微藻生物固碳技术在“双碳”目标中的应用前景

微藻生长速度快、CO₂固定效率高,每生产1 t微藻生物质可固定1.83 t CO₂。同时,微藻还可将固定的CO₂转化为油脂、蛋白质、多糖、色素和不饱和脂肪酸等物质,能够实现CO₂的高值化利用。因此,微藻生物固碳技术在CO₂捕集和利用方面具有极大的发展潜能。本文首先阐述了高效固定CO₂藻株的选育、提高微藻生物固定CO₂的培养策略、微藻处理烟道气化合物技术、微藻高效培养光生物反应器的开发及新兴技术助力微藻碳减排等内容,再结合现阶段微藻生物固碳技术所面临的挑战,展望了微藻生物固定CO₂在“双碳”目标中的应用前景,以期为利用微藻高效固定CO₂、高值化利用CO₂提供参考,从而加速“双碳”目标的实现。     

湖泊生态系统碳汇特征及其潜在碳中和价值研究

为了应对气候挑战,达成碳达峰远景目标,需要正确评估自然资源碳中和价值。湖泊作为具有独特生态、人文价值的地理单元,因碳循环强度高、碳排放总量大,是传统意义上的碳源。通过梳理近期相关研究成果,对比不同类型湖泊碳汇/源状况,湖泊生态系统以一系列碳汇特征表现出潜在的碳中和价值。强烈的光合作用可以使水体CO₂欠饱和,但由呼吸-光合作用、碳酸盐岩溶蚀作用带来的水体碱度、CO₂分压pCO₂提高也有利于湖泊碳汇增益。CO₂在水体中大量溶解,积极参与到湖泊碳循环,将pCO₂高于40 Pa作为判断湖泊为碳源的依据可能忽视了水体碱度上升带来的碳汇。在湖泊沉积物中有机碳的累积受到生态系统光合-呼吸作用的影响,当异养微生物群落能及时分解沉入湖底的衰亡组织、有机质时,沉积物中有机碳不会大量累积,当呼吸对光合的相对滞后,有机碳才会大量累积。湖泊生态系统的生产力决定了固碳能力,是湖泊发挥碳汇效益的重要“碳库”。由水生植物固定下的CO₂总量不如浮游植物,但在过程中发挥了“压舱石”般的稳定作...     

微生物利用CO2的研究进展

微生物利用CO₂,这不仅提高了自然界碳的利用率,而且能将CO₂转化为高附加值产品,可以实现资源的再利用。利用现代生物技术改造微生物以提高CO₂的利用效率对生物制造和实现碳中和有一定的促进作用。本文首先总结了微生物利用CO₂的6个主要天然途径,并分析这些途径的优缺点,这是对相关途径进行改造的基础;其次提供克服天然途径中碳原子利用率低、能耗高、产品得率低等问题的方法;再次阐述了面向CO₂高效利用的合成生物技术研究方向以及途径改造的主要策略和效果;最后展望了微生物利用CO₂的机遇及挑战,以期为改造微生物提高CO₂利用效率和高附加值产品的生产奠定基础。     

亚热带毛竹人工林土壤呼吸组分动态变化及其影响因素

利用Li-8100土壤碳通量测量系统,研究了2013年4月—2014年3月浙江临安市毛竹人工林土壤呼吸、异养呼吸和自养呼吸速率的动态变化规律.结果表明：毛竹人工林土壤总呼吸速率、异养呼吸速率和自养呼吸速率均呈现出明显的季节变化特征,最高值出现在7月,最低值出现在1月,年平均值分别为2.93、1.92和1.01 μmol CO₂·m^-2·s^-1.毛竹林土壤总呼吸、异养呼吸和自养呼吸年累积CO₂排放量分别为37.25、24.61和12.64 t CO₂·hm^-2·a^-1.土壤呼吸各组分均与土壤5 cm温度呈显著指数相关,土壤总呼吸、异养呼吸和自养呼吸的温度敏感系数Q₁₀值分别为2.05、1.95和2.34.土壤总呼吸速率、异养呼吸速率与土壤水溶性有机碳(WSOC)含量均呈显著相关,而自养呼吸与WSOC无显著相关性;土壤呼吸各组分与土壤含水〖JP2〗量以及微生物生物量碳均无显著相关性.土壤温度是影响毛竹人工林土壤呼吸及其组分季节变化的主要驱动因子,土壤WSOC含量是影响土壤总呼吸和异养呼吸的重要环境因子.     

