Natural forest regeneration through fire protection is a less imminent threat for truly stable savannas than afforestation.

African bistable savannas have important biodiversity value and merit conservation. At the same time, forest restoration is a nature-based solution that can be used to increase biodiversity, carbon stocks, and human well-being. Here we describe an experiment based on natural forest regeneration through the exclusion of anthropogenic fire. We show that it is easier to let nature do its work instead of channeling it into an artificial man-made ecosystem through human-induced burning or planting. We emphasize that nature-based solutions must be biome-appropriate and the choice between restoring forests or protecting savannas requires a thorough understanding of the local context.

Protecting an artificial savanna as a nature-based solution to restore carbon and biodiversity in the Democratic Republic of the Congo.

A large share of the global forest restoration potential is situated in artificial 'unstable' mesic African savannas, which could be restored to higher carbon and biodiversity states if protected from human-induced burning. However, uncertainty on recovery rates in protected unstable savannas impedes science-informed forest restoration initiatives. Here, we quantify the forest restoration success of anthropogenic fire exclusion within an 88-ha mesic artificial savanna patch in the Kongo Central province of the Democratic Republic of the Congo (DR Congo). We found that aboveground carbon recovery after 17 years was on average 11.40 ± 0.85 Mg C ha-1 . Using a statistical model, we found that aboveground carbon stocks take 112 ± 3 years to recover to 90% of aboveground carbon stocks in old-growth forests. Assuming that this recovery trajectory would be representative for all unstable savannas, we estimate that they could have a total carbon uptake potential of 12.13 ± 2.25 Gt C by 2100 across DR Congo, Congo and Angola. Species richness recovered to 33.17% after 17 years, and we predicted a 90% recovery at 54 ± 2 years. In contrast, we predicted that species composition would recover to 90% of old-growth forest composition only after 124 ± 3 years. We conclude that the relatively simple and cost-efficient measure of fire exclusion in artificial savannas is an effective nature-based solution to climate change and biodiversity loss. However, more long-term and in situ monitoring efforts are needed to quantify variation in long-term carbon and diversity recovery pathways. Particular uncertainties are spatial variability in socio-economics and growing conditions as well as the effects of projected climate change.

Earlier onset and slower heartwood investment in faster-growing trees of African tropical species.

BACKGROUND AND AIMS: Heartwood plays an important role in maintaining the structural integrity of trees. While, its formation has long be thought to be solely driven by internal ageing processes, more recent hypotheses suggest that heartwood formation acts as a regulator of the tree water balance by modulating sapwood quantities. Testing both hypotheses would shed light on the potential ecophysiological nature of heartwood formation, a very common process in trees. METHODS: We measured heartwood and sapwood quantities, xylem conduits and growth ring width and number on 406 stems of Pericopsis elata with ages ranging from 2 to 237 years. A subset of 17 trees with similar ages but varying growth rate were sampled in a shaded (slower growth) and sun-exposed (faster growth) site. We used regression analysis and structural equations modelling to investigate the dynamics and drivers of heartwood formation. KEY RESULTS: We found a positive effect of growth rate on the probability of heartwood occurrence, suggesting an earlier heartwood onset in faster-growing stems. After this onset age, heartwood area increases with stem diameter and age. Despite the similar heartwood production per unit stem diameter increment, shaded trees produce heartwood faster than sun-exposed trees. Tree age and hydraulics showed similar direct effects on heartwood and sapwood area of sun-exposed trees, suggesting their mutual role in driving the heartwood dynamics of sun-exposed trees. However, for shaded trees, only tree hydraulics showed a direct effect, suggesting its prominent role over age in driving the heartwood dynamics in limited growing conditions. The positive relationship between growth rate and maximum stomatal conductance supports this conclusion. CONCLUSIONS: Heartwood area increases as the tree ages but at a slower rate in trees where water demand is balanced by a sufficient water supply. Our findings suggest that heartwood formation is not only a structural but also a functional process.

Advanced X-ray CT scanning can boost tree ring research for earth system sciences.

