The number of tree species on Earth.

One of the most fundamental questions in ecology is how many species inhabit the Earth. However, due to massive logistical and financial challenges and taxonomic difficulties connected to the species concept definition, the global numbers of species, including those of important and well-studied life forms such as trees, still remain largely unknown. Here, based on global ground-sourced data, we estimate the total tree species richness at global, continental, and biome levels. Our results indicate that there are ∼73,000 tree species globally, among which ∼9,000 tree species are yet to be discovered. Roughly 40% of undiscovered tree species are in South America. Moreover, almost one-third of all tree species to be discovered may be rare, with very low populations and limited spatial distribution (likely in remote tropical lowlands and mountains). These findings highlight the vulnerability of global forest biodiversity to anthropogenic changes in land use and climate, which disproportionately threaten rare species and thus, global tree richness.

Integrative analysis of physiology and genomics provides insights into freeze tolerance adaptations of Acacia koa along an elevational cline.

Natural selection for plant species in heterogeneous environments creates genetic variation for traits such as cold tolerance. While physiological or molecular analyses have been used to evaluate stress tolerance adaptations, combining these approaches may provide deeper insight. Acacia koa (koa) occurs from sea level to 2300 m in Hawai'i, USA. At high elevations, natural koa populations have declined due to deforestation, and freeze tolerance is a limiting factor for tree regeneration. We used physiology and molecular analyses to evaluate cold tolerance of koa populations from low (300-750 m), middle (750-1500 m), and high elevations (1500-2100 m). Half of the seedlings were cold acclimated by exposure to progressively lowered air temperatures for eight weeks (from 25.6/22.2°C to 8/4°C, day/night). Using the whole plant physiology-freezing test and koa C-repeat Binding Factor CBF genes, our results indicated that koa can be cold-acclimated when exposed to low, non-freezing temperatures. Seedlings from high elevations had consistently higher expression of Koa CBF genes associated with cold tolerance, helping to explain variation in cold-hardy phenotypes. Evaluation of the genetic background of 22 koa families across the elevations with low coverage RNA sequencing indicated that high elevation koa had relatively low values of heterozygosity, suggesting that adaptation is more likely to arise in the middle and low elevation sources. This physiology and molecular data for cold tolerance of koa across the elevation gradient of the Hawaiian Islands provides insights into natural selection processes and may help to support guidelines for conservation and seed transfer in forest restoration efforts.

Biochar alleviating heavy metals phytotoxicity in sludge-amended soil varies with plant adaptability.

Recycling sewage sludge (SS) to soil potentially causes soil heavy metal (HM) pollution and plant phytotoxicity. Biochar plays an important role in alleviating HM phytotoxicity, and responses vary with the feedstocks and usage of biochar. However, the effect of plant adaptability on biochar-mediated alleviation is poorly understood. Here, SS-derived biochar (SB) and rice straw-derived biochar (RB) applied at rates of 1.5% and 3% (W/W, SB1.5, SB3, RB1.5, and RB3) were used to improve the properties of soil amended with SS at 50% (W/W). Alleviation of phytotoxicity by biochar was further analyzed with SS-sensitive plant Monstera deliciosa and SS-resistant plant Ruellia simplex. Results revealed that both SB and RB significantly decreased the soil's bulk density and increased water retention. They also changed soil organic matter content and HMs fractionation. The addition of SB or RB alleviated the SS phytotoxicity, and they significantly promoted the growth and the root morphology and physiological index of M. deliciosa. But for R. simplex, these significant changes only synchronously occurred in SB3 treatment. The alleviation in M. deliciosa was more prominent and more closely connected with soil property changes than in R. simplex. Also, more soil property predictors were observed to play an important role in M. deliciosa growth than in R. simplex growth. These results indicated that biochar alleviating HMs phytotoxicity in SS-amended soil is associated with the changes of soil property. Moreover, the alleviation varies more prominently with plant adaptability than with biochar feedstocks and usage.

