Physiological and morphological responses of red alder and sitka alder to flooding

Red alder (Alnus rubra Bong.) and sitka alder (A. viridis ssp. sinuata [Regel] Löve & Löve) are nitrogen-fixing woody species that grow sympatrically along the Pacific coast of North America. Red alder is found in poorly drained lowlands, as well as in soils of moist upland slopes, whereas sitka alder generally colonizes well-drained soils. To identify factors that contribute to flood tolerance, we conducted greenhouse experiments subjecting both species to a 20-day flood and 10-day recovery and red alder to a 50-day flood and 20-day recovery. We determined the effect of this stress on nitrogenase activity, root and nodule alcohol dehydrogenase (ADH) activity, lenticel and adventitious root development, relative growth rate (RGR), and leaf gas exchange. After 24 h of flooding, nitrogenase activity could not be detected in either species. Limited nitrogenase activity did return in red alder at the end of a 10-day recovery following the 20-day flood, but sitka alder showed no recovery of nitrogenase activity. After 50 days of continuous flooding, red alder nitrogenase activity returned to pretreatment levels. Red alder root and nodule ADH activity was more than twice that of sitka alder under flooded conditions. Sitka alder showed extensive root mortality and leaf abscission over the same 20-day flooding period. Flooded red alder exhibited an initial decline in root RGR, but recovered between days 10 and 20 with the formation of adventitious roots. Furthermore, initiation of adventitious roots in red alder coincided with an increase in stomatal conductance without a similar recovery of carbon dioxide exchange rate. Sitka alder formed few adventitious roots, lost much of its root and leaf biomass, and showed no restoration of growth during flooding or recovery. Different responses of red and sitka alder to flooding serve as a partial explanation for the different patterns of distribution of these species and suggest some adaptations of red alder that permit flood tolerance.     

Anaerobic metabolism enzymes as markers of flooding stress in maize seeds

Maize (Zea mays L.) seeds differ in their relative tolerance to the anaerobic environment caused by flooding. Seed tolerance to flooding stress depends on cellular and metabolic processes since gross anatomical responses have not developed at the pre-emergence stage. The study reported here characterizes the activities of four anaerobic respiratory enzymes: pyruvate decarboxylase (PDC), alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), and malic enzyme (ME) in the flood-tolerant A632 and floodsusceptible Mo 17 inbred maize seeds during flooding at 10 and 25°C. Each inbred consisted of two seed lots possessing 95% and 75% germination levels. Flooding increased the activities of all four enzymes. However, no consistent correlation between anaerobic enzyme activity and flood tolerance was observed across genotype, seed quality and flooding temperature. The results indicate that it may not be feasible to use whole-seed anaerobic enzyme activities to predict maize seed performance under flooding stress. Contribution from the Soil Drainage Research Unit, USDA-ARS, Columbus, OH, in cooperation with the Ohio Agricultural Research and Development Center, The Ohio State University. OARDC Journal Article No. 66–86.     

RELATIONSHIPS BETWEEN FLOODING TOLERANCE,LIFE HISTORY,AND SHORT-TERM COMPETITIVE PERFORMANCE IN THREE SPECIES OF POLYGONUM

The objective of this study was to compare the growth and short-term (single season) competitive performance of three species of Polygonum known to differ in flooding tolerance and life history. Polygonum amphibium is a perennial with low sexual reproductive effort and a relatively high degree of flooding tolerance, P. lapathifolium is an annual species with a high sexual reproductive effort and a low tolerance to flooding, and P. hydropiperoides is intermediate to the other two in terms of sexual reproductive effort and flooding tolerance. In order to determine the relative growth and competitive abilities of these species, mixtures and monocultures of plants were grown in pots and maintained under three flooding regimes: 1) flooded, 2) partially drained, and 3) well drained. Both P. hydropiperoides and P. amphibium grew best under flooded and partially drained conditions with reduced growth in the drained treatment. Polygonum lapathifolium, in contrast, grew as well in the drained treatment as in the more flooded treatments. Results from competition experiments were consistent in showing the relative competitive abilities to be P. lapathifolium > P. hydropiperoides > P. amphibium regardless of flooding regime. Thus, short-term competitive performance was found to trade off with flood tolerance rather than with sexual reproductive effort.     

