ALBERT

Description: Plants from arid environments display covarying traits to survive or resist drought. Plant drought resistance and ability to survive long periods of low soil water availability should involve leaf phenology coordination with leaf and stem functional traits related to water status. This study tested correlations between phenology and functional traits involved in plant water status regulation in 10 Sonoran Desert tree species with contrasting phenology. Species seasonal variation in plant water status was defined by calculating their relative positions along the iso/anisohydric regulation continuum based on their hydroscape areas (HA)—a metric derived from the relationship between predawn and midday water potentials—and stomatal and hydraulic traits. Additionally, functional traits associated with plant water status regulation, including lamina vessel hydraulic diameter (DHL), stem-specific density (SSD), and leaf mass per area (LMA) were quantified per species. To characterize leaf phenology, leaf longevity (LL) and canopy foliage duration (FD) were determined. HA was strongly correlated with FD but not with LL; HA was significantly associated with SSD and leaf hydraulic traits (DHL, LMA) but not with stem hydraulic traits (vulnerability index, relative conductivity); and FD was strongly correlated with LMA and SSD. Leaf physiological characteristics affected leaf phenology when it was described as canopy foliage duration, better than when described as leaf longevity. Stem and leaf structure and hydraulic functions were not only relevant for categorizing species along the iso/anisohydric continuum but also allowed identifying different strategies of desert trees within the ‘fast–slow’ plant economics spectrum. The results in this study pinpoint the set of evolutionary pressures that shape the Sonoran Desert Scrub physiognomy.

Description: Drought negatively impacts growth and productivity of plants, particularly in arid and semi-arid regions. Although drought events can take place in summer and winter, differences on the impact of drought on physiological processes between seasons are largely unknown. The aim of this study was to elucidate metabolic strategies of date palms in response to drought in summer and winter season. To identify such differences, we exposed date palm seedlings to a drought-recovery regime, both in simulated summer and winter climate. Leaf hydration, carbon discrimination (∆13C), primary and secondary metabolite composition and contents were analyzed. Depending on season, drought differently affected physiological and biochemical traits of the leaves. In summer, drought induced significantly decreased leaf hydration, concentrations of ascorbate, most sugars, primary and secondary organic acids, as well as phenolic compounds, while thiol, amino acid, raffinose and individual fatty acid contents were increased compared to well-watered plants. In winter, drought had no effect on leaf hydration, ascorbate and fatty acids contents, but resulted in increased foliar thiol and amino acid levels as observed in summer. Compared to winter, foliar traits of plants exposed to drought in summer only partly recovered after re-watering. Memory effects on water relations, primary and secondary metabolites seem to prepare foliar traits of date palms for repeated drought events in summer. Apparently, a well-orchestrated metabolic network, including the anti-oxidative system, compatible solutes accumulation and osmotic adjustment, and maintenance of cell membrane stability strongly reduces the susceptibility of date palms to drought. These mechanisms of drought compensation may be more frequently required in summer.

Description: Predicting tree frost tolerance is critical to select adapted species according to both the current and predicted future climate. The relative change in water to carbohydrate ratio is a relevant trait to predict frost acclimation in branches from many tree species. The objective of this study is to demonstrate the interspecific genericity of this approach across nine tree species. In the studied angiosperm species, frost hardiness dynamics were best correlated to a decrease in water content at the early stage of acclimation (summer and early autumn). Subsequently, frost hardiness dynamics were more tightly correlated to soluble carbohydrate contents until spring growth resumption. Based on different model formalisms, we predicted frost hardiness at different clade levels (angiosperms, family, genus and species) with high to moderate accuracy (1.5–6.0°C RMSE) and robustness (2.8–6.1°C RMSEP). The TOT model, taking all soluble carbohydrate and polyols into account, was more effective and adapted for large scale studies aiming to explore frost hardiness across a wide range of species. The ISC model taking the individual contribution of each soluble carbohydrate molecule into account was more efficient at finer scale such as family or species. The ISC model performance also suggests that the role of solutes cannot be reduced to a 'bulk' osmotic effect as could be computed if all of them were located in a single, common, compartment. This study provides sets of parameters to predict frost hardiness in a wide range of species, and clues in targeting specific carbohydrate molecules to improve frost hardiness.

Description: Mastic trees (Pistacia lentiscus L.) are dioecious perennial plants highly adapted to Mediterranean climates but display a high sensitivity to winter periods. In order to understand how sex, leaf phenology and ecological context could condition sensitivity to winter and associated mechanisms to acclimate to these conditions, photoinhibition and photo-oxidative stress markers were examined in mastic trees (Pistacia lentiscus L.) from a natural population growing in the Garraf Natural Park for a consecutive 12-month period (seasonal study), as well as in three populations naturally growing in the Montseny Natural Park, including the highest altitudes described for this species, during winter (altitudinal study). Results from these studies indicate that both the winter period and higher elevation influenced the degree of photoinhibition, but this was not conditioned by sex. In fact, winter photoinhibition occurred transiently even though it was accompanied by enhanced chlorophyll loss and malondialdehyde contents. Stress acclimation was achieved through biochemical adjustments in chloroplasts, characterized by anthocyanin shielding, increased de-epoxidation state of the xanthophyll cycle as well as tocopherol accumulation, and phenological adaptations, the latter allowing a complete resetting of the physiological performance of leaves. Moreover, although females showed higher lipid peroxidation than males during the coldest winter months, at the highest elevation and during flowering in spring, this oxidative stress was mild and transient with no negative consequences for the physiology of plants. It is concluded that evergreen mastic trees acclimate to winter conditions and higher elevations by activation of antioxidant defenses together with phenological adjustments, altogether playing a crucial role in plant survival. Sexual dimorphism in mastic trees appears as a relevant factor when considering sensitivity to photo-oxidative stress in winter and altitudinal conditions.

Description: The continuously increasing atmospheric carbon dioxide concentration ([CO2]) has substantial effects on plant growth, and on the composition and structure of forests. However, how plants respond to elevated [CO2] (e[CO2]) under intra- and interspecific competition has been largely overlooked. In this study, we employed Abies faxoniana and Picea purpurea seedlings to explore the effects of e[CO2] (700 ppm) and plant–plant competition on plant growth, physiological and morphological traits, and leaf ultrastructure. We found that e[CO2] stimulated plant growth, photosynthesis and nonstructural carbohydrates (NSC), affected morphological traits and leaf ultrastructure, and enhanced water and nitrogen use efficiencies in A. faxoniana and P. purpurea. Under interspecific competition and e[CO2], P. purpurea showed a higher biomass accumulation, photosynthetic capacity and rate of ectomycorrhizal infection, and higher water and nitrogen use efficiencies compared with A. faxoniana. However, under intraspecific competition and e[CO2], the two conifers showed no differences in biomass accumulation, photosynthetic capacity, and water and nitrogen use efficiencies. In addition, under interspecific competition and e[CO2], A. faxoniana exhibited higher NSC levels in leaves as well as more frequent and greater starch granules, which may indicate carbohydrate limitation. Consequently, we concluded that under interspecific competition, P. purpurea possesses a positive growth and adjustment strategy (e.g., a higher photosynthetic capacity and rate of ectomycorrhizal infection, and higher water and nitrogen use efficiencies), while A. faxoniana likely suffers from carbohydrate limitation to cope with rising [CO2]. Our study highlights that plant–plant competition should be taken into consideration when assessing the impact of rising [CO2] on the plant growth and physiological performance.

Description: The Canary Islands, an archipelago east of Morocco's Atlantic coast, present steep altitudinal gradients covering various climatic zones from hot deserts to subalpine Mediterranean, passing through fog-influenced cloud forests. Unlike the majority of the Canarian flora, Pinus canariensis grow along most of these gradients, allowing the study of plant functioning in contrasting ecosystems. Here we assess the water sources (precipitation, fog) of P. canariensis and its physiological behavior in its different natural environments. We analyzed carbon and oxygen isotope ratios of water and organics from atmosphere, soil and different plant organs and tissues (including 10-year annual time series of tree-ring cellulose) of six sites from 480 to 1990 m asl on the Canary island La Palma. We found a decreasing δ18O trend in source water that was overridden by an increasing δ18O trend in needle water, leaf assimilates and tree-ring cellulose with increasing altitude, suggesting site-specific tree physiological responses to relative humidity. Fog-influenced and fog-free sites showed similar δ13C values, suggesting photosynthetic activity to be limited by stomatal closure and irradiance at certain periods. Besides, we observed an 18O-depletion (fog-free and timberline sites) and 13C-depletion (fog-influenced and fog-free sites) in latewood compared to earlywood caused by seasonal differences in: (i) water uptake (i.e., deeper ground water during summer drought, fog water frequency and interception) and (ii) meteorological conditions (stem radial growth and latewood δ18O correlated with winter precipitation). In addition, we found evidence for foliar water uptake and strong isotopic gradients along the pine needle axis in water and assimilates. These gradients are likely the reason for an unexpected underestimation of pine needle water δ18O when applying standard leaf water δ18O models. Our results indicate that soil water availability and air humidity conditions are the main drivers of the physiological behavior of pine along the Canary Island's altitudinal gradients.

Description: Tetrastigma hemsleyanum is a liana plant with promising medicinal and ornamental values. Its calabash-shaped roots (CRs) are served as a traditional Chinese herb. However, it takes a long growth period to form CRs. In the present study, three types of architectural roots, including fine roots (FRs), bar-shaped roots (BRs) and CRs, were employed as materials, and the characteristics of histo-anatomy and digital RNA-seq transcriptome profiles were analyzed. Among the three types of roots, the vascular bundles in FRs were intact, while some of the vascular bundles degenerated in BRs, and only few traces of vascular bundles existed in CRs. Meanwhile, no obvious cell inclusions were found in the cytoplasm of FRs, while a few inclusions were found in BRs, and abundant inclusions were detected in CRs, which might be the main source of medicinal components in roots. The transcriptome profiles and qRT-PCR validation indicated that seven up-regulated genes encoding xyloglucan glycosyltransferase, ACC oxidase, CYP711A1, SHORT-ROOT transcript factor, galacturonosyltransferas, WAT1 and WRKY, and two down-regulated genes encoded LRR receptor-like serine/threonine-protein kinase and CYP83B1, were probably involved in the formation and development of CRs. Besides, GO terms of intrinsic component of membrane, integral component of membrane, cell periphery, membrane part, plasma membrane, membrane, intrinsic component of plasma membrane, cellular chemical homeostasis, and plasma membrane part were probably related to the formation of CRs. KEGG pathways related to the development of CRs probably included MAPK signaling pathway-plant, plant hormone signal transduction, and circadian rhythm-plant. Our finding suggested a probable mode for the formation of CRs.

Description: The mountain birch (Betula pubescens var. pumila (L.)) forest in the Subarctic is periodically exposed to insect outbreaks, which are expected to intensify due to climate change. To mitigate abiotic and biotic stresses, plants have evolved chemical defenses, including volatile organic compounds (VOCs) and non-volatile specialized compounds (NVSCs). Constitutive and induced production of these compounds, however, are poorly studied in Subarctic populations of mountain birch. Here, we assessed the joint effects of insect herbivory, elevation, and season on foliar VOC emissions and NVSC contents of mountain birch. VOCs were sampled in situ by an enclosure technique and analyzed by gas chromatography–mass spectrometry. NVSCs were analyzed by liquid chromatography–mass spectrometry using an untargeted approach. At low elevation, experimental herbivory by winter moth larvae (Operophtera brumata) increased emissions of monoterpenes and homoterpenes over the three-week feeding period, and sesquiterpenes and green leaf volatile in the end of the feeding period. At high elevation, however, herbivory augmented only homoterpene emissions. The more pronounced herbivory effects at low elevation were likely due to higher herbivory intensity. Of the individual compounds, linalool, ocimene, 4,8-dimethylnona-1,3,7-triene, 2-methyl butanenitrile, and benzyl nitrile were among the most responsive compounds in herbivory treatments. Herbivory also altered foliar NVSC profiles at both low and high elevations, with the most responsive compounds likely belonging to fatty acyl glycosides and terpene glycosides. Additionally, VOC emissions from non-infested branches were higher at high than low elevation, particularly during the early season, which was mainly driven by phenological differences. VOC emissions varied substantially over the season, largely reflecting the seasonal variations in temperature and light levels. Our results suggest that if insect herbivory pressure continues to rise in the mountain birch forest with ongoing climate change, it will significantly increase VOC emissions with important consequences for local trophic interactions and climate.

Description: Ground-level ozone (O3) pollution affects the plant carbon and water balance, but the relative contributions of impaired photosynthesis and the loss of stomatal functioning to the O3-induced reductions in water use efficiency (WUE) remain unclear. We combined the leaf stable dual isotopic signatures of carbon (δ13C) and oxygen (δ18O) with related instantaneous gas exchange performance to determine the effects of O3 dose on the net photosynthetic rate (An), stomatal conductance (gs) and intrinsic WUE (iWUE = An/gs) in four tree species (one being a hybrid) exposed to five O3 levels. The iWUE declined for each step increase in O3 level, reflecting progressive loss of the coupling between leaf carbon gain and water loss. In ambient compared to charcoal-filtered air, the decreased iWUE was associated with reductions in both An and gs (i.e., decreased δ13C and increased δ18O). In elevated O3 treatments, however, the iWUE declines were caused by reduced An at constant or increased gs. The results show that the dual isotope approach provides a robust way to gather time-integrated information on how O3 pollution affects leaf gas exchange. Our study highlights that O3-induced decoupling between photosynthesis and stomatal regulation causes large and progressive declines in the WUE of forest trees, demonstrating the need for incorporating this, hitherto unaccounted, effect into vegetation models.

Description: Resprouting plants are distributed in many vegetation communities worldwide. With increasing resprout age post-severe-disturbance, new stems grow rapidly at their early age, and decrease in their growth with gradually decreasing water status thereafter. However, there is little knowledge about how stem hydraulic strategies and anatomical traits vary post-disturbance. In this study, the stem water potential (Ψstem), maximum stem hydraulic conductivity (Kstem-max), water potential at 50% loss of hydraulic conductivity (Kstem  P50), and anatomical traits of Caragana korshinkii resprouts were measured during a 1- to 13-year post-disturbance period. We found that the Kstem-max decreased with resprout age from 1-year-old resprouts (84.2 mol m−1 s−1 MPa−1) to 13-year-old resprouts (54.2 mol m−1 s−1 MPa−1) as a result of decreases in the aperture fraction (Fap) and the sum of aperture area on per unit intervessel wall area (Aap). The Kstem  P50 of the resprouts decreased from 1-year-old resprouts (−1.8 MPa) to 13-year-old resprouts (−2.9 MPa) as a result of increases in vessel implosion resistance (t/b)2, wood density (WD), vessel grouping index (GI), and decreases in Fap, and Aap. These shifts in hydraulic structure and function resulted in an age-based divergence in hydraulic strategies i.e., a change from an acquisitive strategy to a conservative strategy, with increasing resprout age post-disturbance.