Plasticity of phenotype and heteroblasty in contrasting populations of Acacia koa.

BACKGROUND AND AIMS: Heteroblastic plant species, whose morphology or growth habit changes suddenly during development, offer unique opportunities to investigate the role of selection in canalizing development or increasing the adaptive importance of plasticity. Leaf forms of the Hawaiian tree Acacia koa (koa) change morphologically and physiologically during the first year of growth, providing time to study abiotic factors influencing transition rates relative to other Acacia species. METHODS: The roles of light and water availability in triggering transition to the mature leaf form in contrasting (wet/dry) ecotypes of koa were investigated using a novel modelling technique to distinguish between chronological and ontogenetic controls in triggering transition. A light quality treatment was included to test interactions of heterophylly (the presence of multiple leaf forms) with heteroblastic processes on the resulting phenotype at transition. KEY RESULTS: Increased light intensity increased transition rates, but reduced red to far-red light (R:FR) ratios did not affect transition rates, solidifying the current paradigm of heteroblasty. However, evidence was found for earlier transition ontogenetically under water stress, which is not part of the current paradigm and could differentiate the role of heteroblasty in some Acacia species versus other heteroblastic species. Ecotypic responses also indicate that plasticity of development could vary across koa's range and the adaptive significance of heteroblasty could be marginalized or amplified dependent on the disparate selective pressures present across koa's range. CONCLUSIONS: The use of novel survival functions and a species with an elongated transition time helped to elucidate abiotic modifiers of ontogenetic trajectories. Differences in ontogenetic trajectories between contrasting ecotypes suggest that ongoing climate and land use change will have non-uniform effects on koa regeneration and establishment dynamics across its range.

Forecasting effects of tree species reintroduction strategies on carbon stocks in a future without historical analog.

American chestnut (Castanea dentata) was once an important component forests in the central Appalachians (USA), but it was functionally extirpated nearly a century ago. Attempts are underway to reintroduce blight-resistant chestnut to its former range, but it is uncertain how current forest composition, climate, and atmospheric changes and disturbance regimes will interact to determine future forest dynamics and ecosystem services. The combination of novel environmental conditions (e.g. climate change), a reintroduced tree species and new disturbance regimes (e.g. exotic insect pests, fire suppression) have no analog in the past that can be used to parameterize phenomenological models. We therefore used a mechanistic approach within the LANDIS-II forest landscape model that relies on physiological first principles to project forest dynamics as the outcome of competition of tree cohorts for light and water as a function of temperature, precipitation, CO2 concentration, and life history traits. We conducted a factorial landscape simulation experiment to evaluate specific hypotheses about future forest dynamics in two study sites in the center of the former range of chestnut. Our results supported the hypotheses that climate change would favor chestnut because of its optimal temperature range and relative drought resistance, and that chestnut would be less competitive in the more mesic Appalachian Plateau province because competitors will be less stressed. The hypothesis that chestnut will increase carbon stocks was supported, although the increase was modest. Our results confirm that aggressive restoration is needed regardless of climate and soils, and that increased aggressiveness of chestnut restoration increased biomass accumulation. The hypothesis that chestnut restoration will increase both compositional and structural richness was not supported because chestnut displaced some species and age cohorts. Although chestnut restoration did not markedly enhance carbon stocks, our findings provide hope that this formerly important species can be successfully reintroduced and associated ecosystem services recovered.

Plantation performance of chestnut hybrids and progenitors on reclaimed Appalachian surface mines.