Aerenchyma development and elevated alcohol dehydrogenase activity as alternative responses to hypoxic soils in the Piriqueta caroliniana complex

The ability of plants to make morphological or physiological adjustments in response to environmental cues allows them to survive and reproduce under a wide range of conditions. One stress that plants are often exposed to is soil oxygen depletion due to flooding. Plants can respond to hypoxic soils by producing oxygen-conducting aerenchymous tissue or through induction of enzymes in the ethanolic fermentation pathway. Here we use greenhouse experiments to examine flood responses in plants of the Piriqueta caroliniana (Turneraceae) complex, which occupy a range of moisture regimes. Morphotypes and hybrids in this complex exhibited contrasting responses to hypoxic conditions. Genotypes from flooded habitats developed aerenchyma and did not substantially elevate levels of alcohol dehydrogenase (ADH) activity, an enzyme associated with anaerobic respiration. Plants from drier sites, on the other hand, did not develop aerenchyma but had much higher levels of ADH activity. Plants with aerenchymous tissue had substantially higher rates of growth under sustained flooding. Results are consistent with the hypothesis that aerenchyma development is an effective strategy in habitats subject to persistent flooding, while elevating activity of enzymes for ethanolic fermentation is effective only under ephemeral flooding. The range of phenotypic responses observed illustrates contrasting adaptive strategies that can lead to habitat isolation and evolutionary divergence.     

Alcohol dehydrogenase activity in the roots of marsh plants in naturally waterlogged soils

The aim of this work was to discover whether oxygen tensions in the roots of marsh plants in flooded soils are high enough to allow fully acrobic metabolism. Activity of alcohol dehydrogenase (ADH), a protein synthesised in anoxic plants, was measured in roots of marsh plants growing in habitats where the availability of oxygen to the roots would be expected to differ. Roots of Carex riparia in standing water had ADH activities about 2.5 times higher than those of phosphofructokinase, and comparable to ADH activities of Poa trivialis, Urtica dioica and Ranunculus repens roots in dry soil. Removal of the oxygen supply via aerenchyma to Carex roots caused a 30-fold increase in ADH activity relative to that of phosphofructokinase. There was no change in ADH activity with depth in Carex roots in waterlogged soil, but in Filipendula ulmaria roots activity was 14 times higher below 10 cm depth than near the surface. Urtica roots in waterlogged soil had alcohol dehydrogenase activities 26 times higher than roots in dry soil, but for Poa and Ranunculus roots this figure was only 1.7 and 4.2, respectively. These results indicate that the oxygen tensions in the roots of marsh plants in waterlogged soil differ considerably among species. Ethanol was the major product of fermentation in roots of all species studied. There was no correlation between ADH activity and the rate of ethanol production under anoxia of Urtica roots. The physiological significance of high ADH activities in roots is thus unclear.Abbreviations ADH alcohol dehydrogenase - PFK phosphofructokinase - PFP pyrophosphate:fructose 6-phosphate phosphotransferase     

Cryptic genetic variation and adaptation to waterlogging in Caledonian Scots pine,Pinus sylvestris L.