Description: The contribution of amino acids (AAs) to soil nitrogen (N) fluxes is higher than previously thought. The fact that AA uptake is pivotal for N nutrition in boreal ecosystems highlights plant AA transporters as key components of the N cycle. At the same time, very little is known about AA transport and respective transporters in trees. Tree genomes may contain 13 or more genes encoding the lysine histidine transporter (LHT) family proteins, and this complicates the study of their significance for tree N-use efficiency. With the strategy of obtaining a tool to study N-use efficiency, our aim was to identify and characterize a relevant AA transporter in hybrid aspen (Populus tremula L. x tremuloides Michx.). We identified PtrLHT1.2, the closest homolog of Arabidopsis thaliana (L.) Heynh AtLHT1, which is expressed in leaves, stems and roots. Complementation of a yeast AA uptake mutant verified the function of PtrLHT1.2 as an AA transporter. Furthermore, PtrLHT1.2 was able to fully complement the phenotypes of the Arabidopsis AA uptake mutant lht1 aap5, including early leaf senescence-like phenotype, reduced growth, decreased plant N levels and reduced root AA uptake. Amino acid uptake studies finally showed that PtrLHT1.2 is a high affinity transporter for neutral and acidic AAs. Thus, we identified a functional AtLHT1 homolog in hybrid aspen, which harbors the potential to enhance overall plant N levels and hence increase biomass production. This finding provides a valuable tool for N nutrition studies in trees and opens new avenues to optimizing tree N-use efficiency.

Description: Synergistic regulation in leaf architecture and photosynthesis is essential for salt tolerance. However, how plant sex and inorganic nitrogen sources alter salt stress-dependent photosynthesis remains unknown. Leaf anatomical characteristics and photosynthesis of Populus cathayana Rehder females and males were investigated under salt stress conditions combined with nitrate NO3− and ammonium NH4+ supplies to clarify the underlying mechanisms. In salt-stressed females, we observed an increased mesophyll spongy cell density, a reduced chloroplast density, a decreased surface area of chloroplasts adjacent to the intercellular air space (Sc/S) and an increased mesophyll cell area per transverse section width (S/W), consequently causing mesophyll conductance (gm) and photosynthesis inhibition, especially under NH4+ supply. Conversely, males with a greater mesophyll palisade tissue thickness and chloroplast density, but a lower spongy cell density had lower S/W and higher Sc/S, and higher gm and photosynthesis. NH4+-fed females had a lower CO2 conductance through cell wall and stromal conductance perpendicular to the cell wall, but a higher chloroplast conductance from the cell wall (gcyt1) than females supplied with NO3−, whereas males had a higher chloroplast conductance and lower CO2 conductance through cell wall when supplied with NO3− instead of NH4+ under salt stress. These findings indicate sex-specific strategies in coping with salt stress related to leaf anatomy and gm under both types of nitrogen supplies, which may contribute to sex-specific CO2 capture and niche segregation.

Description: DORMANCY-ASSOCIATED MADS-box (DAM) and SHORT VEGETATIVE PHASE (SVP) genes have been implicated in the regulation of winter dormancy in perennials. Ectopic expression of apple (Malus × domestica Borkh. ‘Royal Gala’) DAM and SVP genes delays budbreak and constrains lateral shoot outgrowth. In this study, we used RNA interference (RNAi) to simultaneously target all apple DAM and SVP genes in order to study their role and mode of action in the regulation of bud dormancy, budbreak and flowering. A synthetic construct carrying a hairpin fragment assembled from sequences specific to coding regions of three DAM and two SVP genes was used to generate transgenic lines. Reduced expression of DAM/SVP genes resulted in delayed leaf senescence and abscission in autumn, failure to enter bud dormancy in winter and continual growth of new leaves regardless of the season for over 3 years. Precocious flowering but normal flower morphology, fertility and fruit development were observed. The non-dormant phenotype was associated with modified phytohormone composition. The content of gibberellins (GAs) and jasmonates (JAs) was significantly increased in terminal buds of RNAi lines compared with wildtype plants, accompanied by elevated expression of the key GA biosynthesis pathway gene GIBBERELLIN 20 OXIDASE-2 (MdGA20ox-2) along with the FLOWERING LOCUS T gene MdFT2. The key mediator of plasmodesmatal closure, MdCALLOSE SYNTHASE 1 (MdCALS1), was repressed in RNAi lines. This study provides functional evidence for the role of DAM/SVP genes in vegetative phenology of apple and paves the way for production of low-chill varieties suitable for growth in warming climates.

Description: Species interactions mediate tree responses to water limitation because competition and/or facilitation alter plant physiology and growth. However, because it is difficult to isolate the effects of plant–plant interactions and water limitation from other environmental factors, the mechanisms underlying tree physiology and growth in coexisting plants under drought are poorly understood. We investigated how species interactions and water limitation impact the physiology and growth of trembling aspen (Populus tremuloides), narrowleaf cottonwood (Populus angustifolia) and ponderosa pine (Pinus ponderosa) seedlings in a controlled environment growth chamber, using aspen as a focal species. Seedlings were grown in pots alone or with a con- or hetero-specific seedling, and were subjected to a water limitation treatment. Growth, water status and physiological traits were measured before, during and after the treatment. Under well-watered conditions, the presence of another seedling affected growth or biomass allocation in all species, but did not impact the physiological traits we measured. Under water limitation, the presence of a competing seedling had a marginal impact on seedling growth and physiological traits in all species. Throughout the study, the magnitude and direction of seedling responses were complex and often species-specific. Our study serves as an important step toward testing how species’ interactions modify physiological responses and growth in well-watered and water-limited periods.

Description: Deciduous trees mostly rely on non-structural carbohydrates (NSC—soluble carbohydrates and starch) stored prior to dormancy to sustain both spring bloom and the initial phase of spring growth prior to the transition of leaves from sink to source. Winter management of NSC, their loss due to respiration, reallocation patterns and remobilization during spring, seems to be key to a timely and synchronous bloom. To assess tree dependence on NSC during dormancy, we tested whether the interruption of local branch NSC accumulation prior to dormancy by defoliation and the interruption of NSC translocation by phloem girdling influence spring phenology in three major deciduous Mediterranean nut crop species: Prunus dulcis (Mill.) D.A Webb, a hybrid between Pistacia integerrima (J. L. Stewart ex Brandis) and P. atlantica Desf. (referred to as P. integerrima), and Juglans regia L. Defoliation treatments had different effects on NSC concentration in different species depending on the time of application. However, despite the significant initial impact (increase or decrease of NSC concentration), with time this impact diminished resulting in overall similar concentrations between control and defoliated branches suggesting the presence of NSC reallocation during dormancy. Phloem girdling in P. dulcis and P. integerrima resulted in reduced export activity and greater NSC concentrations, while in J. regia girdling resulted in lower NSC concentrations, indicating that this species requires a net import of NSC during dormancy. Bud break was distinctly delayed by both defoliation and phloem girdling in all the three species, providing evidence of the significant roles that fall NSC accumulation and winter NSC management play in priming trees for spring growth resumption.

Description: Improving the efficiency of fertilizer application is paramount to both the sustainability and profitability of forest plantations. Therefore, developing reliable, cost-effective tools to assess tree nutritional status is of great interest. This investigation sought to assess the use of phloem sap-derived metabolites as an indicator of nutritional status on a background of seasonal water availability of Eucalyptus globulus (Labill) trees grown under field conditions. Phloem is a central conduit for long-distance transport and signaling in plants and offers great promise in reflecting plant-scale resource limitations. Changes in the abundance of solutes and isotopes in phloem sap are sensitive to environmental cues. With a focus on both water and nutrient availability, we characterize patterns in phloem sugars, amino acids and the abundance of carbon isotopes in phloem sap obtained from E. globulus among different seasons and fertilizer treatments. Phloem-derived total amino acid concentration was found to increase with an increasing nitrogen (N) supply; however, this response was lost with the concurrent addition of phosphorus and at the highest level of N supply. Significant seasonal variation in all measured parameters was also detected, highlighting the need for caution in making quantitative relationships with growth. Broader implications of the interactive effects of both water supply and multi-nutrient additions and relationships with growth are discussed.

Description: The scandent shrub plant form is a variant of liana that has upright and self-supporting stems when young but later becomes a climber. We aimed to explore the associations of stem and leaf traits among sympatric lianas, scandent shrubs and trees, and the effects of growth form and leaf habit on variation in stem or leaf traits. We measured 16 functional traits related to stem xylem anatomy, leaf morphology and nutrient stoichiometry in eight liana, eight scandent shrub and 21 tree species co-occurring in a subalpine cold temperate forest at an elevation of 2600–3200 m in Southwest China. Overall, lianas, scandent shrubs and trees were ordered along a fast–slow continuum of stem and leaf functional traits, with some traits overlapping. We found a consistent pattern of lianas 〉 scandent shrubs 〉 trees for hydraulically weighted vessel diameter, maximum vessel diameter and theoretical hydraulic conductivity. Vessel density and sapwood density showed a pattern of lianas = scandent shrubs 〈 trees, and lianas 〈 scandent shrubs = trees, respectively. Lianas had significantly higher specific leaf area and lower carbon concentration than co-occurring trees, with scandent shrubs showing intermediate values that overlapped with lianas and trees. The differentiation among lianas, scandent shrubs and trees was mainly explained by variation in stem traits. Additionally, deciduous lianas were positioned at the fast end of the trait spectrum, and evergreen trees at the slow end of the spectrum. Our results showed for the first time clear differentiation in stem and leaf traits among sympatric liana, scandent shrub and tree species in a subalpine cold temperate forest. This work will contribute to understanding the mechanisms responsible for variation in ecological strategies of different growth forms of woody plants.

Description: Understanding seasonal variation in photosynthesis is important for understanding and modeling plant productivity. Here, we used shotgun sampling to examine physiological, structural and spectral leaf traits of upper canopy, sun-exposed leaves in Quercus coccinea Münchh (scarlet oak) across the growing season in order to understand seasonal trends, explore the mechanisms underpinning physiological change and investigate the impact of extrapolating measurements from a single date to the whole season. We tested the hypothesis that photosynthetic rates and capacities would peak at the summer solstice, i.e., at the time of peak photoperiod. Contrary to expectations, our results reveal a late-season peak in both photosynthetic capacity and rate before the expected sharp decrease at the start of senescence. This late-season maximum occurred after the higher summer temperatures and vapor pressure deficit and was correlated with the recovery of leaf water content and increased stomatal conductance. We modeled photosynthesis at the top of the canopy and found that the simulated results closely tracked the maximum carboxylation capacity of Rubisco. For both photosynthetic capacity and modeled top-of-canopy photosynthesis, the maximum value was therefore not observed at the summer solstice. Rather, in each case, the measurements at and around the solstice were close to the overall seasonal mean, with values later in the season leading to deviations from the mean by up to 41 and 52%, respectively. Overall, we found that the expected Gaussian pattern of photosynthesis was not observed. We conclude that an understanding of species- and environment-specific changes in photosynthesis across the season is essential for correct estimation of seasonal photosynthetic capacity.

Description: Apomixis is a form of reproduction that does not involve the fertilization of female gametes by male gametes but instead involves the production of offspring directly from the female parent. The offspring of apomixis are genetically identical to the female parent and inherit its traits. Therefore, apomixis has great potential for application to agricultural genetic breeding. However, it remains unclear whether apomictic species require pollination, and the impacts of pollination on such species are poorly understood. We investigated the effects of pollination on the apomictic species Zanthoxylum bungeanum Maxim. by analyzing its fertilization process, assembling its transcriptome, and measuring hormone concentrations, fruit setting rate and gene expression levels. Transcriptome sequencing of pollinated and unpollinated fruits resulted in a total of 69,131 PacBio reads. Of these, 7102 genes were up-regulated and 6491 genes were down-regulated. Analysis of the differentially expressed genes (DEGs) and construction of a weighted gene co-expression network showed that many DEGs were involved in plant hormone signal transduction, suggesting that hormonal signaling during development differs between pollinated and unpollinated fruit. The germination rate of Z. bungeanum pollen in vitro was only 11%, and pollen could not germinate in the embryo sac to complete fertilization. Although pollination did not enable Z. bungeanum to complete the sexual reproduction process, it significantly increased abscisic acid (ABA) concentration and fruit setting rate. Spraying 100 μg l−1 ABA also significantly increased the fruit setting rate. Therefore, ABA appears to be a key factor in the regulation of fruit setting in apomictic Z. bungeanum. Based on these results, we suggest that some male plants be cultivated in Z. bungeanum plantations or exogenous ABA be sprayed to increase the likelihood of pollination and thereby increase the fruit setting rate.

Description: Interlocked grain occurs when the orientation of xylem fibres oscillates, alternating between left- and right-handed spirals in successive wood layers. The cellular mechanisms giving rise to interlocked grain, thought to involve the slow rotation of fusiform initials within the vascular cambium, remain unclear. We suggest that observations of wood structure at the cellular level, but over large areas, might reveal these mechanisms. We assayed timber from several commercially important tropical angiosperms from the genus Khaya (African mahogany) that exhibit interlocked grain using X-ray computed microtomography followed by orthogonal slicing and image processing in ImageJ. Reconstructed tangential longitudinal sections were processed with the ImageJ directionality plug-in to directly measure fibre orientation and showed grain deviations of more than 10° from vertical in both left- and right-handed directions. Grain changed at locally constant rates, separated by locations where the direction of grain change sharply reversed. Image thresholding and segmentation conducted on reconstructed cross sections allowed the identification of vessels and measurement of their location, with vessel orientations then calculated in Matlab and, independently, in recalculated tangential longitudinal sections with the directionality plug-in. Vessel orientations varied more than fibre orientations, and on average deviated further from vertical than fibres at the locations where the direction of grain change reversed. Moreover, the reversal location for vessels was shifted ~400 μm towards the pith compared with the fibres, despite both cell types arising from the same fusiform initials within the vascular cambium. We propose a simple model to explain these distinct grain patterns. Were an auxin signal to control both the reorientation of cambial initials, as well as coordinating the end-on-end differentiation and linkage of xylem vessel elements, then it would be possible for fibres and vessels to run at subtly different angles, and to show different grain reversal locations.

Description: Tree growth is generally considered to be temperature limited at upper elevation treelines, yet climate factors controlling tree growth at semiarid treelines are poorly understood. We explored the influence of climate on stem growth and stable isotopes for Polylepis tarapacana Philipi, the world’s highest elevation tree species, which is found only in the South American Altiplano. We developed tree-ring width index (RWI), oxygen (δ18O) and carbon (δ13C) chronologies for the last 60 years at four P. tarapacana stands located above 4400 m in elevation, along a 500 km latitude aridity gradient. Total annual precipitation decreased from 300 to 200 mm from the northern to the southern sites. We used RWI as a proxy of wood formation (carbon sink) and isotopic tree-ring signatures as proxies of leaf-level gas exchange processes (carbon source). We found distinct climatic conditions regulating carbon sink processes along the gradient. Current growing-season temperature regulated RWI at northern-wetter sites, while prior growing-season precipitation determined RWI at arid southern sites. This suggests that the relative importance of temperature to precipitation in regulating tree growth is driven by site water availability. By contrast, warm and dry growing seasons resulted in enriched tree-ring δ13C and δ18O at all study sites, suggesting that similar climate conditions control carbon-source processes along the gradient. Site-level δ13C and δ18O chronologies were significantly and positively related at all sites, with the strongest relationships among the southern drier stands. This indicates an overall regulation of intercellular carbon dioxide via stomatal conductance for the entire P. tarapacana network, with greater stomatal control when aridity increases. This manuscript also highlights a coupling (decoupling) between physiological processes at leaf level and wood formation as a function of similarities (differences) in their climatic sensitivity. This study contributes to a better understanding and prediction of the response of high-elevation Polylepis woodlands to rapid climate changes and projected drying in the Altiplano.

