Comparative Stem Transcriptome Analysis Reveals Pathways Associated with Drought Tolerance in Maritime Pine Grafts.

The maritime pine (Pinus pinaster Ait.) is a highly valuable Mediterranean conifer. However, recurrent drought events threaten its propagation and conservation. P. pinaster populations exhibit remarkable differences in drought tolerance. To explore these differences, we analyzed stem transcriptional profiles of grafts combining genotypes with contrasting drought responses under well-watered and water-stress regimes. Our analysis underscored that P. pinaster drought tolerance is mainly associated with constitutively expressed genes, which vary based on genotype provenance. However, we identified key genes encoding proteins involved in water stress response, abscisic acid signaling, and growth control including a PHD chromatin regulator, a histone deubiquitinase, the ABI5-binding protein 3, and transcription factors from Myb-related, DOF NAC and LHY families. Additionally, we identified that drought-tolerant rootstock could enhance the drought tolerance of sensitive scions by regulating the accumulation of transcripts involved in carbon mobilization, osmolyte biosynthesis, flavonoid and terpenoid metabolism, and reactive oxygen species scavenging. These included genes encoding galactinol synthase, CBL-interacting serine/threonine protein kinase 5, BEL1-like homeodomain protein, dihydroflavonol 4-reductase, and 1-deoxy-D-xylulose-5-phosphate. Our results revealed several hub genes that could help us to understand the molecular and physiological response to drought of conifers. Based on all the above, grafting with selected drought-tolerant rootstocks is a promising method for propagating elite recalcitrant conifer species, such as P. pinaster.

Monitoring Eastern White Pine Health by Using Field-Measured Foliar Traits and Hyperspectral Data.

Canopy foliar traits serve as crucial indicators of plant health and productivity, forming a vital link between plant conditions and ecosystem dynamics. In this study, the use of hyperspectral data and foliar traits for white pine needle damage (WPND) detection was investigated for the first time. Eastern White Pine (Pinus strobus L., EWP), a species of ecological and economic significance in the Northeastern USA, faces a growing threat from WPND. We used field-measured leaf traits and hyperspectral remote sensing data using parametric and non-parametric methods for WPND detection in the green stage. Results indicated that the random forest (RF) model based solely on remotely sensed spectral vegetation indices (SVIs) demonstrated the highest accuracy of nearly 87% and Kappa coefficient (K) of 0.68 for disease classification into asymptomatic and symptomatic classes. The combination of field-measured traits and remote sensing data indicated an overall accuracy of 77% with a Kappa coefficient (K) of 0.46. These findings contribute valuable insights and highlight the potential of both field-derived foliar and remote sensing data for WPND detection in EWP. With an exponential rise in forest pests and pathogens in recent years, remote sensing techniques can prove beneficial for the timely and accurate detection of disease and improved forest management practices.

Thinning followed by slash burning enhances growth and reduces vulnerability to drought for Pinus nigra.

Increasingly frequent severe drought events are pushing Mediterranean forests to unprecedented responses. Lack of management leads to dense forests that are highly susceptible to drought stress, potentially resulting in extensive dieback and increased vulnerability to other disturbances. Forest treatments like thinning and slash burning reduce competition for resources and have the potential to enhance tree growth and vigor and minimize tree vulnerability to drought. Here, we used tree rings to study the growth and physiological response of black pine (Pinus nigra) to drought in northeastern Spain under different treatments, including two thinning intensities (light and heavy, with 10% and 40% basal area reduction, respectively) followed by two understory treatments (clearing alone and in combination with slash burning), resulting in a research design of four treatments plus an untreated control with three replicates. Specifically, we studied basal area increment (BAI), resilience indices, and intrinsic water use efficiency (iWUE) using carbon and oxygen isotope composition (δ13C and δ18O in tree-ring cellulose) before and after treatments. Our results showed that BAI and resistance to drought increased in the heavy-thin (burned and unburned) and light-thin burned units. Resilience increased in the burned units regardless of the thinning intensity, while recovery was not affected by treatment. Slash burning additionally increased BAI in the light-thin and resistance and resilience in the heavy-thin units compared with clearing alone. The stable isotope analysis revealed a minor effect of treatments on δ13C and δ18O. No change in iWUE among treatments was presumably linked to a proportional increase in both net CO2 assimilation and stomatal conductance, which particularly increased in the heavy-thin (burned and unburned) and light-thin burned units, indicating that these trees were the least affected by drought. This study shows that management approaches aimed at reducing wildfire hazard can also increase the vigor of dominant trees under drought stress. By reducing competition both from the overstory and the understory, thinning followed by clearing alone or in combination with slash burning promotes tree growth and vigor and increases its resistance and resilience to drought.

