Foliar water uptake: a common water acquisition strategy for plants of the redwood forest

Evaluations of plant water use in ecosystems around the world reveal a shared capacity by many different species to absorb rain, dew, or fog water directly into their leaves or plant crowns. This mode of water uptake provides an important water subsidy that relieves foliar water stress. Our study provides the first comparative evaluation of foliar uptake capacity among the dominant plant taxa from the coast redwood ecosystem of California where crown-wetting events by summertime fog frequently occur during an otherwise drought-prone season. Previous research demonstrated that the dominant overstory tree species, Sequoia sempervirens, takes up fog water by both its roots (via drip from the crown to the soil) and directly through its leaf surfaces. The present study adds to these early findings and shows that 80% of the dominant species from the redwood forest exhibit this foliar uptake water acquisition strategy. The plants studied include canopy trees, understory ferns, and shrubs. Our results also show that foliar uptake provides direct hydration to leaves, increasing leaf water content by 2–11%. In addition, 60% of redwood forest species investigated demonstrate nocturnal stomatal conductance to water vapor. Such findings indicate that even species unable to absorb water directly into their foliage may still receive indirect benefits from nocturnal leaf wetting through suppressed transpiration. For these species, leaf-wetting events enhance the efficacy of nighttime re-equilibration with available soil water and therefore also increase pre-dawn leaf water potentials.     

Experimental cloud immersion and foliar water uptake in saplings of Abies fraseri and Picea rubens

Key message

Frequent cloud immersion events result in direct uptake of cloud water and improve plant water potentials during daylight hours in saplings of two dominant cloud forest species.

Abstract

In ecosystems with frequent cloud immersion, the influence on plant water balance can be important. While cloud immersion can reduce plant water loss via transpiration, recent advances in methodology have suggested that many species also absorb water directly into leaves (foliar water uptake). The current study examines foliar water uptake and its influence on daily plant water balance in tree species of the endangered spruce–fir forest of the southern Appalachian Mountains, USA. These mountain-top communities are considered relic, boreal forests that may have persisted because of the benefits of frequent cloud immersion. We examined changes in needle water content, xylem water potentials, and stable isotope values in saplings of the two dominant tree species, Abies fraseri and Picea rubens before and after a 24 h period of experimental cloud immersion. Both species exhibited foliar water uptake following immersion, evidenced by substantial changes in stable isotope values of extracted needle water that reflected the composition of the fog water. In addition, total needle water content improved 3.7–6.4 % following experimental submersion and xylem water potentials were significantly greater (up to 0.33 MPa) in cloud-immersed plants over control plants. These results indicate that foliar water uptake may be an adaptive strategy for utilizing cloud water and improving overall tree vigor in these most southerly distributed boreal species.     

Leaf surface properties of Norway spruce needles exposed to sulphur dioxide and ozone in an open-air fumigation system at Liphook

Samples of current-year and 1-year-old foliage were taken from Norway spruce (Picea abies (L.) Karst.) trees in April 1991, 4 months after a 3–4 year controlled fumigation with O₃ and SO₂ in the open at Liphook, south-east England. Trees were grown in seven plots, and treated in a factorial design with three levels of SO₂ and two levels of O₃ (ambient and c. 1.3 × ambient), with an extra ambient air plot. All statistical analyses were made on plot means. Leaf wettability, as measured by the contact angle of water droplets, was significantly affected by needle age and by SO₂ treatment (P≤0–05. in older needles, decreasing with increasing SO₂ concentration. There was no effect of O₃ on wettability, and no effect of any treatment on amounts of surface wax extracted by immersion of needles in chloroform. Electrolyte leakage rates from detached current-year needles were not affected by prior exposure to O₃, but decreased significantly (P= 0.034) with increasing exposure to SO₂. There was no detectable effect of fumigation on the rate of water loss from detached needles. Similarly, there was no effect of fumigation on the dry weight/fresh weight ratio of needles. The total sulphur content of needles increased significantly (P≤0.0001) with exposure to SO₂ and with needle age. Amounts of water-extractable sulphate, however, varied greatly among plots, but with no pattern with respect to fumigation treatment. It is concluded that leaf wettability and electrolyte leakage rates may be good indicators of the persistent effects of SO₂ on Norway spruce growing in the open air, and that the observed changes in leaf surface properties in response to SO₂ fumigation have implications for the processes, both biotic and abiotic, that occur on leaf surfaces.     