Description: Knowledge on variations of drought resistance traits are needed to predict the potential of trees to acclimate to coming severe drought events. Xylem vulnerability to embolism is a key parameter related to such droughts, and its phenotypic variability relies mainly on environmental plasticity. We investigated the structural determinants controlling the plasticity of vulnerability to embolism, focusing on the key elements involved in the air bubble entry in vessels, especially the intervessel pits. Poplar saplings (Populus tremula x alba (Aiton) Sm., 1804) grown in contrasted water availability or light exposure exhibited differences in the vulnerability to embolism (P50) in a range of 0.76 MPa. We then characterized the structural changes in features related to pit quantity and pit structure, from the pit ultrastructure to the organization of xylem vessels, using different microscopy techniques (transmission electron microscopy, scanning electron microscopy, light microscopy). A multispectral combination of X-ray microtomography and light microscopy analysis allowed measuring the vulnerability of each single vessel and testing some of the relationships between structural traits and vulnerability to embolism inside the xylem. The pit ultrastructure did not change, whereas the vessel dimensions increased with the vulnerability to embolism and the grouping index and fraction of intervessel cell wall both decreased with the vulnerability to embolism. These findings hold when comparing between trees or between the vessels inside the xylem of an individual tree. These results evidenced that plasticity of vulnerability to embolism in hybrid poplar occurs through changes in the pit quantity properties such as pit area and vessel grouping rather than changes on the pit structure.

Description: Carbon allocation between source and sink organs determines plant growth and is influenced by environmental conditions. Under water deficit (WD), plant growth is inhibited before photosynthesis and shoot growth tends to be more sensitive than root growth. However, the modulation of the source–sink relationship by rootstocks remains unsolved in citrus trees under WD. Citrus plants grafted on Rangpur lime are drought tolerant, which may be related to a fine coordination of the source–sink relationship for maintaining root growth. Here, we followed 13C allocation and evaluated physiological responses and growth of Valencia orange trees grafted on three citrus rootstocks (Rangpur lime, Swingle citrumelo and Sunki mandarin) under WD. As compared with plants on Swingle and Sunki rootstocks, ones grafted on Rangpur lime showed higher stomatal sensitivity to the initial variation of water availability and less accumulation of non-structural carbohydrates in roots under WD. High 13C allocation found in Rangpur lime roots indicates this rootstock has high sink demand associated with high root growth under WD. Our data suggest that Rangpur lime rootstock used photoassimilates as sources of energy and carbon skeletons for growing under drought, which is likely related to increases in root respiration. Taken together, our data revealed that carbon supply by leaves and delivery to roots are critical for maintaining root growth and improving drought tolerance, with citrus rootstocks showing differential sink strength under WD.

Description: Parenchyma is an important component of the secondary xylem. It has multiple functions and its fraction is known to vary substantially across angiosperm species. However, the physiological significance of this variation is not yet fully understood. Here, we examined how different types of parenchyma (ray parenchyma [RP], axial parenchyma [AP] and AP in direct contact with vessels [APV]) are coordinated with three essential xylem functions: water conduction, storage of non-structural carbohydrate (NSC) and mechanical support. Using branch sapwood of 15 co-occurring drought-adapted woody species from the subtropical Bonin Islands, Japan, we quantified 10 xylem anatomical traits and examined their linkages to hydraulic properties, storage of soluble sugars and starch and sapwood density. The fractions of APV and AP in the xylem transverse sections were positively correlated with the percentage loss of conductivity in the native condition, whereas that of RP was negatively correlated with the maximum conductivity across species. Axial and ray parenchyma fractions were positively associated with concentrations of starch and NSC. The fraction of parenchyma was independent of sapwood density, regardless of parenchyma type. We also identified a negative relationship between hydraulic conductivity and NSC storage and sapwood density, mirroring the negative relationship between the fractions of parenchyma and vessels. These results suggest that parenchyma fraction underlies species variation in xylem hydraulic and carbon use strategies, wherein xylem with a high fraction of AP may adopt an embolism repair strategy through an increased starch storage with low cavitation resistance.

Description: Sichuan pepper (Zanthoxylum armatum DC.) is a popular spice and is often prescribed in traditional Chinese medicine to treat vomiting, diarrhea, ascariasis, and eczema, among others. Volatile oils from Z. armatum leaves contain active ingredients, with terpenoids being one of the main components. In the present study, the combination of sequencing data of Z. armatum from PacBio single molecule real time and Illumina RNA-Seq platforms facilitated an understanding of the gene regulatory network of terpenoid biosynthesis in pepper leaves. The leaves of three developmental stages from two Z. armatum cultivars, ‘Rongchangwuci’ (WC) and ‘Zhuye’ (ZY), were selected as test materials to construct sequencing libraries. A total of 143,122 predictions of unique coding sequences, 105,465 simple sequence repeats, 20,145 transcription factors, and 4,719 lncRNAs were identified, and 142,829 transcripts were successfully annotated. The occurrence of alternative splicing events was verified by reverse transcription PCR, and quantitative real-time PCR was used to confirm the expression pattern of six randomly selected lncRNAs. A total of 96,931 differentially expressed genes were filtered from different samples. According to functional annotation, a total of 560 candidate genes were involved in terpenoid synthesis, of which 526 were DEGs. To identify the key genes involved in terpenoid biosynthesis, the module genes in different samples, including structural and TFs genes, were analyzed using the weighted gene co-expression network method, and the co-expression network of genes was constructed. Thirty-one terpenoids were identified by gas chromatography–mass spectrometry. The correlation between 18 compounds with significantly different contents and genes with high connectivity in the module was jointly analyzed in both cultivars, yielding 12 candidate DEGs presumably involved in the regulation of terpenoid biosynthesis. Our findings showed that full-length transcriptome SMRT and Illumina RNA sequencing can play an important role in studying organisms without reference genomes and elucidating the gene regulation of a biosynthetic pathway.

Description: Flowering Locus T (FT) promotes flowering by integrating six genetic pathways. In Arabidopsis, the FT protein is transported from leaves to shoot apices and induces flowering. However, contradictory conclusions about floral induction via graft-transmitted FT in trees were reported in previous studies. We obtained extremely early-flowering transgenic woody Jatropha curcas by overexpression of J. curcas FT using Arabidopsis thaliana SUC2 promoter (SUC2:JcFT) and non-flowering transgenic J. curcas by RNA interference (RNAi), which were used to investigate the function of graft-transmitted JcFT in floral induction in woody perennials. Scions from five wild-type species of the Jatropha genus and from JcFT-RNAi transgenic J. curcas were grafted onto SUC2:JcFT rootstocks. Most grafted plants produced flowers in 1–2 months, and the flowering percentage and frequency of various grafted plants decreased with increasing scion length. Consistently, FT protein abundance in scions also decreased with increasing distance from graft junctions to the buds. These findings suggest that FT proteins can be transmitted by grafting and can induce the floral transition in woody perennials, and the efficiency of graft-transmitted JcFT for floral induction depends on the scion length, which may help explain previous seemingly contradictory observations regarding floral induction via graft-transmitted FT in trees.

Description: Global climate change scenarios predict an increase in air temperature, precipitation, and air humidity for northern latitudes. Elevated air humidity may significantly reduce the water flux through forest canopies and affect interactions between water and nutrient uptake. However, we have limited understanding of how altered transpiration would affect root respiration and carbon (C) exudation as fine root morphology acclimates to different water flux. We investigated the effects of elevated air relative humidity (eRH) and different inorganic nitrogen sources (NO3− and NH4+) on above and belowground traits in hybrid aspen (Populus × wettsteinii Hämet-Ahti), silver birch (Betula pendula Roth.), and Scots pine (Pinus sylvestris L.) grown under controlled climate chamber conditions. The eRH significantly decreased the transpiration flux in all species, decreased root mass-specific exudation in pine, and increased root respiration in aspen. eRH also affected fine root morphology, with specific root area increasing for birch but decreasing in pine. The species comparison revealed that pine had the highest C exudation, while birch had the highest root respiration rate. Both humidity and nitrogen treatments affected the share of absorptive and pioneer roots within fine roots; however, the response was species-specific. The proportion of absorptive roots was highest in birch and aspen, the share of pioneer roots was greatest in aspen, and the share of transport roots was greatest in pine. Fine roots with lower root tissue density were associated with pioneer root tips and had a higher C exudation rate. Our findings underline the importance of considering species-specific differences in relation to air humidity and soil nitrogen availability that interactively affect the C input–output balance. We highlight the role of changes in the fine root functional distribution as an important acclimation mechanism of trees in response to environmental change.

Description: The rhizobiome is being increasingly acknowledged as a key player in plant health and breeding strategies. The pine pitch canker (PPC), caused by the fungus Fusarium circinatum, affects pine species with varying susceptibility degrees. Our aims were to explore the bacterial rhizobiome of a susceptible (Pinus radiata) and a resistant (Pinus pinea) species together with other physiological traits, and to analyze shifts upon F. circinatum inoculation. Pinus seedlings were stem inoculated with F. circinatum spores and needle gas exchange and antioxidant-related parameters were analyzed in non-inoculated and inoculated plants. Rhizobiome structure was evaluated through 16S rRNA gene massive parallel sequencing. Species (non-inoculated plants) harbored distinct rhizobiomes (

Description: Understanding the changing patterns of vascular cambium during seasonal cycles is crucial to reveal the mechanisms that control cambium activity and wood formation, but this area has been underexplored, especially in conifers. Here, we quantified the changing cellular morphology patterns of cambial zones during the active, transition and dormant stages. With the help of toluidine blue and periodic acid Schiff staining to visualize cell walls and identify their constituents, we observed decreasing cambial cell layers, thickening of newly formed xylem cell walls and increased polysaccharide granules in phloem from June to the following March over the course of our collecting period. Pectin immunofluorescence showed that dormant stage cambium can produce highly abundant de-esterified homogalacturonan and (1–4)-β-D-galactan epitopes, while active cambium can strong accumulate high methylesterified homogalacturonan. Calcofluor white staining and confocal Raman spectroscopy analysis revealed regular changes in the chemical composition of cell walls, such as relative lower cellulose deposition in transition stage in vascular cambium, and higher lignin accumulation was found in dormant stage in secondary xylem. Moreover, RT-qPCR analysis suggested that various IAA (Aux/IAA protein), CesA, CslA and HDZ genes, as well as NAC, PME3 and PME4, may be involved in cambium activities and secondary xylem formation. Taken together, these findings provide new information about cambium activity and cell differentiation in the formation, structure, and chemistry in conifers during the active–dormant transition.

Description: Future climate change predictions for tropical forests highlight increased frequency and intensity of extreme drought events. However, it remains unclear whether large and small trees have differential strategies to tolerate drought due to the different niches they occupy. The future of tropical forests is ultimately dependent on the capacity of small trees (

Description: Differences in plant shade tolerance constitute a major mechanism driving the succession of forest communities in subtropical forests. However, the indirect effects of differences in light requirements on the growth of mid- and late-successional tree species are unclear, and this potential growth effect has not been explained at the transcriptome level. Here, a typical mid-successional dominant tree species, Schima superba Gardn. et Champ, and a typical late-successional dominant tree species, Cryptocarya concinna Hance were used as materials and planted under 100% full light (FL) and 30% FL (low light, LL) to explore the responses of tree species in different successional stages of subtropical forests to different light environments. Transcriptome sequencing was used to analyze the expression changes in genes related to growth and photoprotection under different light environments. The young leaves of S. superba accumulated more malondialdehyde (MDA) and superoxide radicals (${mathrm{O}}_2^{{{}^{ ullet}}^{-}}$) under LL. A lower hormone content (auxin, cytokinin, gibberellin) in the young leaves, a weaker photosynthetic capacity in the mature leaves and significant downregulation of related gene expression were also found under LL, which resulted in the total biomass of S. superba under LL being lower than that under FL. The young leaves of C. concinna had less MDA and ${mathrm{O}}_2^{{{}^{ ullet}}^{-}}$, and a higher hormone contents under LL than those under FL. There was no significant difference in photosynthetic capacity between mature leaves in contrasting light environments. Although the biomass of C. concinna under LL was less than that under FL, the height of C. concinna under LL was higher than that under FL, indicating that C. concinna could grow well under the two light environments. Our results describing the acclimatization of light at the physiological, molecular and transcriptome levels are important for a complete understanding of successional mechanisms.

Description: Vascular plants with secondary growth develop a periderm mostly composed of dead suberized cork cells to face environmental hostile conditions. Cork oak has a highly active and long-living phellogen forming a remarkably thick periderm that is periodically debarked for industrial purposes. This wounding originates the quick formation of a new traumatic periderm, making cork oak an exceptional model to study the first periderm differentiation during normal development in young sprigs and traumatic (wound) periderm formation after debarking. Here, we studied the poorly known first periderm differentiation steps that involve cell wall suberization, polyphenolic accumulation and programmed cell death (PCD) by combining transmission electron microscopy, histochemical and molecular methods in periderms from young sprigs. These processes were further compared with traumatic periderms formed after wounding using molecular and histochemical techniques, such as the polyphenolic accumulation. In the first periderms from young sprigs, four distinct differentiation stages were defined according to the presence of PCD morphological features. First young and traumatic periderms showed an upregulation of genes related to suberin biosynthesis, proanthocyanidins biosynthesis and transport, autophagy, and PCD. Traumatic periderms revealed an overall upregulation of these genes, likely resulting from ontogeny differences and distinct phellogen origin associated with a faster metabolism, highlighting the impact of wounding on phellogen activity after debarking. First periderms from young sprigs showed gradual accumulation of proanthocyanidins in the vacuoles throughout PCD stages until total filled lumens, whereas in traumatic periderms, these compounds were found cell wall linked in already empty cells. This work enabled a comprehensive overview of the cork cells differentiation processes contributing to deepening the knowledge of the fundamental ontogenic program of this protective tissue, which is also a unique forest product, constituting the basis of a sustainable and profitable industry.

Description: The iso/anisohydric continuum has been used to classify tree species’ drought response strategies. The range over which stomata are regulating leaf water potential (ψl) before turgor loss occurs can be described with metrics such as the dependence of ψl on soil water potential (ψsoil) and the size of ‘hydroscape area’ (HA), but corresponding field data from adult trees are scarce. We examined the stomatal conductance (gs)–ψl relationship in its temporal (diurnal vs seasonal and interannual) and spatial (within-crown vs between-site) variation in European beech, using extensive ψl and gs measurements in the canopy of four beech stands across a precipitation gradient, and complemented the data set by published ψl and gs measurements in further Central European beech stands (including the extreme 2018 drought) in order to cover the full water potential operation space of the species. Both metrics characterize beech as a strictly anisohydric species with δψl/δψsoil 〉〉 1 and HA = 4 MPa2. However, stomates close sensitively in response to increasing vapor pressure deficit, disproving the widely assumed dependence of large ψl variation on looser stomatal control. Characterizing the water status regulation mechanisms of trees requires separating diurnal from day-to-day variation in ψl and gs. The large diurnal and seasonal ψl variation in beech leaves is partly caused by a low leaf tissue elasticity, suggesting that a whole-plant perspective with consideration of osmotic and elastic tissue properties and stem and root hydraulics is needed for fully understanding ψl regulation and the drought tolerance strategy of trees.

