Keeping a positive carbon balance under adverse conditions: responses of photosynthesis and respiration to water stress

Drought and salinity (i.e. soil water stress) are the main environmental factors limiting photosynthesis and respiration and, consequently, plant growth. This review summarizes the current status of knowledge on photosynthesis and respiration under water stress. It is shown that diffusion limitations to photosynthesis under most water stress conditions are predominant, involving decreased mesophyll conductance to CO₂, an important but often neglected process. A general failure of photochemistry and biochemistry, by contrast, can occur only when daily maximum stomatal conductance ( g _s) drops below 0.05–0.10 mol H₂O m⁻² s⁻¹. Because these changes are preceded by increased leaf antioxidant activities ( g _s below 0.15–0.20 mol H₂O m⁻² s⁻¹), it is suggested that metabolic responses to severe drought occur indirectly as a consequence of oxidative stress, rather than as a direct response to water shortage. As for respiration, it is remarkable that the electron partitioning towards the alternative respiration pathway sharply increases at the same g _s threshold, although total respiration rates are less affected. Despite the considerable improvement in the understanding of plant responses to drought, several gaps of knowledge are highlighted which should become research priorities for the near future. These include how respiration and photosynthesis interact at severe stress, what are the boundaries and mechanisms of photosynthetic acclimation to water stress and what are the factors leading to different rates of recovery after a stress period.     

Spatial and temporal variation of microbial respiration rates in a blackwater stream

1. The extent of spatial and temporal variation of microbial respiration was determined in a first-order, sand-bottomed, blackwater stream on the coastal plain of south-eastern Virginia, U.S.A.
2. Annual mean respiration rates (as g O₂ m^–3 h^–1) differed significantly among substrata: leaf litter, 12.9; woody debris, 2.4; surface sediment, 0.8; hyporheic sediment, 0.4; water column, 0.003. Rates associated with wood were higher than those with leaves when expressed per unit surface area.
3. Highest respiration rates on leaves, wood and in the water column occurred during the summer, whereas rates in the sediments were greatest during the late autumn and winter. Water temperature, as well as particulate organic matter and nitrogen content of the substrata, was correlated positively with respiration rates.
4. A stepwise multiple regression showed that temperature and nitrogen content together explained 88% of the variation in respiration rates of leaves and wood. In contrast, particulate organic matter content and nitrogen content explained 89–90% of the variation in respiration in the sediments. Although water temperature was a significant factor in the sediment multiple regressions, its addition as an independent variable improved the regression models only slightly.
5. Annual mean respiration in the stream channel, based on the proportional amount of respiration occurring associated with each type of substratum during each month, was 1.1 kg O₂ m^–2 yr^–1. Seventy per cent of respiration in the stream occurred in the hyporheic zone, 8–13% occurred in the surface sediment, leaf litter or woody debris, and < 1% occurred in the water column. Approximately 16% of total detritus, or 40% of non-woody detritus, stored in the stream during the year was lost to microbial respiration.     

Spatial and temporal variation of microbial respiration rates in a blackwater stream

1. The extent of spatial and temporal variation of microbial respiration was determined in a first-order, sand-bottomed, blackwater stream on the coastal plain of south-eastern Virginia, U.S.A.
2. Annual mean respiration rates (as g O₂ m^–3 h^–1) differed significantly among substrata: leaf litter, 12.9; woody debris, 2.4; surface sediment, 0.8; hyporheic sediment, 0.4; water column, 0.003. Rates associated with wood were higher than those with leaves when expressed per unit surface area.
3. Highest respiration rates on leaves, wood and in the water column occurred during the summer, whereas rates in the sediments were greatest during the late autumn and winter. Water temperature, as well as particulate organic matter and nitrogen content of the substrata, was correlated positively with respiration rates.
4. A stepwise multiple regression showed that temperature and nitrogen content together explained 88% of the variation in respiration rates of leaves and wood. In contrast, particulate organic matter content and nitrogen content explained 89–90% of the variation in respiration in the sediments. Although water temperature was a significant factor in the sediment multiple regressions, its addition as an independent variable improved the regression models only slightly.
5. Annual mean respiration in the stream channel, based on the proportional amount of respiration occurring associated with each type of substratum during each month, was 1.1 kg O₂ m^–2 yr^–1. Seventy per cent of respiration in the stream occurred in the hyporheic zone, 8–13% occurred in the surface sediment, leaf litter or woody debris, and < 1% occurred in the water column. Approximately 16% of total detritus, or 40% of non-woody detritus, stored in the stream during the year was lost to microbial respiration.     