Local adaptation occurs as the result of differential selection among populations. Observations made under common environmental conditions may reveal phenotypic differences between populations with an underlying genetic basis; however, exposure to a contrasting novel environment can trigger release of otherwise unobservable (cryptic) genetic variation. We conducted a waterlogging experiment on a common garden trial of Scots pine, Pinus sylvestris (L.), saplings originating from across a steep rainfall gradient in Scotland. A flood treatment was maintained for approximately 1 year; physiological responses were gauged periodically in terms of photochemical capacity as measured via chlorophyll fluorescence. During the treatment, flooded individuals experienced a reduction in photochemical capacity, F_v/F_m, this reduction being greater for material originating from drier, eastern sites. Phenotypic variance was increased under flooding, and this increase was notably smaller in saplings originating from western sites where precipitation is substantially greater and waterlogging is more common. We conclude that local adaptation has occurred with respect to waterlogging tolerance and that, under the flooding treatment, the greater increase in variability observed in populations originating from drier sites is likely to reflect a relative absence of past selection. In view of a changing climate, we note that comparatively maladapted populations may possess considerable adaptive potential, due to cryptic genetic variation, that should not be overlooked.     

Ethanol synthesis and loss from flooded roots of Medicago sativa L. and Lotus corniculatus L.

Abstract Field flooding of established alfalfa (Medicago sativa L.) and birdsfoot trefoil (Lotus corniculatus L.) for up to 12 d resulted in a significant increase in alcohol dehydrogenase activity (ADH) and an increase in the K_m of ADH in both species. Root concentration of ethanol increased throughout the flooding regime in alfalfa roots. No ethanol was detected in any trefoil root samples. Alfalfa plants which had shoots removed 5 d prior to flooding accumulated significantly higher levels of root ethanol and showed flooding injury sooner, indicating a significant effect of shoots on development of flooding injury. Alfalfa and trefoil plants grown in the greenhouse were flooded and ethanol in the transpiration effluent was trapped and measured. Alfalfa transpired measurable quantities of ethanol which peaked just prior to development of shoot injury symptoms. No ethanol was detected in the transpiration effluent from trefoil shoots. Flooded roots of both alfalfa and trefoil excreted ethanol but alfalfa roots synthesized more total ethanol and retained a larger proportion in the roots than did trefoil. While the ethanol accumulation response in alfalfa and trefoil are consistent with the ethanol ‘self-poisoning’ hypothesis of flooding injury, the very small quantities of ethanol found in these roots still raises questions as to its absolute effect in the plant.     

Introduction of Alcohol Dehydrogenase in Lettuce Seedlings by Flooding Stress

Alcohol dehydrogenase (ADH), its isozyme profiles and ethanol concentration in lettuce (Lactuca sativa L.) seedlings subjected to flooding stress were determined. Flooding stress caused increases in ADH activity and ethanol concentration. By 48 h, ADH activity and ethanol concentration in the flooded seedlings increased 3.2- and 7.0-fold, respectively, in comparison with those in non-stressed seedlings. Five electrophoretically separable ADH bands were found in extract of the flooded seedlings, whereas only two or three ADH bands were found in extract of non-stressed seedlings. These results indicate that lettuce ADH may have a system of three-gene and six-isozyme, and the increase in ADH activity in the flooded seedlings may be due to increased synthesis of the enzyme.     

EFFECTS OF FLOODING AND NUTRIENT ENRICHMENT ON BIOMASS ALLOCATION IN ACER RUBRUM SEEDLINGS

A greenhouse experiment was conducted on Acer rubrum seedlings to evaluate the effects of flood frequency on production and allocation of biomass and to test the effects of N and P fertilization on production and allocation. Seedlings from the Dismal Swamp were subjected to three flood treatments (no flooding, intermittent flooding, and continuous flooding) and four enrichment treatments (no enrichment, N additions, P additions, and N + P additions). More continuous flooding resulted in less biomass production. Biomass increased during the study in all treatments except for root mass in the continuously flooded treatment. However, production of abundant adventitious roots compensated for the lack of normal root growth. Root/shoot ratios exhibited the greatest decreases in the continuously flooded plants. Plants with N + P added had significantly more leaf, stem, and total mass than the nonenriched plants four months into the study. The N + P additions had apparently compensated for the effects of flood stress in the continuously flooded plants by the end of the study. The fertilized seedlings accumulated higher concentrations of N and P, but their nutrient use efficiency (biomass production per unit nutrient absorbed) was lower than in the nonenriched plants. Acer rubrum seedlings survive flooded conditions through several adaptations; however, theirgrowth is slowed by continuous flooding.     