Structure of Ground Vegetation and Natural Regeneration of Tree Species in 12- to 15-Year-Old Bilberry Pine Forest-Clear-cut Complex of Middle Taiga Subzone.

Logging in mature stands where part of the forest is harvested in one or several cuts and part is retained (clearcutting and alternate strip cutting) results in the formation of an ecotone complex (EC), which includes a forest (F) zone, a forest edge (FE) as a transition from the forest to the clear-cut under the canopy, a clear-cut edge (CE) as a transition from the forest to the clear-cut outside of the canopy, and the clear-cut proper (C). The composition and structure of ground vegetation and natural regeneration of woody species (Pinus sylvestris L., Picea abies (L.) H. Karst., Betula sp., Populus tremula L., Sorbus aucuparia L., and Juniperus communis L.) were studied in a bilberry pine forest-clear-cut ecotone complex 12-15 years after stand removal. Specific structural features of ground vegetation and undergrowth (including tree regeneration) were observed in each of the four zones of the ecotone complex formed after logging of the mature forest. A typical forest habitat (zone F) showed a minimum number of young regeneration of Pinus sylvestris, Picea abies, Betula sp., Populus tremula, and Sorbus aucuparia and the highest abundance of the lingonberry V. vitis-idaea L. and bilberry Vaccinium myrtillus L. with a maximum height and a maximum yield of bilberry plants. The amount of tree regeneration in the FE zone was much the same as in the F zone. The projective cover, maximum shoot height, and yield of bilberry and the maximum shoot height of lingonberry in the FE zone were significantly lower than in the F zone. The transitional zone on the clear-cut side (CE) and the clear-cut proper (C) strikingly differed from the forest (F and FE) zones of the ecotone complex by a greater number of deciduous and pine regeneration and a low abundance of dwarf shrubs. The clear-cut proper (C) differed from the CE zone by a higher abundance of grasses and forbs and an established tree regeneration layer composed of pine, birch, and aspen.

The genetic architecture of repeated local adaptation to climate in distantly related plants.

Closely related species often use the same genes to adapt to similar environments. However, we know little about why such genes possess increased adaptive potential and whether this is conserved across deeper evolutionary lineages. Adaptation to climate presents a natural laboratory to test these ideas, as even distantly related species must contend with similar stresses. Here, we re-analyse genomic data from thousands of individuals from 25 plant species as diverged as lodgepole pine and Arabidopsis (~300 Myr). We test for genetic repeatability based on within-species associations between allele frequencies in genes and variation in 21 climate variables. Our results demonstrate significant statistical evidence for genetic repeatability across deep time that is not expected under randomness, identifying a suite of 108 gene families (orthogroups) and gene functions that repeatedly drive local adaptation to climate. This set includes many orthogroups with well-known functions in abiotic stress response. Using gene co-expression networks to quantify pleiotropy, we find that orthogroups with stronger evidence for repeatability exhibit greater network centrality and broader expression across tissues (higher pleiotropy), contrary to the 'cost of complexity' theory. These gene families may be important in helping wild and crop species cope with future climate change, representing important candidates for future study.

Watering the trees for the forest: Drought alleviation in oaks and pines by ancestral ditches.

Traditional ditches ("acequias" in Spanish) derive meltwater and infiltrate groundwater providing ecological services downstream in the semi-arid Sierra Nevada range (SE Spain). Therefore, they may act as a nature-based solution by alleviating drought stress in trees growing near ditches by enhancing growth and reducing their intrinsic water-use efficiency (iWUE). Such a mitigation role of acequias is critical given that some oak (Quercus pyrenaica) and pine (Pinus sylvestris) stands reach their xeric distribution limits in Europe. We compared tree-ring width data and wood δ13C, a proxy of iWUE, in oak and pine stands located near or far (control) from ditches with different infiltration capacity in two watersheds. We assessed how trees responded to climate data, drought stress, and vegetation greenness through correlations and resilience indices. Oak trees located near ditches grew more and responded less to precipitation, soil moisture, a drought index, and greenness than control trees. In pines, we did not find this pattern, and ditch trees grew more than control trees only during an extremely dry year (1995). Climate-growth correlations suggested a longer growing season in ditch pines. Growth of ditch oaks from the "Acequia Nueva" (AN), with high infiltration capacity, responded more to autumn soil moisture and showed the lowest δ13C. Growth was enhanced by cool-wet spring conditions in pines and also by warm-wet conditions in the prior winter in the case of oaks. Control trees showed lower resistance to drought. Control trees presented higher wood δ13C values except for old oaks from the "Acequia Grande" (AG) site which may show long-term acclimation. Traditional ditches alleviate drought stress in oak and pine stands subjected to regional xeric climate conditions.