Scaling of oxygen consumption of Lake Magadi tilapia, a fish living at 37°C

Rates of oxygen consumption were measured in the geothermal, hot spring fish, Oreochromis alcalicus grahami by stopped flow respirometry. At 37° C, routine oxygen consumption followed the allometric relationship: V o₂=0.738 M ^0.75, where V o₂ is ml O₂ h ⁻¹ and M is body mass (g). This represents a routine metabolic rate for a 10 g fish at 37° C of 0.415 ml O₂ g⁻¹ h ⁻¹ (16.4 μmol O₂ g ⁻¹ h ⁻¹). Acutely increasing the temperature from 37 to 42° C significantly elevated the rate of O₂ consumption from 0.739 to 0.970 ml O₂ g ⁻¹ h ⁻¹ ( Q ₁₀=l.72). In the field, O. a. grahami was observed to be 'gulping' air from the surface of the water especially in hot springs that exceeded 40° C. O. a. grahami may utilize aerial respiration when O₂ requirements are high.     

A comparative study of the effects of NaCl salinity on respiration, photosynthesis, and leaf extension growth in Beta vulgaris L. (sugar beet)

Abstract. Three parameters influencing the capacity for carbon accumulation, i.e. photosynthesis, respiration, and leaf extension growth, were studied in Beta vulgaris L. (sugar beet) cultured in nutrient solution containing 0.5 to 500 mol m⁻³ NaCl. Leaf extension growth was the parameter most sensitive to salinity: the initial rate of leaf extension and final leaf length each declined linearly with increase in external NaCl concentration. Photosynthetic O₂ evolution of thin leaf slices did not decline until salinity levels reached 350 to 500 mol m⁻³ NaCl, while respiratory O₂ consumption was not affected by salinity throughout the range. The results suggest that the influence of salinity on the capacity for carbon accumulation in B. vulgaris occurs primarily through reduction in the area of photosynthetic surface.     

PHYSIOLOGICAL RESPONSES TO TEMPERATURE AND IRRADIANCE IN SPIROGYRA (ZYGNEMATALES, CHAROPHYCEAE)1

Spirogyra Link (1820) is an anabranched filamentous green alga that forms free-floating mats in shallow waters. It occurs widely in static waters such as ponds and ditches, sheltered littoral areas of lakes, and stow-flowing streams. Field observations of its seasonal distribution suggest that the 70-μm-wide filament form of Spirogyra should have a cool temperature and high irradiance optimum for net photosynthesis. Measurements of net photosynthesis and respiration were marie at 58 combinations of tight and temperature in a controlled environment facility. Optimum conditions were 25°C and 1500 μmol photons m⁻² s⁻¹, at which net photosynthesis averaged 75.7 mg O₂ gdm⁻¹ h⁻¹. Net photosynthesis was positive at temperatures from 5° to 35°C at most irradiances except at combinations of extremely low irradiances and high temperatures (7 and 23 μmol photons m⁻² s⁻¹ at 30°C and 7, 23, and 35 μmol photons m⁻² s⁻¹ at 35°C). Respiration rates increased with both temperature and prior irradiance. Light-enhanced respiration rates were significantly greater than dark respiration rates following irradiances of 750 μmol photons m⁻² s⁻¹ or greater. Polynomials were fitted to the data to generate response surfaces; such response surfaces can be used to represent net photosynthesis and respiration in ecological models. The data indicate that the alga can tolerate the cool water and high irradiances characteristic of early spring but cannot maintain positive net photosynthesis under conditions of high temperature and low light (e.g. when exposed to self-shading ).     

Respiration rate in maize roots is related to concentration of reduced nitrogen and proliferation of lateral roots

The relationship between specific rate of respiration (respiration rate per unit root dry weight) and concentration of reduced nitrogen was examined for maize ( Zea mays L.) roots. Plants with 2 primary nodal root axes were grown for 8 days in a split-root hydroponic system in which NO-₃ was supplied to both axes at 1.0 mol m⁻³, to one axis at 1.0 mol m⁻³ and the other axis at 0.0 mol m⁻³ or to both axes at 0.0 mol m⁻³ Respiration rates and root characteristics were measured at 2-day intervals. Specific rate of respiration was positively correlated in a nonlinear relationship with concentration of reduced nitrogen. The lowest specific rates of respiration occurred when neither axis received exogenous NO⁻³ and the concentration of reduced nitrogen in the axes was less than 9 mg g⁻¹. The greatest rates occurred in axes that were actively absorbing NO⁻³ and contained more than 35 mg g⁻¹ of reduced nitrogen. At 23 mg g⁻¹ of reduced nitrogen, below which initiation of lateral branches was decreased by 30–50%. specific rate of respiration was 17% greater for roots actively absorbing NO⁻³ than for roots not absorbing NO⁻³ Increases in specific rate of respiration associated with concentrations of reduced nitrogen greater than 23 mg g⁻¹ were concluded to be attributable primarily to proliferation of lateral branches.     

