Low temperature effects on leaf physiology and survivorship in the C3 and C4 subspecies of Alloteropsis semialata

The species richness of C(4) grasses is strongly correlated with temperature, with C(4) species dominating subtropical ecosystems and C(3) types predominating in cooler climates. Here, the effects of low temperatures on C(4) and C(3) grasses are compared, controlling for phylogenetic effects by using Alloteropsis semialata, a unique species with C(4) and C(3) subspecies. Controlled environment and common garden experiments tested the hypotheses that: (i) photosynthesis and growth are greater in the C(4) than the C(3) subspecies at high temperatures, but this advantage is reversed below 20 degrees C; and (ii) chilling-induced photoinhibition and light-mediated freezing injury of leaves occur at higher temperature thresholds in the C(4) than the C(3) plants. Measurements of leaf growth and photosynthesis showed the expected advantages of the C(4) pathway over the C(3) type at high temperatures. These declined with temperature, but were not completely lost until 15 degrees C, and there was no evidence of a reversal to give a C(3) advantage. Chronic chilling (5-15 degrees C) or acute freezing events induced a comparable degree of photodamage in illuminated leaves of both subspecies. Similarly, freezing caused high rates of mortality in the unhardened leaves of both subtypes. However, a 2-week chilling treatment prior to these freezing events halved injury in the C(3) but not the C(4) subspecies, suggesting that C(4) leaves lacked the capacity for cold acclimation. These results therefore suggest that C(3) members of this subtropical species may gain an advantage over their C(4) counterparts at low temperatures via protection from freezing injury rather than higher photosynthetic rates.     

C4 photosynthesis evolved in warm climates but promoted migration to cooler ones

C₄ photosynthesis is considered an adaptation to warm climates, where its functional benefits are greatest and C₄ plants achieve their highest diversity and dominance. However, whether inherent physiological barriers impede the persistence of C₄ species in cool environments remains debated. Here, we use large grass phylogenetic and geographical distribution data sets to test whether (1) temperature influences the rate of C₄ origins, (2) photosynthetic types affect the rate of migration among climatic zones, and (3) C₄ evolution changes the breadth of the temperature niche. Our analyses show that C₄ photosynthesis in grasses originated in tropical climates, and that C₃ grasses were more likely to colonise cold climates. However, migration rates among tropical and temperate climates were higher in C₄ grasses. Therefore, while the origins of C₄ photosynthesis were concentrated in tropical climates, its physiological benefits across a broad temperature range expanded the niche into warmer climates and enabled diversification into cooler environments.     

The response of the high altitude C(4) grass Muhlenbergia montana (Nutt.) A.S. Hitchc. to long- and short-term chilling.

The acclimation of C(4) photosynthesis to low temperature was studied in the montane grass Muhlenbergia montana in order to evaluate inherent limitations in the C(4) photosynthetic pathway following chilling. Plants were grown in growth cabinets at 26 degrees C days, but at night temperatures of either 16 degrees C (the control treatment), 4 degrees C for at least 28 nights (the cold-acclimated treatment), or 1 night (the cold-stress treatment). Below a measurement temperature of 25 degrees C, little difference in the thermal response of the net CO(2) assimilation rate (A) was observed between the control and cold-acclimated treatment. By contrast, above 30 degrees C, A in the cold-acclimated treatment was 10% greater than in the control treatment. The temperature responses of Rubisco activity and net CO(2) assimilation rate were similar below 22 degrees C, indicating high metabolic control of Rubisco over the rate of photosynthesis at cool temperatures. Analysis of the response of A to intercellular CO(2) level further supported a major limiting role for Rubisco below 20 degrees C. As temperature declined, the CO(2) saturated plateau of A exhibited large reductions, while the initial slope of the CO(2) response was little affected. This type of response is consistent with a Rubisco limitation, rather than limitations in PEP carboxylase capacity. Stomatal limitations at low temperature were not apparent because photosynthesis was CO(2) saturated below 23 degrees C at air levels of CO(2). In contrast to the response of photosynthesis to temperature and CO(2) in plants acclimated for 4 weeks to low night temperature, plants exposed to 4 degrees C for one night showed substantial reduction in photosynthetic capacity at temperatures above 20 degrees C. Because these reductions were at both high and low CO(2), enzymes associated with the C(4) carbon cycle were implicated as the major mechanisms for the chilling inhibition. These results demonstrate that C(4) plants from climates with low temperature during the growing season can fully acclimate to cold stress given sufficient time. This acclimation appears to involve reversal of injury to the C(4) cycle following initial exposure to low temperature. By contrast, carbon gain at low temperatures generally appears to be constrained by the carboxylation capacity of Rubisco, regardless of acclimation time. The inability to overcome the Rubisco limitation at low temperature may be an inherent limitation restricting C(4) photosynthetic performance in cooler climates.     