Reclamation of surface mined sites to forests is a preferred post-mining land use option, but performance of planted trees on such sites is variable. American chestnut (Castanea dentata (Marsh.) Borkh.) is a threatened forest tree in the eastern USA that may become an important species option for mine reclamation. Chestnut restoration using backcross hybrids that incorporate blight resistance may be targeted to the Appalachian coal mining region, which corresponds closely with the species' native range. Thus, it is important to understand how chestnut hybrids perform relative to progenitors on reclamation sites to develop restoration prescriptions. Seeds of parents and three backcross generations of chestnut (100% American, 100% Chinese, and BC1F3, BC2F3, and BC3F2 hybrids) were planted into mine soils in West Virginia, USA with shelter treatments. Survival for all stock types was 44% after 8 years (American 39%, Chinese 77%, BC1F3 40%, BC2F3 28%, and BC3F2 35%). Height for all stock types was 33 cm after 8 years (American 28 cm, Chinese 67 cm, BC1F3 30 cm, BC2F3 21 cm, and BC3F2 20 cm). At another site a year later, seedlings of the chestnut stock types were planted into brown (pH 4.6) or gray sandstone (pH 6.3) mine soils and seedling survival across all stock types was 58% after 7 years. Chinese had the highest survival at 82%, while the others ranged from 38 to 66%. Height was 63 cm for all stock types after 7 years. More advanced backcross hybrids (BC2F3 and BC3F2) had the lowest vigor ratings at both sites after 7-8 years. Our results indicate that surface mines in Appalachia may provide a land base for planting blight-resistant chestnuts, although Chinese chestnut outperformed American chestnut and later generation backcross hybrids. As blight-resistant chestnuts establish and spread after planting, chestnut trees may become a component of the forest canopy again and possibly occupy its former niche, but their spread may alter future forest stand dynamics.

Effects of woodland slope on heavy metal migration via surface runoff, interflow, and sediments in sewage sludge application.

Sewage sludge (SS) application to forest plantation soils as a fertilizer and/or soil amendment is increasingly adopted in plantation forest management. However, the potential risks of SS-derived heavy metals (HMs) remain a concern. Many factors, including woodland slope may affect the risks, but the understanding of this issue is limited. This research evaluated the HMs migration via surface runoff, interflow, and sediments when SS was applied in woodlands of varying slopes. We conducted indoor rainfall simulations and natural rainfall experiments to clarify the effect of slope on the migration of HMs via runoff (including surface and interflow) and sediments. In the simulated rainfall experiment, HMs lost via sediments increased by 9.79-27.28% when the slope increased from 5° to 25°. However, in the natural rainfall experiment, when the slope of forested land increased from 7° to 23°, HMs lost via surface runoff increased by 2.38% to 6.13%. These results indciate that the surface runoff water on a high slope (25°) posed high water quality pollution risks. The migration of HMs via surface runoff water or interflow increased as the steepness of the slope increased. The total migration of Cu, Zn, Pb, Ni, Cr and Cd via sediment greatly exceeded that via surface runoff and interflow. Particles ≤ 0.05 mm contributed the most to the ecological risks posed by sediments. Cd was the main source of potential ecological risks in sediments under both experimental conditions.

Effects of sewage sludge application methods on the transport of heavy metals with runoff and their mechanisms.

Woodland utilization is a promising disposal method for sewage sludge (SS). However, the potential risk of heavy metals (HMs) transport with runoff must be considered. Among the various factors influencing HMs loss, SS application methods (Holing application, HA; Broadcasting and mixing application, BM; Broadcasting application, BA) are likely to cause significant effects by altering soil erosion and soil aggregates. This study aimed to determine how SS application methods affect HMs loss, soil aggregates erosion, and how they are related. Accordingly, the losses of HMs in surface runoff, interflow, and sediment were quantified during six simulated rainfalls. The results demonstrated that all methods reduced surface runoff, but BA was the most effective. Additionally, BA significantly reduced the total sediment yield and the total proportion of the <0.05 mm fraction aggregates. Moreover, BA had the smallest cumulative losses of Pb and Cd through surface runoff and Cu, Pb, and Cd through sediment. Sediment was the most important pathway for HMs loss, through which over 76.56 % of HMs were lost. In BA, the <0.05 mm fraction aggregates had the lowest HMs load, whereas in other treatments had the highest (54.33 %-80.33 %). The potential ecological risk coefficient of Cd was beyond "moderate" in all the pathways of BM and "high" in the interflow of each SS treatment. Nonetheless, when the multi-elements were evaluated collectively, the potential ecological risk index for each SS treatment was categorized as "low". Overall, BA not only reduced soil erosion but also posed no risk of HMs pollution. It should be noted that the loss of Cd in the interflow had a great impact, while the <0.05 mm fraction aggregates played a significant role in the HMs load. Thus, the current study not only provides an effective approach for the environmentally safe disposal of SS but also proposes a scientific method for the application of SS in woodlands.