BACKGROUND AND AIMS: Tree rings, as archives of the past and biosensors of the present, offer unique opportunities to study influences of the fluctuating environment over decades to centuries. As such, tree-ring-based wood traits are capital input for global vegetation models. To contribute to earth system sciences, however, sufficient spatial coverage is required of detailed individual-based measurements, necessitating large amounts of data. X-ray computed tomography (CT) scanning is one of the few techniques that can deliver such data sets. METHODS: Increment cores of four different temperate tree species were scanned with a state-of-the-art X-ray CT system at resolutions ranging from 60 µm down to 4.5 µm, with an additional scan at a resolution of 0.8 µm of a splinter-sized sample using a second X-ray CT system to highlight the potential of cell-level scanning. Calibration-free densitometry, based on full scanner simulation of a third X-ray CT system, is illustrated on increment cores of a tropical tree species. KEY RESULTS: We show how multiscale scanning offers unprecedented potential for mapping tree rings and wood traits without sample manipulation and with limited operator intervention. Custom-designed sample holders enable simultaneous scanning of multiple increment cores at resolutions sufficient for tree ring analysis and densitometry as well as single core scanning enabling quantitative wood anatomy, thereby approaching the conventional thin section approach. Standardized X-ray CT volumes are, furthermore, ideal input imagery for automated pipelines with neural-based learning for tree ring detection and measurements of wood traits. CONCLUSIONS: Advanced X-ray CT scanning for high-throughput processing of increment cores is within reach, generating pith-to-bark ring width series, density profiles and wood trait data. This would allow contribution to large-scale monitoring and modelling efforts with sufficient global coverage.

Asynchronous leaf and cambial phenology in a tree species of the Congo Basin requires space-time conversion of wood traits.

BACKGROUND AND AIMS: Wood traits are increasingly being used to document tree performance. In the Congo Basin, however, weaker seasonality causes asynchrony of wood traits between trees. Here, we monitor growth and phenology data to date the formation of traits. METHODS: For two seasons, leaf and cambial phenology were monitored on four Terminalia superba trees (Mayombe) using cameras, cambial pinning and dendrometers. Subsequently, vessel lumen and parenchyma fractions as well as high-resolution isotopes (δ13C/δ18O) were quantified on the formed rings. All traits were dated and related to weather data. KEY RESULTS: We observed between-tree differences in green-up of 45 d, with trees flushing before and after the rainy season. The lag between green-up and onset of xylem formation was 59 ± 21 d. The xylem growing season lasted 159 ± 17 d with between-tree differences of up to 53 d. Synchronized vessel, parenchyma and δ13C profiles were related to each other. Only parenchyma fraction and δ13C were correlated to weather variables, whereas the δ18O pattern showed no trend. CONCLUSIONS: Asynchrony of leaf and cambial phenology complicates correct interpretation of environmental information recorded in wood. An integrated approach including high-resolution measurements of growth, stable isotopes and anatomical features allows exact dating of the formation of traits. This methodology offers a means to explore the asynchrony of growth in a rainforest and contribute to understanding this aspect of forest resilience.

A field-to-desktop toolchain for X-ray CT densitometry enables tree ring analysis.

BACKGROUND AND AIMS: Disentangling tree growth requires more than ring width data only. Densitometry is considered a valuable proxy, yet laborious wood sample preparation and lack of dedicated software limit the widespread use of density profiling for tree ring analysis. An X-ray computed tomography-based toolchain of tree increment cores is presented, which results in profile data sets suitable for visual exploration as well as density-based pattern matching. METHODS: Two temperate (Quercus petraea, Fagus sylvatica) and one tropical species (Terminalia superba) were used for density profiling using an X-ray computed tomography facility with custom-made sample holders and dedicated processing software. KEY RESULTS: Density-based pattern matching is developed and able to detect anomalies in ring series that can be corrected via interactive software. CONCLUSIONS: A digital workflow allows generation of structure-corrected profiles of large sets of cores in a short time span that provide sufficient intra-annual density information for tree ring analysis. Furthermore, visual exploration of such data sets is of high value. The dated profiles can be used for high-resolution chronologies and also offer opportunities for fast screening of lesser studied tropical tree species.