Climate, phylogeny and the ecological distribution of C4 grasses

'C4 photosynthesis' refers to a suite of traits that increase photosynthesis in high light and high temperature environments. Most C4 plants are grasses, which dominate tropical and subtropical grasslands and savannas but are conspicuously absent from cold growing season climates. Physiological attributes of C4 photosynthesis have been invoked to explain C4 grass biogeography; however, the pathway evolved exclusively in grass lineages of tropical origin, suggesting that the prevalence of C4 grasses in warm climates could be due to other traits inherited from their non-C4 ancestors. Here we investigate the relative influences of phylogeny and photosynthetic pathway in determining the ecological distributions of C4 grasses in Hawaii. We find that the restriction of C4 grasses to warmer areas is due largely to their evolutionary history as members of a warm-climate grass clade, but that the pathway does appear to confer a competitive advantage to grasses in more arid environments.     

Cold acclimation in warmer extended autumns impairs freezing tolerance of perennial ryegrass (Lolium perenne) and timothy (Phleum pratense)

The effect of variable autumn temperatures in combination with decreasing irradiance and daylength on photosynthesis, growth cessation and freezing tolerance was investigated in northern‐ and southern‐adapted populations of perennial ryegrass (Lolium perenne) and timothy (Phleum pratense) intended for use in regions at northern high latitudes. Plants were subjected to three different acclimation temperatures; 12, 6 and 9/3°C (day/night) for 4 weeks, followed by 1 week of cold acclimation at 2°C under natural light conditions. This experimental setup was repeated at three different periods during autumn with decreasing sums of irradiance and daylengths. Photoacclimation, leaf elongation and freezing tolerance were studied. The results showed that plants cold acclimated during the period with lowest irradiance and shortest day had lowest freezing tolerance, lowest photosynthetic activity, longest leaves and least biomass production. Higher acclimation temperature (12°C) resulted in lower freezing tolerance, lower photosynthetic activity, faster leaf elongation rate and higher biomass compared with the other temperatures. Photochemical mechanisms were predominant in photoacclimation. The northern‐adapted populations had a better freezing tolerance than the southern‐adapted except when grown during the late autumn period and at the highest temperature; then there were no differences between the populations. Our results indicate that the projected climate change in the north may reduce freezing tolerance in grasses as acclimation will take place at higher temperatures and shorter daylengths with lower irradiance.     

The after-effects of temperature and irradiance during early growth of winter oilseed rape (Brassica napus L. var. oleifera, cv. Górczański) seedlings on the progress of their cold acclimation

