17Drought threatens tropical rainforests over seasonal to decadal timescales [1][2][3][4] , but the drivers 18 of tree mortality following drought remain poorly understood 5,6 . It has been suggested that 19 reduced availability of non-structural carbohydrates (NSC) critically increases mortality risk 20 through insufficient carbon supply to metabolism ('carbon starvation') 7,8 . However little is 21 known about how NSC stores are affected by drought, especially over the long term, and 22 whether they are more important than hydraulic processes in determining drought-induced 23 mortality. Using data from the world's longest-running experimental drought study in tropical 24 rainforest (in the Brazilian Amazon), we test whether carbon starvation or deterioration of the 25 water-conducting pathways from soil to leaf trigger tree mortality. Biomass loss from 26 mortality in the experimentally-droughted forest increased substantially after >10 years of 27 reduced soil moisture availability. The mortality signal was dominated by the death of large 28 trees, which were at a much greater risk of hydraulic deterioration than smaller trees. 29However, we find no evidence that the droughted trees suffered carbon starvation, as their 30 NSC concentrations were similar to those of un-droughted trees, and growth rates did not 31 decline in either living or dying individuals. Our results indicate that hydraulics, rather than 32 carbon starvation, triggers tree death from drought in tropical rainforest. 34Drought-response observations from both field-scale experiments and natural droughts have 35 demonstrated increased mortality over the short-term (1-3 years), with notably higher 36 vulnerability for some taxa, and for larger trees 6,9,10 . After several years of drought, 37 recovering growth rates in smaller trees, dbh (diameter at breast height) <40 cm, and reduced 38 mortality have been recorded at different locations 6,11,12 . However, the long-term (>10 yr) 39 sensitivity of tropical forests to predicted prolonged and repeated water deficit [1][2][3] we synthesise these data to test whether long-term soil moisture deficit alters NSC storage 64 and use in tropical rainforest trees, and if this, or hydraulic processes, are most strongly 65 associated with increased mortality rates. 66By 2014, following 13 years of the TFE treatment, cumulative biomass loss through mortality 67 was 41.0±2.7% relative to pre-treatment values (Fig. 1a), and the rate of loss had increased 68 substantially since the previous reported value of 17.2±0.8%, after 7 years of TFE 6 . 69Accelerating biomass loss and failure to recover substantially, or to reach a new 70 equilibrium 13 , has led to a committed flux to the atmosphere from decomposing necromass of 71 101.9±19.1 Mg C ha -1 (Fig. 1a). This biomass loss has been driven by elevated mortality in 72 the largest trees (Fig. 1b), as previously observed over shorter timescales 6 , and has created a 73 canopy that has had a persistently lower average leaf area index during 2010-2014 74 (12.0±1...

Determining climate change feedbacks from tropical rainforests requires an understanding of how carbon gain through photosynthesis and loss through respiration will be altered. One of the key changes that tropical rainforests may experience under future climate change scenarios is reduced soil moisture availability. In this study we examine if and how both leaf photosynthesis and leaf dark respiration acclimate following more than 12 years of experimental soil moisture deficit, via a through‐fall exclusion experiment (TFE) in an eastern Amazonian rainforest. We find that experimentally drought‐stressed trees and taxa maintain the same maximum leaf photosynthetic capacity as trees in corresponding control forest, independent of their susceptibility to drought‐induced mortality. We hypothesize that photosynthetic capacity is maintained across all treatments and taxa to take advantage of short‐lived periods of high moisture availability, when stomatal conductance (g s) and photosynthesis can increase rapidly, potentially compensating for reduced assimilate supply at other times. Average leaf dark respiration (R d) was elevated in the TFE‐treated forest trees relative to the control by 28.2 ± 2.8% (mean ± one standard error). This mean R d value was dominated by a 48.5 ± 3.6% increase in the R d of drought‐sensitive taxa, and likely reflects the need for additional metabolic support required for stress‐related repair, and hydraulic or osmotic maintenance processes. Following soil moisture deficit that is maintained for several years, our data suggest that changes in respiration drive greater shifts in the canopy carbon balance, than changes in photosynthetic capacity.

[1] Changes in land-use and climate are likely to alter moisture and substrate availability in tropical forest soils, but quantitative assessment of the role of resource constraints as regulators of soil trace gas fluxes is rather limited. The primary objective of this study was to quantify the effects of moisture and substrate availability on soil trace gas fluxes in an Amazonian regrowth forest. We measured the efflux of carbon dioxide (CO 2 ), nitric oxide (NO), nitrous oxide (N 2 O), and methane (CH 4 ) from soil in response to two experimental manipulations. In the first, we increased soil moisture availability during the dry season by irrigation; in the second, we decreased substrate availability by continuous removal of aboveground litter. In the absence of irrigation, soil CO 2 efflux decreased during the dry season while irrigation maintained soil CO 2 efflux levels similar to the wet season. Large variations in soil CO 2 efflux consistent with a significant moisture constraint on respiration were observed in response to soil wet-up and dry-down events. Annual soil C efflux for irrigated plots was 27 and 13% higher than for control plots in 2001 and 2002, respectively. Litter removal significantly reduced soil CO 2 efflux; annual soil C efflux in 2002 was 28% lower for litter removal plots compared to control plots. The annual soil C efflux:litterfall C ratio for the control treatment (4.0-5.2) was consistent with previously reported values for regrowth forests that indicate a relatively large belowground C allocation. In general, fluxes of N 2 O and CH 4 were higher during the wet season and both fluxes increased during dry-season irrigation. There was no seasonal effect on NO fluxes. Litter removal had no significant impact on N oxide or CH 4 emissions. Net soil nitrification did not respond to dry-season irrigation, but was somewhat reduced by litter removal. Overall, these results demonstrate significant soil moisture and substrate constraints on soil trace gas emissions, particularly for CO 2 , and suggest that climate and land-use changes that alter moisture and substrate availability are therefore likely to have an impact on atmosphere chemistry.

