Profile and concentration of the low molecular weight organic acids and phenolic compounds created by two-year-old Acer platanoides seedlings growing under different As forms.

Two-year-old seedlings of Acer platanoides were cultivated during a three-month hydroponic experiment in modified Knop solution enriched with inorganic (As(III), As(V)) and organic (dimethylarsinic acid - DMA) arsenic forms at 0.06 mM, 0.6 mM and their combinations. The profile and content of low molecular weight organic acids (LMWOAs) and phenolic compounds were also determined in the rhizosphere, roots and leaves. Arsenic (As) treatment caused an elevated creation of the above mentioned metabolites, which was higher in leaves than in the rhizosphere or roots, and their overall content was correlated with the concentration of As in A. platanoides organs. The addition of all As forms strongly induced the exudation of citric and oxalic acids into the rhizosphere, while malonic, acetic, citric and malic acids were formed in the roots. The most differential profile of roots was confirmed for As(V) 0.06 mM (4-hydroxybenzoic (4-HBA), syringic, 2,5 dihydroxybenzoic (2,5-DHBA), caffeic, chlorogenic, ferulic, p-coumaric and sinapic acids and catechin). The obtained results indicate that the presence of particular As forms has a significant impact on the content and profile of exuded and created LMWOAs and phenolic compounds, and can also have a decisive influence on the activation of appropriate detoxification mechanisms.

Wetland macrophyte community response to and recovery from direct application of glyphosate-based herbicides.

Community-scale impacts of glyphosate-based herbicides on wetland plant communities and the magnitude of those impacts that should be considered biologically relevant are poorly understood. We contrast three different thresholds for setting biologically meaningful critical effect sizes for complex ANOVA study designs. We use each of the of the critical effect sizes to determine optimal α levels for assessment of how different concentrations of glyphosate-based herbicides affect wetland plant communities over two years of herbicide application (alone and in combination with agricultural fertilizers) and two subsequent years without herbicide (or fertilizer) application. The application of glyphosate-based herbicides was found to result in a decrease in macrophyte species richness, an increase in macrophyte species evenness, a decrease in macrophyte cover and a reduction in community similarity. There was little evidence that nutrient additions directly or indirectly affected plant community endpoints. The glyphosate effects were evident in the first year of herbicide application in 2009, and became more pronounced in the second year of herbicide application in 2010. However, when herbicides were not applied in 2011, recovery was observed in most endpoints, with the exception being species evenness, for which partial recovery was not observed until 2012. Optimal α levels differed among the three critical effect sizes for each ANOVA term and endpoint combination, however regardless of differences in α levels, conclusions were generally consistent across all critical effect sizes.

Differences of Acer platanoides L. and Tilia cordata Mill. Response patterns/survival strategies during cultivation in extremely polluted mining sludge - A pot trial.

The study aimed to evaluate the physiological mechanisms underlying differences in metals and metalloid uptake and tolerance of two tree species cultivated in mining waste material. Two-year old Acer platanoides L. and Tilia cordata Mill. were cultivated in mining sludge characterized by high pH, salinity and an extremely high concentration of As. Both species were able to develop leaves from leafless seedlings, however, their total biomass was greatly reduced in comparison to control plants, following the severe disturbances in chlorophyll content. Phytoextraction abilities were observed for T. cordata for Ba, Nb, Rb and Se, and phytostabilisation was stated for Pd, Ru, Sc and Sm for both species, Ba and Nd for A. platonoides and Be for T. cordata only. Metal exclusion was observed for the majority of detected elements indicating an intense limitation of metal transport to photosynthetic tissue. A diversified uptake of elements was accompanied by a species-specific pattern of physiological reaction during the cultivation in sludge. Organic ligands (glutatnione and low-molecular-weight organic acids) were suppressed in A. platanoides, and enhanced biosynthesis of phenolic compounds was observed for both species, being more pronounced in T. cordata. Despite its higher accumulation of key metabolites for plant reaction to oxidative stress, such as phenolic acids, flavonoids and organic ligands, T. cordata exhibited relatively lower tolerance to sludge, probably due to the increased uptake and translocation rate of toxic metal/loids to aerial organs and/or restricted accumulation of salicylic acid which is known to play a decisive role in mechanisms of plant tolerance.