Description: The role of carbon (C) and nutrient uptake, allocation, storage and especially their interactions in survival and recovery of trees under increased frequencies and intensities of drought events is not well understood. A full factorial experiment with four soil water content regimes ranging from extreme drought to well-watered conditions and two fertilization levels was carried out. We aimed to investigate whether nutrient addition mitigates drought effects on downy oak (Quercus pubescens Willd.) and whether storage pools of non-structural carbohydrates (NSC) are modified to enhance survival after 2.5 years of drought and recovery after drought relief. Physiological traits, such as photosynthesis, predawn leaf water potential as well as tissue biomass together with pools and dynamics of NSC and nutrients at the whole-tree level were investigated. Our results showed that fertilization played a minor role in saplings’ physiological processes to cope with drought and drought relief, but reduced sapling mortality during extreme drought. Irrespective of nutrient supply, Q. pubescens showed increased soluble sugar concentration in all tissues with increasing drought intensity, mostly because of starch degradation. After 28 days of drought relief, tissue sugar concentrations decreased, reaching comparable values to those of well-watered plants. Only during the recovery process from extreme drought, root NSC concentration strongly declined, leading to an almost complete NSC depletion after 28 days of rewetting, simultaneously with new leaves flushing. These findings suggest that extreme drought can lead to root C exhaustion. After drought relief, the repair and regrowth of organs can even exacerbate the root C depletion. We concluded that under future climate conditions with repeated drought events, the insufficient and lagged C replenishment in roots might eventually lead to C starvation and further mortality.

Description: Auxin is involved in various developmental processes of plants, including cell division in cambium and xylem differentiation. However, most studies linking auxin and xylem cell production are performed in environments with a strong seasonality (i.e., temperate and boreal climates). The temporal dynamics of auxin and cambial activity of subtropical trees remain basically unknown. In this study, we sampled four microcores weekly in three individuals of Chinese red pine (Pinus massoniana Lamb.) from February to December 2015–16 to compare xylem formation with auxin concentration in subtropical China. During the entire period of sampling, the number of cambial cells varied from 2 to 7, while the number of cells in the enlarging zone ranged from 1 to 4 and from 1 to 5 in the wall-thickening zone. In 2015, the average auxin concentration was 3.46 ng g−1, with 33 xylem cells being produced at the end of the year. In 2016, a lower auxin concentration (2.59 ng g−1) corresponded to a reduced annual xylem production (13.7 cells). No significant relationship between auxin concentration and number of xylem cells in differentiation was found at the weekly scale. Unlike in boreal and temperate forests, the lack of wood formation seasonality in subtropical forests makes it more difficult to reveal the relationship between auxin concentration and number of xylem cells in differentiation at the intra-annual scale. The frequent and repeated samplings might have reduced auxin concentration in the developing cambium and xylem, resulting in a lower xylem cell production.

Description: In an attempt to comprehensively study the dynamics of non-structural carbon compounds (NCCs), we measured the seasonal changes of soluble sugars, starch, lipids and sugar alcohols in the leaves, branches, stem and roots of the fast-growing Pinus contorta (Loudon) (pine) and slow-growing Picea glauca (Moench) Voss (spruce) trees growing in a boreal climate. In addition to measuring the seasonal concentrations of these compounds, the relative contribution of these compounds to the total NCC pool within the organs of trees (~8 m tall) was estimated and compared across different phenological and growth stages. Both species showed large seasonal shifts from starch to sugars from spring to fall in nearly all organs and tissues; most likely an adaptation to the cold winters. For both species, the total fluctuation of sugar + starch across the year (i.e., the difference between the minimum and maximum observed across collection times) was estimated to be between 1.6 and 1.8 kg for all NCCs. The fluctuation, however, was 1.40 times greater than the minimum reserves in pine, while only 0.72 times the minimum reserves in spruce. By tissue type, NCC fluctuations were greatest in the roots of both species. Roots showed a large build-up of reserves in late spring, but these reserves were depleted over summer and fall. Storage reserves in needles and branches declined over the summer, and this decline may be linked to the sink strength of the stem during diameter growth. Some notable highlights of this holistic study: a late winter build-up of sugars in the stem xylem of both species, but especially spruce; and an increase in sugar alcohols in the bark of spruce in very late winter, which could indicate mobilization to support early growth in spring and high lipid reserves in the bark of pine, which appeared not to be impacted by seasonal changes between summer and winter. Collectively, these observations point toward a more conservative NCC reserve strategy in spruce compared with pine, which is consistent with its stress tolerance and greater longevity.

Description: Noncoding RNAs (ncRNAs) play pivotal roles in various biological processes in plants. However, the role of ncRNAs in tapping panel dryness (TPD) of rubber tree (Hevea brasiliensis) is largely unknown. Here, the whole transcriptomes of bark tissues from healthy and TPD trees were performed to identify differentially expressed long ncRNAs (DELs), microRNAs/miRNA (DEMs), genes (DEGs), and their regulatory networks involved in TPD. A total of 263 DELs, 174 DEMs, and 1,574 DEGs were identified in the bark of TPD tree compared with that of healthy tree. KEGG analysis revealed that most of the DEGs and targets of DELs and DEMs were mainly enriched in metabolic pathways, biosynthesis of secondary metabolites, and plant hormone signal transduction. Additionally, the majority of DEGs and DELs related to rubber biosynthesis were down-regulated in TPD trees. Furthermore, 98 DEGs and 44 DELs were targeted by 54 DEMs, 190 DEGs were identified as putative targets of 56 DELs, and two and 44 DELs were predicted as precursors and endogenous target mimics (eTMs) of two and six DEMs, respectively. Based on these, the DEL-DEM-DEG regulatory network involved in TPD was constructed, and 13 hub DELs, three hub DEMs and two hub DEGs were identified. The results provide novel insights into the regulatory roles of ncRNAs underlying TPD and lay a foundation for future functional characterization of lncRNAs, miRNAs, and genes involved in TPD in rubber tree.

Description: The use of size-controlling rootstocks is central to modern high-density fruit production systems. While biological mechanisms responsible for vigor control are not fully understood, differences in water relations and carbohydrate storage ability have been suggested as two potential factors. To better understand the processes that control growth vigor, we analyzed the trunk radial variation at seasonal and diurnal timescales and measured the midday leaf water potential (ΨMD), leaf gas exchange and concentrations of non-structural carbohydrates (NSC) in apple trees of variety ‘Jonagold’ grafted on two rootstocks of contrasting growth vigor (dwarfing J-TE-G vs invigorating J-TE-H). The measurements were conducted during an exceptionally hot and dry summer. We found that smaller annual trunk radial increments in dwarfed trees were primarily due to an earlier cessation of trunk secondary growth. The interdiurnal trunk circumference changes (ΔC) were slightly lower in dwarfed trees, and these trees also had fewer days with positive ΔC values, particularly during the driest summer months. The trunks of dwarfed trees shrank gradually during the drought, showed less pronounced diurnal variation of trunk circumference and the maximum trunk daily shrinkage was only weakly responsive to the vapor pressure deficit. These results indicated that lower turgidity in the cambial region may have limited the trunk radial expansion in dwarfed trees during the hot and dry days. Dwarfed trees also maintained lower ΨMD and leaf gas exchange rates during the summer drought. These parameters decreased in parallel for both rootstock combinations, suggesting their similar drought sensitivity. Similar concentrations and seasonal dynamics of NSC in both rootstock combinations, together with their similar spring growth rates, suggest that NSC reserves were not directly limiting for growth. Our results support the prominent role of water relations in rootstock-induced size-controlling mechanisms and highlight the complexity of this topic.

Description: Hickory (Carya cathayensis Sarg.) is an extraordinary nut-bearing deciduous arbor with high content of oil in its embryo. However, the molecular mechanism underlying high oil accumulation is mostly unknown. Here, we reported that the lipid droplets and oil accumulation gradually increased with the embryo development and the oil content was up to ~76% at maturity. Furthermore, transcriptome and proteome analysis of developing hickory embryo identified 32,907 genes and 9857 proteins. Time-series analysis of gene expressions showed that these genes were divided into 12 clusters and lipid metabolism-related genes were enriched in Cluster 3, with the highest expression levels at 95 days after pollination (S2). Differentially expressed genes and proteins indicated high correlation, and both were enriched in the lipid metabolism. Notably, the genes involved in biosynthesis, transport of fatty acid/lipid and lipid droplets formation had high expression levels at S2, while the expression levels of other genes required for suberin/wax/cutin biosynthesis and lipid degradation were very low at all the sampling time points, ultimately promoting the accumulation of oil. Quantitative reverse-transcription PCR analysis also verified the results of RNA-seq. The co-regulatory networks of lipid metabolism were further constructed and WRINKLED1 (WRI1) was a core transcriptional factor located in the nucleus. Of note, CcWRI1A/B could directly activate the expression of some genes (CcBCCP2A, CcBCCP2B, CcFATA and CcFAD3) required for fatty acid synthesis. These results provided in-depth evidence for revealing the molecular mechanism of high oil accumulation in hickory embryo.

Description: Leaf mass per area (Ma), nitrogen content per unit leaf area (Narea), maximum carboxylation capacity (Vcmax) and the ratio of leaf-internal to ambient CO2 partial pressure (χ) are important traits related to photosynthetic function, and they show systematic variation along climatic and elevational gradients. Separating the effects of air pressure and climate along elevational gradients is challenging due to the covariation of elevation, pressure and climate. However, recently developed models based on optimality theory offer an independent way to predict leaf traits and thus to separate the contributions of different controls. We apply optimality theory to predict variation in leaf traits across 18 sites in the Gongga Mountain region. We show that the models explain 59% of trait variability on average, without site- or region-specific calibration. Temperature, photosynthetically active radiation, vapor pressure deficit, soil moisture and growing season length are all necessary to explain the observed patterns. The direct effect of air pressure is shown to have a relatively minor impact. These findings contribute to a growing body of research indicating that leaf-level traits vary with the physical environment in predictable ways, suggesting a promising direction for the improvement of terrestrial ecosystem models.

Description: Functional structural plant models of tree crops are useful tools that were introduced more than two decades ago. They can represent the growth and development of a plant through the in silico simulation of the 3D architecture in connection with physiological processes. In tree crops, physiological processes such as photosynthesis, carbon allocation, and growth are usually integrated into these models although other functions such as water and nutrient uptake are often disregarded. The implementation of the 3D architecture involves different techniques such as L-system frameworks, pipe model concepts, and Markovian models to simulate branching processes, bud fates, and elongation of stems based on the production of metamers. The simulation of root architecture is still a challenge for researchers due to a limited amount of information and experimental issues in dealing with roots because root development is not based on the production of metamers. This review aims to focus on functional-structural models of fruit tree crops, highlighting their physiological components. The potential and limits of these tools are reviewed to point out the topics that still need more attention.

Description: The dependence of trees on carbon and nutrient storage is critical to predicting the forest vulnerability under climate change, but whether evergreen and deciduous species differ in their use and allocation of stored resources during spring phenology is unclear. Using a high temporal resolution, we evaluated the role of spring phenology and shoot growth as determinants of the carbon and nutrient storage dynamics in contrasting leaf habits. We recorded the phenology and shoot elongation and determined the concentrations of total non-structural carbohydrates (NSC), starch, soluble carbohydrates (SC), nitrogen (N) and phosphorus (P) in buds, expanding shoots and previously formed shoots of two sympatric Nothofagus species with contrasting leaf habit. Species reached similar shoot lengths, though shoot expansion started 35 days earlier and lasted c. 40 days more in the deciduous species. Thus, while the deciduous species had a relatively constant shoot growth rate, the evergreen species experienced a conspicuous growth peak for c. 20 days. In the evergreen species, the greatest decreases in NSC concentrations of previously formed shoots and leaves coincided with the maximum shoot expansion rate and fruit filling, with minimums of 63% and 65% relative to values at bud dormancy, respectively. In contrast, minimum NSC concentrations of the previously formed shoots of the deciduous species were only 73% and occurred prior to the initiation of shoot expansion. Bud N and P concentrations increased during budbreak, while previously formed shoots generally did not decrease their nutrient concentrations. Late spring phenology and overlapping of phenophases contributed to the greater dependence on storage of proximal tissues in the studied evergreen compared to deciduous species, suggesting that phenology is a key determinant of the contrasting patterns of storage use in evergreen and deciduous species.

Description: Plants constitute 80% of the biomass on earth, and almost two thirds of this biomass is found in wood. Wood formation is a carbon demanding process and relies on carbon transport from photosynthetic tissues. Thus, understanding the transport process is of major interest for understanding terrestrial biomass formation. Here we review the molecules and mechanisms used to transport and allocate carbon in trees. Sucrose is the major form in which carbon is transported, found in the phloem sap of all so far investigated tree species. However, in several tree species sucrose is accompanied by other molecules, notably polyols and the raffinose family of oligosaccharides. We describe the molecules that constitute each of these transport groups, and their distribution across different tree species. Further, we detail the metabolic reactions for their synthesis, the mechanisms by which trees load and unload these compounds in and out of the vascular system, and how they are radially transported in the trunk and finally catabolized during wood formation. We also address a particular carbon recirculation process between phloem and xylem that occurs in trees during the annual cycle of growth and dormancy. A search of possible evolutionary drivers behind the diversity of C carrying molecules in trees reveals no consistent differences in carbon transport mechanisms between angiosperm and gymnosperm trees. Furthermore, the distribution of C forms across species suggests that climate related environmental factors will not either explain the diversity of carbon transport forms. However, the consideration of C transport mechanisms in relation to tree—rhizosphere coevolution deserves further attention. To conclude the review, we identify possible future lines of research in this field.

Description: To better understand hydraulic adaptations of coastal Douglas-fir (P. menziesii var. menziesii (Mirb.) Franco) to local climate, we examined genetic (G) and environmental (E) responses of branch hydraulic architecture of 7-year-old saplings from dry and wet climates of origin grown at a relatively dry and a relatively wet common garden site in western Oregon. We sampled two years of branch growth from three dry-source and three wet-source families grown at both sites (72 branches, total). Overall, only 4 of the 11 traits had significant genetic (G) effects, whereas 9 traits had significant environmental (E) effects (P 

Description: Considering the temporal responses of carbon isotope discrimination (Δ13C) to local water availability in the spatial analysis of Δ13C is essential for evaluating the contribution of environmental and genetic facets of plant Δ13C. Using tree-ring Δ13C from years with contrasting water availability at 76 locations across the natural range of loblolly pine, we decomposed site-level Δ13C signals to maximum Δ13C in well-watered conditions (Δ13Cmax) and isotopic drought sensitivity (m) as a change in Δ13C per unit change of Palmer’s Drought Severity Index (PDSI). Site water status, especially the tree lifetime average PDSI, was the primary factor affecting Δ13Cmax. The strong spatial correlation exhibited by m was related to both genetic and environmental factors. The long-term average water availability during the period relevant to trees as indicated by lifetime average PDSI correlated with Δ13Cmax, suggesting acclimation in tree gas-exchange traits, independent of incident water availability. The positive correlation between lifetime average PDSI and m indicated that loblolly pines were more sensitive to drought at mesic than xeric sites. The m was found to relate to a plant’s stomatal control, and may be employed as a genetic indicator of efficient water use strategies. Partitioning Δ13C to Δ13Cmax and m provided a new angle for understanding sources of variation in plant Δ13C, with several fundamental and applied implications.