Optimistic growth of marginal region plantations under climate warming: Assessing divergent drought resilience.

Given the context of significant global warming and the intensification of extreme climate events in the last century, large-scale reforestation and afforestation have been recognized as effective strategies to mitigate the climate crisis. Since the 1970s, China has launched several afforestation programs aimed at regional ecological protection, playing an important role in reaching carbon neutrality by 2060. This study provided a detailed analysis of the growth suitability of the main planted conifers (Pinus sylvestris var. mongolica and Pinus tabulaeformis) and broadleaves (Populus spp., Robinia pseudoacacia) in the semi-arid northern China. We compared the radial growth trends of plantations and their responses to extreme droughts from 1980 to 2018. Growth of most plantations has significantly increased over time, but broadleaves showed recent growth reductions in the past decade, which may be related to tree age and reduced soil moisture. Nevertheless, under warmer climate scenarios, the growth of plantations is forecasted to continue increasing. Broadleaves showed a better post-drought recovery, probably linked to their anisohydric behavior, than conifers, which presented a better resistance to drought. Growth of conifers depended more on warmer temperature and better precipitation conditions during the growing season, whereas broadleaves mainly reacted to warm temperature. Additionally, pre-drought growth levels weakened resilience components, while post-drought precipitation compensated for drought-induced growth deficit. Growth and resilience were negatively related to tree age, while higher stand density reduced growth. This assessment and projections of growth and drought resilience indicate the sustainability of most plantations in semi-arid regions, but future warmer and drier conditions may lead to an uncertain future regarding forest health and reduce their carbon sink potential.

Hydrothermal environments lead to differences in the radial growth response of Pinus tabulaeformis to climate in the monsoon marginal zone, Northwestern China.

Regional climatic differences increase the complexity of tree radial growth responses to climate change in the monsoon marginal zones and may alter the carbon sequestration capacity of forests. In this study, we collected cores of Pinus tabulaeformis trees at nine sampling sites across different regions. We analysed the relationship between tree-ring width chronology and climatic factors at different sites using dendroecological methods. We used the tree-ring index to calculate resistance, recovery, and resilience as well as to explore the capacity of radial growth to cope with drought events. The results indicate that (1) Drought was the primary factor limiting tree growth, and tree-ring climate response patterns varied across three regions. Tree growth was sensitive to both temperature and precipitation in the eastern Qilian Mountains, while it was more sensitive to temperature in the Hassan Mountains and more sensitive to precipitation in the Helan Mountains. (2) The tree-ring climate response pattern remained unstable over time, and the relative influence of current climate on tree growth increased. (3) The ecological resilience of trees to extreme events varies across three regions, which could be attributed to regional moisture conditions and the duration of drought. In the context of the management and protection of trees in the study area in the future, more attention should be paid to the elasticity of tree growth after drought events.

Trunk distortion weakens the tree productivity revealed by half-sib progeny determination of Pinus yunnanensis.