Experiments performed under controlled conditions showed that level of PPFD (photosynthetic photon flux density) during early seedlings growth (preceding cold acclimation at +2 °C) was not the key factor for the development of frost resistance. It did not modify the beneficial effects of prehardening (Rapacz 1997, in this issue) at moderately low (+12 °C) day temperature. Now I have shown that the increase of PPFD may replace to some extent prehardening in the development of frost resistance. It was particularly seen in non-prehardened plants, which had been grown under warm-day (+20 °C) conditions. Prehardening performed under controlled conditions, as well as seedlings growth under natural autumn conditions in the field, allowed to maintain a high net-photosynthesis rate at chilling temperatures. A net-photosynthesis rate during cold acclimation at +2 °C corresponded well with higher frost resistance. As a result, seedlings non subjected to prehardening and grown before cold acclimation under low PPFD acclimated better, if the cold treatment was applied only at nights (+20/2 °C day/night). Only under such conditions the photosynthetic rate was sufficiently high to allow plants to reach a higher level of frost resistance. All other plants acclimated better when they were exposed to the hardening temperature continuously during days and nights (+2/2 °C day/night).     

低温锻炼后桑树幼苗光合作用和抗氧化酶对冷胁迫的响应

以桑树品种“秋雨”为试验材料,研究了桑树幼苗在低温锻炼、冷胁迫和常温恢复期间的光合作用和抗氧化酶活性的变化.结果表明: 12 ℃3 d低温锻炼明显提高了桑树幼苗的抗冷性.3 ℃3 d冷胁迫下,12 ℃3 d低温锻炼后的桑树幼苗叶片净光合速率(P_n)、气孔导度(G_s)和PSⅡ 最大光化学效率(F_v/F_m)明显高于对照(未经低温锻炼)处理的桑树幼苗,而且其在常温下的恢复也较对照桑树幼苗迅速.在12 ℃ 3 d低温锻炼和3 ℃ 3 d冷胁迫期间,桑树幼苗叶片脯氨酸和可溶性糖含量明显增加,而经低温锻炼的桑树幼苗叶片丙二醛(MDA)含量明显低于未经低温锻炼的桑树幼苗,经低温锻炼的桑树幼苗叶片抗坏血酸过氧化物酶(APX)活性则明显高于未经低温锻炼的桑树幼苗.说明渗透调节物质含量增加和APX活性提高在低温锻炼诱导桑树幼苗的抗冷性上发挥着重要的作用.     

PHOTOSYNTHETIC SENSITIVITY TO TEMPERATURE IN POPULATIONS OF TWO C4 BOUTELOUA (POACEAE) SPECIES NATIVE TO DIFFERENT ALTITUDES

The relationships between photosynthesis, flowering, and growth temperatures were examined experimentally in four populations of the C₄ grass genus Bouteloua. Field-collected plants were grown under two temperature regimes, cool (20 C day/6 C night) and warm (30/16), representative of the extreme populations. Populations collected from the warm climates had significantly lower photosynthetic capacity when grown in the cool chamber relative to the warm chamber, while photosynthetic capacity in the cool climate populations did not differ between the growth conditions. Additionally, exposure to a 2-day cold temperature treatment (10/-2), representative of late-season frosts in high altitude sites, resulted in further reductions in photosynthesis in the warm climate plants, but not in the cool climate plants. This effect was greater for plants grown in the cool growth chamber. Flowering was reduced by 70% in the warm climate plants grown in the cool chamber, and was correlated with photosynthetic inhibition following the short-term cold temperature treatment. These results indicate that genetic differentiation for photosynthetic temperature sensitivity has occurred in the cool climate populations, and that long-term exposure to cool temperatures coupled with short-term relatively extreme low temperatures results in greater photosynthetic inhibition in nontolerant populations.     

Is photosynthetic acclimation to low temperature controlled by capacities for storage and growth at low temperature? Results from comparative studies of grasses and trees

The ability of warm-grown leaves to acclimate their photosynthetic machinery to low, non-freezing temperature was compared for contrasting species of grasses and trees. All trees ( Betula pubescens , Salix sp. and Picea abies ), and young plants of one of the grasses ( Hordeum vulgare ) showed acclimation of photosynthesis while the other two grasses ( Phalaris arundinacea and Festuca ovina ) did not. It was those species that maintained leaf sugar concentrations essentially unchanged that showed acclimation. Trees maintained leaf sugar concentrations essentially unchanged by effectively converting leaf sugar surpluses into storage compounds. Grasses were, by comparison, less effective. However, very young plants of Hordeum maintained leaf sugar concentrations unchanged by continued growth rather than by increased storage. This diversity of low-temperature responses are discussed in relation to possible different priorities of trees and grasses: for grasses to undergo cold hardening by allowing sugars to rise, and for trees to store sugars to allow photosynthesis to operate independently of growth as growth varies with growth rhythm and air temperature.     