Physiological adaptation of waxapple to waterlogging

Waxapple (Syzygium samarangense Men. et. Perry) plants receiving up to 40d of continuous flooding treatment showed no symptoms of physiological disorder, but the treatment resulted in early flowering. In this report, several physiological parameters of flooded plants are compared with those of nonflooded plants. Both control plants and 9-d-flooding-treated plants exhibited aerenchyma formation in the cortex tissue beginning 5 cm from the root tip. After 7d flooding treatment, the oxygen consumption rate of the root section was only 20% of that of the controls. Following flooding treatment, the roots showed an increase in alcohol dehydrogenase activity as well as an increase in three isozymes. However, malate dehydrogenase activity was decreased, and no significant change of NADP-malic enzyme activity was observed. There were no significant differences in levels of ethylene, 1-aminocyclopropane-1-carboxylic acid and 1-(malonylamino) cyclopropane-1-carboxylic acid in petiole and roots of flooded and non-flooded plants during the stage examined. It is inferred that the presence of aerenchyma in the root cortex allows a higher level of internal gas exchange, and thus, makes waxapple surprisingly flood tolerant. However, reduced root oxygen consumption rate may have limited root respiration rate and vegetative growth.     

Effects of flood pre-conditioning on responses of three bottomland tree species to soil waterlogging

The potential for improving the performance of seedlings of woody species under flood stress was evaluated. Seedlings of baldcypress (Taxodium distichum), nuttall oak (Quercus nuttallii), and swamp chestnut oak (Quercus michauxii) were subjected to a two-phase study in which soil moisture regime was controlled. During Phase I, plants were randomly assigned to either: Control, well-watered and drained conditions; or intermittent flooding, alternating flooding and drained conditions. Following completion of Phase I, seedlings in each treatment were assigned randomly to one of two treatments: well-watered but drained or continuously flooded. Thus during Phase II, plants were divided into four groups: Control (A), intermittent flooding during Phase I and draining during Phase II (B), control during Phase I and continuously flooding during Phase II (C), and intermittently flooded during Phase I and continuously flooded during Phase II (D). Stomatal conductance (gw) in baldcypress increased in flooded treatments ranging from 112 to 128 percnt; of controls. Net photosynthesis (Pn) in baldcypress was not significantly affected by flooding while in oaks, Pn and gw decreased significantly in response to flooding. In oaks, flood pre-conditioning did not affect growth or physiological responses during phase II significantly. However, stomatal conductance of flood-hardened baldcypress was increased compared to seedlings that were not hardened. The study species developed significantly higher root porosity under flooded conditions compared to controls. Flood pre-conditioning did not appear to increase root porosity although it may have accelerated the process.     

The response of certainTrifolium spp. to calcium in sand culture

Summary The response to calcium ofTrifolium incarnatum, T. pratense, T. repens andT. hybridum was determined over the range 4 to 128 ppm calcium in sand culture.T. incarnatum showed the greatest response to calcium and was severely affected by low levels of calcium.T. hybridum was the least responsive to calcium and showed no significant response over the range tested, although a gradual increase in yield with increasing calcium was apparent. The responses ofT. pratense andT. repens were intermediate and differed significantly from those of bothT. incarnatum andT. hybridum. There was no significant difference between the response ofT. pratense andT. repens.The results are considered in the light of the ecologically observed edaphic tolerances of the species, and it is concluded that there is a marked relationship between response to calcium and edaphic tolerance of the species.In view of the differences in response to calcium recently demonstrated within the speciesT. repens, the magnitude of the differences between species shown here may not strictly apply to all varieties or populations of those species.     