A conceptual framework for restoration of threatened plants: the effective model of American chestnut (Castanea dentata) reintroduction.

We propose a conceptual framework for restoration of threatened plant species that encourages integration of technological, ecological, and social spheres. A sphere encompasses ideas relevant to restoration and the people working within similar areas of influence or expertise. Increased capacity within a sphere and a higher degree of coalescing among spheres predict a greater probability of successful restoration. We illustrate this with Castanea dentata, a foundation forest tree in North America that was annihilated by an introduced pathogen; the species is a model that effectively merges biotechnology, reintroduction biology, and restoration ecology. Because of C. dentata's ecological and social importance, scientists have aggressively pursued blight resistance through various approaches. We summarize recent advancements in tree breeding and biotechnology that have emerged from C. dentata research, and describe their potential to bring new tools to bear on socio-ecological restoration problems. Successful reintroduction of C. dentata will also depend upon an enhanced understanding of its ecology within contemporary forests. We identify a critical need for a deeper understanding of societal influences that may affect setting and achieving realistic restoration goals. Castanea dentata may serve as an important model to inform reintroduction of threatened plant species in general and foundation forest trees in particular.

Cultivation Using Coir Substrate and P or K Enriched Fertilizer Provides Higher Resistance to Drought in Ecologically Diverse Quercus Species.

Nursery cultivation practices can be modified to increase resistance to water stress in forest seedlings following field establishment, which may be increasingly important under climate change. We evaluated the morphological (survival, growth) and physiological (chlorophyll fluorescence, leaf water potential) responses to water stress for three ecologically diverse Quercus species (Q. robur, Q. pubescens, and Q. ilex) with varying traits resulting from the combination of growing media (peat, coir) and fertilization (standard, P-enriched, K-enriched). For all species under water stress, seedlings grown in coir had generally higher growth than those grown in peat. Seedlings fertilized with P performed better, particularly for survival; conversely, K fertilization resulted in inconsistent findings. Such results could be explained by a combination of factors. P fertilization resulted in higher P accumulation in seedlings, while no K accumulation was observed in K fertilized seedlings. As expected, the more drought-sensitive species, Q. robur, showed the worst response, while Q. pubescens had a drought resistance equal or better to Q. ilex despite being classified as intermediate in drought resistance in Mediterranean environments.

Co-limitation towards lower latitudes shapes global forest diversity gradients.

The latitudinal diversity gradient (LDG) is one of the most recognized global patterns of species richness exhibited across a wide range of taxa. Numerous hypotheses have been proposed in the past two centuries to explain LDG, but rigorous tests of the drivers of LDGs have been limited by a lack of high-quality global species richness data. Here we produce a high-resolution (0.025° × 0.025°) map of local tree species richness using a global forest inventory database with individual tree information and local biophysical characteristics from ~1.3 million sample plots. We then quantify drivers of local tree species richness patterns across latitudes. Generally, annual mean temperature was a dominant predictor of tree species richness, which is most consistent with the metabolic theory of biodiversity (MTB). However, MTB underestimated LDG in the tropics, where high species richness was also moderated by topographic, soil and anthropogenic factors operating at local scales. Given that local landscape variables operate synergistically with bioclimatic factors in shaping the global LDG pattern, we suggest that MTB be extended to account for co-limitation by subordinate drivers.