Summary The tropics are predicted to become warmer and drier, and understanding the sensitivity of tree species to drought is important for characterizing the risk to forests of climate change. This study makes use of a long‐term drought experiment in the Amazon rainforest to evaluate the role of leaf‐level water relations, leaf anatomy and their plasticity in response to drought in six tree genera.The variables (osmotic potential at full turgor, turgor loss point, capacitance, elastic modulus, relative water content and saturated water content) were compared between seasons and between plots (control and through‐fall exclusion) enabling a comparison between short‐ and long‐term plasticity in traits. Leaf anatomical traits were correlated with water relation parameters to determine whether water relations differed among tissues.The key findings were: osmotic adjustment occurred in response to the long‐term drought treatment; species resistant to drought stress showed less osmotic adjustment than drought‐sensitive species; and water relation traits were correlated with tissue properties, especially the thickness of the abaxial epidermis and the spongy mesophyll.These findings demonstrate that cell‐level water relation traits can acclimate to long‐term water stress, and highlight the limitations of extrapolating the results of short‐term studies to temporal scales associated with climate change.

The fate of tropical forests under future climate change is dependent on the capacity of their trees to adjust to drier conditions. The capacity of trees to withstand drought is likely to be determined by traits associated with their hydraulic systems. However, data on whether tropical trees can adjust hydraulic traits when experiencing drought remain rare. We measured plant hydraulic traits (e.g. hydraulic conductivity and embolism resistance) and plant hydraulic system status (e.g. leaf water potential, native embolism and safety margin) on >150 trees from 12 genera (36 species) and spanning a stem size range from 14 to 68 cm diameter at breast height at the world's only longrunning tropical forest drought experiment. Hydraulic traits showed no adjustment following 15 years of experimentally imposed moisture deficit. This failure to adjust resulted in these drought-stressed trees experiencing significantly lower leaf water potentials, and higher, but variable, levels of native embolism in the branches. This result suggests that hydraulic damage caused by elevated levels of embolism is likely to be one of the key drivers of drought-induced mortality following long-term soil moisture deficit. We demonstrate that some hydraulic traits changed with tree size, however, the direction and magnitude of the change was controlled by taxonomic identity. Our results suggest that Amazonian trees, both small and large, have limited capacity to acclimate their hydraulic systems to future droughts, potentially making them more at risk of drought-induced mortality.

The absorption of atmospheric water directly into leaves enables plants to alleviate the water stress caused by low soil moisture, hydraulic resistance in the xylem and the effect of gravity on the water column, while enabling plants to scavenge small inputs of water from leaf‐wetting events. By increasing the availability of water, and supplying it from the top of the canopy (in a direction facilitated by gravity), foliar uptake (FU) may be a significant process in determining how forests interact with climate, and could alter our interpretation of current metrics for hydraulic stress and sensitivity. FU has not been reported for lowland tropical rainforests; we test whether FU occurs in six common Amazonian tree genera in lowland Amazônia, and make a first estimation of its contribution to canopy–atmosphere water exchange. We demonstrate that FU occurs in all six genera and that dew‐derived water may therefore be used to “pay” for some morning transpiration in the dry season. Using meteorological and canopy wetness data, coupled with empirically derived estimates of leaf conductance to FU (kfu), we estimate that the contribution by FU to annual transpiration at this site has a median value of 8.2% (103 mm/year) and an interquartile range of 3.4%–15.3%, with the biggest sources of uncertainty being kfu and the proportion of time the canopy is wet. Our results indicate that FU is likely to be a common strategy and may have significant implications for the Amazon carbon budget. The process of foliar water uptake may also have a profound impact on the drought tolerance of individual Amazonian trees and tree species, and on the cycling of water and carbon, regionally and globally.

Transpiration from the Amazon rainforest generates an essential water source at a global and local scale. However, changes in rainforest function with climate change can disrupt this process, causing significant reductions in precipitation across Amazonia, and potentially at a global scale. We report the only study of forest transpiration following a long-term (>10 year) experimental drought treatment in Amazonian forest. After 15 years of receiving half the normal rainfall, droughtrelated tree mortality caused total forest transpiration to decrease by 30%. However, the surviving droughted trees maintained or increased transpiration because of reduced competition for water and increased light availability, which is consistent with increased growth rates. Consequently, the amount of water supplied as rainfall reaching the soil and directly recycled as transpiration increased to 100%. This value was 25% greater than for adjacent nondroughted forest. If these drought conditions were accompanied by a modest increase in temperature (e.g., 1.5°C), water demand would exceed supply, making the forest more prone to increased tree mortality. K E Y W O R D Sdrought, sap flux, transpiration, tree mortality, tropical forest, water cycling

Summary Fine root dynamics is widely recognized as an important biogeochemical process, but there are few data on fine root growth and its response to soil resource availability, especially for tropical forests. We evaluated the response of fine root dynamics to altered availability of soil water and nutrients in a 20‐yr‐old forest regrowth in eastern Amazonia. In one experiment the dry season reduction in soil moisture was alleviated by irrigation. In the other experiment, nutrient supply was reduced by litter removal. We used the ingrowth core technique to measure fine root mass growth, length growth, mortality and specific root length. Dry‐season irrigation had no significant effect on mass and length of live and dead roots, whereas litter removal reduced mass and length of live roots. For both irrigation and litter removal experiments, root growth was significantly greater in the dry season than in the wet season. Increased root growth was associated with decreased soil water availability. However, root growth did not increase in response to nutrient reduction in litter removal plots. Overall, our results suggest that belowground allocation may differ according to the type of soil resource limitation.