黄河三角洲典型滨海盐沼湿地土壤CO2和CH4排放对水盐变化的响应

滨海盐沼湿地是重要的“蓝碳”碳汇,研究水盐变化对土壤碳矿化(CO₂和CH₄排放)的影响,对理解滨海盐沼湿地的碳汇稳定机制具有重要意义。该研究选取黄河三角洲典型盐沼湿地土壤为研究对象,通过水盐梯度模拟实验,研究土壤碳矿化、理化性质、微生物生物量及群落结构对不同土壤水分和盐分含量的响应。主要结果:(1)水盐变化对土壤CO₂、CH₄排放量以及CH₄:CO₂的影响均不存在交互作用,CO₂排放量随土壤含水量增加呈先升后降的单峰型变化趋势,盐分含量升高则显著抑制CO₂排放;水分含量升高对CH₄排放具有显著促进作用,盐分升高则显著抑制CH₄排放。(2)水盐变化对土壤可溶性有机碳(DOC)含量具有弱交互作用,在低水分处理下,DOC随着土壤盐分的增加呈减少趋势,但在高水分处理下呈增加趋势;CO₂排放与DOC含量呈显著正相关关系,而CH₄排放与D...     

湿润亚热带峰丛洼地岩溶土壤系统中碳分布及其转移

以桂林丫吉村岩溶试验场为例,研究了湿润亚热带峰丛洼地表层岩溶带生物量碳库、凋落物碳库、土壤有机碳库(SOM)及其活泼性、有机碳分解速率、土壤中CO₂浓度和土壤呼吸CO₂排放,表明岩溶系统中丰富的碳库提供了系统中CO₂的来源,并用D¹³C证实春夏岩溶活跃季节中岩溶输出C约60%来自土壤CO₂.由此认为,驱动岩溶作用的CO₂并非直接来自大气CO₂,而是大气-植物-土壤-水碳素转移的结果,因而揭示了土壤作为碳环境地球化学界面对于表生带岩溶作用的驱动意义.     

蒙古栎叶片及其土壤碳、氮同位素自然丰度对大气CO2浓度升高的响应

大气CO₂浓度升高对土壤氮素转化过程产生重要影响,研究其变化有助于更好地预测陆地生态系统的固碳潜力.氮同位素自然丰度作为生态系统氮素循环过程的综合指标能够有效地指示CO₂浓度升高对土壤氮素转化过程的影响.本研究采用开顶箱CO₂ 熏蒸法研究连续10年的大气CO₂ 浓度升高对我国东北地区蒙古栎及其土壤和微生物生物量碳、氮同位素自然丰度的影响.结果表明: 大气CO₂浓度升高改变了土壤氮循环过程,增加了土壤微生物和植物叶片δ¹⁵N;促进了富¹³C土壤有机碳分解,中和了贫¹³C植物光合碳输入的效果,导致土壤可溶性有机碳和微生物碳δ¹³C在CO₂升高条件下没有发生显著变化.这些结果表明,CO₂浓度升高很可能促进了土壤有机质矿化过程,并加剧了系统氮限制的状态.     

马占相思人工林净二氧化碳交换和碳同位素通量

应用稳定碳同位素技术,对马占相思人工林冠层受光和遮荫叶片的碳同化率(A_net)和叶面积指数(L)进行加权,将叶片水平的¹³C甄别率(Δ_i)扩展至冠层光合甄别率(Δ_canopy),测定光合固定和呼吸释放的碳同位素通量及其净交换通量.结果表明：Δ_canopy的日变化明显,日出前和中午出现较低值(18.47‰和19.87‰),而日落前达到最大(21.21‰);秋季末期(11月)至翌年夏季,Δ_canopy逐步升高,年平均为(20.37±0.29)‰.不同季节自养呼吸(日间叶片呼吸除外)和异养呼吸释放CO₂的碳同位素比率(δ¹³C)平均值分别为(-28.70±0.75)‰和(-26.75±1.3)‰,春季林冠夜间呼吸CO₂的δ¹³C最低(-30.14‰),秋季末期最高(-28.01‰).马占相思林与大气的CO₂碳同位素通量在春季和夏季中午时峰值分别为178.5和217 μmol·m^-2 ·s^-1·‰,日均值分别为638.4 和873.2 μmol·m^-2·s^-1·‰.冠层叶片吸收CO₂的碳同位素通量较呼吸释出CO₂的碳同位素通量高1.6～2.5倍,表明马占相思林日间吸收大量CO₂,降低空气CO₂浓度,具有改善环境的良好生态服务功能.     