Charcoal-inferred Holocene fire and vegetation history linked to drought periods in the Democratic Republic of Congo.

The impact of Holocene drought events on the presumably stable Central African rainforest remains largely unexplored, in particular the significance of fire. High-quality sedimentary archives are scarce, and palynological records mostly integrate over large regional scales subject to different fire regimes. Here, we demonstrate a direct temporal link between Holocene droughts, palaeofire and vegetation change within present-day Central African rainforest, using records of identified charcoal fragments extracted from soil in the southern Mayumbe forest (Democratic Republic of Congo). We find three distinct periods of local palaeofire occurrence: 7.8-6.8 ka BP, 2.3-1.5 ka BP, 0.8 ka BP - present. These periods are linked to well-known Holocene drought anomalies: the 8.2 ka BP event, the 3rd millennium BP rainforest crisis and the Mediaeval Climate Anomaly. During and after these Holocene droughts, the Central African rainforest landscape was characterized by a fragmented pattern with fire-prone open patches. Some fires occurred during the drought anomalies although most fires seem to lag behind them, which suggests that the open patches remained fire-prone after the actual climate anomalies. Charcoal identifications indicate that mature rainforest patches did persist through the Early to Mid-Holocene climatic transition, the subsequent Holocene thermal optimum and the third millennium BP rainforest crisis, until 0.8 ka BP. However, disturbance and fragmentation were probably more prominent near the boundary of the southern Mayumbe forest. Furthermore, the dominance of pioneer and woodland savanna taxa in younger charcoal assemblages indicates that rainforest regeneration was hampered by increasingly severe drought conditions after 0.8 ka BP. These results support the notion of a dynamic forest ecosystem at multicentury time scales across the Central African rainforest.

Heading for a fall: The fate of old wind-thrown beech trees (Fagus sylvatica) is detectable in their growth pattern.

Common beech (Fagus sylvatica) is one of the most important deciduous tree species in European forests. However, climate-change-induced drought may threaten its dominant position. The Sonian Forest close to Brussels (Belgium) is home to some of the largest beech trees in the world. This UNESCO world heritage site is famous for its high density of very large beech trees as a result of its climatic suitability, fertile soil conditions, and past management. Here we utilized tree-ring data from increment cores to investigate the growth of these old and monumental beech trees, evaluating their growth trends, response to past climate, and the effect of mast years on 39 living and 16 recently wind-thrown trees. Our analysis reveals that the sampled trees were generally sensitive to spring and summer droughts but recovered quickly after such an extreme climatic event. The growth trend of living trees has remained high and only shows a slight, statistically insignificant, decline over the past 50 years. Although the overall growth rate remains strong (BAI 50 cm2/year), the past five decades have shown strong inter-annual growth variations due to frequent and more intense droughts combined with an increased frequency of mast years. We also found notable differences in growth patterns between the living trees and those that had recently been wind-thrown. While there were no significant differences between living and wind-thrown trees in response to droughts, heatwaves, or mast years when examining year-to-year growth changes, the wind-thrown trees did exhibit considerably lower overall growth rates and a significant downward trend in growth (BAI -0.57 cm2/year). This difference in growth trends has been apparent since at least the 1980s. Overall, the findings of this study can provide valuable insights for understanding the long-term dynamics of lowland beech forests and their responses to climate change.

SmartWoodID-an image collection of large end-grain surfaces to support wood identification systems.