Description: Pine wood nematodes (PWNs; Bursaphelenchus xylophilus) infect pine trees and cause serious pine wilt disease. Eastern white pine (Pinus strobus) has resistance to PWN. However, the detailed defense mechanisms of P. strobus against PWN are not well known. When P. strobus plants were infected with PWNs, the accumulation of stilbenoids, dihydropinosylvin monomethyl ether (DPME) and pinosylvin monomethyl ether (PME) was increased remarkably. Both DPME and PME had high nematicidal activity. The nematicidal activity of the two compounds was resulted in a developmental stage-dependent manner. Pinosylvin monomethyl ether was more toxic to adult PWNs than juveniles, whereas DPME was found more toxic to juvenile PWNs than the adults. The genes involved in PME and DPME biosynthesis such as phenylalanine ammonia-lyase (PAL), 4-coumarate-CoA ligase (4CL), pinosylvin synthase (STS) and pinosylvin O-methyltransferase (PMT) were isolated using de novo sequencing of the transcriptome in P. strobus. In addition, transcription factors (TFs; bHLH, MYB and WRKY) related to stilbene biosynthesis were isolated. qPCR analyses of the selected genes (PAL, 4CL, STS and PMT) including TFs (bHLH, MYB and WRKY) revealed that the expression level of the selected genes highly enhanced after PWN infection. Our results suggest that pinosylvin-type stilbenoid biosynthesis is highly responsive to PWN infection and plays an important role in PWN resistance of P. strobus trees.

Description: Dormancy release and reactivation of temperate-zone trees involve the temperature-modulated expression of cell-cycle genes. However, information on the detailed regulatory mechanism is limited. Here, we compared the transcriptomes of the stems of active and dormant larch trees, emphasizing the expression patterns of cell-cycle genes and transcription factors and assessed their relationships and responses to temperatures. Twelve cell-cycle genes and 31 transcription factors were strongly expressed in the active stage. Promoter analysis suggested that these 12 genes might be regulated by transcription factors from 10 families. Altogether, 73 cases of regulation between 16 transcription factors and 12 cell-cycle genes were predicted, while the regulatory interactions between LaMYB20 and LaCYCB1;1, and LaRAV1 and LaCDKB1;3 were confirmed by yeast one-hybrid and dual-luciferase assays. Last, we found that LaRAV1 and LaCDKB1;3 had almost the same expression patterns during dormancy release and reactivation induced naturally or artificially by temperature, indicating that the LaRAV1-LaCDKB1;3 module functions in the temperature-modulated dormancy release and reactivation of larch trees. These results provide new insights into the link between temperature and cell-cycle gene expression, helping to understand the temperature control of tree growth and development in the context of climate change.

Description: Variable winter temperatures cause a year-to-year discrepancy in the phenology of deciduous trees. This implies that an intrinsic ‘winter clock’ synchronizes bloom with the progression of winter to spring. The carbohydrate–temperature (C–T) model established a mechanistic association between carbohydrate metabolism in dormant trees and hourly winter temperatures. Using historical winter temperature and bloom times of Prunus dulcis (Mill.) D. A. Webb (almond), Malus domestica L. (apple), Pistachia vera L. (pistachio) and Juglans regia L. (walnut) in California and Washington states, we parametrized species-specific metabolic parameters to the C–T model. There was a sound fit between actual and projected bloom dates with a deviation (root mean square error) of 4–7 days in all species. The parameterized model enabled us to study how the observed variability in soluble carbohydrate concentrations at senescence (SC0) could affect bloom time. The C–T model projected that low SC0 could advance, while high SC0 possibly delays, the bloom of the early blooming almond trees. In contrast, high SC0 would advance the bloom of apple, pistachio and walnut trees. These novel projections suggest that after experimental validation, SC0 could guide post-harvest farming applications that affect fall carbohydrate accumulation to mediate the effects of climate shifts.

Description: Productivity of fruit tree crops depends on the interaction between plant physiology, environmental conditions and agricultural practices. We develop a mechanistic model of fruit tree crops that reliable simulates the dynamics of variables of interest for growers and consequences of agricultural practices while relying on a minimal number of inputs and parameters. The temporal dynamics of carbon content in the different organs (i.e., shoots—S, roots—R and fruits—F) are the result of photosynthesis by S, nutrient supply by R, respiration by S, R and F, competition among different organs, photoperiod and initial system conditions partially controlled by cultural practices. We calibrate model parameters and evaluate model predictions using unpublished data from a peach (Prunus persica) experimental orchard with trees subjected to different levels of branch pruning and fruit thinning. Fiinally, we evaluate the consequences of different combinations of pruning and thinning intensities within a multi-criteria analysis. The predictions are in good agreement with the experimental measurements and for the different conditions (pruning and thinning). Our simulations indicate that thinning and pruning practices actually used by growers provide the best compromise between total shoot production, which impacts next year’s abundance of shoots and fruits, and current year’s fruit production in terms of quantity (yield) and quality (average fruit size). This suggests that growers are not only interested in maximizing current year’s yield but also in its quality and its durability. The present work provides for modelers a system of equations based on acknowledged principles of plant science easily modifiable for different purposes. For horticulturists, it gives insights on the potentialities of pruning and thinning. For ecologists, it provides a transparent quantitative framework that can be coupled with biotic and abiotic stressors.

Description: Tracking wood formation in semiarid regions during the seasonal march of precipitation extremes has two important applications. It can provide (i) insight into the adaptive capacities of trees to drought and (ii) a basis for a richer interpretation of tree-ring data, assisting in a deeper understanding of past and current climate. In the southwestern USA, the anatomical signature of seasonally bimodal precipitation is the ‘false ring’—a band of latewood-like cells in the earlywood. These occur when a particularly deep drought during the early growing season ends abruptly with timely, mid-growing season monsoonal rains. Such conditions presented in southern Arizona in 2014, enabling us to explore false-ring formation in ponderosa pine (Pinus ponderosa Lawson and C. Lawson) and Douglas-fir (Pseudotsuga menziesii Mirb. Franco) in mixed-conifer forest at 2573 m above sea level. We ask: what were the cell-by-cell timings and durations in the phases of wood cell development in 2014? How do these seasonal patterns relate to strongly fluctuating environmental conditions during the growing season? We took weekly microcores from March through November from six ponderosa pine and seven Douglas-fir trees at a well-instrumented flux tower site. Thin sections were prepared, and we counted cells in cambial, expansion, cell wall thickening and mature phases. For ponderosa pine trees forming a false ring, the first impact of intensifying seasonal drought was seen in the enlarging phase and then, almost a month later, in cambial activity. In this species, recovery from drought was associated with recovery first in cambial activity, followed by cell enlargement. This timing raised the possibility that cell division may be affected by atmospheric moisture increases before soil recharge. In both species, the last false-ring cells matured during the summer rainy season. Bimodal cambial activity coincident with moisture availability was observed in both species, whether or not they formed a false ring. This deeper knowledge of the precise timing of both developmental and environmental events should help define mechanistic connections among these factors in creating bimodal growth patterns.

Description: Low temperatures and drought are the main environmental factors affecting plant growth and productivity across most of the terrestrial biomes. The objective of this study was to analyze the effects of water deficits before the onset of low temperatures in winter to enhance freezing resistance in olive trees. The study was carried out near the coast of Chubut, Argentina. Plants of five olive cultivars were grown outdoor in pots and exposed to different water deficit treatments. We assessed leaf water relations, ice nucleation temperature (INT), cell damage (LT50), plant growth and leaf nitrogen content during summer and winter in all cultivars and across water deficit treatments. Leaf INT and LT50 decreased significantly from summer to winter within each cultivar and between treatments. We observed a trade-off between resources allocation to freezing resistance and vegetative growth, such that an improvement in resistance to sub-zero temperatures was associated with lower growth in tree height. Water deficit applied during summer increased the amount of osmotically active solutes and decreased the leaf water potentials. This type of legacy effect persists during the winter after the water deficit even when treatment was removed by natural rainfalls.

Description: Elevated atmospheric CO2 concentration (eCO2) commonly stimulates net leaf assimilation, decreases stomatal conductance and has no clear effect on leaf respiration. However, effects of eCO2 on whole-tree functioning and its seasonal dynamics remain far more uncertain. To evaluate temporal and spatial variability in eCO2 effects, 1-year-old European aspen trees were grown in two treatment chambers under ambient (aCO2, 400 p.p.m.) and elevated (eCO2, 700 p.p.m.) CO2 concentrations during an early (spring 2019) and late (autumn 2018) seasonal experiment. Leaf (net carbon assimilation, stomatal conductance and leaf respiration) and whole-tree (stem growth, sap flow and stem CO2 efflux) responses to eCO2 were measured. Under eCO2, carbon assimilation was stimulated during the early (1.63-fold) and late (1.26-fold) seasonal experiments. Stimulation of carbon assimilation changed over time with largest increases observed in spring when stem volumetric growth was highest, followed by late season down-regulation, when stem volumetric growth ceased. The neutral eCO2 effect on stomatal conductance and leaf respiration measured at leaf level paralleled the unresponsive canopy conductance (derived from sap flow measurements) and stem CO2 efflux measured at tree level. Our results highlight that seasonality in carbon demand for tree growth substantially affects the magnitude of the response to eCO2 at both leaf and whole-tree level.

Description: Drought can lead to important shifts in population dynamics if it occurs during seedling establishment. With the aim of elucidating the underlying mechanisms of drought tolerance and resilience, here we monitored the survival of seedlings of the Mediterranean shrub Cistus albidus L. throughout a year growing in the natural Park of the Montserrat Mountains (Spain) and, additionally, we studied the response to severe drought and subsequent recovery after rewatering of seedlings grown in growth chambers. To find possible mechanisms explaining how seedlings respond to drought, growth and survival together with physiological-related parameters such as chlorophyll contents, vitamin E and stress-related phytohormones were measured. We found that survival decreased by 30% at the end of summer and that the main proxy of seedling survival was total chlorophyll. This proxy was further confirmed in the growth chambers, where we found that seedlings that recovered from drought had higher levels of total chlorophyll compared with the seedlings that did not recover. Furthermore, modulation of vitamin E and jasmonates contents appeared to be crucial in the drought response of C. albidus seedlings. We propose a prediction model of survival that includes total chlorophyll height, leaf mass area and maximum photosystem II efficiency with chlorophyll contents being a good long-term predictor of C. albidus seedling survival under severe stress, which, in turn, could help to better foresee population fluctuations in the field.

Description: Spring drought episodes are becoming more frequent and intensive in European temperate forests. To study tree resilience to spring drought, Norway spruce seedlings were exposed to three levels of drought stress (well-watered (W), moderately stressed (M) and severely stressed (S)) for 42 days and then fully irrigated for 14 days. Drought strongly reduced gas exchange parameters for both M and S seedlings. After 42 days, stomatal conductance was lower by 83 and 97% in M and S, respectively, than in W seedlings. Respiration prevailed over photosynthesis in S seedlings at the end of the drought period. Drought mostly reduced longitudinal growth, especially in shoots and needles. Xylem growth reduction was caused mainly by a lower number of newly produced tracheids, not by changes in their size. Norway spruce seedlings showed good resilience to spring drought, as the observed physiological parameters started to recover after rewatering and seedlings started to sprout and form new tracheids. In M seedlings, all physiological traits recovered to the level of W seedlings during the 14-day irrigation period but the recovery took longer in S seedlings. Shoots and needles did not regrow in length but leaf mass per area increased during the recovery phase. To conclude, Norway spruce seedlings showed good resilience to spring single-drought event, but time necessary to full recovery from stress could make seedlings more vulnerable to recurrent drought events.

Description: Extreme weather events are increasing in frequency and intensity due to global climate change. We hypothesized that tree carbon reserves are crucial for resilience of beech, buffering the source–sink imbalance due to late frosts and summer droughts, and that different components of non-structural carbohydrates (NSCs) play specific roles in coping with stressful situations. To assess the compound effects on mature trees of two extreme weather events, first a late frost in spring 2016 and then a drought in summer 2017, we monitored the phenology, radial growth and the dynamics of starch and soluble sugars in a Mediterranean beech forest. A growth reduction of 85% was observed after the spring late frost, yet not after the drought event. We observed a strong impact of late frost on starch, which also affected its dynamic at the beginning of the subsequent vegetative season. In 2017, the increase of soluble sugars, associated with starch hydrolysis, played a crucial role in coping with the severe summer drought. Non-structural carbohydrates helped to counteract the negative effects of both events, supporting plant survival and buffering source–sink imbalances under stressful conditions. Our findings indicate a strong trade-off between growth and NSC storage in trees. Overall, our results highlight the key role of NSCs on beech trees, response to extreme weather events, confirming the resilience of this species to highly stressful events. These insights are useful for assessing how forests may respond to the potential impacts of climate change on ecosystem processes in the Mediterranean area.

Description: Polyploidy generally provides an advantage in phenotypic variation and growth vigor. However, the underlying mechanisms remain poorly understood. The tetraploid Liriodendron sino-americanum (Liriodendron × sinoamericanum P.C Yieh ex C.B. Shang & Zhang R.Wang) exhibits altered morphology compared with its diploid counterpart, including larger, thicker and deeper green leaves, bigger stomata, thicker stems and increased tree height. Such characteristics can be useful in ornamental and industrial applications. To elucidate the molecular mechanisms behind this variation, we performed a comparative transcriptome and proteome analysis. Our transcriptome data indicated that some photosynthesis genes and pathways were differentially altered and enriched in tetraploid L. sino-americanum, mainly related to F-type ATPase, the cytochrome b6/f complex, photosynthetic electron transport, the light harvesting chlorophyll protein complexes, and photosystem I and II. Most of the differentially expressed proteins we could identify are also involved in photosynthesis. Our physiological results showed that tetraploids have an enhanced photosynthetic capacity, concomitant with great levels of sugar and starch in leaves. This suggests that tetraploid L. sino-americanum might experience comprehensive transcriptome reprogramming of genes related to photosynthesis. This study has especially emphasized molecular changes involved in photosynthesis that accompany polyploidy, and provides a possible explanation for the altered phenotype of polyploidy plants in comparison with their diploid form.

Description: Adventitious rooting of walnut species (Juglans L.) is known to be rather difficult, especially for mature trees. The adventitious root formation (ARF) capacities of mature trees can be significantly improved by rejuvenation. However, the underlying gene regulatory networks (GRNs) of rejuvenation remain largely unknown. To characterize such regulatory networks, we carried out the transcriptomic study using RNA samples of the cambia and peripheral tissues on the bottom of rejuvenated and mature walnut (Juglans hindsii × J. regia) cuttings during the ARF. The RNA sequencing data suggested that zeatin biosynthesis, energy metabolism and substance metabolism were activated by rejuvenation, whereas photosynthesis, fatty acid biosynthesis and the synthesis pathways for secondary metabolites were inhibited. The inter- and intra-module GRNs were constructed using differentially expressed genes. We identified 35 hub genes involved in five modules associated with ARF. Among these hub genes, particularly, beta-glucosidase-like (BGLs) family members involved in auxin metabolism were overexpressed at the early stage of the ARF. Furthermore, BGL12 from the cuttings of Juglans was overexpressed in Populus alba × P. glandulosa. Accelerated ARF and increased number of ARs were observed in the transgenic poplars. These results provide a high-resolution atlas of gene activity during ARF and help to uncover the regulatory modules associated with the ARF promoted by rejuvenation.