The role of the photosynthetic apparatus in cold acclimation of <Emphasis Type="Italic">Lolium multiflorum</Emphasis>. Characteristics of novel genotypes low-sensitive to PSII over-reduction

During cold acclimation by higher plants, temperature perception via changes in redox state of Photosystem II (PSII) and subsequent acclimation of the photosynthetic apparatus to cold is very important for achieving freezing tolerance. These properties were studied in two groups (A and B) of the same backcross 3 (BC₃) progeny derived from a triploid hybrid of Festuca pratensis (2×) × Lolium multiflorum (4×) backcrossed three times onto diploid L. multiflorum cultivars. Leaves of Group A plants formed at 20°C at medium-low light were unable to acclimate their photosynthetic apparatus to cold. Compared to Group B, the Group A plants were also more frost sensitive. This acclimation ability correlated with the freezing tolerance of the plants. However, leaves of the same Group A plants developed at 20°C, but under higher-light conditions had increased ability to acclimate their photosynthetic apparatus to cold. It was concluded that Group A plants may have impaired PSII temperature perception, and this then resulted in their poor capability to cold acclimate.     

Are budburst dates,dormancy and cold acclimation in walnut trees (<Emphasis Type="Italic">Juglans regia</Emphasis> L.) under mainly genotypic or environmental control?

As observed for most stresses, tree frost resistance can be split into two main processes: avoidance and tolerance. Avoidance of freezing is achieved by introducing species only in the climatic context in which the probability of freezing events is very low for the sensitive stages of buds or stems; i.e., when good synchronism exists between the annual cycle and the critical climatic periods. Buds become able to grow only after chilling requirements have been satisfied (endodormancy released) during winter; they subsequently break after heat requirements have been completed (end of ecodormancy) in early spring. Actually, this period is often subject to more or less severe freezing events. Trees are also able to adjust their freezing tolerance by increasing their capacity of extracellular freezing and decreasing the possibility of intracellular freezing through the process of frost acclimation. Both freezing resistance processes (avoidance and tolerance) are environmentally driven (by photoperiod and temperature), but there are also genotypic effects among species or cultivars. Here, we evaluated the degree to which differences in dormancy release and frost acclimation were related to environmental and genetic influences by comparing trees growing in common garden conditions. This investigation was carried out for two winters in lowland and mountain locations on different walnut genotypes differing significantly for budburst dates. Chilling requirement for endodormancy release and heat requirement during ecodormancy were evaluated in all situations. In addition, frost acclimation was assessed by the electrolyte leakage method on stems from the same trees before leaf fall through budburst. No significant differences were observed in chilling requirements among genotypes. Moreover, frost acclimation dynamics were similar between genotypes or locations when expressed depending on chilling units accumulated since 15 September as a time basis instead of Julian day. The only exception was for maximal frost hardiness observed during winter with the timber-oriented being significantly more resistant than fruit-oriented genotypes. Heat requirement was significantly different among genotypes. Thus, growth was significantly faster in fruit-oriented than in wood-oriented genotypes. Furthermore, among wood-oriented genotypes, differences in growth rate were observed only at cold temperatures. Frost acclimation changes differed significantly between fruit- and wood- walnuts from January through budburst. In conclusion, from September through January, the acclimation dynamic was driven mainly by environmental factors whereas from January through budburst a significant genotype effect was identified in both frost tolerance and avoidance processes.     