Wood identification is a key step in the enforcement of laws and regulations aimed at combatting illegal timber trade. Robust wood identification tools, capable of distinguishing a large number of timbers, depend on a solid database of reference material. Reference material for wood identification is typically curated in botanical collections dedicated to wood consisting of samples of secondary xylem of lignified plants. Specimens from the Tervuren Wood Collection, one of the large institutional wood collections around the world, are used as a source of tree species data with potential application as timber. Here, we present SmartWoodID, a database of high-resolution optical scans of the end-grain surfaces enriched with expert wood anatomical descriptions of macroscopic features. These can serve as annotated training data to develop interactive identification keys and artificial intelligence for computer vision-based wood identification. The first edition of the database consists of images of 1190 taxa, with a focus on potential timber species from the Democratic Republic of the Congo with at least four different specimens per species included. Database URL https://hdl.handle.net/20.500.12624/SmartWoodID_first_edition.

Fluctuations of cambial activity in relation to precipitation result in annual rings and intra-annual growth zones of xylem and phloem in teak (Tectona grandis) in Ivory Coast.

BACKGROUND AND AIMS: Teak forms xylem rings that potentially carry records of carbon sequestration and climate in the tropics. These records are only useful when the structural variations of tree rings and their periodicity of formation are known. METHODS: The seasonality of ring formation in mature teak trees was examined via correlative analysis of cambial activity, xylem and phloem formation, and climate throughout 1·5 years. Xylem and phloem differentiation were visualized by light microscopy and scanning electron microscopy. KEY RESULTS: A 3 month dry season resulted in semi-deciduousness, cambial dormancy and formation of annual xylem growth rings (AXGRs). Intra-annual xylem and phloem growth was characterized by variable intensity. Morphometric features of cambium such as cambium thickness and differentiating xylem layers were positively correlated. Cambium thickness was strongly correlated with monthly rainfall (R(2) = 0·7535). In all sampled trees, xylem growth zones (XGZs) were formed within the AXGRs during the seasonal development of new foliage. When trees achieved full leaf, the xylem in the new XGZs appeared completely differentiated and functional for water transport. Two phloem growth rings were formed in one growing season. CONCLUSIONS: The seasonal formation pattern and microstructure of teak xylem suggest that AXGRs and XGZs can be used as proxies for analyses of the tree history and climate at annual and intra-annual resolution.

Whole genome duplication of wild-type and CINNAMYL ALCOHOL DEHYDROGENASE1-downregulated hybrid poplar reduces biomass yield and causes a brittle apex phenotype in field-grown wild types.

The potential of whole genome duplication to increase plant biomass yield is well-known. In Arabidopsis tetraploids, an increase in biomass yield was accompanied by a reduction in lignin content and, as a result, a higher saccharification efficiency was achieved compared with diploid controls. Here, we evaluated whether the results obtained in Arabidopsis could be translated into poplar and whether the enhanced saccharification yield upon alkaline pretreatment of hairpin-downregulated CINNAMYL ALCOHOL DEHYDROGENASE1 (hpCAD) transgenic poplar could be further improved upon a whole genome duplication. Using a colchicine treatment, wild-type (WT) Populus tremula x P. alba cv. INRA 717-1B4, a commonly used model clone in tree biotechnology research, and hpCAD tetraploids were generated and grown in the greenhouse. In parallel, WT tetraploid poplars were grown in the field. In contrast to Arabidopsis, a whole genome duplication of poplar had a negative impact on the biomass yield of both greenhouse- and field-grown trees. Strikingly, field-grown WT tetraploids developed a brittle apex phenotype, i.e., their tip broke off just below the apex. In addition, the chromosome doubling altered the biomass composition of field-grown, but not of greenhouse-grown tetraploid poplars. More specifically, the lignin content of field-grown tetraploid poplars was increased at the expense of matrix polysaccharides. This increase in lignin deposition in biomass is likely the cause of the observed brittle apex phenotype, though no major differences in stem anatomy or in mechanical properties could be found between di- and tetraploid WT poplars grown in the field. Finally, without biomass pretreatment, the saccharification efficiency of greenhouse- and field-grown WT diploids was not different from that of tetraploids, whereas that of greenhouse-grown hpCAD tetraploids was higher than that of greenhouse-grown diploids. Upon alkaline pretreatment, the saccharification yield of diploids was similar to that of tetraploids for all genotypes and growth conditions tested. This study showed that a whole genome duplication in hybrid WT and hpCAD poplar did neither result in further improvements in biomass yield, nor in improved biomass composition and, hence, saccharification performance.