The influence of summertime fog and overcast clouds on the growth of a coastal Californian pine: a tree-ring study

The coast of California is home to numerous rare, endemic conifers and other plants that are limited in distribution by drought sensitivity and the summer-dry climate that prevails across most of the state. Ecologists have long assumed that some coastal plant populations survived the early Pleistocene transition to a warmer and drier environment because they benefit from frequent fog and stratus clouds that provide water and shade during the rainless summer. One such population is that of Torrey pine (Pinus torreyana ssp. Insularis) on Santa Rosa Island in Channel Islands National Park. Here we report that the tree-ring width record from this population indicates strong growth sensitivities to summer fog drip and cloud shading. We quantified the effects of summer cloud cover by comparing ring-width indices to coastal airport cloud-frequency records (1944–2004). For the first time observed, summertime cloud frequency correlated positively with ring-width indices, regardless of whether the effect of rainfall was first removed from the ring-width record. The effect of ground-level fog was strongest in July early mornings (03:00 PST, R ² = 0.262, P < 0.0002). The effect of clouds high enough to provide shade but not fog water was also strongest in July, but climbed steadily throughout the day before becoming strongest in late afternoon (16:00–18:00 PST, R ² = 0.148, P < 0.004). Correlations were substantially stronger in years with higher soil moisture, suggesting that growth response to summer clouds is strongly affected by pre-summer rainfall. A change in the height and/or timing of coastal cloud formation with climate change would likely affect this and other populations of California’s coastal vegetation.     

Bark water uptake promotes localized hydraulic recovery in coastal redwood crown

Coastal redwood (Sequoia sempervirens), the world's tallest tree species, rehydrates leaves via foliar water uptake during fog/rain events. Here we examine if bark also permits water uptake in redwood branches, exploring potential flow mechanisms and biological significance. Using isotopic labelling and microCT imaging, we observed that water entered the xylem via bark and reduced tracheid embolization. Moreover, prolonged bark wetting (16 h) partially restored xylem hydraulic conductivity in isolated branch segments and whole branches. Partial hydraulic recovery coincided with an increase in branch water potential from about ?5.5 ± 0.4 to ?4.2 ± 0.3 MPa, suggesting localized recovery and possibly hydraulic isolation. As bark water uptake rate correlated with xylem osmotic potential (R² = 0.88), we suspect a symplastic role in transferring water from bark to xylem. Using historical weather data from typical redwood habitat, we estimated that bark and leaves are wet more than 1000 h per year on average, with over 30 events being sufficiently long (>24 h) to allow for bark‐assisted hydraulic recovery. The capacity to uptake biologically meaningful volumes of water via bark and leaves for localized hydraulic recovery throughout the crown during rain/fog events might be physiologically advantageous, allowing for relatively constant transpiration.     

Wetting of the upper needle surface of Abies grandis: influence of pH, wax chemistry and epiphyllic microflora on contact angles

Wetting of the upper needle surface of Abies grandis Lindl. by aqueous solutions of different pH values was investigated. With increasing needle age, contact angles decreased significantly from about 75° on current-year needles to values lower than 30° on 4-year-old needles. On older needles, contact angles were significantly lower, by more than 10°, when aqueous solutions of pH9-0 were used compared with those of pH3-0. On the surfaces of older needles, contact angle titrations were carried out, contact angles being measured with aqueous solutions covering a pH range from 3.0 to 11.0. Measured titration curves showed clear inflection points around pH 7.0, indicating the existence of ionizable carboxylie groups in the interface between needle surface and atmosphere. The evidence seems convincing that the pronounced pH dependence of wetting is mainly due to the presence and/or activity of epiphyllic micro-organisms, whereas the cuticular wax composition of Abies grandis needles does not appear to contribute significantly to this phenomenon. Thus, the results presented here allow the general conclusion that changes of contact angles measured on leaf surfaces may not always be due to changes in the leaf surface chemistry and/or the fine structure of leaf surface waxes, but may also be due to increased amounts of epiphyllic micro-organisms significantly altering the leaf surface wetting properties.     