Subtle topographical differences along a floodplain promote different plant strategies among Paspalum dilatatum subspecies and populations

It was hypothesised that subtle topographical differences might cause the existence of ecotypes along a floodplain. The apomict grass Paspalum dilatatum subspecies dilatatum inhabits flood‐prone lowlands as well as nearby uplands in the floodplains of Argentina, while the sexual P. dilatatum subspecies flavescens almost exclusively inhabits the uplands. The aim of the present study was to identify the different traits that allow these P. dilatatum populations to inhabit different habitats. Plants of P. dilatatum were reciprocally transplanted between uplands and lowlands. Morphophysiological traits related to flooding tolerance were measured during a flood. Subspecies dilatatum from the uplands and subspecies flavescens showed a high physiological performance in the uplands but a considerable decrease in stomatal conductance, net photosynthesis rates and tiller number in the flooded lowlands. In contrast, the subspecies dilatatum from the lowlands showed relatively lower and stable stomatal conductance, photosynthesis rates and leaf water potential at both sites. Subspecies dilatatum from the lowlands outperformed upland populations at the lowland site with respect to tillering. Leaves of subspecies dilatatum from the lowlands that had grown at the lowland habitat had a lower blade/sheath proportion than leaves of plants transplanted to the uplands. This behavior did not occur in both upland populations. Results suggest that dilatatum Lowland plants have the typical strategy of stress‐tolerant genotypes and that the upland populations are adapted to habitats where competitive species are selected. In conclusion, habitats with subtle differences in topographic level can favour both ecotypic differentiations within an apomict subspecies but also the maintenance of morphophysiological similitudes between coexisting upland populations belonging to different subspecies.     

Is the field water use of Eucalyptus largiflorens F. Muell. affected by short-term flooding?

Abstract Transpiration of and water sources for Eucalyptus largiflorens F. Muell trees at three sites on a semi-arid floodplain of the lower River Murray in southern Australia were investigated during the course of a flood. Two of the sites were flooded for 32 days while the third was not flooded. At both flooded sites transpiration was not suppressed during the period when the sites were flooded, indicating that anoxia did not occur. Available literature suggests that E. largiflorens has very low O² consumption, and that lateral roots can absorb O² from soil water at very low partial pressures. The oxygen stored in the unsaturated region of the soil appeared sufficient, in this case, to meet the requirements of the trees. Transpiration at the flooded sites did not increase in comparison with that at the non-flooded site in the period following flooding. We suggest that this was possibly due to the above-average rainfall experienced in the months preceding the flood and/or physiological adaptations to the highly saline conditions that prevented the trees taking full advantage of the additional water. The sources of water used by the trees were investigated using the naturally occurring stable isotopes of water and measurements of soil water suction. These showed that prior to the flood shallow rain-derived soil water was being used at all three sites. Ten days after flooding ceased trees at both flooded sites were using shallow flood-derived soil water while at the non-flooded sites they were using groundwater. Results from the flooded sites 7 and 22 weeks after flooding ceased showed that as the flood- and rain-derived water became limited the trees supplemented their supplies with groundwater from the capillary fringe. The non-flooded trees used various combinations of rain-derived soil water and groundwater during this period.     

ANATOMICAL AND METABOLIC RESPONSES TO WATERLOGGING AND SALINITY IN SPARTINA ALTERNIFLORA AND S. PATENS (POACEAE)

