The Precious Potential of the Sacred Tree Chamaecyparis obtusa (Siebold & Zucc.) Endl. as a Source of Secondary Metabolites with Broad Biological Applications.

Chamaecyparis obtusa (Siebold & Zucc.) Endl., which belongs to the Cupressaceae family, occurs naturally in North America and Asia, especially in Korea, Taiwan and Japan, where it is an evergreen, coniferous, sacred, ethnic tree. It has many useful varieties that are widespread throughout the world and grown for decorative purposes. It is most commonly used as an ornamental plant in homes, gardens or parks. It is also widely used in many areas of the economy; for example, its wood is used in architecture as well as furniture production. In addition, oil extracted from Chamaecyparis obtusa is increasingly used in cosmetology for skin care. Due to its wide economic demand, mainly in Japan, it represents the largest area of plantation forest. Despite this, it is on the red list of endangered species. Its use in ethnopharmacology has led to more and more research in recent years in an attempt to elucidate the potential mechanisms of its various biological activities, such as antimicrobial, antioxidant, anticancer, antidiabetic, antiasthmatic, anti-inflammatory, antiallergic, analgesic and central nervous system effects. It has also been shown that Chamaecyparis obtusa can be used as an insect repellent and an ingredient in plant disease treatment. This thesis provides a comprehensive review of the biological studies to date, looking at different areas of the economic fields of potential use of Chamaecyparis obtusa.

Exploration capacity versus specific enzymatic activity of ectomycorrhizas in response to primary productivity and soil phosphorus availability in Bornean tropical rainforests.

Ectomycorrhizal (ECM) fungi are functionally important in biogeochemical cycles in tropical ecosystems. Extracellular enzymatic activity of ECM on a ground-area basis is the product of two attributes; exploration capacity (ECM surface-area) and specific enzymatic activity. Here, we elucidated which attribute better explained the ECM enzymatic activity in response to different levels of soil phosphorus (P) and Nitrogen (N) availability in five Bornean tropical rainforests. We determined the surface area of ECM root tips as well as the enzymatic activities per ECM surface area for carbon (C), N and P degrading enzymes in each site. We evaluated the relationship of ECM enzyme activities with the resource availabilities of C (Above-ground net primary production; ANPP), N, and P of ECM by a generalized linear mixed model. The ECM enzymatic activities on a ground-area basis were more significantly determined by specific enzymatic activity than by the exploration capacity. Specific enzymatic activities were generally negatively affected by C (ANPP) and soil P availability. ECM fungi enhance the specific enzyme activity rather than the exploration capacity to maintain the capacity of nutrient acquisition. The less dependence of ECM fungi on the exploration capacity in these forests may be related to the limitation of C supply from host trees. We highlighted the adaptive mechanisms of ECM fungi on nutrient acquisition in tropical ecosystems through the response of enzymatic activity to nutrient availability across the elements.

Underlying plant trait strategies for understanding the carbon sequestration in Banj oak Forest of Himalaya.

Plant functional attributes are subjected to environmental adjustments, which lead to modulations in forest processes under environmental changes. However, a comprehensive assessment of the relationships between plant traits and carbon stock remains subtle. The present study attempted to accomplish the gap of knowledge by examining the linkages between forest carbon with plant traits within the Banj Oak forest in the Garhwal Himalaya. Twelve individuals from three major species in the Banj Oak forest were randomly selected for trait measurements, and soil samples were collected randomly across the area for evaluation of soil nutrients and carbon. Forest biomass and soil carbon were estimated following standard protocols. A Structural Equation Model (SEM) was applied to establish the relationship between above ground carbon (AGC) and soil organic carbon (SOC) with leaf and stem traits, and soil nutrients. Stem traits were tree height and tree diameter; whereas leaf morphological traits were leaf area, specific leaf area, leaf dry matter content; leaf physiological traits were photosynthesis rate, stomatal conductance, and transpiration rate; and leaf biochemical traits were leaf carbon concentration, leaf nitrogen concentration, and leaf phosphorus concentration. Soil nutrients were available nitrogen, available phosphorus, and exchangeable potassium. Based on SEM results, AGC of the forest was positively correlated with stem traits and leaf physiological traits, while negatively correlated with leaf morphological traits. SOC was positively correlated with soil nutrients and leaf biochemical traits, whereas negatively correlated with stem traits. These findings may support for precise quantification of forest carbon and modeling of forest carbon stocks besides providing inputs to forest managers for devising effective forest management strategies.