Description: Abies alba (Mill.) has a high potential for mitigating climate change in European mountain forests; yet, its natural regeneration is severely limited by ungulate browsing. Here, we simulated browsing in a common garden experiment to study growth and physiological traits, measured from bulk needles, using a randomized block design with two levels of browsing severity and seedlings originating from 19 populations across Switzerland. Genetic factors explained most variation in growth (on average, 51.5%) and physiological traits (10.2%) under control conditions, while heavy browsing considerably reduced the genetic effects on growth (to 30%), but doubled those on physiological traits related to carbon storage. While browsing reduced seedling height, it also lowered seedling water-use efficiency (decreased $delta ^{13}$C) and increased their $delta ^{15}$N. Different populations reacted differently to browsing stress, and for seedling height, starch concentration and $delta ^{15}$N, population differences appeared to be the result of natural selection. First, we found that populations originating from the warmest regions recovered the fastest from browsing stress, and they did so by mobilizing starch from their needles, which suggests a genetic underpinning for a growth-storage trade-off across populations. Second, we found that seedlings originating from mountain populations growing on steep slopes had a higher $delta ^{15}$N in the common garden than those originating from flat areas, indicating that they have been selected to grow on N-poor, potentially drained, soils. This finding was corroborated by the fact that nitrogen concentration in adult needles was lower on steep slopes than on flat ground, strongly indicating that steep slopes are the most N-poor environments. These results suggest that adaptation to climate and soil nitrogen availability, as well as ungulate browsing pressure, co-determine the regeneration and range limit of silver fir.

Description: Drought stress is an environmental factor that seriously threatens plant growth, development and yield. VQ proteins are transcriptional regulators that have been reported to be involved in plant growth, development and the responses to biotic and abiotic stressors. However, the relationship between VQ proteins and drought stress has not been well documented in plants. In this study, overexpressing the apple VQ motif-containing protein (MdVQ37) gene in apple plants markedly reduced the tolerance to drought. Physiological and biochemical studies further demonstrated lower enzymatic activities and decreased photosynthetic capacity in transgenic lines compared with wild-type (WT) plants under drought stress. Ultrastructural analysis of leaves showed that the leaves and palisade tissues from the transgenic lines were significantly thinner than those from WT plants. Salicylic acid (SA) analysis indicated that overexpression of MdVQ37 increased the accumulation of 2,5-DHBA by up-regulating the expression of the SA catabolic gene, which ultimately resulted to a significant reduction in endogenous SA content and the disruption of the SA-dependent signaling pathway under drought stress. Applying SA partially increased the survival rate of the transgenic lines under drought stress. These results demonstrate that the regulatory function of apple MdVQ37 is implicated in drought stress, through a change in leaf development and SA homeostasis. This study provides novel insight into understanding the multiple functions of VQ proteins.

Description: Light and water availability are likely to vary over the lifespan of closed-canopy forest trees with understory trees experiencing greater limitations to growth by light and canopy trees greater limitation due to drought. As drought and shade have opposing effects on isotope discrimination (Δ13C), paired measurement of ring width and Δ13C can potentially be used to differentiate between water and light limitations on tree growth. We tested this approach for Cedrela trees from three tropical forests in Bolivia and Mexico that differ in rainfall and canopy structure. Using lifetime ring width and Δ13C data for trees of up to and over 200 years old, we assessed how controls on tree growth changed from understory to the canopy. Growth and Δ13C are mostly anti-correlated in the understory but this anti-correlation disappeared or weakened when trees reached the canopy, especially at the wettest site. This indicates that understory growth variation is controlled by photosynthetic carbon assimilation (A) due to variation in light levels. Once trees reached the canopy, inter-annual variation in growth and Δ13C at one of the dry sites showed positive correlations, indicating inter-annual variation in growth is driven by variation in water stress affecting stomatal conductance (gs). Paired analysis of ring widths and carbon isotopes provides significant insight to discerning between environmental factors controlling growth over trees’ life; strong light limitations for understory trees in closed-canopy moist forests switched to drought stress for (sub)canopy trees in dry forests. We show that combined isotope and ring width measurements can significantly improve our insights in tree functioning and be used to disentangle limitations due to shade from those due to drought.

Description: Canopy structure—the size and distribution of tree crowns and the spatial and temporal distribution of leaves within them—exerts dominant control over primary productivity, transpiration and energy exchange. Stand structure—the spatial arrangement of trees in the forest (height, basal area and spacing)—has a strong influence on forest growth, allocation and resource use. Forest response to elevated atmospheric CO2 is likely to be dependent on the canopy and stand structure. Here, we investigated elevated CO2 effects on the forest structure of a Liquidambar styraciflua L. stand in a free-air CO2 enrichment experiment, considering leaves, tree crowns, forest canopy and stand structure. During the 12-year experiment, the trees increased in height by 5 m and basal area increased by 37%. Basal area distribution among trees shifted from a relatively narrow distribution to a much broader one, but there was little evidence of a CO2 effect on height growth or basal area distribution. The differentiation into crown classes over time led to an increase in the number of unproductive intermediate and suppressed trees and to a greater concentration of stand basal area in the largest trees. A whole-tree harvest at the end of the experiment permitted detailed analysis of canopy structure. There was little effect of CO2 enrichment on the relative leaf area distribution within tree crowns and there was little change from 1998 to 2009. Leaf characteristics (leaf mass per unit area and nitrogen content) varied with crown depth; any effects of elevated CO2 were much smaller than the variation within the crown and were consistent throughout the crown. In this young, even-aged, monoculture plantation forest, there was little evidence that elevated CO2 accelerated tree and stand development, and there were remarkably small changes in canopy structure. Questions remain as to whether a more diverse, mixed species forest would respond similarly.

Description: Microbes living in plant tissues—endophytes—are mainly studied in crop plants where they typically colonize the root apoplast. Trees—a large carbon source with a high capacity for photosynthesis—provide a variety of niches for endophytic colonization. We have earlier identified a new type of plant–endophyte interaction in buds of adult Scots pine, where Methylorubrum species live inside the meristematic cells. The endosymbiont Methylorubrum extorquens DSM13060 significantly increases needle and root growth of pine seedlings without producing plant hormones, but by aggregating around host nuclei. Here, we studied gene expression and metabolites of the pine host induced by M. extorquens DSM13060 infection. Malic acid was produced by pine to potentially boost M. extorquens colonization and interaction. Based on gene expression, the endosymbiont activated the auxin- and ethylene (ET)-associated hormonal pathways through induction of CUL1 and HYL1, and suppressed salicylic and abscisic acid signaling of pine. Infection by the endosymbiont had an effect on pine meristem and leaf development through activation of GLP1-7 and ALE2, and suppressed flowering, root hair and lateral root formation by downregulation of AGL8, plantacyanin, GASA7, COW1 and RALFL34. Despite of systemic infection of pine seedlings by the endosymbiont, the pine genes CUL1, ETR2, ERF3, HYL, GLP1-7 and CYP71 were highly expressed in the shoot apical meristem, rarely in needles and not in stem or root tissues. Low expression of MERI5, CLH2, EULS3 and high quantities of ononitol suggest that endosymbiont promotes viability and protects pine seedlings against abiotic stress. Our results indicate that the endosymbiont positively affects host development and stress tolerance through mechanisms previously unknown for endophytic bacteria, manipulation of plant hormone signaling pathways, downregulation of senescence and cell death-associated genes and induction of ononitol biosynthesis.

Description: Quantifying inter-specific variations of tree resilience to drought and revealing the underlying mechanisms are of great importance to the understanding of forest functionality particularly in water-limited regions. So far, comprehensive studies incorporating investigations in inter-specific variations of long-term growth patterns of trees and the underlying physiological mechanisms are very limited. Here, in a semi-arid site of northern China, tree radial growth rate, inter-annual tree-ring growth responses to climate variability, as well as physiological characteristics pertinent to xylem hydraulics, carbon assimilation and drought tolerance were analyzed in seven pine species growing in a common environment. Considerable inter-specific variations in radial growth rate, growth response to drought and physiological characteristics were observed among the studied species. Differently, the studied species exhibited similar degrees of resistance to drought-induced branch xylem embolism with water potential corresponding to 50% loss hydraulic conductivity ranged from −2.31 to −2.96 MPa. We found that higher branch hydraulic efficiency is related to greater leaf photosynthetic capacity, smaller hydraulic safety margin and lower woody density (P  0.05). Rather, species with higher hydraulic conductivity and photosynthetic capacity was more sensitive to drought stress and tended to show weaker growth resistance to extreme drought events as quantified by tree-ring analyses, which is at least partially due to a trade-off between hydraulic efficiency and safety across species. This study thus demonstrates the importance of drought resilience rather than instantaneous water and carbon flux capacity in determining tree growth in water-limited environments.

Description: Boreal trees are capable of taking up organic nitrogen (N) as effectively as inorganic N. Depending on the abundance of soil N forms, plants may adjust physiological and morphological traits to optimize N uptake. However, the link between these traits and N uptake in response to soil N sources is poorly understood. We examined Pinus sylvestris seedlings’ biomass growth and allocation, transpiration, and N uptake in response to additions of organic (the amino acid arginine) or inorganic N (ammonium-nitrate). We also monitored in-situ soil N fluxes in the pots following an addition of N, using a microdialysis system. Supplying organic N resulted in a stable soil N flux, whereas the inorganic N resulted in a sharp increase of nitrate flux followed by a rapid decline, demonstrating a fluctuating N supply and a risk for loss of nitrate from the growth medium. Seedlings supplied with organic N achieved a greater biomass with a higher N content, thus reaching a higher N recovery compared with those supplied inorganic N. In spite of a higher N concentration in organic N seedlings, root-to-shoot ratio and transpiration per unit leaf area were similar to those of inorganic N seedlings. We conclude that enhanced seedlings’ nutrition and growth under the organic N source may be attributed to a stable supply of N, owing to a strong retention rate in the soil medium.

Description: Natural rubber, a strategically essential raw material used in manufacturing throughout the world, is produced from coagulated and refined latex of rubber tree (Hevea brasiliensis). It is known that phytohormone jasmonate (JA) plays an essential role in regulating latex biosynthesis. However, it is unclear how the JA signal is sensed in a rubber tree. Here, we showed that Hevea brasiliensis CORONATINE-INSENSITIVE 1 (HbCOI1) acts as a receptor that perceives JA to recruit HbJAZ1 for signal transduction. We found that HbCOI1 restores male sterility and JA responses of the coi1–1 mutant in Arabidopsis. The identification of a JA receptor in the rubber tree is essential for elucidating the molecular mechanisms underlying JA-regulated latex biosynthesis. Our results elucidate the mechanism of JA perception in Hevea brasiliensis and also provide an efficient strategy to identify JA receptors in woody plants.

Description: Nervonic acid (24:1) is a major component in nerve and brain tissues and it has important applications in food and pharmaceutical industries. Malania oleifera seeds contain about 40% nervonic acid. However, the mechanism of nervonic acid biosynthesis and accumulation in seeds of this endangered tree species remains unknown. In this study, developmental changes in fatty acid composition within embryos and their pericarps were investigated. Nervonic acid proportions steadily increased in developing embryos but 24:1 was not detected in pericarps at any stage. Two 3-ketoacyl-CoA synthase (KCS) homologs have been isolated from M. oleifera developing seeds by homologous cloning methods. Both KCSs are expressed in developing embryos but not detected in pericarps. Based on a phylogenetic analysis, these two KCSs were named as MoKCS4 and MoKCS11. Seed-specific expression of the MoKCS11 in Arabidopsis led to about 5% nervonic acid accumulation, while expression of the MoKCS4 did not show an obvious change in fatty acid composition. It is noteworthy that the transformation of the same MoKCS11 construct into two Brassica. napus cultivars with high erucic acid did not produce the expected accumulation of nervonic acid, although expression of MoKCS11 was detected in the developing embryos of transgenic lines. In contrast, overexpression of MoKCS11 results in similar level of nervonic acid accumulation in camelina, a species which contains a similar level of 11Z-eicosenoic acid as does Arabidopsis. Taken together, the MoKCS11 may have a substrate preference for 11Z-eicosenoic acid, but not for erucic acid, in planta.

Description: In temperate trees, optimal timing and quality of flowering directly depend on adequate winter dormancy progression, regulated by a combination of chilling and warm temperatures. Physiological, genetic and functional genomic studies have shown that hormones play a key role in bud dormancy establishment, maintenance and release. We combined physiological and transcriptional analyses, quantification of abscisic acid (ABA) and gibberellins (GAs), and modeling to further investigate how these signaling pathways are associated with dormancy progression in the flower buds of two sweet cherry cultivars. Our results demonstrated that GA-associated pathways have distinct functions and may be differentially related with dormancy. In addition, ABA levels rise at the onset of dormancy, associated with enhanced expression of ABA biosynthesis PavNCED genes, and decreased prior to dormancy release. Following the observations that ABA levels are correlated with dormancy depth, we identified PavUG71B6, a sweet cherry UDP-GLYCOSYLTRANSFERASE gene that up-regulates active catabolism of ABA to ABA glucosyl ester (ABA-GE) and may be associated with low ABA content in the early cultivar. Subsequently, we modeled ABA content and dormancy behavior in three cultivars based on the expression of a small set of genes regulating ABA levels. These results strongly suggest the central role of ABA pathway in the control of dormancy progression and open up new perspectives for the development of molecular-based phenological modeling.

Description: The carbon and oxygen isotopic composition of water and assimilates in plants reveals valuable information on plant response to climatic conditions. Yet, the C and O uptake, incorporation, and allocation processes determining isotopic compositions are not fully understood. We carried out a dual-isotope labelling experiment at high humidity with 18O-enriched water (H218O) and 13C-enriched CO2 (13CO2) with attached Scots pine (Pinus sylvestris) branches and detached twigs of hemi-parasitic mistletoes (Viscum album ssp. austriacum) in a naturally dry coniferous forest, where also a long-term irrigation takes place. After 4 hours of label exposure, we sampled previous and recent year leaves, twig phloem, and twig xylem over 192 h for the analysis of isotope ratios in water and assimilates. For both species, the uptake into leaf water and the incorporation of the 18O-label into leaf assimilates was not influenced by soil moisture, while the 13C-label incorporation into assimilates was significantly higher under irrigation compared to control dry conditions. Species-specific differences in leaf morphology or needle age did not affect 18O-label uptake into leaf water, but the incorporation of both tracers into assimilates was two times lower in mistletoe than in pine. The 18O-label allocation in water from pine needles to twig tissues was two times higher for phloem than for xylem under both soil moisture conditions. In contrast, the allocation of both tracers in pine assimilates were similar and not affected by soil moisture, twig tissue or needle age. Soil moisture effects on 13C-label but not on 18O-label incorporation into assimilates can be explained by the stomatal responses at high humidity, non-stomatal pathways for water, and isotope exchange reactions. Our results suggest that non-photosynthetic 18O-incorporation processes may have masked prevalent photosynthetic processes. Thus, isotopic variation in leaf water could also be imprinted on assimilates when photosynthetic assimilation rates are low.

Description: For trees in forests, striving for light is matter of life and death, either by growing taller towards brighter conditions or by expanding the crown to capture more of the available light. Here we present a mechanistic model for the development path of stem height and crown size, accounting for light capture and growth, as well as mortality risk. We determine the optimal growth path among all possible trajectories using dynamic programming. The optimal growth path follows a sequence of distinct phases: (i) initial crown size expansion, (ii) stem height growth towards the canopy, (iii) final expansion of the crown in the canopy, and (iv) seed production without further increase in size. The transition points between these phases can be optimized by maximizing fitness, defined as expected life-time reproductive production. The results imply that to reach the canopy in an optimal way, trees must consider the full profile of expected increasing light levels towards the canopy. A short-sighted maximization of growth based on initial light conditions can result in arrested height growth, preventing the tree from reaching the canopy. The previous result can explain canopy stratification, and why canopy species often get stuck at certain size under a shading canopy. The model explains why trees with lower wood density have a larger diameter at a given tree height and grow taller than trees with higher wood density. The model can be used to implement plasticity in height versus diameter growth in individual based vegetation and forestry models.