蒙古栎叶片及其土壤碳、氮同位素自然丰度对大气CO2浓度升高的响应

大气CO₂浓度升高对土壤氮素转化过程产生重要影响,研究其变化有助于更好地预测陆地生态系统的固碳潜力.氮同位素自然丰度作为生态系统氮素循环过程的综合指标能够有效地指示CO₂浓度升高对土壤氮素转化过程的影响.本研究采用开顶箱CO₂ 熏蒸法研究连续10年的大气CO₂ 浓度升高对我国东北地区蒙古栎及其土壤和微生物生物量碳、氮同位素自然丰度的影响.结果表明: 大气CO₂浓度升高改变了土壤氮循环过程,增加了土壤微生物和植物叶片δ¹⁵N;促进了富¹³C土壤有机碳分解,中和了贫¹³C植物光合碳输入的效果,导致土壤可溶性有机碳和微生物碳δ¹³C在CO₂升高条件下没有发生显著变化.这些结果表明,CO₂浓度升高很可能促进了土壤有机质矿化过程,并加剧了系统氮限制的状态.     

红壤和风沙土添加不同化学结构有机碳对外源碳去向及累积贡献的影响

植物凋落物碳输入显著影响陆地生态系统土壤CO₂排放和有机碳(SOC)形成,然而,针对不同质地土壤添加不同化学结构外源碳去向依然不清楚。本研究将¹³C标记的葡萄糖、淀粉和纤维素添加至红壤和风沙土,比较2种质地土壤添加不同化学结构外源碳在土壤释放的CO₂、SOC、可溶性有机碳(DOC)和微生物生物量碳(MBC)库的净累积量、回收率及贡献比例上的差异。结果表明: 添加外源有机碳显著提高了CO₂、SOC、DOC和MBC的δ¹³C值,且随着外源有机碳化学结构复杂性的增加,CO₂的δ¹³C峰值依次延迟出现;外源有机碳种类、土壤类型和培养时间均显著改变外源碳去向及其在各碳库的贡献比例;在风沙土中,外源有机碳更多被矿化为CO₂,且CO₂库的外源碳净累积量和回收率大小依次为葡萄糖>淀粉>纤维素;红壤添加外源碳转变为SOC的累积量和回收率显著高于风沙土,且红壤SOC库的外源碳净累积量和回收率大小顺序也为葡萄糖>淀粉>纤维素。可见,外源有机碳化学结构和土壤质地共同调控外源碳去向及累积贡献。     

Non-phototrophic CO 2 fixation by soil microorganisms

Although soils are generally known to be a net source of CO₂ due to microbial respiration, CO₂ fixation may also be an important process. The non-phototrophic fixation of CO₂ was investigated in a tracer experiment with ¹⁴CO₂ in order to obtain information about the extent and the mechanisms of this process. Soils were incubated for up to 91 days in the dark. In three independent incubation experiments, a significant transfer of radioactivity from ¹⁴CO₂ to soil organic matter was observed. The process was related to microbial activity and could be enhanced by the addition of readily available substrates such as acetate. CO₂ fixation exhibited biphasic kinetics and was linearly related to respiration during the first phase of incubation (about 20–40 days). The fixation amounted to 3–5% of the net respiration. After this phase, the CO₂ fixation decreased to 1–2% of the respiration. The amount of carbon fixed by an agricultural soil corresponded to 0.05% of the organic carbon present in the soil at the beginning of the experiment, and virtually all of the fixed CO₂ was converted to organic compounds. Many autotrophic and heterotrophic biochemical processes result in the fixation of CO₂. However, the enhancement of the fixation by addition of readily available substrates and the linear correlation with respiration suggested that the process is mainly driven by aerobic heterotrophic microorganisms. We conclude that heterotrophic CO₂ fixation represents a significant factor of microbial activity in soils.     