Twisted trunks are not uncommon in trees, but their effects on tree growth are still unclear. Among coniferous tree species, the phenomenon of trunk distortion is more prominent in Pinus yunnanensis. To expand the germplasm of genetic resources, we selected families with excellent phenotypic traits to provide material for advanced generation breeding. The progeny test containing 93 superior families (3240 trees) was used as the research material. Phenotypic measurements and estimated genetic parameters (family heritability, realistic gain and genetic gain) were performed at 9, 15, and 18 years of age, respectively. The genetic evaluation yielded the following results (1) The intra-family variance component of plant height (PH) was greater than that of the inter-family, while the inter-family variance components of other traits (diameter at breast height (DBH), crown diameter (CD), height under branches (HUB), degree of stem-straightness (DS)) were greater than that of the intra-family, indicating that there was abundant variation among families and potential for selection. (2) At half rotation period (18 years old), there was a significant correlation among the traits. The proportion of trees with twisted trunks (level 1-3 straightness) reached 48%. The DS significantly affected growth traits, among which PH and DBH were the most affected. The volume loss rate caused by twisted trunk was 18.06-56.75%, implying that trunk distortion could not be completely eliminated after an artificial selection. (3) The influence of tree shape, crown width, and trunk on volume increased, and the early-late correlation between PH, DBH and volume was extremely significant. The range of phenotypic coefficient of variation, genetic variation coefficient and family heritability of growth traits (PH, DBH, and volume) were 44.29-127.13%, 22.88-60.87%, and 0.79-0.83, respectively. (4) A total of 21 superior families were selected by the method of membership function combined with independent selection. Compared with the mid-term selection (18 years old), the accuracy of early selection (9 years old) reached 77.5%. The selected families' genetic gain and realistic gain range were 5.79-19.82% and 7.12-24.27%, respectively. This study can provide some useful reference for the breeding of coniferous species.

PmHs1pro-1 monitors Bsursaphelenchus xylophilus infection and activates defensive response in resistant Pinus massoniana.

Plant resistance (R) genes play a crucial role in the detection of effector proteins secreted by pathogens, either directly or indirectly, as well as in the subsequent activation of downstream defence mechanisms. However, little is known about how R genes regulate the defence responses of conifers, particularly Pinus massoniana, against the destructive pine wood nematode (PWN; Bursaphelenchus xylophilus). Here, we isolated and characterised PmHs1pro-1, a nematode-resistance gene of P. massoniana, using bioinformatics, molecular biology, histochemistry and transgenesis. Tissue-specific expressional pattern and localisation of PmHs1pro-1 suggested that it was a crucial positive regulator in response to PWN attack in resistant P. massoniana. Meanwhile, overexpression of PmHs1pro-1 was found to activate reactive oxygen species (ROS) metabolism-related enzymes and the expressional level of their key genes, including superoxide dismutase, peroxidase and catalase. In addition, we showed that PmHs1pro-1 directly recognised the effector protein BxSCD1of PWN, and induced the ROS burst responding to PWN invasion in resistant P. massoniana. Our findings illustrated the molecular framework of R genes directly recognising the effector protein of pathology in pine, which offered a novel insight into the plant-pathogen arms race.

Exploring behavioural and physiological adaptations in mountain pine beetle in response to elevated ozone concentrations.

Elevated ozone (eO3) concentrations pose a threat to insect populations by potentially altering their behaviour and physiology. This study investigates the effects of eO3 concentrations on the mountain pine beetle which is a major tree-killing species of conifers in northwestern North America. We are particularly interested in understanding the effects of eO3 concentrations on beetle behaviour and physiology and possible transgenerational impacts on bark beetle broods. We conducted O3-enrichment experiments in a controlled laboratory setting using different O3 concentrations (100-200 ppb; projected for 2050-2100) and assessed various beetle responses, including CO2 respiration, mating behaviour, survival probability, locomotion, and attraction behaviour. Transgenerational impacts on the first and second generations were also analyzed by studying brood morphology, mating behaviour, survival, and pheromone production. We found that beetles exposed to eO3 concentrations had shorter oviposition galleries and reduced brood production. Beetle pheromones were also degraded by eO3 exposure. However, exposure to eO3 also prompted various adaptive responses in beetles. Despite reduced respiration, eO3 improved locomotor activity and the olfactory response of beetles. Surprisingly, beetle survival probability was also improved both in the parents and their broods. We also observed transgenerational plasticity in the broods of eO3-exposed parents, suggesting potential stress resistance mechanisms. This was evident by similar mating success, oviposition gallery length, and brood numbers produced in both control and eO3 concentration treatments. This study demonstrates the sensitivity of mountain pine beetles to increased O3 concentrations, contributing crucial insights into the ecological implications of eO3 concentrations on their populations. Overall, the outcome of this study contributes to informed climate change mitigation strategies and adaptive management practices for the development of resilient forests in response to emerging forest insect pests worldwide.

Tree rings reveal the transient risk of extinction hidden inside climate envelope forecasts.