A neotropical perspective on the uniqueness of the Holocene among interglacials.

Understanding how tropical systems have responded to large-scale climate change, such as glacial-interglacial oscillations, and how human impacts have altered those responses is key to current and future ecology. A sedimentary record recovered from Lake Junín, in the Peruvian Andes (4085 m elevation) spans the last 670,000 years and represents the longest continuous and empirically-dated record of tropical vegetation change to date. Spanning seven glacial-interglacial oscillations, fossil pollen and charcoal recovered from the core showed the general dominance of grasslands, although during the warmest times some Andean forest trees grew above their modern limits near the lake. Fire was very rare until the last 12,000 years, when humans were in the landscape. Here we show that, due to human activity, our present interglacial, the Holocene, has a distinctive vegetation composition and ecological trajectory compared with six previous interglacials. Our data reinforce the view that modern vegetation assemblages of high Andean grasslands and the presence of a defined tree line are aspects of a human-modified landscape.

Long-term ecological responses of a lowland dipterocarp forest to climate changes and nutrient availability.

Understanding the long-term impact of projected climate change on tropical rainforests is critical given their central role in the Earth's system. Palaeoecological records can provide a valuable perspective on this problem. Here, we examine the effects of past climatic changes on the dominant forest type of Southeast Asia - lowland dipterocarp forest. We use a range of proxies extracted from a 1400-yr-old lacustrine sedimentary sequence from north-eastern Philippines to determine long-term vegetation responses of lowland dipterocarp forest, including its dominant tree group dipterocarps, to changes in precipitation, fire and nutrient availability over time. Our results show a positive relationship between dipterocarp pollen accumulation rates (PARs) and leaf wax hydrogen isotope values, which suggests a negative effect of drier conditions on dipterocarp abundance. Furthermore, we find a positive relationship between dipterocarp PARs and the proxy for phosphorus availability, which suggests phosphorus controls the productivity of these keystone trees on longer time scales. Other pollen taxa show widely varying relationships with the abiotic factors, demonstrating a high diversity of plant functional responses. Our findings provide novel insights into lowland dipterocarp forest responses to changing climatic conditions in the past and highlight potential impacts of future climate change on this globally important ecosystem.

Leaf nutrient resorption of two life-form tree species in urban gardens and their response to soil nutrient availability.

Background: Leaf nutrient resorption is a key strategy in plant conservation that minimizes nutrient loss and enhances productivity. However, the differences of the nutrient resorption among garden tree species in urban ecosystems were not clearly understood, especially the differences of nitrogen resorption efficiency (NRE) and phosphorous resorption efficiency (PRE) between evergreen and deciduous trees. Methods: We selected 40 most generally used garden tree specie belonged two life forms (evergreen and deciduous) and investigated the nitrogen (N) and phosphorus (P) concentrations in green and senesced leaves and soil nutrient concentrations of nine samples trees for each species. Then, the nutrient concentrations and resorption efficiency were compared, and the soil nutrients utilization strategies were further analyzed. Results: The results showed that the N concentration was significantly higher in the green and senesced leaves of deciduous trees than in the leaves of evergreen trees. The two life-form trees were both N limited and evergreen trees were more sensitive to N limitation. The NRE and PRE in the deciduous trees were significantly higher than those in the evergreen trees. The NRE was significantly positively correlated with the PRE in the deciduous trees. As the soil N and P concentrations increased, the nutrient resorption efficiency (NuRE) of the evergreen trees increased, but that of the deciduous trees decreased. Compared with the deciduous trees, the evergreen trees were more sensitive to the feedback of soil N and P concentrations. These findings reveal the N and P nutrient resorption mechanism of evergreen and deciduous trees and fill a gap in the understanding of nutrient resorption in urban ecosystems.

Tree islands enhance biodiversity and functioning in oil palm landscapes.