自养微生物同化CO_2的分子生态研究及同化碳在土壤中的转化

大气中CO2浓度持续升高和全球气候变暖是亟待解决的重大环境问题。自养微生物在环境中广泛分布,能直接参与CO2的同化,因此研究自养微生物同化CO2的分子生态学机制具有重大的科学意义。以往对自养微生物的研究多针对基因组DNA,从DNA水平揭示了不同生态系统中碳同化自养微生物的种群结构和多样性,但这些微生物在生态系统中的具体功能有待进一步的研究。近年来,随着转录组学研究技术和稳定同位素探针技术(SIP)的发展,自养微生物同化CO2的生态机理研究不断深入,这些研究明确揭示了碳同化自养微生物是河流、湖泊和海洋生态系统中CO2固定作用的驱动者,并新发现了一些具有CO2同化功能的微生物群落。基于国内外有关研究进展,从DNA和RNA水平上对自养微生物同化CO2的分子机理以及稳定同位素探针技术(SIP)在碳同化微生物研究中的应用进行了分析和总结,初步展望了RNA-SIP技术在陆地生态系统碳同化微生物分子生态学研究中的前景。同时,探讨了陆地生态系统同化碳的转化和稳定性机理,以期为深入了解生态系统碳循环过程和应对气候变化提供理论依据。     

Inhibitory effect of organic carbon on CO₂ fixing by non-photosynthetic microbial community isolated from the ocean

The inhibitory effect of organic carbon on CO₂ fixation (CF) by the non-photosynthetic microbial community (NPMC) and its mechanism were studied. The results showed that different concentrations of glucose inhibited CF to some extent. However, when these microorganisms pre-cultured with glucose were re-cultured without organic carbon, their CF efficiency differed significantly from the control based on the glucose concentration in the pre-culture. ATP as bioenergy and NADH as reductant had no obvious inhibitory effect on CF; conversely, they improved CF efficiency to some extent, especially when both were present simultaneously. These results implied that not all organic materials inhibited CF by NPMC, and only those that acted as good carbon sources, such as glucose, inhibited CF. Moreover, some metabolites generated during the catabolism of glucose by heterotrophic metabolism of NPMC might inhibit CF, while other cumulated materials present in the cell interior, such as ATP and NADH, might improve CF.     

Photoautotrophic growth of soybean cells in suspension culture III. Characterization of carbon fixation products under high and low CO2 levels

A photoautotrophic soybean suspension culture (SB-P) was used to study CO₂ assimilation while exposed to elevated or ambient CO₂ levels. These studies showed that under elevated CO₂ (5% v/v) malate is the dominant fixation product, strongly suggesting that phosphoenolpyruvate carboxylase (PEPCase) is the primary enzyme involved in carbon fixation in these cells under their normal growth conditions. Citrate and [aspartate + glutamate] were also significant fixation products during fifteen minutes of exposure to ¹⁴CO₂. During the ten minute unlabeled CO₂ chase however, ¹⁴C-malate continued to increase while citrate and [aspartate + glutamate] declined. Fixation of ¹⁴CO₂ under ambient CO₂ levels (0.037%) showed a very different product pattern as 3-phosphoglycerate was very high in the first one to two minutes followed by increases in [serine + glycine] and [aspartate + glutamate]. Hexose phosphates were also quite high initially but then declined relatively rapidly. Thus, the carbon fixation pattern at ambient CO₂ levels resembles somewhat that seen in C3 leaf cells while that seen at elevated CO₂ levels more closely resembles that of a C₄ plant. The initial fixation product of C₃ plants, 3-PGA, was never detectable under high CO₂ conditions. These data suggest that an in vitro photoautotrophic system would be suitable for studying carbon fixation physiology during photosynthetic and non-photosynthetic growth.Abbreviations SB-P photoautotrophic soybean cells - PEPCase phosphoenol-pyruvate carboxylase - RuBPCase ribulose bisphosphate carboxylase/oxygenase - 3-PGA 3-phosphoglycerate     