The effects of waterlogging and salinity (25 or 325 mol m ³ NaCl) stressors on the anatomy and metabolism of the marsh grasses 5. alterniflora Loisel. and S. patens Aiton (Muhl.) were investigated in a V factorial greenhouse experiment over 30 d. Waterlogging and salinity in combination resulted in anatomical and metabolic responses in both species. Waterlogging reduced soil redox potential and decreased root-specific gravity significantly in both species. The inadequacy of aerenchyma development under hypoxia to support aerobic root respiration in S. patens was indicated by significant increases in root alcohol dehydrogenase (ADH) activity of 1,752% and 420%, respectively, in the low and high salinity treatments. ADH activity was not increased significantly by flooding of S. alterniflora. Proline concentrations in roots and leaves were low at low salinities and increased significantly at high salinities in both species, but only under drained conditions. Decrease in leaf elongation by high salinity occurred in drained, but not flooded treatments in both species. Under flooded conditions, leaf elongation was significantly greater in S. alterniflora than S. patens. Greatest leaf elongation occurred in flooded low salinity S. alterniflora plants that had the least proline. Although both species are adapted to waterlogging and salinity, S. alterniflora appears to be more tolerant of reducing soil conditions and less responsive to higher salinity than S. patens.     

Early physiological flood tolerance is followed by slow post‐flooding root recovery in the dryland riparian tree Eucalyptus camaldulensis subsp. refulgens

We investigated physiological and morphological responses to flooding and recovery in Eucalyptus camaldulensis subsp. refulgens, a riparian tree species from a dryland region prone to intense episodic floods. Seedlings in soil flooded for 88 d produced extensive adventitious roots, displayed stem hypertrophy (stem diameter increased by 93%) and increased root porosity owing to aerenchyma formation. Net photosynthesis (P_n) and stomatal conductance (g_s) were maintained for at least 2 weeks of soil flooding, contrasting with previous studies of other subspecies of E. camaldulensis. Gradual declines followed in both g_s (30% less than controls) and P_n (19% less). Total leaf soluble sugars did not differ between flooded and control plants. Root mass did not recover 32 d after flooding ceased, but g_s was not lower than controls, suggesting the root system was able to functionally compensate. However, the limited root growth during recovery after flooding was surprising given the importance of extensive root systems in dryland environments. We conclude that early flood tolerance could be an adaptation to capitalize on scarce water resources in a water‐limited environment. Overall, our findings highlight the need to assess flooding responses in relation to a species' fitness for particular flood regimes or ecological niches.     

Ecophysiological adaptations of black spruce (<Emphasis Type="Italic"> Picea mariana</Emphasis>) and tamarack (<Emphasis Type="Italic"> Larix laricina</Emphasis>) seedlings to flooding

Black spruce [ Picea mariana (Mill.) B.S.P.] and tamarack [ Larix laricina (Du Roi) K. Koch] are the predominant tree species in boreal peatlands. The effects of 34 days of flooding on morphological and physiological responses were investigated in the greenhouse for black spruce and tamarack seedlings in their second growing season (18 months old). Flooding resulted in reduced root hydraulic conductance, net assimilation rate and stomatal conductance and increased needle electrolyte leakage in both species. Flooded tamarack seedlings maintained a higher net assimilation rate and stomatal conductance compared to flooded black spruce. Flooded tamarack seedlings were also able to maintain higher root hydraulic conductance compared to flooded black spruce seedlings at a comparable time period of flooding. Root respiration declined in both species under flooding. Sugar concentration increased in shoots while decreasing in roots in both species under flooding. Needles of flooded black spruce appeared necrotic and electrolyte leakage increased over time with flooding and remained significantly higher than in flooded tamarack seedlings. No visible damage symptoms were observed in flooded tamarack seedlings. Flooded tamarack seedlings developed adventitious roots beginning 16 days after the start of flooding treatment. Adventitious roots exhibited significantly higher root hydraulic conductivity than similarly sized flooded tamarack roots. Flooded black spruce lacked any such morphological adaptation. These results suggest that tamarack is better able to adjust both morphologically and physiologically to prolonged soil flooding than black spruce seedlings.     