Intraspecific Variation Along an Elevational Gradient Alters Seed Scarification Responses in the Polymorphic Tree Species Acacia koa.

Physical dormancy in seeds can challenge restoration efforts where scarification conditions for optimal germination and seedling vigor are unknown. For species that occur along wide environmental gradients, optimal scarification conditions may also differ by seed source. We examined intraspecific variation in optimal scarification conditions for germination and seedling performance in koa (Acacia koa), which occurs across a wide range of environmental conditions. To evaluate scarification responses, we recorded imbibition percentage, germination percentage, germination time, seedling abnormalities, early mortality, seedling growth, and seedling survivorship. From these, we developed a scarification index (SI) that integrates these measures simultaneously. We hypothesized that seeds from lower elevation sources exposed to higher temperatures would have harder seed coats and would require more intense scarification treatments. To test this hypothesis, we repeatedly exposed seeds to hot water differing in temperature and time until seeds imbibed. Supporting the hypothesis, seeds from lower elevation sources generally required more intense scarification, although we found substantial variation among sources. Koa seeds germinated in about a week following imbibition. Boiling seeds (i.e., maintaining at 100°C) was effective for imbibing seeds but it also substantially reduced germination percentages. Repeated exposure to 90 to 100°C water did not reduce germination percentage but decreased seedling performance and increased early mortality. No seeds remained unimbibed after six attempts of boiling germinated whereas seeds remaining unimbibed after 15 attempts of exposure to 90 to 100°C water showed high germination percentages. Abnormalities in seedling development were rare but increased with treatment intensity. Exposure to 100°C water for 1 min overall generated the best SI values but the best treatment differed by elevation, and the treatment with the best SI was rarely predicted from the highest germination percentages. Seeds that imbibed without any treatment germinated at the same level as manually filed seeds but produced poor seedling quality. Variation in mother tree environments along an elevational gradient can lead to differences in seed coat characteristics, which may explain differing responses to treatments. Scarification treatments affected processes beyond imbibition and germination and using an index like SI may improve efficiency by identifying optimal scarification treatments while reducing seed waste.

Diversified effects of co-planting landscape plants on heavy metals pollution remediation in urban soil amended with sewage sludge.

Recycling sewage sludge (SS) as a soil amendment potentially causes soil heavy metals (HMs) contamination. This study investigated the potential roles of landscape plants co-planting in SS-amended soil remediation. Three landscape trees Mangifera persiciforma, Bischofia javanica, and Neolamarckia cadamba (NC), and three ground cover plants Dianella ensifolia, Syngonium podophyllum, and Schefflera odorata (SO) were selected for the tree-ground cover co-planting. Species in different co-planting treatments exhibited diversified effects on the growth, root morphology, HMs uptake, and HMs accumulation. Five plant characteristics including total root length, total surface of roots (diameter <2 mm), specific root length, shoot dry weight and root dry weight played crucial roles in plant HMs uptake. Structural equation modeling analysis revealed that different co-planting treatments drive species to develop an active, passive, or avoidance strategy to accumulate HMs, resulting in a diversity of HMs removal efficiency. Co-planting of NC with SO promoted NC and SO HMs accumulation and resulted in the greatest HMs contents decline (48.0% for Cd, 24.9% for Cu, 33.8% for Zn, and 27.2% for Ni) and the lowest potential ecological risk. Co-planting of landscape tree and ground cover plants with an active strategy can be a potential candidate for HMs phytoremediation of SS-amended soil.

Normalization criteria determine the interpretation of nitrogen effects on the root hydraulics of pine seedlings.