Uptake and utilization of inorganic carbon by cyanobacteria

In the cyanobacteria, mechanisms exist that allow photosynthetic CO₂ reduction to proceed efficiently even at very low levels of inorganic carbon. These inducible, active transport mechanisms enable the cyanobacteria to accumulate large internal concentrations of inorganic carbon that may be up to 1000-fold higher than the external concentration. As a result, the external concentration of inorganic carbon required to saturate cyanobacterial photosynthesis in vivo is orders of magnitude lower than that required to saturate the principal enzyme (ribulose bisphosphate carboxylase) involved in the fixation reactions. Since CO₂ is the substrate for carbon fixation, the cyanobacteria somehow perform the neat trick of concentrating this small, membrane permeable molecule at the site of CO₂ fixation. In this review, we will describe the biochemical and physiological experiments that have outlined the phenomenon of inorganic carbon accumulation, relate more recent genetic and molecular biological observations that attempt to define the constituents involved in this process, and discuss a speculative theory that suggests a unified view of inorganic carbon utilization by the cyanobacteria.Abbreviations C_i Inorganic carbon - H-cells Cells grown under high CO₂ - L-cells Cells grown under low CO₂ - RuBP Ribulose-1,5-bisphosphate - WT Wild type     

Feedback of carbon and nitrogen cycles enhances carbon sequestration in the terrestrial biosphere

The efforts to explain the ‘missing sink’ for anthropogenic carbon dioxide (CO₂) have included in recent years the role of nitrogen as an important constraint for biospheric carbon fluxes. We used the Nitrogen Carbon Interaction Model (NCIM) to investigate patterns of carbon and nitrogen storage in different compartments of the terrestrial biosphere as a consequence of a rising atmospheric CO₂ concentration, in combination with varying levels of nitrogen availability. This model has separate but closely coupled carbon and nitrogen cycles with a focus on soil processes and soil–plant interactions, including an active compartment of soil microorganisms decomposing litter residues and competing with plants for available nitrogen. Biological nitrogen fixation is represented as a function of vegetation nitrogen demand. The model was validated against several global datasets of soil and vegetation carbon and nitrogen pools. Five model experiments were carried out for the modeling periods 1860–2002 and 2002–2100. In these experiments we varied the nitrogen availability using different combinations of biological nitrogen fixation, denitrification, leaching of soluble nitrogen compounds with constant or rising atmospheric CO₂ concentrations. Oversupply with nitrogen, in an experiment with nitrogen fixation, but no nitrogen losses, together with constant atmospheric CO₂, led to some carbon sequestration in organismic pools, which was nearly compensated by losses of C from soil organic carbon pools. Rising atmospheric CO₂ always led to carbon sequestration in the biosphere. Considering an open nitrogen cycle including dynamic nitrogen fixation, and nitrogen losses from denitrification and leaching, the carbon sequestration in the biosphere is of a magnitude comparable to current observation based estimates of the ‘missing sink.’ A fertilization feedback between the carbon and nitrogen cycles occurred in this experiment, which was much stronger than the sum of separate influences of high nitrogen supply and rising atmospheric CO₂. The demand‐driven biological nitrogen fixation was mainly responsible for this result. For the modeling period 2002–2100, NCIM predicts continued carbon sequestration in the low range of previously published estimates, combined with a plausible rate of CO₂‐driven biological nitrogen fixation and substantial redistribution of nitrogen from soil to plant pools.     

Heterotrophic Fixation of CO2 in Soil

The occurrence of heterotrophic CO₂ fixation by soil microorganisms was tested in several mineral soils differing in pH and two artificial soils (a mixture of silica sand, alfalfa powder, and nutrient medium inoculated with a soil suspension). Soils were incubated at ambient (∼0.05 vol%) and elevated (∼5 vol%) CO₂ concentrations under aerobic conditions for up to 21 days. CO₂ fixation was detected using either a technique for determining the natural abundance of ¹³C or by measuring the distribution of labeled ¹⁴C-CO₂ in soil and bacteria. The effects of elevated CO₂ on microbial biomass (direct counts, chloroform fumigation extraction method), composition of microbial community (phospholipid fatty acids), microbial activity (respiration, dehydrogenase activity), and turnover rate were also measured. Heterotrophic CO₂ fixation was proven in all soils under study, being higher in neutral soils. The main portion of the fixed CO₂ (98–99%) was found in extracellular metabolites while only ∼1% CO₂ was incorporated into microbial cells. High CO₂ concentration always induced an increase in microbial activity, changes in the composition of the microbial community, and a decrease in microbial turnover. The results suggest that heterotrophic CO₂ fixation could be a widespread process in soils.     