In the United Nations Decade on Ecosystem Restoration1, large knowledge gaps persist on how to increase biodiversity and ecosystem functioning in cash crop-dominated tropical landscapes2. Here, we present findings from a large-scale, 5-year ecosystem restoration experiment in an oil palm landscape enriched with 52 tree islands, encompassing assessments of ten indicators of biodiversity and 19 indicators of ecosystem functioning. Overall, indicators of biodiversity and ecosystem functioning, as well as multidiversity and ecosystem multifunctionality, were higher in tree islands compared to conventionally managed oil palm. Larger tree islands led to larger gains in multidiversity through changes in vegetation structure. Furthermore, tree enrichment did not decrease landscape-scale oil palm yield. Our results demonstrate that enriching oil palm-dominated landscapes with tree islands is a promising ecological restoration strategy, yet should not replace the protection of remaining forests.

Patterns of physical, chemical, and metabolic characteristics of sugar maple leaves with depth in the crown and in response to nitrogen and phosphorus addition.

Few previous studies have described the patterns of leaf characteristics in response to nutrient availability and depth in the crown. Sugar maple has been studied for both sensitivity to light, as a shade-tolerant species, and sensitivity to soil nutrient availability, as a species in decline due to acid rain. To explore leaf characteristics from the top to bottom of the canopy, we collected leaves along a vertical gradient within mature sugar maple crowns in a full-factorial nitrogen (N) by phosphorus (P) addition experiment in three forest stands in central New Hampshire, USA. Thirty-two of the 44 leaf characteristics had significant relationships with depth in the crown, with the effect of depth in the crown strongest for leaf area, photosynthetic pigments and polyamines. Nitrogen addition had a strong impact on the concentration of foliar N, chlorophyll, carotenoids, alanine and glutamate. For several other elements and amino acids, N addition changed patterns with depth in the crown. Phosphorus addition increased foliar P and boron (B); it also caused a steeper increase of P and B with depth in the crown. Since most of these leaf characteristics play a direct or indirect role in photosynthesis, metabolic regulation or cell division, studies that ignore the vertical gradient may not accurately represent whole-canopy performance.

Nitrogen and light regulate symbiotic nitrogen fixation by a temperate forest tree.

Symbiotic nitrogen fixation (SNF) is a critical mechanism of ecosystem recovery, and in forests of the eastern United States, the most common tree species that supports SNF is black locust (Robinia pseudoacacia L.). Despite its prevalence, black locust's fixation strategy-whether it maintains fixation at a constant rate (obligate fixation) or reduces its fixation rate (facultative fixation)-is unknown. Here, we examined how nitrogen and light control SNF by black locust, by growing seedlings under two nitrogen levels and across four levels of light. Seedlings were harvested after 12 weeks to determine biomass changes, nodule activity, and photosynthetic rates. Black locust seedlings increased biomass growth with increasing light, but only in the absence of nitrogen addition, while seedling root:shoot (biomass) modestly declined with increasing light regardless of nitrogen level. We found that black locust behaved like a facultative fixer, and regulated fixation by excising or maintaining nodules, and by controlling nodule biomass and activity. Specifically, nitrogen addition reduced seedling investment in nodule biomass (g g-1) by 63%, and reduced seedling allocation to nitrogen fixation (µmol C2H4 g-1 h-1) by 66%. In contrast, light affected nitrogen fixation through two indirect pathways. First, light increased plant growth, and hence nitrogen demands, which caused an increase in nitrogen fixation proportional to biomass. Second, light increasd photosynthetic activity, which was positively associated with nodule activity, but only in the absence of nitrogen addition. Our findings for how black locust regulates SNF can improve predictions of ecosystem SNF under the changing environmental conditions.

Differential responses and mechanistic controls of soil phosphorus transformation in Eucalyptus plantations with N fertilization and introduced N2 -fixing tree species.

Introducing N2 -fixing tree species into Eucalyptus plantations could replace nitrogen (N) fertilization to maintain high levels of N consumption and productivity. However, N enrichment may exacerbate phosphorus (P) limitation as Eucalyptus robusta Smith is extensively planted in P-poor tropical and subtropical soils. We conducted a field experiment in a pure plantation of Eucalyptus urophylla × grandis to investigate the impacts of N fertilization and introduced an N2 -fixing tree of Dalbergia odorifera T. Chen on soil P transformation. Nitrogen fertilization significantly enhanced soil occluded P pool and reduced the other P pools due to acidification-induced pH-sensitive geochemical processes, lowering Eucalyptus leaf P concentration with higher N : P ratio. By contrast, introduced N2 -fixing tree species did not change soil pH, labile inorganic P pool, and Eucalyptus leaf N : P ratio, even enhanced organic P pools and reduced occluded P pool probably due to altering microbial community composition particularly stimulating arbuscular mycorrhiza fungal abundance. Our results revealed differential responses and mechanistic controls of soil P transformation in Eucalyptus plantations with N fertilization and introduced N2 -fixing tree species. The dissolution of occluded P pool along with organic P accumulation observed in the mixed plantations may represent a promising future to better manage soil P availability.