Plant hydraulics is key for plant survival and growth because it is linked to gas exchange and drought resistance. Although the environment influences plant hydraulics, there is no clear consensus on the effect of nitrogen (N) supply, which may be, in part, due to different hydraulic conductance normalization criteria and studied species. The objective of this study was to compare the variation of root hydraulic properties using several normalization criteria in four pine species in response to three contrasting N fertilization regimes. We studied four closely related, yet ecologically distinct species: Pinus nigra J.F. Arnold, Pinus pinaster Ait., Pinus pinea L. and Pinus halepensis Mill. Root hydraulic conductance (Kh) was measured with a high-pressure flow meter, and values were normalized by total leaf area (leaf specific conductance, Kl), xylem cross-section area (xylem specific conductance, Ks), total root area (root specific conductance, Kr) and the area of fine roots (fine root specific conductance, Kfr). Controlling for organ size differences allowed comparison of the hydraulic efficiency of roots to supply or absorb water among fertilization treatments and species. The effect of N on the root hydraulic efficiency depended on the normalization criteria. Increasing N availability reduced Kl and Ks, but increased Kh, Kr and especially Kfr. The positive effect of N on Kr and Kfr was positively related to seedling relative growth rate and was also consistent with published results at the interspecific level, whereby plant hydraulics is positively linked to photosynthesis and transpiration rate and fast growth. In contrast, normalization by leaf area and xylem cross-sectional area (Kl and Ks) reflected opposite responses to Kr and Kfr. This indicates that the normalization criteria determine the interpretation of the effect of N on plant hydraulics, which can limit species and treatment comparisons.

Hybrid Breeding for Restoration of Threatened Forest Trees: Evidence for Incorporating Disease Tolerance in Juglans cinerea.

Hybridization is a potential tool for incorporating stress tolerance in plants, particularly to pests and diseases, in support of restoration and conservation efforts. Butternut (Juglans cinerea) is a species for which hybridization has only recently begun being explored. This North American hardwood tree is threatened due to Ophiognomonia clavigignenti-juglandacearum (Ocj), the causal fungus of butternut canker disease (BCD), first observed in 1967. Observational evidence in some wild J. cinerea populations indicates that naturalized hybrids of J. cinerea with Japanese walnut (Juglans ailantifolia) may be more tolerant to BCD than non-admixed J. cinerea, but this has not been formally tested in a controlled trial. We aimed to examine potential BCD tolerance within and between J. cinerea and J. cinerea × J. ailantifolia hybrids and to determine if there is a difference in canker growth between BCD fungal isolates. Five-year-old J. cinerea and hybrid trees were inoculated with two Ocj fungal isolates collected from natural infections found in two different sites in Indiana, United States, and a blank control (agar only). Measurements of both artificially induced and naturally occurring cankers were taken at 8-, 12-, 20-, 24-, and 32-month post-inoculation. Differences in canker presence/absence and size were observed by fungal isolate, which could help explain some of the differences in BCD severity seen between J. cinerea populations. Smaller and fewer cankers and greater genetic gains were seen in hybrid families, demonstrating that hybrids warrant further evaluation as a possible breeding tool for developing BCD-resistant J. cinerea trees.

Drought tolerance and acclimation in Pinus ponderosa seedlings: the influence of nitrogen form.

Drought is a limiting factor to forest regeneration and restoration, which is likely to increase in intensity and duration under future climates. Nitrogen (N) nutrition is related to drought-resistance mechanisms in trees. However, the influence of chemical N form (inorganic and organic N) on physiological traits related to drought resistance has been sparsely studied in conifer seedlings. We investigated the effect of N forms on morpho-physiological traits of Pinus ponderosa Dougl. ex Laws. seedlings and subsequent influences in drought tolerance and acclimation. One-year-old seedlings were fertilized during 10 weeks at 9 mM N with different N forms [either NH4+, NO3- or organic N (amino acids mixture)] in their second year of growth. After fertilization, we measured traits associated with intrinsic drought tolerance (shoot water relations, osmotic regulation, photosynthesis and cell membrane stability). Seedlings were then subjected to an 8-week drought period at varying drought intensities to evaluate plant acclimation mechanisms. We demonstrated that P. ponderosa seedlings could efficiently use amino acids as a primary N source, showing similar performance to those grown with inorganic N forms. Nitrogen form influenced mainly drought-acclimation mechanisms rather than intrinsic drought tolerance. Osmotic potential at saturation (Ψπsat) was marginally affected by N form, and a significant relationship between proline concentration in needles and Ψπsat was found. During acclimation, seedlings fertilized with organic N minimized needle senescence, retained more nutrients in the oldest needles, had maximum increments in proline concentration and hastened the development of water-use efficiency mechanisms compared with those fertilized with inorganic N sources. Our results suggest an improved physiological drought acclimation of organic N-fertilized seedlings.