The contribution of fog to the water relations of Sequoia sempervirens (D. Don): foliar uptake and prevention of dehydration

Fog is a defining feature of the coastal California redwood forest and fog inputs via canopy drip in summer can constitute 30% or more of the total water input each year. A great deal of occult precipitation (fog and light rain) is retained in redwood canopies, which have some of the largest leaf area indices known (Westman & Whittaker, Journal of Ecology 63, 493–520, 1975). An investigation was carried out to determine whether some fraction of intercepted fog water might be directly absorbed through leaf surfaces and if so, the importance of this to the water relations physiology of coast redwood, Sequoia sempervirens. An array of complimentary techniques were adopted to demonstrate that fog is absorbed directly by S. sempervirens foliage. Xylem sap transport reversed direction during heavy fog, with instantaneous flow rates in the direction of the soil peaking at approximately 5–7% of maximum transpiration rate. Isotopic analyses showed that up to 6% of a leaf's water content could be traced to a previous night's fog deposition, but this amount varied considerably depending on the age and water status of the leaves. Old leaves, which appear most able to absorb fog water were able to absorb distilled water when fully submersed at an average rate of 0.90 mmol m² s^?1, or about 80% of transpiration rates measured at the leaf level in the field. Sequoia sempervirens has poor stomatal control in response to a drying atmosphere, with rates of water loss on very dry nights up to 40% of midday summer values and rates above 10% being extremely common. Owing to this profligate water use behaviour of S. sempervirens, it appears that fog has a greater role in suppressing water loss from leaves, and thereby ameliorating daily water stress, than in providing supplemental water to foliar tissues per se. Although direct foliar absorption from fog inputs represents only a small fraction of the water used each day, fog's in reducing transpiration and rehydrating leaf tissues during the most active growth periods in summer may allow for greater seasonal carbon fixation and thus contribute to the very fast growth rates and great size of this species.     

Seasonal and age-related variation in the needle quality of five conifer species

Summary Age changes of foliage resource quality (water, nitrogen, fibre, phenolics and toughness) were studied in five species of conifer (Pinus sylvestris L.), Picea abies (L.) Karsten, Tsuga heterophylla (Rafinesque) Sargent (all Pinaceae), Chamaecyparis lawsonian (Murray) Parlatore and Thuja plicata D. Don (both Cupressaceae) over a 2-year period in an English forest.Mature foliage of Pinus sylvestris was characterized by higher levels of nitrogen, fibre and toughness, and lower phenolics, and that of Tsuga heterophylla by higher levels of phenolics, and lower fibre and toughness levels, than the mature needles of the other species studied. Immature needles had higher levels of water and nitrogen, and lower levels of fibre and toughness, than older needles. Immature needles of Picea abies and Tsuga heterophylla had a high concentration of phenolics, which decreased with needle maturity. By mid-August, the levels of most of the foliar constituents in current-year needles had stabilized at levels maintained for the next year. Sampling revealed a fall in the concentration of phenolics, fibre and water in mature needles between March and June. Possible reasons for this seasonal trend are discussed. The levels of conifer foliar constituents were compared with levels recorded in broadleaf trees. Conifers had greater concentrations of all measured foliar constituents, but, with the exception of the six fold greater toughness of conifer needles, the differences between broadleaves and conifers were less than those between the immature and mature conifer needles. Previous studies have related the abundance of Lepidoptera on conifers to hostplant taxonomic relationships. However, the foliar constituents measured in this study were poor predictors of taxonomic relationships between the conifers. It is suggested that the abundance of Lepidoptera on conifers is not determined by levels of general foliar constituents and the role of other hostplant factors in shaping lepidopteran utilization of conifers is discussed.     

Leaf Surface Microflora May Significantly Affect Studies on Foliar Uptake of Chemicals

Uptake and desorption kinetics of methylglucose and pentachlorophenol (PCP) by needles of Picea abies (L.) Karst. were studied. Uptake of methylglucose was very rapid, equilibrium was obtained within 2 h and the amounts of methylglucose associated with the needles could be completely desorbed within 8 h. In contrast, PCP uptake was steady for several hours and the amounts of PCP taken up could only partially be desorbed again. Needles treated with NaN₃ prior to uptake measurements, showed no methylglucose uptake at all, whereas PCP uptake was not affected. Thus it is concluded that the hydrophilic methylglucose was taken up solely by epiphytic microorganisms, whereas the lipophilic PCP was taken up into the needle interior in significant amounts. Consequences of these results on studies of foliar uptake of chemicals are pointed out.     

Determination of foliar uptake of chemicals: influence of leaf surface microflora

Analysing uptake and desorption kinetics of methylglucose in needles of Picea abies (L.) Karst., the effect of NaN₃ on methylglucose uptake, and the correlation between different degrees of colonization of needle surfaces with epiphytes and equilibrium uptake of methylglucose, showed that methylglucose was taken up only by epiphytes but not by the needles themselves. Thus, amounts of methylglucose taken up are a measure of the amounts of epiphytic microorganisms. Uptake of methylglucose into epiphytes was very rapid and equilibrium was obtained within 2h. Amounts taken up could completely be desorbed again from epiphytes within 2-6 h. It is pointed out that the possible influence of leaf surface microorganisms must be considered in studies of foliar uptake of chemicals, in order to avoid erroneous conclusions.     