Hydration, protons and onset of physiological activities in maize seeds

Dry maize ( Zea mays L.) seed components, namely, embryo and endosperm, provide model materials for studies on water-dependent mechanisms in cellular function. We explored the thermodynamics of hydration for both tissues, along with their dielectric behavior, as a function of water content. In addition, we evaluated the direct current (DC) conductivity due to water protons. Our data on embryo tissue show large enthalpic and entropic peaks at water content [h, in g H₂O (g dry sampie)⁻¹] around 0.08 g g⁻¹, indicating very tight binding and ordering of water molecules. With increasing water content both enthalpy and entropy decrease, and the completion of primary hydration requires h ∼ 0.26 g g⁻¹. Data for endosperm tissue show the absence of such an enthalpic peak and a reduced degree of ordering for h < 0.10 g g⁻¹. The DC protonic conductivity shows explosive growth above a threshold hydration level h_c= 0.082 g g⁻¹ and h_c= 0.12 g g⁻¹, for embryo and endosperm, respectively. Protonic conduction can be considered within the framework of a percolation modell characterized by a hydration threshold and by a power law increase in conductivity with further hydration. The critical exponent of the power law is in agreement with theory for a two-dimensional percolative process. This percolative water-assisted behavior reflects the presence of an extended network of water molecules adsorbed on the surface of proteins and/or membranes inside cells. We consider this percolative protonic conduction as being a prerequisite to respiration processes.     

Activity of methanotrophic bacteria in Green Bay sediments

Abstract Sediment pore water samples obtained from a 19 m station in Green Bay in Lake Michigan were examined for levels of ambient dissolved methane and copper, and for the potential for in situ methane oxidation by methanotrophs found within surface sediments. The in situ methane concentration in the upper oxic sediment layer ranged from 20–150 μmol · 1⁻¹ at this station. The activity of methanotrophs and the kinetics of methane oxidation in these sediments were demonstrated by the uptake of radiolabeled methane. K_s values varied between 4.1–9.6 nmol · cm³ of sediment slurry. High V_max values (12.7–35.2 nmol · cm⁻³ · h⁻¹) suggest a large population of methanotrophs in the sediments. An average methane flux to the oxic sediments of 0.24 mol · m⁻² · year⁻¹ was calculated from the pore water methane gradients. Pore water concentrations of copper in the upper sediment layer ranged from 10–120 nmol · 1⁻¹. Based upon the copper concentration, other measured parameters, and equilibrium conditions defined by WATEQF4, an estimate for dissolved free Cu²⁺ concentration of 5–38 nmol · 1⁻¹ pore water was obtained. Several factors control the rate of methane oxidation, including oxygen, methane, and the bioavailability of free Cu²⁺.     

The respiration of young coho Oncorhynchus kisutch at gradually declining oxygen levels and during recovery from oxygen deprivation

The respiration of coho salmon, Oncorhynchus kisutch , weighing between 15 and 50 g was measured at gradually declining oxygen levels and at temperatures ranging between 14 and 17°C. The maximum and minimum oxygen concentrations tested were 250 and 40 μmol L⁻¹, respectively. Respiration rates were measured for 1 h periods before oxygen concentration was lowered by 12.5 or 25.0 μmol oxygen L⁻¹. At the end of these endurance tests the oxygen level was returned to normoxic conditions and respiration rates were determined for the recovery period. Under normoxic conditions (> 200 μmol L⁻¹) the respiration of coho levelled around 5.1 μmol g⁻¹ wet weight h⁻¹. At intermediate levels between 150 and 200 μmol oxygen L⁻¹, the average rate increased to 5.8 μmol g⁻¹ h⁻¹, which could be attributed to higher spontaneous activity of the test animals. At low oxygen levels (< 150 μmol⁻¹) average respiration rates dropped to values between 5.5 and 5.7 μmol g⁻¹ h⁻¹, reaching a minimum of 3.8 μmol g⁻¹ h⁻¹ at oxygen levels below 50 μmol L^μ. First mortality was observed in this range. After exposure to reduced oxygen levels the fish maintained a higher respiration rate when again exposed to normoxic oxygen levels above 200 μmol L⁻¹. Increased respiration rates were observed for a recovery period of 6 h.     