Biological N2 fixation by heterotrophic and phototrophic bacteria in association with straw

Much of the crop residues, including cereal straw, that are produced worldwide are lost by burning. Plant residues, and in particular straw, contain large amounts of carbon (cellulose and hemicellulose) which can serve as substrates for the production of microbial biomass and for biological N₂ fixation by a range of free-living, diazotrophic bacteria. Microorganisms with the dual ability to utilise cellulose and fix N₂ are rate, but some strains that utilize hemicellulose and fix N₂ have been found. Generally, cellulolysis and diazotrophy are carried out by a mixed microbial community in which N₂-fixing bacteria utilise cellobiose and glucose produced from straw by cellulolytic microorganisms. N₂-fixing bacteria include heterotrophic and phototrophic organisms and the latter are apparently more prominent in flooded soils such as rice paddies than in dryland soils. The relative contributions of N₂ fixed by heterotrophic diazotrophic bacteria compared with cyanobacteria and other phototrophic bacteria depend on the availability of substrates from straw decomposition and on environmental pressures. Measurements of asymbiotic N₂ fixation are limited and variable but, in rice paddy systems, rates of 25 kg N ha^-1 over 30 days have been found, whereas in dryland systems with wheat straw, in situ measurements have indicated up to 12 kg N ha^-1 over 22 days. Straw-associated N₂ fixation is directly affected by environmental factors such as temperature, moisture, oxygen concentration, soil pH and clay content as well as farm management practices. Modification of managements and use of inoculants offer ways of improving asymbiotic N₂ fixation.In laboratory culture systems, inoculation of straws with cellulolytic and diazotrophic microorganisms has resulted in significant increases in N₂ fixation in comparison to uninoculated controls and gains of N of up to 72 mg N fixed g^-1 straw consumed have been obtained, indicating the potential of inoculation to improve N gains in composts that can then be used as biofertilisers. Soils, on the other hand, contain established, indigenous microbial populations which tend to exclude inoculant microorganisms by competition. As a consequence, improvements in straw-associated N₂ fixation in soils have been achieved mostly by specific straw-management practices which encourage microbial activity by straw-decomposing and N₂-fixing microorganisms.Further research is needed to quantify more accurately the contribution of asymbiotic N₂ fixation to cropping systems. New strains of inoculants, including those capable of both cellulolytic and N₂-fixing activity, are needed to improve the N content of biofertilisers produced from composts. Developments of management practices in farming systems may result in further improvements in N₂ fixation in the field.     

Metabolism of homoacetogens

Homoacetogenic bacteria are strictly anaerobic microorganisms that catalyze the formation of acetate from C₁ units in their energy metabolism. Most of these organisms are able to grow at the expense of hydrogen plus CO₂ as the sole energy source. Hydrogen then serves as the electron donor for CO₂ reduction to acetate. The methyl group of acetate is formed from CO₂ via formate and reduced C₁ intermediates bound to tetrahydrofolate. The carboxyl group is derived from carbon monoxide, which is synthesized from CO₂ by carbon monoxide dehydrogenase. The latter enzyme also catalyzes the formation of acetyl-CoA from the methyl group plus CO. Acetyl-CoA is then converted either to acetate in the catabolism or to cell carbon in the anabolism of the bacteria. The homoacetogens are very versatile anaerobes, which convert a variety of different substrates to acetate as the major end product.     

ENHANCED TRANSPORT OF INORGANIC CARBON INTO ALGAL CELLS AND ITS IMPLICATIONS FOR THE BIOLOGICAL FIXATION OF CARBON1,2

Kinetics of uptake of inorganic carbon by the freshwater green alga Chlamydomonas reinhardtii Dang. suggest that rates of fixation may be enhanced at low tensions of CO₂ by transport of bicarbonate from the cell surface to the chloroplast. Results are evaluated in the context of models that treat diffusion and reaction of dissolved inorganic carbon across a 3 dimensional finite boundary layer, and they are consistent with the claim that CO₂ alone is the substrate used during carbon fixation. An alternative hypothesis, which presumes that both CO₂ and bicarbonate are used as substrates, yields predictions which are inconsistent with the data. Instead, bicarbonate seems to act only as a vehicle for the transport of inorganic carbon into the cell, thereby adding its flux to that of CO₂, and enhancing rates of synthesis that would otherwise be restricted by uptake of CO₂ alone.