Detecting thin adhesive coatings in wood fiber materials with laboratory-based dual-energy computed tomography (DECT).

The distribution and good spreading of adhesive resins is critical for the wood-based panels industry. Full 3D non-destructive characterization is necessary, but methods are limited due to the chemical similarities between the resins and the wood fibers. For X-ray microtomography ([Formula: see text]CT), the doping of the resin with a highly attenuating contrast agent is necessary to visualize the resin distribution. However, the attenuation signal remains hard to segment clearly due to partial volume effects in the image, and phase mixing in the material. To help in the identification of the doped resin, dual-energy X-ray CT (DECT) is used to exploit the contrast agent's K-edge, based on simulations which take into account the polychromatic properties of the X-ray tube and detector response. The contrast agent's identification with DECT is validated with elemental mapping using scanning electron microscopy combined with energy-dispersive spectroscopy (SEM-EDX) on the surface of a wood-based panel sample, using data fusion between DECT and SEM-EDX. Overall, DECT results here in the first 3D identification of doped resin inside wood fiberboards, guiding the industry's efforts in further improving the durability of wood-based panels.

Moisture Dynamics of Wood-Based Panels and Wood Fibre Insulation Materials.

Moisture performance is an important factor determining the resistance of wood-based building materials against fungal decay. Understanding how material porosity and chemistry affect moisture performance is necessary for their efficient use, as well as for product optimisation. In this study, three complementary techniques (X-ray computed tomography, infrared and low-field NMR spectroscopy) are applied to elucidate the influence of additives, manufacturing process and material structure on the liquid water absorption and desorption behaviour of a selection of wood-based panels, thermally modified wood and wood fibre insulation materials. Hydrophobic properties achieved by thermal treatment or hydrophobic additives such as paraffin and bitumen, had a major influence on water absorption and desorption rates. When hydrophobic additives did not play a role, pore distributions and manufacturing process had a decisive influence on the amount and rate of absorption and desorption. In that case, a higher porosity resulted in a higher water absorption rate. Our results show that there is a clear potential for tailoring materials towards specific moisture performance by better understanding the influence of different material characteristics. This is useful both for achieving desired moisture buffering as well as to increase service life of wood-based materials. From a sustainability perspective, fit-for-purpose moisture performance is often easier to achieve and preferred than wood protection by biocide preservative treatments.

Improved wood species identification based on multi-view imagery of the three anatomical planes.

BACKGROUND: The identification of tropical African wood species based on microscopic imagery is a challenging problem due to the heterogeneous nature of the composition of wood combined with the vast number of candidate species. Image classification methods that rely on machine learning can facilitate this identification, provided that sufficient training material is available. Despite the fact that the three main anatomical sections contain information that is relevant for species identification, current methods only rely on transverse sections. Additionally, commonly used procedures for evaluating the performance of these methods neglect the fact that multiple images often originate from the same tree, leading to an overly optimistic estimate of the performance. RESULTS: We introduce a new image dataset containing microscopic images of the three main anatomical sections of 77 Congolese wood species. A dedicated multi-view image classification method is developed and obtains an accuracy (computed using the naive but common approach) of 95%, outperforming the single-view methods by a large margin. An in-depth analysis shows that naive accuracy estimates can lead to a dramatic over-prediction, of up to 60%, of the accuracy. CONCLUSIONS: Additional images from non-transverse sections can boost the performance of machine-learning-based wood species identification methods. Additionally, care should be taken when evaluating the performance of machine-learning-based wood species identification methods to avoid an overestimation of the performance.

High-resolution proxies for wood density variations in Terminalia superba.