Formation of the enveloping layer of the chum salmon egg in isotonic salt solutions

After stimulation in a hypotonic solution (9.4 mOsm kg⁻¹), inseminated eggs of the chum salmon Oncorhynchus keta initiate cleavages in isotonic salmon Ringer's solution (267.3 mOsm kg⁻¹) containing 3.2 mM Ca²⁺ ions. Blastomeres of these eggs, however, separate from each other and the enveloping layer is not observed at the blastula stage. An increase in external divalent cations rescues the separation; the concentration of CaCl₂ in the external medium should be 25 mM or more to induce close contact of blastomeres and the formation of an enveloping layer in isotonic salt solutions. The effectiveness of Ca²⁺ ions can be substituted by Mg²⁺, Sr²⁺ and Zn²⁺ ions; the same results are obtained in isotonic MgCl₂ and SrCl₂ solutions (100 mM) or in isotonic salmon Ringer's solution containing Zn ions (6.2 mM). The close contact of blastomeres and the formation of an enveloping layer are also observed in a low Ca²⁺ concentration (< 0.1 mM) in a hypotonic salt solution (9.4 mOsm kg⁻¹). The Ca²⁺ level in the external medium to induce the enveloping layer formation seems to be correlated with the salinity of the incubation medium. It is suggested that adhesion molecules on the surface of blastomeres in the chum salmon eggs are different in properties from those found in sea urchin and other fish species.     

Effects of boundary layer conductance on substomatal pressures of carbon dioxide

Abstract. Gas exchange measurements were made on single leaves of three C₃ and one C₄ species at air speeds of 0.4 and 4.0 m s⁻¹ to determine if boundary layer conductance substantially affected the substomatal pressure of carbon dioxide. Boundary layer conductances to water vapour were 0.4 to 0.5 mol m⁻² s⁻¹ at the lower air speed, and 1.2 to 1.5 mol m⁻² s⁻¹ at the higher air speed. Substomatal carbon dioxide pressures were about 5 Pa lower at low boundary layer conductance in the C₃ species, and about 3 Pa lower in the C₄ species when measurements were made at high and moderate photosynthetic photon flux densities. No evidence of stomatal adjustment to altered boundary layer conductance was found. Photosynthetic rates at high photon flux densities were reduced by about 20% at the low air speed in the C₃ species. The commonly reported values of substomatal carbon dioxide pressure for C₃ and C₄ species were found to occur only when measurements were made at the higher air speed.     

Metabolism of a desert stream

SUMMARY. Rates of photosynthesis and community respiration were determined for benthic assemblages in Sycamore Creek, a Sonoran Desert stream in Arizona. Benthos in this stream can be separated into (1) mats of Cladophora glomerata and associated epiphytes and (2) assemblages of epipelic diatoms and blue-green algae. Community respiration and net photosynthesis were measured for these assemblages using submerged light-dark chambers in situ . Multiple regression analysis was used to predict (1) gross photosynthesis as a function of photosynthetically active radiation, temperature and chlorophyll-α concentration; and (2) community respiration as a function of temperature and biomass.
Calculations suggest that Sycamore Creek is autotrophic during the summer ( P/R = 1.7) and that the rates of gross photosynthesis ( P =8.5 g O₂ m⁻² day⁻¹) and community respiration ( R = 5.1 g O₂ m⁻² day⁻¹) are high for a small stream. Considerable difference exists between the Cladophora mat assemblages, in which mean P is 12.5gO₂m⁻² day⁻¹and the P/R ratio is 2.3, and the epipelic assemblages in which mean P is 4.4 g O₂m⁻² day⁻¹ and P/R is 0.96. The high rate of gross photosynthesis, low litter inputs, high biomass of algae and the intermittent but severe floods that characterize Sycamore Creek indicate that this stream and other similar desert streams are net exporters of organic matter and are, thereby, truly autotrophic stream ecosystems.     