Physiological,morphological, and growth responses ofPlatanus occidentalis seedlings to flooding

Summary Flooding ofPlatanus occidentalis L. seedlings for up to 40 days induced several changes including early stomatal closure, greatly accelerated ethylene production by stems, formation of hypertrophied lenticels and adventitious roots on submerged portions of stems, and marked growth inhibition. Poor adaptation ofPlatanus occidentalis seedlings to soil inundation was shown in stomatal closure during the entire flooding period, inhibition of root elongation and branching, and death of roots. Some adaptation to flooding was indicated by (1) production of hypertrophied lenticels which may assist in exchange of dissolved gases in flood water and in release of toxic compounds, and (2) production of adventitious roots on stems which may increase absorption of water. These adaptations appeared to be associated with greatly stimulated ethylene production in stems of flooded plants. The greater reduction of root growth over shoot growth in flooded seedlings will result in decreased drought tolerance after the flood waters recede. The generally low tolerance to flooding of seedlings of species that are widely rated as highly flood tolerant is emphasized.     

Seasonal Dependent Effects of Flooding on Plant Species Survival and Zonation: a Comparative Study of 10 Terrestrial Grassland Species

Past research has provided compelling evidence that variation in flooding duration is the predominant factor underlying plant species distribution along elevation gradients in river floodplains. The role of seasonal variation in flooding, however, is far from clear. We addressed this seasonal effect for 10 grassland species by testing the hypothesis that all species can survive longer when flooded in winter than when flooded in summer. We carried out an inundation experiment under simulated conditions of summer and winter flooding in the greenhouse. The results showed that all species survived longer under winter floods than under summer floods. However, responses upon flooding were species-specific. All summer flood-tolerant species had high tolerance for winter floods as well, but summer flood sensitive species survived either a little longer, or dramatically longer when flooded under simulated winter conditions. Next, we examined whether winter or summer survival best predicted the lower distribution limits of the species as measured in a natural flooding gradient after an extremely long winter flood. We found a strong significant relationship between the lower distribution limits of species in the field and their tolerance to summer floods, although we measured the lower limits 14 years after the latest major summer flood. In contrast, no such significant relationship existed with species tolerance to winter floods. Some relatively intolerant species occurred at much higher floodplain elevations as was expected from their tolerance to winter inundation in the experiments. This suggests that zonation patterns as created by occasional summer floods may be maintained for a long time, probably due to the limited ability of species to re-colonise lower positions in the floodplain.     

Physiological and biochemical responses of Theobroma cacao L. genotypes to flooding

Flooding is common in lowlands and areas with high rainfall or excessive irrigation. A major effect of flooding is the deprivation of O₂ in the root zone, which affects several biochemical and morphophysiological plant processes. The objective of this study was to elucidate biochemical and physiological characteristics associated with tolerance to O₂ deficiency in two clonal cacao genotypes. The experiment was conducted in a greenhouse with two contrasting clones differing in flood tolerance: TSA-792 (tolerant) and TSH-774 (susceptible). Leaf gas exchange, chlorophyll (Chl) fluorescence, chemical composition and oxidative stress were assessed during 40 d for control and flooded plants. Flooding induced a decrease in net photosynthesis, stomatal conductance and transpiration of both genotypes. In flood conditions, the flood-susceptible clone showed changes in chlorophyll fluorescence, reductions in chlorophyll content and increased activity of peroxidase and polyphenol oxidase. Flooding also caused changes in macro- and micronutrients, total soluble sugars and starch concentrations in different plant organs of both genotypes. Response curves for the relationship between photosynthetically active radiation (PAR) and net photosynthetic rate (P _N) for flooded plants were similar for both genotypes. In flood conditions, the flood-susceptible clone exhibited (1) nonstomatal limitations to photosynthesis since decreased in maximum potential quantum yield of PSII (F_v/F_m) values indicated possible damage to the PSII light-harvesting complex; (2) oxidative stress; (3) increased leaf chlorosis; and (4) a reduction in root carbohydrate levels. These stresses resulted in death of several plants after 30 d of flooding.