Given the importance of climate in shaping species' geographic distributions, climate change poses an existential threat to biodiversity. Climate envelope modeling, the predominant approach used to quantify this threat, presumes that individuals in populations respond to climate variability and change according to species-level responses inferred from spatial occurrence data-such that individuals at the cool edge of a species' distribution should benefit from warming (the "leading edge"), whereas individuals at the warm edge should suffer (the "trailing edge"). Using 1,558 tree-ring time series of an aridland pine (Pinus edulis) collected at 977 locations across the species' distribution, we found that trees everywhere grow less in warmer-than-average and drier-than-average years. Ubiquitous negative temperature sensitivity indicates that individuals across the entire distribution should suffer with warming-the entire distribution is a trailing edge. Species-level responses to spatial climate variation are opposite in sign to individual-scale responses to time-varying climate for approximately half the species' distribution with respect to temperature and the majority of the species' distribution with respect to precipitation. These findings, added to evidence from the literature for scale-dependent climate responses in hundreds of species, suggest that correlative, equilibrium-based range forecasts may fail to accurately represent how individuals in populations will be impacted by changing climate. A scale-dependent view of the impact of climate change on biodiversity highlights the transient risk of extinction hidden inside climate envelope forecasts and the importance of evolution in rescuing species from extinction whenever local climate variability and change exceeds individual-scale climate tolerances.

Factors limiting the potential range expansion of lodgepole pine in Interior Alaska.

Understanding the factors influencing species range limits is increasingly crucial in anticipating migrations due to human-caused climate change. In the boreal biome, ongoing climate change and the associated increases in the rate, size, and severity of disturbances may alter the distributions of boreal tree species. Notably, Interior Alaska lacks native pine, a biogeographical anomaly that carries implications for ecosystem structure and function. The current range of lodgepole pine (Pinus contorta var. latifolia) in the adjacent Yukon Territory may expand into Interior Alaska, particularly with human assistance. Evaluating the potential for pine expansion in Alaska requires testing constraints on range limits such as dispersal limitations, environmental tolerance limits, and positive or negative biotic interactions. In this study, we used field experiments with pine seeds and transplanted seedlings, complemented by model simulations, to assess the abiotic and biotic factors influencing lodgepole pine seedling establishment and growth after fire in Interior Alaska. We found that pine could successfully recruit, survive, grow, and reproduce across our broadly distributed network of experimental sites. Our results show that both mammalian herbivory and competition from native tree species are unlikely to constrain pine growth and that environmental conditions commonly found in Interior Alaska fall well within the tolerance limits for pine. If dispersal constraints are released, lodgepole pine could have a geographically expansive range in Alaska, and once established, its growth is sufficient to support pine-dominated stands. Given the impacts of lodgepole pine on ecosystem processes such as increases in timber production, carbon sequestration, landscape flammability, and reduced forage quality, natural or human-assisted migration of this species is likely to substantially alter responses of Alaskan forest ecosystems to climate change.

Carbon fluxes and water-use efficiency in a Pinus tabuliformis plantation in Northeast China and their relationship to drought.

The impact of extreme weather events on carbon fluxes and water-use efficiency (WUE) in revegetated areas under water-limited conditions is poorly understood. We analyzed changes in carbon fluxes and WUE over three years of eddy-covariance measurements in a Pinus tabuliformis plantation in Northeast China to investigate carbon fluxes and WUE responses to drought events at different time scales. Mean annual net ecosystem exchange (NEE), gross primary production (GPP), and ecosystem respiration (Re) were -368.48, 1042.42, and 673.94 g C m-2, respectively. Drought events increased NEE, as GPP was more sensitive to water stress than Re at different growing stages. Mean annual WUE was 2.46 g C kg-1 H2O, and plant phenology played a key role in WUE responses to drought. Water stress had negative and positive effects on daily WUE at the early and late growing stages, respectively, and daily WUE was generally insensitive to drought at the mid growing stage. A lagged effect existed in the carbon fluxes and WUE dynamics after drought events at various time scales. Water stress at the early growing stage was more important than that at other growing stages on annual carbon sequestration and WUE, as it dominated canopy growth in the current year. The annual mean normalized difference vegetation index controlled interannual variations in carbon fluxes and WUE in the plantation. Our findings contribute to the prediction of possible changes in carbon and water fluxes under climate warming in the afforested areas of Northeast China.

Diverging growth trends and climate sensitivities of individual pine trees after the 1976 extreme drought.