Change in the Respiratory Rate of Sea Urchin Spermatozoa following Sperm-egg Interaction in the Absence of Mg2+

Spermatozoa of the sea urchin, Hemicentrotus pulcherrimus (10⁸ cells/ml), preincubated with unfertilized eggs deprived of jelly coats (more than l0⁵ cells/ml) at 20°C for 20min in Mg²⁺ free artificial sea water containing 1 mM Ca²⁺ (MFASW), exhibited very low respiration, which was enhanced by 2, 4 dinitrophenol (DNP). The fertilization rate in MFASW was usually less than 5% and was about 25% at most. Preincubation with fertilized eggs (with and without a fertilization membrane) in MFASW did not reduced the respiratory rate of spermatozoa. The rate of sperm respiration was lower in MFASW than in artificial sea water (ASW), but was higher than the respiratory rate of spermatozoa preincubated in MFASW with unfertilized eggs. Sperm respiration in MFASW or in ASW was not stimulated by 2, 4 dinitrophenol. Almost complete inhibition of sperm respiration was obtained with unfertilized eggs fixed with glutaraldehyde at concentrations of above 10⁵ cells/ml in MFASW and of about l0⁴ cells/ml in ASW. The respiratory rate of spermatozoa treated with fixed eggs was enhanced by DNP. It is concluded that the respiratory rate of the spermatozoa is reduced by their interaction with unfertilized eggs before their penetration into the eggs.     

The effect of increased deposition of atmospheric nitrogen on Calluna vulgaris in upland Britain

Regular (monthly) additions of NH₄NO₃ (4–12 g N m⁻² yr⁻¹) were made over a period of 8 yr (1989–98) to areas of moorland in North Wales dominated by the ericaceous shrub Calluna vulgaris . Results from the early stages of the experiment (1990–94) have shown marked and dose-related increases in shoot extension and canopy height in response to the nitrogen treatments, with significantly higher shoot nitrogen contents. The nitrogen-related stimulation in the growth of the C. vulgaris canopy over this period has resulted in large accumulations of litter on the high-nitrogen-treated plots (6.6 kg m⁻² in plots treated with 12 g N m⁻² yr⁻¹, compared with 3.8 kg m⁻² for the water controls). Litter nitrogen concentrations were also significantly increased at the higher rates of nitrogen addition, leading to a doubling of the total return of nitrogen to the litter layer over the experimental period. These changes in vegetation structure were associated with large reductions in the abundance of the bryophyte and lichen species normally present under the untreated canopy. Results since 1994, however, show little increase in shoot extension in response to the nitrogen treatments, with no clear dose response to increasing levels of addition. These findings are associated with a dose-related increase in the susceptibility of the nitrogen-treated areas of the C. vulgaris canopy to late winter injury, characterized as browning of the shoot tips in early to late spring. These results indicate that deleterious effects are now accumulating as a result of the long-term addition of nitrogen to these moorland plots.     

Water vapour uptake by diapausing eggs of a tropical walking stick

Abstract. This study is the first to demonstrate the capacity of an arthropod egg, that of a tropical walking stick Extatosoma tiaratum (Macleuy), to absorb water vapour from the air. This species diapauses both as an early embryo and then again as a pharate first instar larva, and both stages are capable of absorbing water vapour. Water vapour absorption occurs at lower humidities and at a lower rate for an egg in early embryonic diapause (c. a_v 0.30, 0.516 mg h^-1δ_v^-1) than in the diapausing pharate first instar (c. a_v 0.60, 0.725 mgh^-1δa_v^-1) at 25C. In addition to having the capacity to gain water at very low vapour activities, water is efficiently conserved as indicated by the low rate of water loss (0.015% h^-1 in the early embryo and 0.046% h^-1 in the pharate larva at 25C). Eggs that have been killed lose water when held at a hydrating vapour activity, thus implying that active uptake contributes to net absorption. Wax block experiments suggest that water is absorbed over the entire chorionic surface. Eggs of five other insect species that were examined [Lymantria dispar (L.), Bombyx mori (L.), Antheraea polyphemus (Cram.), Oncopeltus fasciatus (Dallas) and Diaferomera femorata (Say)] lacked the ability to absorb atmospheric water.     