Morpho-Physiological and Genomic Evaluation of Juglans Species Reveals Regional Maladaptation to Cold Stress.

Climate change may have unpredictable effects on the cold hardiness of woody species planted outside of their range of origin. Extreme undulations in temperatures may exacerbate susceptibility to cold stress, thereby interfering with productivity and ecosystem functioning. Juglans L. and their naturally occurring interspecific F1 hybrids, are distributed natively across many temperate regions, and J. regia has been extensively introduced. Cold hardiness, an environmental and genetic factor yet to be evaluated in many native and introduced Juglans species, may be a limiting factor under future climate change and following species introductions. We evaluated cold hardiness of native North American and Eastern Asian Juglans along with J. regia genotypes using field data from the Midwestern United States (Indiana), controlled freezing tests, and genome sequencing with close assessment of Juglans cold hardy genes. Many Juglans species previously screened for cold-hardiness were genotypes derived from the Midwest, California, and Europe. In 2014, despite general climate adaptation, Midwestern winter temperatures of -30°C killed J. regia originating from California; however, naturalized Midwestern J. regia survived and displayed low damage. Hybridization of J. regia with black walnut (J. nigra) and butternut (J. cinerea) produced F1s displaying greater cold tolerance than pure J. regia. Cold hardiness and growth are variable in Midwestern J. regia compared to native Juglans, East Asian Juglans, and F1 hybrids. Phylogeny analyses revealed that J. cinerea sorted with East Asian species using the nuclear genome but with North American species using the organellar genome. Investigation of selected cold hardy genes revealed that J. regia was distinct from other species and exhibited less genetic diversity than native Juglans species Average whole genome heterozygosity and Tajima's D for cold hardy genes was low within J. regia samples and significantly higher for hybrid as well as J. nigra. We confirmed that molecular and morpho-physiological data were highly correlated and thus can be used effectively to characterize cold hardiness in Juglans species. We conclude that the genetic diversity within local J. regia populations is low and additional germplasm is needed for development of more regionally adapted J. regia varieties.

Unraveling Shade Tolerance and Plasticity of Semi-Evergreen Oaks: Insights From Maritime Forest Live Oak Restoration.

Quercus spp. (oaks) are generally intermediate in shade tolerance, yet there is large variation within the genus in shade tolerance and plasticity in response to varying resource availability. Ecophysiological knowledge specific to semi-evergreen Quercus spp. from subtropical maritime forests is lacking relative to temperate deciduous oaks. We studied the influence of light availability and plant competition on leaf physiology and performance of semi-evergreen Quercus virginiana on a barrier island along the US southern Atlantic coast. Seedlings were underplanted in pine (Pinus taeda) plantation stands with varying overstory density (clear-cut, heavy thin, light thin, and non-thinned; creating a gradient of understory light availability) and vegetation (no competition removal or herbaceous competition removal) treatments. After 2 years, seedling survival was higher with increasing light availability (clear-cut = heavy thin > light thin > non-thinned). Seedling growth (i.e., diameter, height, and crown width) increased similarly with increasing thinning intensity, while vegetation control was mainly beneficial to seedling growth in clear-cuts. These responses were partially explained by foliar nitrogen and leaf trait measurements, which followed the same pattern. Q. virginiana seedlings demonstrated high plasticity in their ability to acclimate to varying resource availability, as indicated by light response curves, specific leaf area, stomatal density, stomatal pore index, and maximum theoretical stomatal conductance. Light compensation and saturation points illustrated seedling capacity to increase net CO2 assimilation with increased light availability. Leaves on trees in the high light environment had the highest net CO2 assimilation, stomatal density, stomatal pore index, maximum theoretical stomatal conductance, and lowest specific leaf area. Although we demonstrated the relative shade tolerance of Q. virginiana in lower light environments (i.e., heavy and light thin plots), this semi-evergreen species shows high plasticity in capacity to respond to varying resource availability, similar to other Quercus spp. from mesic and Mediterranean environments.