Seasonal differences in photosynthesis between the C3 and C4 subspecies of Alloteropsis semialata are offset by frost and drought

The regional abundance of C₄ grasses is strongly controlled by temperature, however, the role of precipitation is less clear. Progress in elucidating the direct effects of photosynthetic pathway on these climate relationships is hindered by the significant genetic divergence between major C₃ and C₄ grass lineages. We addressed this problem by examining seasonal climate responses of photosynthesis in Alloteropsis semialata , a unique grass species with both C₃ and C₄ subspecies. Experimental manipulation of rainfall in a common garden in South Africa tested the hypotheses that: (1) photosynthesis is greater in the C₄ than C₃ subspecies under high summer temperatures, but this pattern is reversed at low winter temperatures; and (2) the photosynthetic advantage of C₄ plants is enhanced during drought events. Measurements of leaf gas exchange over 2 years showed a significant photosynthetic advantage for the C₄ subspecies under irrigated conditions from spring through autumn. However, the C₄ leaves were killed by winter frost, while photosynthesis continued in the C₃ plants. Unexpectedly, the C₄ subspecies also lost its photosynthetic advantage during natural drought events, despite greater water-use efficiency under irrigated conditions. This study highlights previously unrecognized roles for climatic extremes in determining the ecological success of C₃ and C₄ grasses.     

Is C4 photosynthesis less phenotypically plastic than C3 photosynthesis?

C4 photosynthesis is a complex specialization that enhances carbon gain in hot, often arid habitats where photorespiration rates can be high. Certain features unique to C4 photosynthesis may reduce the potential for phenotypic plasticity and photosynthetic acclimation to environmental change relative to what is possible with C3 photosynthesis. During acclimation, the structural and physiological integrity of the mesophyll-bundle sheath (M-BS) complex has to be maintained if C4 photosynthesis is to function efficiently in the new environment. Disruption of the M-BS structure could interfere with metabolic co-ordination between the C3 and C4 cycles, decrease metabolite flow rate between the tissues, increase CO2 leakage from the bundle sheath, and slow enzyme activity. C4 plants have substantial acclimation potential, but in most cases lag behind the acclimation responses in C3 plants. For example, some C4 species are unable to maintain high quantum yields when grown in low-light conditions. Others fail to reduce carboxylase content in shade, leaving substantial over-capacity of Rubisco and PEP carboxylase in place. Shade-tolerant C4 grasses lack the capacity for maintaining a high state of photosynthetic induction following sunflecks, and thus may be poorly suited to exploit subsequent sunflecks compared with C3 species. In total, the evidence indicates that C4 photosynthesis is less phenotypically plastic than C3 photosynthesis, and this may contribute to the more restricted ecological and geographical distribution of C4 plants across the Earth.     

Chilling sensitivity of the frost-tolerant potato Solanum commersonii

Frost tolerance has been reported in the shoots of wild, tuberiferous potato species such as Solanum commersonii when the plants are grown in either field or controlled conditions. However, these plants can survive as underground tubers and avoid unfavorable environmental conditions altogether. As such, leaf growth and photosynthesis at low temperature may not be required for survival of the plants. In order to determine the temperature sensitivity of S. commersonii shoots, we examined leaf growth, development and photosynthesis in plants raised at 20/16°C (day/night). 12/9°C and 5/2°C. S. commersonii leaves grown at 5°C exhibited a marked decrease in leaf area and in total chlorophyll (Chl) content per leaf area when compared with leaves grown at 20°C. Furthermore, leaves grown at 5°C did not exhibit the expected decrease in either water content or susceptibility to low-temperature-induced photoinhibition that normally characterizes cold acclimation in frost-tolerant plants. Measurements of CO₂-saturated O₂ evolution showed that the photosynthetic apparatus of 5°C plants was functional, even though the efficiency of photosystem II photochemistry was reduced by growth at 5°C. A decrease in the resolution of the M-peak in the slow transients for Chl a fluorescence in leaves grown at 12 and 5°C and in all leaves exposed to high light at 5°C indicated that low temperature significantly affected processes on the reducing side of Q_A, the primary quinone electron acceptor in photosystem II. Thus S. commarsonii exhibits the characteristics of a plant that is limited by chilling temperatures. Although S. commersonii can tolerate light frosts, its sensitivity to chilling temperatures may result in shoot dieback in winter in its native habitat. The plants may avoid both chilling and freezing temperatures by overwintering as underground tubers.     