Description: Several studies have suggested that CO2 transport in the transpiration stream can considerably bias estimates of root and stem respiration in ring-porous and diffuse-porous tree species. Whether this also happens in species with tracheid xylem anatomy and lower sap flow rates, such as conifers, is currently unclear. We infused 13C-labeled solution into the xylem near the base of two 90-year-old Pinus sylvestris trees. A custom-built gas exchange system and an online isotopic analyzer were used to sample the CO2 efflux and its isotopic composition continuously from four positions along the bole and one upper canopy shoot in each tree. Phloem and needle tissue 13C enrichment was also evaluated at these positions. Most of the 13C label was lost by diffusion within a few meters of the infusion point indicating rapid CO2 loss during vertical xylem transport. No 13C enrichment was detected in the upper bole needle tissues. Furthermore, mass balance calculations showed that c. 97% of the locally respired CO2 diffused radially to the atmosphere. Our results support the notion that xylem CO2 transport is of limited magnitude in conifers. This implies that the concerns that stem transport of CO2 derived from root respiration biases chamber-based estimates of forest carbon cycling may be unwarranted for mature conifer stands.

Description: Wood density (WD) is often used as a proxy for hydraulic traits such as vulnerability to drought-induced xylem cavitation and maximum water transport capacity, with dense-wooded species generally being more resistant to drought-induced xylem cavitation, having lower rates of maximum water transport and lower sapwood capacitance than light-wooded species. However, relationships between WD and the hydraulic traits that they aim to predict have not been well established in tropical forests, where modeling is necessary to predict drought responses for a high diversity of unmeasured species. We evaluated WD and relationships with stem xylem vulnerability by measuring cavitation curves, sapwood water release curves and minimum seasonal water potential (Ψmin) on upper canopy branches of six tree species and three liana species from a single wet tropical forest site in Panama. The objective was to better understand coordination and trade-offs among hydraulic traits and the potential utility of these relationships for modeling purposes. We found that parameters from sapwood water release curves such as capacitance, saturated water content and sapwood turgor loss point (Ψtlp,x) were related to WD, whereas stem vulnerability curve parameters were not. However, the water potential corresponding to 50% loss of hydraulic conductivity (P50) was related to Ψtlp,x and sapwood osmotic potential at full turgor (πo,x). Furthermore, species with lower Ψmin showed lower P50, Ψtlp,x and πo,x suggesting greater drought resistance. Our results indicate that WD is a good easy-to-measure proxy for some traits related to drought resistance, but not others. The ability of hydraulic traits such as P50 and Ψtlp,x to predict mortality must be carefully examined if WD values are to be used to predict drought responses in species without detailed physiological measurements.

Description: As an important economic crop in tropical areas, Areca catechu L. affects the livelihood of millions of farmers. The Areca yellow leaf phenomenon (AYLP) leads to severe crop losses and plant death. To better understand the relationship of microbes and chlorotic Areca leaves, microbial community structure as well as its correlation with differential metabolites was investigated by high-throughput sequencing and metabolomic approaches. High-throughput sequencing of the internal transcribed spacer 1 and 16S rRNA gene revealed that fungal diversity was dominated by Ascomycota and the bacterial community consisted of Proteobacteria as well as Actinobacteria. The microbiota structure on chlorotic Areca leaves exhibited significant changes based on non-metric multidimensional scaling analysis, which were attributed to 477 bacterial genera and 183 fungal genera. According to the results of the Kruskal–Wallis test, several potential pathogens were enriched on chlorotic Areca leaves. Further analysis based on metabolic pathways predicted by Phylogenetic Investigation of Communities by Reconstruction of Unobserved States revealed the metabolism of half-yellow leaves and yellow leaves microbiota were significantly elevated in amino acid metabolism, carbohydrate metabolism, glycan biosynthesis and metabolism, metabolism of cofactors and vitamins, partial xenobiotics biodegradation and metabolism. Furthermore, 22 significantly variable metabolites in Areca leaves were identified by ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry and statistical analysis. Moreover, we further investigated the correlation between the predominant microbes and differential metabolites. Taken together, the association between AYLP and microbiome of Areca leaves was explored from the microecological perspective by omics techniques, and these findings provide new insights into possible prevention, monitoring and control of AYLP in the future.

Description: Insect outbreaks of increasing frequency and severity in forests are predicted due to climate change. Insect herbivory is known to promote physiological changes in forest trees. However, little is known about whether these plant phenotypic adjustments have cascading effects on tree microbial symbionts such as fungi in roots and foliage. We studied the impact of defoliation by the pine processionary moth in two infested Pinus nigra forests through a multilevel sampling of defoliated and non-defoliated trees. We measured tree growth, nutritional status and carbon allocation to chemical defenses. Simultaneously, we analysed the putative impact of defoliation on the needle endophytes and on the soil fungal communities. Higher concentrations of chemical defenses were found in defoliated trees, likely as a response to defoliation; however, no differences in non-structural carbohydrate reserves were found. In parallel to the reductions in tree growth and changes in chemical defenses, we observed shifts in the composition of needle endophytic and soil fungal communities in defoliated trees. Defoliated trees consistently corresponded with a lower biomass of ectomycorrhizal fungi in both sites, and a higher alpha diversity and greater relative abundance of belowground saprotrophs and pathogens. However, ectomycorrhizal alpha diversity was similar between non-defoliated and defoliated trees. Specific needle endophytes in old needles were strongly associated with non-defoliated trees. The potential role of these endophytic fungi in pine resistance should be further investigated. Our study suggests that lower biomass of ectomycorrhizal fungi in defoliated trees might slow down tree recovery since fungal shifts might affect tree-mycorrhizal feedbacks and can potentially influence carbon and nitrogen cycling in forest soils.

Description: NAC (NAM, AFAT and CUC) proteins play necessary roles in plant response to environmental stresses. However, the functional roles of NAC genes in moso bamboo (Phyllostachys edulis), an essential economic perennial woody bamboo species, are not well documented. In this study, we retrieved 152 PeNAC genes from the moso bamboo V2 genome, and PeSNAC-1 was isolated and functionally characterized. PeSNAC-1 was localized in the nucleus and had no transactivation activity in yeast. PeSNAC-1 extremely expressed in rhizome and young roots (0.1 and 0.5 cm) and was significantly induced by drought and salt treatments but repressed by abscisic acid (ABA), methyl jasmonate and high temperature (42 °C) in moso bamboo. Under water shortage and salinity conditions, survival ratios, Fv/Fm values, physiological indexes such as activities of superoxide dismutase, peroxidase and catalase and contents of malondialdehyde, H2O2 and proline were significantly higher in transgenic rice than the wild type, which suggests enhanced tolerance to drought and salt stress in PeSANC-1 overexpressed plants. Transcript levels of Na+/H+ antiporter and Na+ transporter genes (OsSOS1, OsNHX1 and OsHKT1;5), ABA signaling and biosynthesis genes (OsABI2, OsRAB16, OsPP2C68, OsLEA3-1, OsLEA3, OsNCED3, OsNCED4 and OsNCED5) and ABA-independent genes (OsDREB1A, OsDREB1B and OsDREB2A) were substantially higher in transgenic as compared with the wild type. Moreover, protein interaction analysis revealed that PeSNAC-1 could interact with stress responsive PeSNAC-2/4 and PeNAP-1/4/5 in both yeast and plant cells, which indicates a synergistic effect of those proteins in regulating the moso bamboo stress response. Our data demonstrate that PeSNAC-1 likely improved salt and drought stress tolerance via modulating gene regulation in both ABA-dependent and independent signaling pathways in transgenic rice. In addition, PeSNAC-1 functions as an important positive stress regulator in moso bamboo, participating in PeSNAC-1 and PeSNAC-2/4 or PeSNAC-1 and PeNAP-1/4/5 interaction networks.

Description: Callus differentiation is a key developmental process in plant regeneration from cells. A better understanding of the genetic architecture of callus differentiation timing can help improve tissue transformation and the efficiency of artificial propagation. In this study, we investigated genotypic variation in callus differentiation capacity among 297 diverse P. euphratica trees sampled from a natural population. We employed a genome-wide association study (GWAS) of binary and growth-based parameters to identify loci and characterize the genetic architecture and genetic network underlying regulation of callus differentiation in P. euphratica. The results of this GWAS experiment suggested potential associations controlling whether the callus could differentiate and the process of callus differentiation. We identified multiple significant quantitative trait loci (QTLs), including the genes LOG1 and LOG7 and a locus containing WOX1. We reconstructed a genetic network that visualizes how each QTL interacts uniquely with other variants, and several core QTLs were detected that are involved in the degree of callus differentiation, providing potential targets for selection. This study represents one of the first to identify genetic variants affecting callus differentiation in a forest tree. Our results suggest that callus differentiation may be a typical qualitative-quantitative trait controlled by a major gene as well as polygenes across the genome of P. euphratica. This GWAS will help to design more complex and specific molecular tools for systematically manipulating organ regeneration.

Description: For a better understanding of plant nutrition processes, it is important to study the flux of nutrients within plants. However, existing xylem sap sampling methods are typically destructive and do not allow for repeated, highly frequent measurements of nutrient concentration. In this paper, we present a novel use of microdialysis (MD) for characterizing xylem sap phosphate (PO43−) concentration as a possible alternative to destructive sampling. First, MD probes were tested under laboratory conditions in vitro, in a stirred solution test, and in vivo, using beech tree stem segments. Exponential decline in the relative recovery (RR) with an increasing MD pumping rate allows for determining an optimal sampling interval (i.e., the maximum amount of sample volume with the minimum required concentration). The RR changed only minimally, with a change in the simulated sap flow velocity during the in vivo stem segment test. This suggests that MD can be applied over a range of naturally occurring sap flow velocities. Differences in the ionic strength between the xylem sap and the perfusate pumped through the MD did not influence the RR. Then, MD was successfully applied in a 24 h field campaign in two beech trees of different ages and allowed for in situ assessments of the diurnal variation of PO43− concentration and (together with xylem flow measurements) flux variability in living trees. Both beech trees exhibited the same diurnal pattern in PO43− concentrations with higher concentrations in the younger tree. The xylem PO43− concentration measured with MD was in the same order of magnitude as that received through destructive sampling in the younger tree. The MD probes did not show a decline in RR after the field application. We showed that MD can be applied to capture the PO43− concentration dynamics in the xylem sap with bihourly resolution under field conditions.

Description: Apple cuticular wax can protect plants from environmental stress, determine fruit luster and improve postharvest fruit storage quality. In recent years, dry weather, soil salinization and adverse environmental conditions have led to declines in apple fruit quality. However, few studies have reported the molecular mechanisms of apple cuticular wax biosynthesis. In this study, we identified a long-chain acyl-CoA synthetase MdLACS2 gene from apple. The MdLACS2 protein contained an AMP-binding domain and demonstrated long-chain acyl-CoA synthetase activity. MdLACS2 transgenic Arabidopsis exhibited reductions in epidermal permeability and water loss; change in the expression of genes related to cuticular wax biosynthesis, transport and transcriptional regulation; and differences in the composition and ultrastructure of cuticular wax. Moreover, the accumulation of cuticular wax enhanced the resistance of MdLACS2 transgenic plants to drought and salt stress. The main protein functional interaction networks of LACS2 were predicted, revealing a preliminary molecular regulation pathway for MdLACS2-mediated wax biosynthesis in apple. Our study provides candidate genes for breeding apple varieties and rootstocks with better fruit quality and higher stress resistance.

Description: Soil alkalization affects apple production in northwest China. Autophagy is a highly conserved degradative protein pathway in eukaryotes. Autophagy in plants can be activated by various abiotic factors. We previously identified the positive role of the autophagy-related gene MdATG18a in drought, nitrogen deficiency and resistance to Diplocarpon mali infection in apple. However, it is still unclear whether ATG18a is related to alkaline stress. In this study, we used hydroponic culture to simulate alkaline stress and found that the overexpression of MdATG18a significantly improved the tolerance of apple to alkaline stress. The overexpression of MdATG18a increased biomass, photosynthetic rate and antioxidant capacity of transgenic plants compared with wild-type plants under alkaline stress. The overexpression of MdATG18a promoted γ-aminobutyric acid (GABA) shunt via an increase in glutamate (GABA precursor) and GABA contents and upregulation of GABA shunt-related genes. In addition, the overexpression of MdATG18a significantly upregulated the expression of other core ATG genes and increased the formation of autophagosomes under alkaline stress. In conclusion, these results suggest that the overexpression of MdATG18a in apple enhances alkaline tolerance and the GABA shunt, which may be owing to the increase in autophagic activity.

Description: Conifers have evolved different chemical and anatomical defences against a wide range of antagonists. Resin ducts produce, store and translocate oleoresin, a complex terpenoid mixture that acts as both a physical and a chemical defence. Although resin duct characteristics (e.g., number, density, area) have been positively related to biotic resistance in several conifer species, the literature reporting this association remains inconclusive. Axial resin ducts recorded in annual growth rings are an archive of annual defensive investment in trees. This whole-life record of defence investment can be analysed using standard dendrochronological procedures, which allows us to assess interannual variability and the effect of understudied drivers of phenotypic variation on resin-based defences. Understanding the sources of phenotypic variation in defences, such as genetic differentiation and environmental plasticity, is essential for assessing the adaptive potential of forest tree populations to resist pests under climate change. Here, we reviewed the evidence supporting the importance of resin ducts in conifer resistance, and summarized current knowledge about the sources of variation in resin duct production. We propose a standardized methodology to measure resin duct production by means of dendrochronological procedures. This approach will illuminate the roles of resin ducts in tree defence across species, while helping to fill pivotal knowledge gaps in plant defence theory, and leading to a robust understanding of the patterns of variation in resin-based defences throughout the tree’s lifespan.

Description: Plant respiration can acclimate to changing environmental conditions and vary between species as well as biome types, although belowground respiration responses to ongoing climate warming are not well understood. Understanding the thermal acclimation capacity of root respiration (Rroot) in relation to increasing temperatures is therefore critical in elucidating a key uncertainty in plant function in response to warming. However, the degree of temperature acclimation of Rroot in rainforest trees and how root chemical and morphological traits are related to acclimation is unknown. Here we investigated the extent to which respiration of fine roots (≤2 mm) of four tropical and four warm-temperate rainforest tree seedlings differed in response to warmer growth temperatures (control and +6 °C), including temperature sensitivity (Q10) and the degree of acclimation of Rroot. Regardless of biome type, we found no consistent pattern in the short-term temperature responses of Rroot to elevated growth temperature: a significant reduction in the temperature response of Rroot to +6 °C treatment was only observed for a tropical species, Cryptocarya mackinnoniana, whereas the other seven species had either some stimulation or no alteration. Across species, Rroot was positively correlated with root tissue nitrogen concentration (mg g−1), while Q10 was positively correlated with root tissue density (g cm−3). Warming increased root tissue density by 20.8% but did not alter root nitrogen across species. We conclude that thermal acclimation capacity of Rroot to warming is species-specific and suggest that root tissue density is a useful predictor of Rroot and its thermal responses in rainforest tree seedlings.