Integrated application of sewage sludge, earthworms and Jatropha curcas on abandoned rare-earth mine land soil.

Improving soil fertility is a critical component of abandoned rare-earth mine land (ARL) revegetation. To study the effects of sewage sludge (SS), earthworms, and Jatropha curcas in ARL revegetation, SS (40% in mass ratio) and earthworms (0, 40, 60, and 80 individual adult Eisenia fetida kg-1) were applied to abandoned rare-earth mine land soil (ARLS) and then J. curcas was grown in a potting experiment. The organic carbon, nutrients (N, P, K) and heavy metals (HMs; Cd, Cu, Zn) contents in ARLS and the biomass and nutrients uptake in J. curcas were significantly increased by SS amendment. Application of 80 individual E. fetida kg-1 significantly increased availability of P and K in SS-amended ARLS relative to other treatments. Earthworms increased the height, ground diameter and biomass of J. curcas, but the promotion of biomass became weaker as earthworm density increased. After J. curcas harvest, the contents of Cd, Cu and Zn in SS-amended ARLS were reduced by 15%, 23%, and 19%, respectively. With the joint application of J. curcas and earthworms, a much larger decrease in contents of Cd (34-40%), Cu (31-44%) and Zn (24-29%) in SS-amended ARLS were observed, and the HMs ecological risks were reduced from "moderate potential" to "low potential". Moreover, J. curcas and earthworms together exerted more reduction in the exchangeable fraction HMs in SS-amended ARLS than J. curcas alone. Our results suggest that the integrated application of SS, earthworms and J. curcas is an effective approach for ARL revegetation.

Carbon translocation patterns associated with new root proliferation during episodic growth of transplanted Quercus rubra seedlings.

Patterns of carbon allocation in northern red oak (Quercus rubra L.), characterized by episodic growth through recurrent single-season flushing, vary by growth stage. To examine post-transplant timing and carbohydrate sources for new root growth, dormant, bare-root, half-sibling northern red oak seedlings were transplanted to pots and placed in a favorable growth chamber environment. Unlabeled seedlings were harvested at transplant and at the bud swell stage. After leaf emergence, seedlings were exposed to (14)CO(2) at the linear shoot, linear leaf or lag growth stages. Seedlings were then placed in a growth room for 48 h to allow for translocation of (14)C-labeled current photosynthate and its stabilization in sink component plant parts. Seedlings were subsequently harvested and tissue (14)C:(12)C ratio analyzed. New root growth began during the linear shoot growth stage. However, no increase in (14)C:(12)C ratio was found in new roots until the linear leaf and lag growth stages, indicating a downward shift in translocation of current photosynthate to fuel new root growth. In old roots, (14)C:(12)C ratio increased at the lag stage. Our results indicate that both stored carbohydrates and current photosynthate contribute to new root growth of transplanted northern red oak seedlings; stored carbohydrates promote initial new root proliferation, whereas current photosynthate assumes a greater role as new leaves mature and the flush terminates. Optimizing nursery practices to increase carbohydrate reserves may reduce the time required to establish root-soil contact and facilitate early post-planting survival.