Crown closure affects endophytic leaf mycobiome compositional dynamics over time in Pseudotsuga menziesii var. menziesii

Old-growth Pseudotsuga menziesii var. menziesii forests produce complex environmental and spatial gradients along which biota assemble. Given this, it has been proposed that changes in the crown microenvironment are associated with different community assembly outcomes for needle fungi. Using high-throughput sequencing, the endophytic mycobiomes of needles were characterized for increasing ages of needles sampled along the boles of eight coastal Douglas-fir trees. Leveraging airborne light detection and ranging (LiDAR) data to create three-dimensional “point cloud” representations of tree crowns revealed that crown closure accounted for more fungal compositional variation than height in crown, and fungal richness and diversity were positively correlated with increasing crown closure. Supplementing the point clouds of each climbed tree with clouds from >5,000 randomly selected trees in the study area showed that fungal communities from closed portions of the crown were increasingly structured with needle age. These findings highlight the importance of the crown microenvironment in the development of foliar fungal communities for a foundation tree species.     

Effects of climate change on labile and structural carbon in Douglas-fir needles as estimated by δ13C and Carea measurements

Models of photosynthesis, respiration, and export predict that foliar labile carbon (C) should increase with elevated CO₂ but decrease with elevated temperature. Sugars, starch, and protein can be compared between treatments, but these compounds make up only a fraction of the total labile pool. Moreover, it is difficult to assess the turnover of labile carbon between years for evergreen foliage. Here, we combined changes in foliar C_area (C concentration on an areal basis) as needles aged with changes in foliar isotopic composition (δ¹³C) caused by inputs of ¹³C‐depleted CO₂ to estimate labile and structural C in needles of different ages in a four‐year, closed‐chamber mesocosm experiment in which Douglas‐fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings were exposed to elevated temperature (ambient + 3.5 °C) and CO₂ (ambient + 179 ppm). Declines in δ ¹³C of needle cohorts as they aged indicated incorporation of newly fixed labile or structural carbon. The δ ¹³C calculations showed that new C was 41 ± 2% and 28 ± 3% of total needle carbon in second‐ and third‐year needles, respectively, with higher proportions of new C in elevated than ambient CO₂ chambers (e.g. 42 ± 2% vs. 37 ± 6%, respectively, for second‐year needles). Relative to ambient CO₂, elevated CO₂ increased labile C in both first‐ and second‐year needles. Relative to ambient temperature, elevated temperature diminished labile C in second‐year needles but not in first‐year needles, perhaps because of differences in sink strength between the two needle age classes. We hypothesize that plant‐soil feedbacks on nitrogen supply contributed to higher photosynthetic rates under elevated temperatures that partly compensated for higher turnover rates of labile C. Strong positive correlations between labile C and sugar concentrations suggested that labile C was primarily determined by carbohydrates. Labile C was negatively correlated with concentrations of cellulose and protein. Elevated temperature increased foliar %C, possibly due to a shift of labile constituents from low %C carbohydrates to relatively high %C protein. Decreased sugar concentrations and increased nitrogen concentrations with elevated temperature were consistent with this explanation. Because foliar constituents that vary in isotopic signature also vary in concentrations with leaf age or environmental conditions, inferences of c_i/c_a values from δ ¹³C of bulk leaf tissue should be done cautiously. Tracing of ¹³C through foliar carbon pools may provide new insight into foliar C constituents and turnover.     

Interaction of soil filamentous fungi affects needle composition and nutrition of Norway spruce seedlings