Description: The present study aimed to explore the possible functions of radial oxygen loss (ROL) on mangrove nutrition. A field survey was conducted to explore the relations among ROL, root anatomy and leaf N in different mangrove species along a continuous tidal gradient. Three mangroves with different ROL (Avicennia marina [A. marina], Kandelia obovata and Rhizophora stylosa) were then selected to further explore the dynamics of N at the root-soil interface. The results showed that seaward pioneer mangrove species such as A. marina appeared to exhibit higher leaf N despite growing under poorer nutrient conditions. Greater leaf N in pioneer mangroves coincided with their special root structure (e.g., high porosity together with a thin lignified/suberized exodermis) and powerful ROL. An interesting positive relation was observed between ROL and leaf N in mangroves. Moreover, rhizo-box data further showed that soil nitrification was also strongly correlated with ROL. A. marina, which had the highest ROL among the three mangrove species studied, consistently possessed the highest levels of NO3−, nitrification and ammonia-oxidizing bacteria and archaea gene copies in the rhizosphere. Besides, both NO3− and NH4+ influxes were found to be higher in the roots of A. marina when compared to those of K. obovata and R. stylosa. In summary, greater N acquisition by pioneer mangroves such as A. marina was strongly correlated with ROL which would regulate N transformation and translocation at the root-soil interface. The implications of this study may be significant in mangrove nutrition and the mechanisms involved in mangrove zonation.

Description: The Dynamic model has been described as one of the most accurate models to quantify chill accumulation based on hourly temperatures in nuts and temperate fruits. This model considers that a dynamic process occurs at a biochemical level that determines the endodormancy breaking through the accumulation of the so-called portions. The kinetic parameters present in the model should reflect how the fruit trees integrate chilling exposure and thus they should be characteristic for each species. However, the original parameter values, reported in the late 1980s, are still being used. Even if the use of such parameter values is useful to compare among chilling requirements (CRs) for different species or cultivars, it is not the optimal choice when one intends to explain the CR variations in different years for a given cultivar. In this work we propose a data-based model calibration that makes use of phenological data for different apricot cultivars within different years to obtain model parameters, which minimize the variations among years and that have, at the same time, physical meaning to characterize the incumbent species. Results reveal that the estimation not only reduces the accumulated portion dispersion within the considered time periods but also allows to improve the CR predictions for subsequent years. We propose a set of model parameter values to predict endodormancy breaking dates in the apricot cultivars studied here.

Description: Dark septate endophytes (DSEs) are one of the most studied groups of root fungal endophytes in recent years. However, the effects of DSE on host plant are still under debate, and the molecular mechanisms are poorly understood. In this study, we identified a DSE fungus of the genus Anteaglonium, named T010, from the wild blueberry. When inoculated into Vaccinium corymbosum L. plants, T010 could enhance root growth and promote shoot branching, leading to increased plant growth. By comparative transcriptome analysis, we obtained 1948 regulated differentially expressed genes (DEGs) from the V. corymbosum plants treated by T010. Further functional enrichment analysis identified a series of DEGs enriched in transcriptional regulation, material transport, phytohormone biosynthesis and flavonoid biosynthesis. Moreover, the comparative analysis of liquid chromatography–mass spectrometry verified that T010 treatment induced the changes in the contents of various phytohormones and flavonoids. This is the first report on the isolation of DSE fungi of the genus Anteaglonium from blueberry roots. Moreover, our results suggested that T010 colonization could result in a series of changes in cell metabolism, biosynthesis and signal pathways, thereby promoting plant growth. Particularly, the changes of phytohormone and flavonoid metabolism induced by T010 colonization might contribute to the promotion of blueberry growth. Our results will provide new insights into understanding of the interaction of DSE fungi and host plants, as well as the development and utilization of DSE preparations.

Description: An increased frequency of fire events on the Slovenian Karst is in line with future climate change scenarios for drought-prone environments worldwide. It is therefore of the utmost importance to better understand tree–fire–climate interactions for predicting the impact of changing environment on tree functioning. To this purpose, we studied the post-fire effects on leaf development, leaf carbon isotope composition (δ13C), radial growth patterns and the xylem and phloem anatomy in undamaged (H-trees) and fire-damaged trees (F-trees) of Quercus pubescens Willd. with good resprouting ability in spring 2017, the growing season after a rangeland fire in August 2016. We found that the fully developed canopy of F-trees reached only half of the leaf area index values measured in H-trees. Throughout the season, F-trees were characterized by higher water potential and stomatal conductivity and achieved higher photosynthetic rates compared to unburnt H-trees. The foliage of F-trees had more negative δ13C values than those of H-trees. This reflects that F-trees less frequently meet stomatal limitations due to reduced transpirational area and more favourable leaf-to-root ratio. In addition, the growth of leaves in F-trees relied more on the recent photosynthates than on reserves due to the fire disturbed starch accumulation in the previous season. Cambial production stopped 3 weeks later in F-trees, resulting in 60 and 22% wider xylem and phloem increments, respectively. A novel approach by including phloem anatomy in the analyses revealed that fire caused changes in conduit dimensions in the early phloem but not in the earlywood. However, premature formation of the tyloses in the earlywood vessels of the youngest two xylem increments in F-trees implies that xylem hydraulic integrity was also affected by heat. Analyses of secondary tissues showed that although xylem and phloem tissues are interlinked changes in their transport systems due to heat damage are not necessarily coordinated.

Description: Many non-native, invasive woody species in mesic forests of North America are both shade tolerant and more productive than their native counterparts, but their ability to tolerate disturbances remains unclear. In particular, complete defoliation associated with herbivory and extreme weather events may have larger impacts on invaders if natives maintain greater resource reserves to support regrowth. On the other hand, invaders may be more resilient to partial defoliation by means of upregulation of photosynthesis or may be better able to take advantage of canopy gaps to support refoliation. Across a light gradient, we measured radial growth, new leaf production, non-structural carbohydrates (NSCs), chlorophyll content and survival in response to varying levels of defoliation in saplings of two native and two invasive species that commonly co-occur in deciduous forests of Eastern North America. Individuals were subjected to one of the four leaf removal treatments: no-defoliation controls, 50% defoliation over three growing seasons, 100% defoliation over one growing season and 100% defoliation over two growing seasons. Contrary to our hypothesis, native and invasive species generally did not differ in defoliation responses, although invasive species experienced more pronounced decreases in leaf chlorophyll following full defoliation and native species’ survival was more dependent on light availability. Radial growth progressively decreased with increasing defoliation intensity, and refoliation mass was largely a function of sapling size. Survival rates for half-defoliated saplings did not differ from controls (90% of saplings survived), but survival rates in fully defoliated individuals over one and two growing seasons were reduced to 45 and 15%, respectively. Surviving defoliated saplings generally maintained control NSC concentrations. Under high light, chlorophyll concentrations were higher in half-defoliated saplings compared with controls, which may suggest photosynthetic upregulation. Our results indicate that native and invasive species respond similarly to defoliation, despite the generally faster growth strategy of invaders.

Description: Due to the increase in atmospheric CO2 concentrations, the ratio of carbon fixed by assimilation to water lost by transpiration through stomatal conductance (intrinsic water-use efficiency, iWUE) shows a long-term increasing trend globally. However, the drivers of short-term (inter-annual) variability in iWUE of tropical trees are poorly understood. We studied the inter-annual variability in iWUE of three South Asian tropical moist forest tree species (Chukrasia tabularis A.Juss., Toona ciliata M. Roem. and Lagerstroemia speciosa L.) derived from tree-ring stable carbon isotope ratio (δ13C) in response to variations of environmental conditions. We found a significantly decreasing trend in carbon discrimination (Δ13C) and an increasing trend in iWUE in all the three species, with a species-specific long-term trend in intercellular CO2 concentration (Ci). Growing season temperatures were the main driver of inter-annual variability of iWUE in C. tabularis and L. speciosa, whereas previous year temperatures determined the iWUE variability in T. ciliata. Vapor pressure deficit was linked with iWUE only in C. tabularis. Differences in shade tolerance, tree stature and canopy position might have caused this species-specific variation in iWUE response to climate. Linear mixed effect modeling successfully simulated iWUE variability, explaining 41–51% of the total variance varying with species. Commonality analysis revealed that temperatures had a dominant influence on the inter-annual iWUE variability (64–77%) over precipitation (7–22%) and atmospheric CO2 concentration (3–6%). However, the long-term variations in iWUE were explicitly determined by the atmospheric CO2 increase (83–94%). Our results suggest that the elevated CO2 and concomitant global warming might have detrimental effects on gas exchange and other physiological processes in South Asian tropical moist forest trees.

Description: Conifers are, for the most part, competitively excluded from tropical rainforests by angiosperms. Where they do occur, conifers often occupy sites that are relatively infertile. To gain insight into the physiological mechanisms by which angiosperms outcompete conifers in more productive sites, we grew seedlings of a tropical conifer (Podocarpus guatemalensis Standley) and an angiosperm pioneer (Ficus insipida Willd.) with and without added nutrients, supplied in the form of a slow-release fertilizer. At the conclusion of the experiment, the dry mass of P. guatemalensis seedlings in fertilized soil was approximately twofold larger than that of seedlings in unfertilized soil; on the other hand, the dry mass of F. insipida seedlings in fertilized soil was ~20-fold larger than seedlings in unfertilized soil. The higher relative growth rate of F. insipida was associated with a larger leaf area ratio and a higher photosynthetic rate per unit leaf area. Higher overall photosynthetic rates in F. insipida were associated with an approximately fivefold larger stomatal conductance than in P. guatemalensis. We surmise that a higher whole-plant hydraulic conductance in the vessel bearing angiosperm F. insipida enabled higher leaf area ratio and higher stomatal conductance per unit leaf area than in the tracheid bearing P. guatemalensis, which enabled F. insipida to capitalize on increased photosynthetic capacity driven by higher nitrogen availability in fertilized soil.

Description: Salix matsudana Koidz is a low cadmium (Cd)-accumulating willow, whereas its cultivated variety, Salix matsudana var. matsudana f. umbraculifera Rehd., is a high Cd-accumulating and tolerant willow (HCW). The physiological and molecular mechanisms underlying differential Cd accumulation and tolerance in the two Salix species are poorly understood. Here, we confirmed that the differential Cd translocation capacity from roots to the shoots leads to the differential Cd accumulation in their aboveground parts between these two willow genotypes. Cadmium accumulation happens preferentially in the transport pathway, and Cd is mainly located in the vacuolar, cell wall and intercellular space in HCW bark by cadmium location analysis at tissue and subcellular levels. Comparative transcriptome analysis revealed that higher expressions of several metal transporter genes (ATP-binding cassette transporters, K+ transporters/channels, yellow stripe-like proteins, zinc-regulated transporter/iron-regulated transporter-like proteins, etc.) are involved in root uptake and translocation capacity in HCW; meanwhile, ascorbate–glutathione metabolic pathways play essential roles in Cd detoxification and higher tolerance of the Cd-accumulator HCW. These results lay the foundation for further understanding the molecular mechanisms of Cd accumulation in woody plants and provide new insights into molecular-assisted-screening woody plant varieties for phytoremediation.

Description: Herbivory is one of the most globally distributed disturbances affecting carbon (C)-cycling in trees, yet our understanding of how it alters tree C-allocation to different functions such as storage, growth or rhizodeposition is still limited. Prioritized C-allocation to storage replenishment vs growth could explain the fast recovery of C-storage pools frequently observed in growth-reduced defoliated trees. We performed continuous 13C-labeling coupled to clipping to quantify the effects of simulated browsing on the growth, leaf morphology and relative allocation of stored vs recently assimilated C to the growth (bulk biomass) and non-structural carbohydrate (NSC) stores (soluble sugars and starch) of the different organs of two tree species: diffuse-porous (Betula pubescens Ehrh.) and ring-porous (Quercus petraea [Matt.] Liebl.). Carbon-transfers from plants to bulk and rhizosphere soil were also evaluated. Clipped birch and oak trees shifted their C-allocation patterns above-ground as a means to recover from defoliation. However, such increased allocation to current-year stems and leaves did not entail reductions in the allocation to the rhizosphere, which remained unchanged between clipped and control trees of both species. Betula pubescens and Q. petraea showed differences in their vulnerability and recovery strategies to clipping, the ring-porous species being less affected in terms of growth and architecture by clipping than the diffuse-porous. These contrasting patterns could be partly explained by differences in their C cycling after clipping. Defoliated oaks showed a faster recovery of their canopy biomass, which was supported by increased allocation of new C, but associated with large decreases in their fine root biomass. Following clipping, both species recovered NSC pools to a larger extent than growth, but the allocation of 13C-labeled photo-assimilates into storage compounds was not increased as compared with controls. Despite their different response to clipping, our results indicate no preventative allocation into storage occurred during the first year after clipping in either of the species.

Description: During the growing season, trees allocate photoassimilates to increase their aboveground woody biomass in the stem (ABIstem). This ‘carbon allocation’ to structural growth is a dynamic process influenced by internal and external (e.g., climatic) drivers. While radial variability in wood formation and its resulting structure have been intensively studied, their variability along tree stems and subsequent impacts on ABIstem remain poorly understood. We collected wood cores from mature trees within a fixed plot in a well-studied temperate Fagus sylvatica L. forest. For a subset of trees, we performed regular interval sampling along the stem to elucidate axial variability in ring width (RW) and wood density (ρ), and the resulting effects on tree- and plot-level ABIstem. Moreover, we measured wood anatomical traits to understand the anatomical basis of ρ and the coupling between changes in RW and ρ during drought. We found no significant axial variability in ρ because an increase in the vessel-to-fiber ratio with smaller RW compensated for vessel tapering towards the apex. By contrast, temporal variability in RW varied significantly along the stem axis, depending on the growing conditions. Drought caused a more severe growth decrease, and wetter summers caused a disproportionate growth increase at the stem base compared with the top. Discarding this axial variability resulted in a significant overestimation of tree-level ABIstem in wetter and cooler summers, but this bias was reduced to ~2% when scaling ABIstem to the plot level. These results suggest that F. sylvatica prioritizes structural carbon sinks close to the canopy when conditions are unfavorable. The different axial variability in RW and ρ thereby indicates some independence of the processes that drive volume growth and wood structure along the stem. This refines our knowledge of carbon allocation dynamics in temperate diffuse-porous species and contributes to reducing uncertainties in determining forest carbon fixation.

Description: Anthocyanin pigmentation is an important consumption trait of apple (Malus domestica Borkh.). In this study, we focused on the identification of NAC (NAM, ATAF1/2 and CUC2) proteins involved in the regulation of anthocyanin accumulation in apple flesh. A group of MdNACs was selected for comparison of expression patterns between the white-fleshed cultivar ‘Granny Smith’ and red-fleshed ‘Redlove’. Among them, MdNAC42 was screened, which exhibited a higher expression level in red-fleshed than in white-fleshed fruit, and has a positive correlation with anthocyanin content as fruits ripened. Moreover, overexpression of MdNAC42 in apple calli resulted in the up-regulation of flavonoid pathway genes, including MdCHS, MdCHI, MdF3H, MdDFR, MdANS and MdUFGT, thereby increasing the accumulation of anthocyanins, which confirmed the roles of MdNAC42 in anthocyanin biosynthesis. Notably, MdNAC42 was demonstrated to have an obvious interaction with MdMYB10 either in vitro or in vivo by yeast two-hybrid combined with bimolecular fluorescence complementation, further suggesting that MdNAC42 is an important part of the regulatory network controlling the anthocyanin pigmentation of red-fleshed apples. To the best of our knowledge, this is the first report identifying the MdNAC gene as related to anthocyanin accumulation in red-fleshed apples. This study provides valuable information for improving the regulatory model of anthocyanin biosynthesis in apple fruit.