Atmospheric phosphorus deposition amplifies carbon sinks in simulations of a tropical forest in Central Africa.

Spatial redistribution of nutrients by atmospheric transport and deposition could theoretically act as a continental-scale mechanism which counteracts declines in soil fertility caused by nutrient lock-up in accumulating biomass in tropical forests in Central Africa. However, to what extent it affects carbon sinks in forests remains elusive. Here we use a terrestrial biosphere model to quantify the impact of changes in atmospheric nitrogen and phosphorus deposition on plant nutrition and biomass carbon sink at a typical lowland forest site in Central Africa. We find that the increase in nutrient deposition since the 1980s could have contributed to the carbon sink over the past four decades up to an extent which is similar to that from the combined effects of increasing atmospheric carbon dioxide and climate change. Furthermore, we find that the modelled carbon sink responds to changes in phosphorus deposition, but less so to nitrogen deposition. The pronounced response of ecosystem productivity to changes in nutrient deposition illustrates a potential mechanism that could control carbon sinks in Central Africa. Monitoring the quantity and quality of nutrient deposition is needed in this region, given the changes in nutrient deposition due to human land use.

Phosphorus limitation of early growth differs between nitrogen-fixing and nonfixing dry tropical forest tree species.

Tropical forests are often characterized by low soil phosphorus (P) availability, suggesting that P limits plant performance. However, how seedlings from different functional types respond to soil P availability is poorly known but important for understanding and modeling forest dynamics under changing environmental conditions. We grew four nitrogen (N)-fixing Fabaceae and seven diverse non-N-fixing tropical dry forest tree species in a shade house under three P fertilization treatments and evaluated carbon (C) allocation responses, P demand, P-use, investment in P acquisition traits, and correlations among P acquisition traits. Nitrogen fixers grew larger with increasing P addition in contrast to non-N fixers, which showed fewer responses in C allocation and P use. Foliar P increased with P addition for both functional types, while P acquisition strategies did not vary among treatments but differed between functional types, with N fixers showing higher root phosphatase activity (RPA) than nonfixers. Growth responses suggest that N fixers are limited by P, but nonfixers may be limited by other resources. However, regardless of limitation, P acquisition traits such as mycorrhizal colonization and RPA were nonplastic across a steep P gradient. Differential limitation among plant functional types has implications for forest succession and earth system models.

Litter quality and decomposition responses to drought in a northeastern US deciduous forest.

Even though drought impacts on tree physiology have been identified, whether drought affects leaf litter chemistry that, in turn, influences litter decay rates is still poorly understood. We compared litter quality and decomposition for two cohorts of leaves from five co-occurring seasonally deciduous tree species: Acer saccharum, Tilia americana, Quercus rubra, Quercus alba, and Ostrya virginiana. One cohort experienced a growing-season drought, and the other cohort came from the same trees in the ensuing, post-drought growing season. Leaf litter production was greater for drought litter than post-drought litter for all five species. Specific leaf area and nitrogen concentrations were 20% greater for the drought cohort than the post-drought cohort. Concentrations of non-structural carbohydrates were about 14% greater for the drought cohort, except for greater values for post-drought A. saccharum litter. Pectin in the middle lamella of leaf litter was 31% lower for the drought cohort compared to post-drought cohort. We found few differences in litter decay rates between drought and post-drought cohorts, although Q. rubra litter had more decomposition for the post-drought cohort than the drought cohort, whereas A. saccharum litter had more decomposition for the drought cohort than the post-drought cohort. Leaf litter decay rates for the drought cohort were related to litter nitrogen and lignin concentrations, whereas decay rates for the post-drought cohort were related to litter carbohydrate concentrations. Our findings suggest that the role of drought events on seasonally deciduous forest ecosystems must recognize species-specific, idiosyncratic responses in leaf litter quality and decomposition.

Convergence in phosphorus constraints to photosynthesis in forests around the world.