Summer droughts are affecting the productivity and functioning of central European forests, with potentially lasting consequences for species composition and carbon sequestration. Long-term recovery rates and individual growth responses that may diverge from species-specific and population-wide behaviour are, however, poorly understood. Here, we present 2052 pine (Pinus sylvestris) ring width series from 19 forest sites in south-west Germany to investigate growth responses of individual trees to the exceptionally hot and dry summer of 1976. This outstanding drought event presents a distinctive test case to examine long-term post-drought recovery dynamics. We have proposed a new classification approach to identify a distinct sub-population of trees, referred to as "temporarily affected trees", with a prevalence ranging from 9 to 33 % across the forest stands. These trees exhibited an exceptionally prolonged growth suppression, lasting over a decade, indicating significantly lower resilience to the 1976 drought and a 50 % reduced capacity to recover to pre-drought states. Furthermore, shifts in resilience and recovery dynamics are accompanied by changing climate sensitivities, notably an increased response to maximum temperatures and summer droughts in post-1976 affected pines. Our findings underscore the likely interplay between individual factors and micro-site conditions that contribute to divergent tree responses to droughts. Assessing these factors at the individual tree level is recommended to advancing our understanding of forest responses to extreme drought events. By analyzing sub-population growth patterns, our study provides valuable insights into the impacts of summer droughts on central European forests in context of increasing drought events.

Context-dependent benefits of forest soil addition on Aleppo pine seedling performance under drought and grass competition.

Seedling establishment under natural conditions is limited by numerous interacting factors. Here, we tested the combined effects of drought, herbaceous competition, and ectomycorrhizal inoculation on the performance of Aleppo pine seedlings grown in a net-house. The roots of all pine seedlings were strongly dominated by Geopora, a fungal genus known to colonize seedlings in dry habitats. Ectomycorrhizal fungi (EMF) inoculum significantly increased seedling height, biomass, and the number of side branches. However, under either competition or drought, the positive effect of EMF on seedling biomass and height was greatly reduced, while the effect on shoot branching was maintained. Further, under a combination of drought and competition, EMF had no influence on either plant growth or shape. The discrepancy in pine performance across treatments highlights the complexity of benefits provided to seedlings by EMF under ecologically relevant settings.

Pathophysiology and transcriptomic responses of Pinus armandii defenses to ophiostomatoid fungi.

Pinus armandii Franch. is an ecologically and economically important evergreen tree species native to western China. Dendroctonus armandi Tsai and Li and pathogenic ophiostomatoid fungi pose substantial threats to P. armandii. With the interplay between species, the defense mechanisms of P. armandii have evolved to withstand external biotic stressors. However, the interactions between P. armandii and pathogenic ophiostomatoid fungal species/strains remain poorly understood. We aimed to analyze the pathophysiological and molecular changes in P. armandii following artificial inoculation with four ophiostomatoid species (Graphilbum parakesiyea, Leptographium qinlingense, Ophiostoma shennongense and Ophiostoma sp. 1). The study revealed that L. qinlingense produced the longest necrotic lesions, and G. parakesiyea produced the shortest. All strains induced monoterpenoid release, and monoterpene levels of P. armandii were positively correlated with fungal virulence (R2 = 0.93, P < 0.01). Co-inoculation of two dominant highly (L. qinlingense) and weakly virulent (O. shennongense) pathogens reduced the pathogenicity of the highly virulent fungi. Transcriptomic analysis of P. armandii (LQ: L. qinlingense treatments, QS: co-inoculation treatments and OS: O. shennongense treatments) showed that the expression pattern of differentially expressed genes (DEGs) between QS and OS was similar, but different from that of LQ. The DEGs (LQ vs QS) involved in flavonoid biosynthesis and phenylpropanoid biosynthesis were downregulated. Notably, compared with LQ, QS significantly decreased the expression of host defense-related genes. This study provides a valuable theoretical basis for managing infestations of D. armandi and associated ophiostomatoid fungi.

Isohydricity and hydraulic isolation explain reduced hydraulic failure risk in an experimental tree species mixture.