The effects of the interactions of soil filamentous fungi (including saprotrophic, mycorrhizal and endophytic fungi) on several morphological and physiological parameters of Norway spruce seedlings [Picea abies (L.) Karst.] were studied in a pot experiment. Both mycorrhizal variants (Hebeloma bryogenes and Cadophora finlandica) were slightly inhibited with respect to the accumulation of aboveground biomass. However, these variants exhibited significantly improved foliar content of N, P, and Ca (H. bryogenes variant also K), compared to controls. The presence of the saprotrophic fungus Setulipes androsaceus attenuated the positive effect of mycorrhizal fungi on Ca, P, and K content, but did not reduce the positive effect of mycorrhizal fungi on the N content of the seedlings’ needles. Raman spectroscopy revealed that both mycorrhizal fungi increased the foliar content of carotenoids compared to the controls, but the effect diminished in the presence of S. androsaceus. In contrast, in the presence of the dark septate endophyte Phialocephala fortinii, needles exhibited significantly higher wax content and lower carotenoid content compared to the mycorrhizal variants. The presence of the saprotrophic isolate Serpula himantoides resulted in a decrease in needle waxes in comparison to controls. The needles’ carotenoid concentration was positively correlated with the levels of needle nutrients (N, P, K, Ca, Mg), while correlation of needle nutrients (N, P) with the needle wax concentration was negative. We conclude that not only individual fungi but also their interactions profoundly affect the nutrition and needle composition of Norway spruce seedlings.     

Inferring foliar water uptake using stable isotopes of water

A growing number of studies have described the direct absorption of water into leaves, a phenomenon known as foliar water uptake. The resultant increase in the amount of water in the leaf can be important for plant function. Exposing leaves to isotopically enriched or depleted water sources has become a common method for establishing whether or not a plant is capable of carrying out foliar water uptake. However, a careful inspection of our understanding of the fluxes of water isotopes between leaves and the atmosphere under high humidity conditions shows that there can clearly be isotopic exchange between the two pools even in the absence of a change in the mass of water in the leaf. We provide experimental evidence that while leaf water isotope ratios may change following exposure to a fog event using water with a depleted oxygen isotope ratio, leaf mass only changes when leaves are experiencing a water deficit that creates a driving gradient for the uptake of water by the leaf. Studies that rely on stable isotopes of water as a means of studying plant water use, particularly with respect to foliar water uptake, must consider the effects of these isotopic exchange processes.     

Colonization,Species Composition,and Conidial Production of Aquatic Hyphomycetes on Chirpine Needle Litter in a Freshwater Kumaun Himalayan Stream

Aquatic hyphomycetes colonizing the submerged chirpine (pinus roxburghii SARG .) needle litter in a high altitude, Kumaun Himalayan stream were studied. 15 species belonging to different genera of aquatic Hyphomycetes have been recognized as the colonizers of chirpine needle litter. Clavariopsis aquatica, Heliscus lugdunensis, Lunulospora cymbiformis, Triscelophorus acuminatus and T. monosporus were found with a high frequency of occurrence. The conidial production was highest in Flagellospora penicillioides, however, Campylospora chaetocladia, L. cymbiformis and T. acuminatus had less number of conidia per unit area of pine needles. The chirpine needle litter decomposition in the freshwater habitat is also discussed.     

Foggy days and dry nights determine crown‐level water balance in a seasonal tropical montane cloud forest

The ecophysiology of tropical montane cloud forest (TMCF) trees is influenced by crown‐level microclimate factors including regular mist/fog water inputs, and large variations in evaporative demand, which in turn can significantly impact water balance. We investigated the effect of such microclimatic factors on canopy ecophysiology and branch‐level water balance in the dry season of a seasonal TMCF in Veracruz, Mexico, by quantifying both water inputs (via foliar uptake, FU) and outputs (day‐ and night‐time transpiration, NT). Measurements of sap flow, stomatal conductance, leaf water potential and pressure–volume relations were obtained in Quercus lanceifolia, a canopy‐dominant tree species. Our results indicate that FU occurred 34% of the time and led to the recovery of 9% (24 ± 9.1 L) of all the dry‐season water transpired from individual branches. Capacity for FU was independently verified for seven additional common tree species. NT accounted for approximately 17% (46 L) of dry‐season water loss. There was a strong correlation between FU and the duration of leaf wetness events (fog and/or rain), as well as between NT and the night‐time vapour pressure deficit. Our results show the clear importance of fog and NT for the canopy water relations of Q. lanceifolia.     

Relevance of crustacean carapace wettability for fouling

Carapace wettability and density of fouling organisms (bacteria, diatoms, protozoa, fungi, macro-organisms) were investigated for 45 crustacean species (Hoplocarida, Decapoda) from 15 families in the Gulf of Thailand. The results show that crustaceans can create and maintain characteristic carapace wettabilities. About 21 species (47%) possess highly wettable carapaces with contact angles below 20°. Contact angles between 20° and 40° were recorded for four species (2%), angles between 40° and 60° for eight species (4%) and from 60° to 70° for 11 (24%) species. One species, Alpheus euphrosyne (Alpheidae, Decapoda), exhibited an extremely low surface wettability (contact angle: 91°). Densities of colonisers and contact angles did not correlate. Very low wettability by water ( > 90°) may only contribute little to fouling reduction in A. euphrosyne which showed the most hydrophobic carapace surface and was colonised by the lowest numbers of bacteria among all species and no other colonisers at all. We conclude that surface wettability is of little relevance for antifouling defence in crustaceans.     