Time course of photosynthetic temperature acclimation in Carex eleocharis Bailey

Abstract Photosynthetic temperature acclimation in Carex eleocharis has been demonstrated in a previous study in which warm grown (35/15°C) plants were shown to have photosynthetic temperature optima approximately 14°C higher than cool grown (20/15°C) plants (Monson, Littlejohn & Williams, 1983). The current study examined the time course of this acclimation by determining photo-synthetic temperature optima as a function of time, of cool grown plants moved to warm growing conditions. Leaves which had developed under cool conditions were capable of an upward adjustment of 6–8°C of their optimum photosynthetic temperature within a time span of 6–14 d. For greatest photosynthetic temperature acclimation it was necessary for leaves to form and develop entirely under warm conditions. These leaves exhibited a 14–15°C upward adjustment of their optimum temperature for photosynthesis within 20–31 d since moving plants from cool to warm growing conditions. Thus, the time course of this acclimation is of short enough duration to be significant during the growing season and presumably contributes toward the ability of this species to maintain active growth during the cool and warm portions of the growing season. It is also noted that the plant with its capacity to form new leaves, has a much wider acclimation capacity than any single leaf.     

Temperature response of photosynthesis in C3, C4, and CAM plants: temperature acclimation and temperature adaptation

Most plants show considerable capacity to adjust their photosynthetic characteristics to their growth temperatures (temperature acclimation). The most typical case is a shift in the optimum temperature for photosynthesis, which can maximize the photosynthetic rate at the growth temperature. These plastic adjustments can allow plants to photosynthesize more efficiently at their new growth temperatures. In this review article, we summarize the basic differences in photosynthetic reactions in C₃, C₄, and CAM plants. We review the current understanding of the temperature responses of C₃, C₄, and CAM photosynthesis, and then discuss the underlying physiological and biochemical mechanisms for temperature acclimation of photosynthesis in each photosynthetic type. Finally, we use the published data to evaluate the extent of photosynthetic temperature acclimation in higher plants, and analyze which plant groups (i.e., photosynthetic types and functional types) have a greater inherent ability for photosynthetic acclimation to temperature than others, since there have been reported interspecific variations in this ability. We found that the inherent ability for temperature acclimation of photosynthesis was different: (1) among C₃, C₄, and CAM species; and (2) among functional types within C₃ plants. C₃ plants generally had a greater ability for temperature acclimation of photosynthesis across a broad temperature range, CAM plants acclimated day and night photosynthetic process differentially to temperature, and C₄ plants was adapted to warm environments. Moreover, within C₃ species, evergreen woody plants and perennial herbaceous plants showed greater temperature homeostasis of photosynthesis (i.e., the photosynthetic rate at high-growth temperature divided by that at low-growth temperature was close to 1.0) than deciduous woody plants and annual herbaceous plants, indicating that photosynthetic acclimation would be particularly important in perennial, long-lived species that would experience a rise in growing season temperatures over their lifespan. Interestingly, across growth temperatures, the extent of temperature homeostasis of photosynthesis was maintained irrespective of the extent of the change in the optimum temperature for photosynthesis (T _opt), indicating that some plants achieve greater photosynthesis at the growth temperature by shifting T _opt, whereas others can also achieve greater photosynthesis at the growth temperature by changing the shape of the photosynthesis–temperature curve without shifting T _opt. It is considered that these differences in the inherent stability of temperature acclimation of photosynthesis would be reflected by differences in the limiting steps of photosynthetic rate.     