Tropical forests take up more carbon (C) from the atmosphere per annum by photosynthesis than any other type of vegetation. Phosphorus (P) limitations to C uptake are paramount for tropical and subtropical forests around the globe. Yet the generality of photosynthesis-P relationships underlying these limitations are in question, and hence are not represented well in terrestrial biosphere models. Here we demonstrate the dependence of photosynthesis and underlying processes on both leaf N and P concentrations. The regulation of photosynthetic capacity by P was similar across four continents. Implementing P constraints in the ORCHIDEE-CNP model, gross photosynthesis was reduced by 36% across the tropics and subtropics relative to traditional N constraints and unlimiting leaf P. Our results provide a quantitative relationship for the P dependence for photosynthesis for the front-end of global terrestrial C models that is consistent with canopy leaf measurements.

1.36 million years of Mediterranean forest refugium dynamics in response to glacial-interglacial cycle strength.

The sediment record from Lake Ohrid (Southwestern Balkans) represents the longest continuous lake archive in Europe, extending back to 1.36 Ma. We reconstruct the vegetation history based on pollen analysis of the DEEP core to reveal changes in vegetation cover and forest diversity during glacial-interglacial (G-IG) cycles and early basin development. The earliest lake phase saw a significantly different composition rich in relict tree taxa and few herbs. Subsequent establishment of a permanent steppic herb association around 1.2 Ma implies a threshold response to changes in moisture availability and temperature and gradual adjustment of the basin morphology. A change in the character of G-IG cycles during the Early-Middle Pleistocene Transition is reflected in the record by reorganization of the vegetation from obliquity- to eccentricity-paced cycles. Based on a quantitative analysis of tree taxa richness, the first large-scale decline in tree diversity occurred around 0.94 Ma. Subsequent variations in tree richness were largely driven by the amplitude and duration of G-IG cycles. Significant tree richness declines occurred in periods with abundant dry herb associations, pointing to aridity affecting tree population survival. Assessment of long-term legacy effects between global climate and regional vegetation change reveals a significant influence of cool interglacial conditions on subsequent glacial vegetation composition and diversity. This effect is contrary to observations at high latitudes, where glacial intensity is known to control subsequent interglacial vegetation, and the evidence demonstrates that the Lake Ohrid catchment functioned as a refugium for both thermophilous and temperate tree species.

Animal wastes as fertilizers enhance growth of young walnut trees under soil drought conditions.

BACKGROUND: Using nutrient-rich animal wastes as organic fertilizers in agricultural practices is a sustainable method for soil amendment and avoiding environmental pollution. In order to evaluate their practical effect, we applied different proportions of animal waste as fertilizers to wet or dry soils that were either planted or not planted with young walnut trees. RESULTS: The results showed that animal waste could increase soil C accumulation and carbon to nitrogen (C/N) ratio and reduce soil organic nitrogen and total nitrogen contents as well as the nitrogen to phosphorus (N/P) ratio in the planted group soil. This framework of soil C and N composition (a high C/N ratio) resulted in high N and Mg contents as well as high Cu and Zn contents in the leaves of the young trees as well as a high dry matter weight/leaf N ratio, causing increased leaf photosynthesis, reduced transpiration and relatively high water use efficiency under soil drought conditions. Also, animal wastes as fertilizers caused the branching of walnut to switch from elongation growth to thickening growth under soil drought conditions. CONCLUSIONS: Principal component analysis and redundancy analysis demonstrated the mechanism by which the soil C/N ratio mediates the flux of available nutrients from the soil to the plant and thereby regulates plant dry matter accumulation and branching architecture under soil drought conditions. The results of this study provide new insights into the improvement of hilly soils using animal waste. © 2020 Society of Chemical Industry.

An analysis of mating biases in trees.