Oxygen and nitrate respiration in a reed swamp sediment from a eutrophic lake

Seasonal measurements of the oxygen and nitrate uptake by a reed swamp sediment were carried out in a shallow, eutrophic Danish lake, Arreskov Sø. The oxidation of organic carbon in the sediment by aerobic and nitrate respiration was 290 and 188 g C m⁻² yr⁻¹ respectively. During winter, nitrate respiration amounted to 94% of the total carbon oxidation, whereas it was zero during summer. On an annual basis nitrate respiration constituted 39% of total respiration. Sediment nitrate uptake was correlated to nitrate concentration. In consequence of this the nitrate uptake rates varied during the year from zero in summer to 55 mg N m⁻² d⁻¹ in spring.
Oxygen uptake rates varied from 30 to 250 mg O₂ m⁻² h⁻¹ during the year, with a maximum uptake in August. The oxygen uptake per year was calculated to 860 g O₂ m⁻². The oxygen uptake rate was correlated to lake temperature and Kjeldahl nitrogen content of the sediment. The oxygen uptake rate, however, showed no correlation with loss on ignition of the sediment. A Q₁₀-value of 2.2 was found for lake measurements in the temperature interval of 5–15°C. The corresponding O₁₀-value in the laboratory was 2.6. A high microbial biomass indicated by the maximum content of Kjeldahl nitrogen and the lowest ratio of loss on ignition on Kjeldahl nitrogen appeared in late August, when the maximum oxygen uptake occurred. The oxygen uptake rate increased during the time interval from sampling to the start of the experiments.     

A carbon budget for nematodes, rotifers and tardigrades in a Swedish coniferous forest soil

Nematodes, rotifers and tardigrades from a Swedish pine forest soil were investigated in a monthly sampling programme lasting for one year. The monthly mean values of number, biomass and oxygen consumption were estimated. All groups fluctuated rather much over the year with a summer minimum (1.1 ˙ 10⁶ animals m^-2) and a winter maximum (6.3 ˙ 10⁶ animals m^-2) for the nematodes. The reasons for these fluctuations are discussed in relation to fluctuations of water content and temperature of the soil. On an annual basis a carbon budget was calculated which gave the following values for this fauna; consumption 4.3, production 0.6, respiration 0.9 and defecation 2.8 g carbon m^-2.     

Probable participation of phospholipase A2 reaction in the process of fertilization-induced activation of sea urchin eggs

In sea urchin eggs activated by sperm, A23187 or melittin, BPB (4-bromophenacyl bromide, a phospholipase A₂ inhibitor) blocked fertilization envelope formation and transient CN⁻-insensitive respiration in a concentration-dependent manner. BPB had virtually no effect on the increase in [Ca²⁺]_i, (cytosolic Ca²⁺ level), the activity of phosphorylase a and the rate of protein synthesis, as well as acid production and augmentation of CN⁻-sensitive respiration. BPB also inhibited fertilization envelope formation and augmentation of CN⁻-insensitive respiration induced by melittin. Melittin, known to be an activator of phospholipase A₂, induced the envelope formation, acid production, augmentation of CN⁻-insensitive and sensitive respiration, but did not cause any increase in [Ca²⁺]_i, the phosphorylase a activity and the rate of protein synthesis. An activation of phospholipase A₂ induced by Ca²⁺ or melittin seems to result in cortical vesicle discharge and production of fatty acids, which are to be utilized in CN⁻-insensitive lipid peroxidase reactions. Activation of other examined cell functions in eggs activated by sperm or A23187, probably results from Ca²⁺-triggered sequential reactions other than Ca²⁺-caused activation of phospholipase A₂.     

The occurrence of aerial respiration in Rhinelepis strigosa during progressive hypoxia

Rhinelepis strigosa did not surface for air breathing in normoxic or moderate hypoxic water. This species initiated air breathing when the P _io₂ in the water reached 22 ± 1 mmHg. Once begun, the air-breathing frequency increased with decreasing P _io₂. Aquatic oxygen consumption was 21·0 ± 1·9ml O₂ kg⁻¹h⁻¹ in normoxic water, and was almost constant during progressive hypoxia until the P _io₂ reached 23·9 mmHg, considered the critical oxygen tension (P_co₂). Gill ventilation increased until close to the P _co₂ (7·9-fold) as a consequence of a greater increase in ventilatory volume than in breathing frequency. Gill oxygen extraction was 42 ± 5% and decreased with hypoxia, but under severe hypoxia returned to values characteristic of normoxic. The critical threshold for air breathing was coincident with the P_co₂ during aquatic respiration. This suggests that the air-breathing response is evoked by the aquatic oxygen tension at which the respiratory mechanisms fail to compensate for environmental hypoxia, and the gill O₂ uptake becomes insufficient to meet O₂ requirements.