Low temperature acclimation of root fatty acid composition, leaf water potential, gas exchange and growth of soybean seedlings

Abstract Root fatty acid composition, photosynthesis, leaf water potentials, stomatal resistances, leaf specific weights, and root: shoot ratios of soybean were measured in two temperature regimes. Groups of soybean plants were grown in controlled chambers of the Duke University Phytotron under two thermoperiods. One group of the plants was grown from seed for 3 weeks in either 29/23°C or 17/11°C thermoperiods, and another group was grown for 2 weeks in 29/23°C and then transferred to the 17/11°C thermoperiod where it remained for 8 days. Broccoli was also grown in either 29/23°C or 17/11°C thermoperiods. Soybean roots contained more unsaturated fatty acids than broccoli roots, although broccoli roots showed a larger increase in unsaturation than soybean roots with decreased temperature. The fatty acid unsaturation in the roots of soybean began to increase rapidly after the temperature regime was changed. The increase was in the new roots produced in the cold regime rather than in the pre-existing roots. The soybean leaf water and osmotic potentials decreased about 0.4 MPa, beginning one day after the transfer from 29/23°C to 17/11°C, but recovered significantly after 8 d. Plants grown at 17/11 °C had lower rates of photosynthesis and adaxial stomatal resistances, but higher root: shoot ratios and specific leaf weights compared to plants grown at 29/23°C. Plants grown and maintained at 29/23°C showed a steady increase in photosynthetic rates over the 8-d experimental period, whether rates were measured in 1 mol m^?3 or 9 mol m^?3 oxygen. Plants transferred to 17/11°C however maintained constant rates of photosynthesis at 1 mol m^?3 O₂, whereas at 9 mol m^?3 rates declined for 2 d then were constant for the remaining 6 d of the experimental period. These results suggest that changes in membrane fatty acid unsaturation is an important aspect of plant acclimation to chilling temperatures in terms of maintaining root permeability and water uptake. However, the degree of unsaturation is not a good indicator of differences in chilling tolerance among species. The apparent acclimation of photorespiration to a constant percentage of photosynthesis suggests a role of photorespiration in the plant.     

Low-temperature photosynthetic performance of a C4 grass and a co-occurring C3 grass native to high latitudes

The photosynthetic performance of C₄ plants is generally inferior to that of C₃ species at low temperatures, but the reasons for this are unclear. The present study investigated the hypothesis that the capacity of Rubisco, which largely reflects Rubisco content, limits C₄ photosynthesis at suboptimal temperatures. Photosynthetic gas exchange, chlorophyll a fluorescence, and the in vitro activity of Rubisco between 5 and 35 °C were measured to examine the nature of the low‐temperature photosynthetic performance of the co‐occurring high latitude grasses, Muhlenbergia glomerata (C₄) and Calamogrostis canadensis (C₃). Plants were grown under cool (14/10 °C) and warm (26/22 °C) temperature regimes to examine whether acclimation to cool temperature alters patterns of photosynthetic limitation. Low‐temperature acclimation reduced photosynthetic rates in both species. The catalytic site concentration of Rubisco was approximately 5.0 and 20 µmol m^?2 in M. glomerata and C. canadensis, respectively, regardless of growth temperature. In both species, in vivo electron transport rates below the thermal optimum exceeded what was necessary to support photosynthesis. In warm‐grown C. canadensis, the photosynthesis rate below 15 °C was unaffected by a 90% reduction in O₂ content, indicating photosynthetic capacity was limited by the capacity of P_i‐regeneration. By contrast, the rate of photosynthesis in C. canadensis plants grown at the cooler temperatures was stimulated 20–30% by O₂ reduction, indicating the P_i‐regeneration limitation was removed during low‐temperature acclimation. In M. glomerata, in vitro Rubisco activity and gross CO₂ assimilation rate were equivalent below 25 °C, indicating that the capacity of the enzyme is a major rate limiting step during C₄ photosynthesis at cool temperatures.