BACKGROUND AND AIMS: Density is a crucial variable in forest and wood science and is evaluated by a multitude of methods. Direct gravimetric methods are mostly destructive and time-consuming. Therefore, faster and semi- to non-destructive indirect methods have been developed. METHODS: Profiles of wood density variations with a resolution of approx. 50 µm were derived from one-dimensional resistance drillings, two-dimensional neutron scans, and three-dimensional neutron and X-ray scans. All methods were applied on Terminalia superba Engl. & Diels, an African pioneer species which sometimes exhibits a brown heart (limba noir). KEY RESULTS: The use of X-ray tomography combined with a reference material permitted direct estimates of wood density. These X-ray-derived densities overestimated gravimetrically determined densities non-significantly and showed high correlation (linear regression, R(2) = 0·995). When comparing X-ray densities with the attenuation coefficients of neutron scans and the amplitude of drilling resistance, a significant linear relation was found with the neutron attenuation coefficient (R(2) = 0·986) yet a weak relation with drilling resistance (R(2) = 0·243). When density patterns are compared, all three methods are capable of revealing the same trends. Differences are mainly due to the orientation of tree rings and the different characteristics of the indirect methods. CONCLUSIONS: High-resolution X-ray computed tomography is a promising technique for research on wood cores and will be explored further on other temperate and tropical species. Further study on limba noir is necessary to reveal the causes of density variations and to determine how resistance drillings can be further refined.

Toxic hazard of leachates from furfurylated wood: comparison between two different aquatic organisms.

Environmental concern regarding the use of toxic preservatives such as chromated copper arsenate (CCA) has been put forward. In the European Union, United States, and Japan, CCA has been phased out for residential and water-contact applications. Ecotoxicological studies of wood treated with conventional preservatives were carried out in the late 1990s, and it was concluded that the main impact is to water and aquatic organisms. Today, alternatives to conventional preservation methods, marketed as "environmentally friendly" or "nontoxic," are emerging. Examples of such alternatives are modified wood, e.g., thermally modified, furfurylated, and acetylated wood. To date, not enough hazard characterization has been performed. In the present study, the Microtox assay with the marine bacterium Vibrio fischeri and the Daphtox procedure with the crustacean Daphnia magna were used as screening methods in an effect assessment. Both organisms were exposed to water leachates from furfurylated wood using two different leaching procedures. The results indicate that Microtox is more sensitive to the toxic components from furfurylated wood than Daphtox. Furthermore, the toxicity of treated Pinus radiata was higher than that of treated Pinus sylvestris. The toxicity did not diminish over the test period, as is the case for preservative-treated wood. The present study found that treatment conditions can influence the toxicity considerably, so toxicity studies should be included in the development of new treatment process. The present study also shows that using an intermediate vacuum-drying step, leading to a more efficient curing/polymerization, results in slightly less hydrophobic oligomers in the product, such that the leachates become less toxic to bacteria.

Three-dimensional x-ray imaging and analysis of fungi on and in wood.

As wood is prone to fungal degradation, fundamental research is necessary to increase our knowledge aiming at product improvement. Several imaging modalities are capable of visualizing fungi, but the X-ray equipment presented in this article can envisage fungal mycelium in wood nondestructively in three dimensions with submicron resolution. Four types of wood subjected to the action of the white rot fungus Coriolus versicolor (Linnaeus) Quélet (CTB 863 A) were scanned using an X-ray-based approach. Comparison of wood volumes before and after fungal exposure, segmented manually or semiautomatically, showed the presence of the fungal mass on and in the wood samples and therefore demonstrated the usefulness of computed X-ray tomography for mycological and wood research. Further improvements to the experimental setup are necessary to resolve individual hyphae and enhance segmentation.

Distinct growth responses to drought for oak and beech in temperate mixed forests.