Assortative mating is a deviation from random mating based on phenotypic similarity. As it is much better studied in animals than in plants, we investigate for trees whether kinship of realized mating pairs deviates from what is expected from the set of potential mates and use this information to infer mating biases that may result from kin recognition and/or assortative mating. Our analysis covers 20 species of trees for which microsatellite data is available for adult populations (potential mates) as well as seed arrays. We test whether mean relatedness of observed mating pairs deviates from null expectations that only take pollen dispersal distances into account (estimated from the same data set). This allows the identification of elevated as well as reduced kinship among realized mating pairs, indicative of positive and negative assortative mating, respectively. The test is also able to distinguish elevated biparental inbreeding that occurs solely as a result of related pairs growing closer to each other from further assortativeness. Assortative mating in trees appears potentially common but not ubiquitous: nine data sets show mating bias with elevated inbreeding, nine do not deviate significantly from the null expectation, and two show mating bias with reduced inbreeding. While our data sets lack direct information on phenology, our investigation of the phenological literature for each species identifies flowering phenology as a potential driver of positive assortative mating (leading to elevated inbreeding) in trees. Since active kin recognition provides an alternative hypothesis for these patterns, we encourage further investigations on the processes and traits that influence mating patterns in trees.

Water-soluble phosphorus contributes significantly to shaping the community structure of rhizospheric bacteria in rocky desertification areas.

Microorganisms play important roles in soil improvement. Therefore, clarifying the contribution of environmental factors in shaping the microbial community structure is beneficial to improve soil fertility in karst rocky desertification areas. Here, the bacterial community structures of eight rhizospheric soil samples collected from perennial fruit plantations were analysed using an Illumina HiSeq2500 platform. The diversity and abundance of bacteria in rocky desertification areas were significantly lower than those in non-rocky desertification areas, while the bacterial community structure was not significantly different between root surface and non-root surface soils in the same rhizospheric soil samples. Proteobacteria predominated in rocky desertification areas, while Actinobacteria predominated in non-rocky desertification areas. Correlation analysis revealed that water-soluble phosphorus content (r2 = 0.8258), latitude (r2 = 0.7556), altitude (r2 = 0.7501), and the age of fruit trees (r2 = 0.7321) were positively correlated with the bacterial community structure, while longitude, pH, and total phosphorus content did not significantly influence the soil bacterial community structure. As water-soluble phosphorus content is derived from insoluble phosphorus minerals, supplementing phosphorus-solubilising bacteria to soils in rocky desertification areas is a feasible strategy for accelerating the dissolution of insoluble phosphorus minerals and improving agricultural production and environment ecology.

Dispersal of allergenic pollen from Cryptomeria japonica and Chamaecyparis obtusa: characteristic annual fluctuation patterns caused by intermittent phase synchronisations.

Trees produce pollen during specific times of the year. Pollen can induce pollinosis, a type of allergic rhinitis, in humans. In Japan, allergenic pollen is mainly dispersed from February to May. Using data collected at 120 observation sites managed by the Japanese Ministry of the Environment, we studied the annual patterns of airborne allergenic pollen. The allergenic pollen showed an alternating ON-OFF cycle, but the length of the cycle differed among regions. We used an in-phase/out-of-phase analysis to quantify two characteristic features of the synchronisation. The degrees of phase synchronisation were strong in eastern and weak in western Japan. The pattern of allergenic pollen dispersal throughout Japan is typical intermittent synchronisation. This is the first study to evaluate allergenic pollen's distribution from a phase synchronisation viewpoint.

Mycorrhizal types differ in ecophysiology and alter plant nutrition and soil processes.

Mycorrhizal fungi benefit plants by improved mineral nutrition and protection against stress, yet information about fundamental differences among mycorrhizal types in fungi and trees and their relative importance in biogeochemical processes is only beginning to accumulate. We critically review and synthesize the ecophysiological differences in ectomycorrhizal, ericoid mycorrhizal and arbuscular mycorrhizal symbioses and the effect of these mycorrhizal types on soil processes from local to global scales. We demonstrate that guilds of mycorrhizal fungi display substantial differences in genome-encoded capacity for mineral nutrition, particularly acquisition of nitrogen and phosphorus from organic material. Mycorrhizal associations alter the trade-off between allocation to roots or mycelium, ecophysiological traits such as root exudation, weathering, enzyme production, plant protection, and community assembly as well as response to climate change. Mycorrhizal types exhibit differential effects on ecosystem carbon and nutrient cycling that affect global elemental fluxes and may mediate biome shifts in response to global change. We also note that most studies performed to date have not been properly replicated and collectively suffer from strong geographical sampling bias towards temperate biomes. We advocate that combining carefully replicated field experiments and controlled laboratory experiments with isotope labelling and -omics techniques offers great promise towards understanding differences in ecophysiology and ecosystem services among mycorrhizal types.