Species mixture is promoted as a crucial management option to adapt forests to climate change. However, there is little consensus on how tree diversity affects tree water stress, and the underlying mechanisms remain elusive. By using a greenhouse experiment and a soil-plant-atmosphere hydraulic model, we explored whether and why mixing the isohydric Aleppo pine (Pinus halepensis, drought avoidant) and the anisohydric holm oak (Quercus ilex, drought tolerant) affects tree water stress during extreme drought. Our experiment showed that the intimate mixture strongly alleviated Q. ilex water stress while it marginally impacted P. halepensis water stress. Three mechanistic explanations for this pattern are supported by our modeling analysis. First, the difference in stomatal regulation between species allowed Q. ilex trees to benefit from additional soil water in mixture, thereby maintaining higher water potentials and sustaining gas exchange. By contrast, P. halepensis exhibited earlier water stress and stomatal regulation. Second, P. halepensis trees showed stable water potential during drought, although soil water potential strongly decreased, even when grown in a mixture. Model simulations suggested that hydraulic isolation of the root from the soil associated with decreased leaf cuticular conductance was a plausible explanation for this pattern. Third, the higher predawn water potentials for a given soil water potential observed for Q. ilex in mixture can-according to model simulations-be explained by increased soil-to-root conductance, resulting from higher fine root length. This study brings insights into the mechanisms involved in improved drought resistance of mixed species forests.

Drought legacies in mixed Mediterranean forests: Analysing the effects of structural overshoot, functional traits and site factors.

Previous favorable climate conditions stimulate tree growth making some forests more vulnerable to hotter droughts. This so-called structural overshoot may contribute to forest dieback, but there is little evidence on its relative importance depending on site conditions and tree species because of limited field data. Here, we analyzed remote sensing (NDVI) and tree-ring width data to evaluate the impacts of the 2017 drought on canopy cover and growth in mixed Mediterranean forests (Fraxinus ornus, Quercus pubescens, Acer monspessulanum, Pinus pinaster) located in southern Italy. Legacy effects were assessed by calculating differences between observed and predicted basal area increment (BAI). Overall, the growth response of the study stands to the 2017 drought was contingent on site conditions and species characteristics. Most sites presented BAI and canopy cover reductions during the drought. Growth decline was followed by a quick recovery and positive legacy effects, particularly in the case of F. ornus. However, we found negative drought legacies in some species (e.g., Q. pubescens, A. monspessulanum) and sites. In those sites showing negative legacies, high growth rates prior to drought in response to previous wet winter-spring conditions may have predisposed trees to drought damage. Vice versa, the positive drought legacy found in some F. ornus site was linked to post-drought growth release due to Q. pubescens dieback and mortality. Therefore, we found evidences of structural drought overshoot, but it was restricted to specific sites and species. Our findings highlight the importance of considering site settings such as stand composition, pre-drought conditions and different tree species when studying structural overshoot. Droughts contribute to modify the composition and dynamics in mixed forests.

The response of physiological and xylem anatomical traits under cadmium stress in Pinus thunbergii seedlings.

Studying the response of physiological and xylem anatomical traits under cadmium stress is helpful to understand plants' response to heavy metal stress. Here, seedlings of Pinus thunbergii Parl. were treated with 50, 100 and 150 mg kg-1 Cd2+ for 28 days. Cadmium and nonstructural carbohydrate content of leaves, stems and roots, root Cd2+ flux, cadmium distribution pattern in stem xylem and phloem, stem xylem hydraulic traits, cell wall component fractions of stems and roots, phytohormonal content such as abscisic acid, gibberellic acid 3, molecule -indole-3-acetic acid, and jasmonic acid from both leaves and roots, as well as xylem anatomical traits from both stems and roots were measured. Root Cd2+ flux increased from 50 to 100 mmol L-1 Cd2+ stress, however it decreased at 150 mmol L-1 Cd2+. Cellulose and hemicellulose in leaves, stems and roots did not change significantly under cadmium stress, while pectin decreased significantly. The nonstructural carbohydrate content of both leaves and stems showed significant changes under cadmium stress while the root nonstructural carbohydrate content was not affected. In both leaves and roots, the abscisic acid content significantly increased under cadmium stress, while the gibberellic acid 3, indole-3-acetic acid and jasmonic acid methylester content significantly decreased. Both xylem specific hydraulic conductivity and xylem water potential decreased with cadmium stress, however tracheid diameter and double wall thickness of the stems and roots were not affected. High cadmium intensity was found in both the stem xylem and phloem in all cadmium stressed treatments. Our study highlighted the in situ observation of cadmium distribution in both the xylem and phloem, and demonstrated the instant response of physiological traits such as xylem water potential, xylem specific hydraulic conductivity, root Cd2+ flux, nonstructural carbohydrate content, as well as phytohormonal content under cadmium stress, and the less affected traits such as xylem anatomical traits, cellulose and hemicellulose.