PURPOSE: Droughts are expected to become more intense and frequent. Mixed forests can be more resilient to extreme events, but are the individual trees in mixed forests also more resilient to drought? METHODS: We sampled 275 trees in 53 temperate forest stands in northern Belgium: monocultures, two-species mixtures, and the three-species mixture of Fagus sylvatica, Quercus robur, and Q. rubra. We related the annual basal area increment of individual trees to drought severity from 1955 to 2015 and calculated growth resistance, recovery, and resilience for six contrasting drought episodes (spring, summer, or full-year drought). RESULTS: Tree growth of the diffuse-porous F. sylvatica was more sensitive to drought, summer drought in particular. The ring-porous Q. robur and Q. rubra were mainly affected by spring drought. In general, a tree's growth response to drought was not affected by tree species diversity, but some identity effects emerged. CONCLUSION: The asynchrony in drought responses among the tree species (a large and immediate decrease in growth followed by swift recovery in F. sylvatica vs a smaller delayed response in Quercus) might stabilize productivity in forests in which both are present. The impact of the predicted increasing drought frequency will depend on the timing of the droughts (spring vs summer).

The stability enigma of hydraulic vulnerability curves: addressing the link between hydraulic conductivity and drought-induced embolism.

Maintaining xylem water transport under drought is vital for plants, but xylem failure does occur when drought-induced embolisms form and progressively spread through the xylem. The hydraulic method is widely considered the gold standard to quantify drought-induced xylem embolism. The method determines hydraulic conductivity (Kh) in cut branch samples, dehydrated to specific drought levels, by pushing water through them. The technique is widely considered for its reliable Kh measurements, but there is some uncertainty in the literature over how to define stable Kh and how that relates to the degree of xylem embolism formation. Therefore, the most common setup for this method was extended to measure four parameters: (i) inlet Kh, (ii) outlet Kh, (iii) radial flow from xylem to surrounding living tissue and (iv) the pressure difference across the sample. From a strictly theoretical viewpoint, hydraulic steady state, where inflow equals outflow and radial flow is zero, will result in stable Kh. Application of the setup to Malus domestica Borkh. branches showed that achieving hydraulic steady state takes considerable time (up to 300 min) and that time to reach steady state increased with declining xylem water potentials. During each experimental run, Kh and xylem water potentials dynamically increased, which was supported by X-ray computed microtomography visualizations of embolism refilling under both high- (8 kPa) and low-pressure (2 kPa) heads. Supplying pressurized water can hence cause artificial refilling of vessels, which makes it difficult to achieve a truly stable Kh in partially embolized xylem.

Analysis of spatio-temporal fungal growth dynamics under different environmental conditions.

Traditionally, fungal growth dynamics were assessed manually, limiting the research to a few environmental conditions and/or fungal species. Fortunately, more automated ways of measurement are gaining momentum due to the availability of cheap imaging and processing equipment and the development of dedicated image analysis algorithms. In this paper, we use image analysis to assess the impact of environmental conditions on the growth dynamics of two economically important fungal species, Coniophora puteana and Rhizoctonia solani. Sixteen environmental conditions combining four temperatures (15, 20, 25 and 30 °C) and four relative humidity (RH) conditions (65, 70, 75 and 80% RH) were tested. Fungal growth characteristics were extracted from images of the growing fungi, taken at regular points in time. Advanced time series analysis was applied to quantitatively compare the effect of the environmental conditions on these growth characteristics. The evolution of the mycelial area and the number of tips over time resulted in typical sigmoidal growth curves. Other growth characteristics such as the mean hyphal segment length did not vary significantly over time. Temperature and RH usually had a combined effect on the growth dynamics of the mycelial area and the number of tips. When defining optimal growth conditions for a fungus, it is therefore of primordial importance that the effect of temperature and RH is assessed simultaneously. At the most extreme conditions we tested, the mycelium most probably experienced water stress when developing over the inert Petri dish surface. An RH of 65% (independent of temperature) for C. puteana and a temperature of 30 °C (independent of RH) for both C. puteana and R. solani therefore always resulted in limited fungal growth, while the optimal growing conditions were at 20 °C and 75% RH and at 25 °C and 80% RH for R. solani and at 20 °C and 75% RH for C. puteana. The method applied in this study offers an updated and broader alternative to classical and narrowly focused studies on fungal growth dynamics, and is well suited to efficiently assess the effect of environmental conditions on fungal growth.