Polystichum munitum (Dryopteridaceae) varies geographically in its capacity to absorb fog water by foliar uptake within the redwood forest ecosystem

? Premise of the study: Fog provides a critical water resource to plants around the world. In the redwood forest ecosystem of northern California, plants depend on fog absorbed through foliar uptake to stay hydrated during the rainless summer. In this study, we identified regions within the redwood ecosystem where the fern Polystichum munitum canopy most effectively absorbs fog drip that reaches the forest floor. ? Methods: We measured the foliar uptake capacity of P. munitum fronds at seven sites along 700 km of the redwood forest ecosystem. We quantified the canopy cover of P. munitum at each site and estimated how much water the fern canopy can acquire aboveground through fog interception and absorption. ? Key results: Throughout the ecosystem, nocturnal foliar uptake increased the leaf water content of P. munitum by 7.2%, and we estimated that the P. munitum canopy can absorb 5 ± 3% (mean ± SE) of intercepted fog precipitation. Strikingly, P. munitum had the highest foliar uptake capacity in the center of the ecosystem and may absorb 10% more of the fog its canopy intercepts in this region relative to other regions studied. Conversely, P. munitum had no foliar uptake capacity in the southern end of the ecosystem. ? Conclusions: This study shows the first evidence that foliar uptake varies within species at the landscape scale. Our findings suggest that the P. munitum at the southern tip of the redwood ecosystem may suffer most from low summertime water availability because it had no potential to acquire fog as an aboveground water subsidy.     

The incidence and implications of clouds for cloud forest plant water relations

Although clouds are the most recognisable and defining feature of tropical montane cloud forests, little research has focussed on how clouds affect plant functioning. We used satellite and ground‐based observations to study cloud and leaf wetting patterns in contrasting tropical montane and pre‐montane cloud forests. We then studied the consequences of leaf wetting for the direct uptake of water accumulated on leaf surfaces into the leaves themselves. During the dry season, the montane forest experienced higher precipitation, cloud cover and leaf wetting events of longer duration than the pre‐montane forest. Leaf wetting events resulted in foliar water uptake in all species studied. The capacity for foliar water uptake differed significantly between the montane and pre‐montane forest plant communities, as well as among species within a forest. Our results indicate that foliar water uptake is common in these forest plants and improves plant water status during the dry season.     

Cell surface hydrophobicity of colistin-susceptible vs resistant Acinetobacter baumannii determined by contact angles: methodological considerations and implications

Contact angle analysis of cell surface hydrophobicity (CSH) describes the tendency of a water droplet to spread across a lawn of filtered bacterial cells. Colistin‐induced disruption of the Gram‐negative outer membrane necessitates hydrophobic contacts with lipopolysaccharide (LPS). We aimed to characterize the CSH of Acinetobacter baumannii using contact angles, to provide insight into the mechanism of colistin resistance. Contact angles were analysed for five paired colistin‐susceptible and resistant Ac. baumannii strains. Drainage of the water droplet through bacterial layers was demonstrated to influence results. Consequently, measurements were performed 0·66 s after droplet deposition. Colistin‐resistant cells exhibited lower contact angles (38·8±2·8–46·8±1·3°) compared with their paired colistin‐susceptible strains (40·7±3·0–48·0±1·4°; anova ; P < 0·05). Contact angles increased at stationary phase (50·3±2·9–61·5±2·5° and 47·4±2·0–50·8±3·2°, susceptible and resistant, respectively, anova ; P < 0·05) and in response to colistin 32 mg l^?1 exposure (44·5±1·5–50·6±2·8° and 43·5±2·2–48·0±2·2°, susceptible and resistant, respectively; anova ; P < 0·05). Analysis of complemented strains constructed with an intact lpxA gene, or empty vector, highlighted the contribution of LPS to CSH. Compositional outer‐membrane variations likely account for CSH differences between Ac. baumannii phenotypes, which influence the hydrophobic colistin–bacterium interaction. Important insight into the mechanism of colistin resistance has been provided. Greater consideration of contact angle methodology is necessary to ensure accurate analyses are performed.