无穷大,而忽略了其对光合作用的限制.但近来的研究表明,g_m是有限的,并随着环境条件的改变而发生变化,此外,g_m的大小直接决定了CO₂在叶片内的扩散量,进而影响到植物光合效率的高低.因此,g_m的估算对于植物光合能力的评估意义重大.目前,气体交换与叶绿素荧光相结合法、曲线拟合法以及瞬时碳同位素(¹³CO₂)辨别法已经成为植物g_m估测的3种常用方法,但国内针对这3种方法原理及其优缺点介绍的文献极少,阐述这3种方法的原理、过程并比较分析其优缺点就显得尤为必要.本文综合了相关文献,从原理、推导过程及优缺点3方面对上述3种g_m估测方法进行了详细介绍.结果表明: 曲线拟合法虽然易于理解,便于操作,但其拟合模型因光合作用的发生状态不同而不同,需要研究者对光合作用不同状态进行严格划分,不利于广泛推广应用;瞬时碳同位素(¹³CO₂)辨别法虽然提高了结果的准确性,但其测定过程比较复杂,对试验操作的要求比较严格,同时该方法对试验误差的敏感性较差,可靠性不高.相较上述两种方法,气体交换与叶绿素荧光相结合法的可操作性更强,可靠性更高,更利于多处理多重复的大样本的观测分析,叶绿素荧光技术的使用,既简化了试验步骤,又降低了试验过程的偶然误差,增加了观测结果的科学性;此外,叶绿素荧光技术还能为叶片提供饱和脉冲活化能,从而最大限度地激发叶片的光合潜能,但该方法也存在很多问题,比如,为了提高叶片叶绿素荧光参数的准确程度,试验中需要使用较低的气体流速,而流速的降低又会增大气体扩散泄漏的风险,所以该方法对选择合理气体流速的要求很高. 综合来看,气体交换与叶绿素荧光相结合法在植物g_m的实际测定中的认可度最高,使用最广泛.     

根际CO2浓度升高对网纹甜瓜光合特性及产量和品质的影响

采用雾培植株根际通CO₂处理方式,研究了开花结果期根际CO₂浓度升高对网纹甜瓜光合作用及产量和品质的影响.结果表明：在网纹甜瓜果实发育期间,与350 μL·L^-1（对照）处理相比,根际2500和5000 μL CO₂·L^-1处理的叶片光合色素含量、净光合速率（P_n）、气孔导度（g_s）、胞间CO₂浓度（C_i）及PSⅡ最大光化学效率（F_v/F_m）均不同程度降低,而气孔限制值（L_s）显著提高,且5000 μL CO₂·L^-1处理的变化幅度高于2500 μL CO₂·L^-1处理;单株产量、果实中维生素C和可溶性糖含量显著降低,有机酸含量显著提高.可见,网纹甜瓜果实发育期间根际CO₂浓度超过2500 μL·L^-1时,其光合作用及果实发育会受到显著抑制,从而导致产量和品质降低.
     

Effect of elevated CO2, temperature and drought on dry matter partitioning and photosynthesis before and after cutting of nodulated alfalfa

The rising atmospheric CO₂ concentration resulting from industrial development may enhance photosynthesis and plant growth. However, there is a lack of research concerning the effect of combined factors such as CO₂, temperature and water availability on plant regrowth following cutting or grazing, which represent the usual methods of managing forage legumes like alfalfa. Elevated CO₂, temperature and drought can interact with cutting factors (e.g. cutting frequency or height), and source-sink balance differences before and after defoliation can modify photosynthetic behaviour and dry matter accumulation, as well as dry matter partitioning between above- and belowground organs. The aim of our study was to determine the interactive effect of CO₂ (ambient, around 350 μmol mol⁻¹ versus 700 μmol mol⁻¹), temperature (ambient versus ambient + 4 °C) and water availability (well-irrigated versus partially irrigated) on dry matter partitioning and photosynthesis in nodulated alfalfa after vegetative normal growth and during regrowth. At the end of vegetative normal growth, CO₂ enhanced dry matter accumulation despite photosynthesis being down-regulated at the end of this period. Photosynthesis was stimulated by elevated CO₂ and resulted in greater dry matter accumulation during the regrowth period. Aboveground organs were affected more by drought than belowground organs during the entire experiment, particularly during vegetative normal growth. The higher drought tolerance (greater growth) observed during the regrowth period may be related to higher mass and greater reserves accumulated in the roots of plants.