Vegetation dynamics associated with changes in atmospheric nitrogen deposition and climate in hardwood forests of Shenandoah and Great Smoky Mountains National Parks, USA.

Ecological effects of atmospheric nitrogen (N) and sulfur (S) deposition on two hardwood forest sites in the eastern United States were simulated in the context of a changing climate using the dynamic coupled biogeochemical/ecological model chain ForSAFE-Veg. The sites are a mixed oak forest in Shenandoah National Park, Virginia (Piney River) and a mixed oak-sugar maple forest in Great Smoky Mountains National Park, Tennessee (Cosby Creek). The sites have received relatively high levels of both S and N deposition and the climate has warmed over the past half century or longer. The model was used to evaluate the composition of the understory plant communities, the alignment between plant species niche preferences and ambient conditions, and estimate changes in relative species abundances as reflected by plant cover under various scenarios of future atmospheric N and S deposition and climate change. The main driver of ecological effects was soil solution N concentration. Results of this research suggested that future climate change might compromise the capacity for the forests to sustain habitat suitability. However, vegetation results should be considered preliminary until further model validation can be performed. With expected future climate change, preliminary estimates suggest that sustained future N deposition above 7.4 and 5.0 kg N/ha/yr is expected to decrease contemporary habitat suitability for indicator plant species located at Piney River and Cosby Creek, respectively.

Influence of biochar and nitrogen on fine root morphology, physiology, and chemistry of Acer mono.

Fine roots play an important role in the overall functions of individual plants. Previous studies showed that fertilization and available soil resources have a notably profound effect on fine root, but there is lack of study centered on how fine root morphology, physiology, and chemistry respond to biochar with N additions. Different levels of biochar (0, 10, 15, and 20 g) and N (0, 2, 4 and 6 g) were applied to Acer mono seedling plants in a field nursery. The root system morphology and root chemistry and physiology were evaluated in line with root length, root diameter, SRL, N and N: C and root respiration. Biochar and N significantly affected root morphology, chemistry and root respiration. Morphological, chemical and physiological parameters were found to be at their maximum with 20 g biochar and 6 g N; however, no significant effect was noted on fourth- and fifth-order roots. Furthermore, a significant increase in root respiration was recognized with the increase in root tissue N concentration and the negative relationship of root respiration with higher branch order. Thus, overall, study parameters indicate that biochar and nitrogen positively influence the Acer mono fine root, and therefore should be used to improve fine root health.

Effects of a common insecticide on wetland communities with varying quality of leaf litter inputs.

Chemical contamination of aquatic systems often co-occurs with dramatic changes in surrounding terrestrial vegetation. Plant leaf litter serves as a crucial resource input to many freshwater systems, and changes in litter species composition can alter the attributes of freshwater communities. However, little is known how variation in litter inputs interacts with chemical contaminants. We investigated the ecological effects resulting from changes in tree leaf litter inputs to freshwater communities, and how those changes might interact with the timing of insecticide contamination. Using the common insecticide malathion, we hypothesized that inputs of nutrient-rich and labile leaf litter (e.g., elm [Ulmus spp.] or maple [Acer spp.]) would reduce the negative effects of insecticides on wetland communities relative to inputs of recalcitrant litter (e.g., oak [Quercus spp.]). We exposed artificial wetland communities to a factorial combination of three litter species treatments (elm, maple, and oak) and four insecticide treatments (no insecticide, small weekly doses of 10 µg L-1, and either early or late large doses of 50 µg L-1). Communities consisted of microbes, algae, snails, amphipods, zooplankton, and two species of tadpoles. After two months, we found that maple and elm litter generally induced greater primary and secondary production. Insecticides induced a reduction in the abundance of amphipods and some zooplankton species, and increased phytoplankton. In addition, we found interactive effects of litter species and insecticide treatments on amphibian responses, although specific effects depended on application regime. Specifically, with the addition of insecticide, elm and maple litter induced a reduction in gray tree frog survival, oak and elm litter delayed tree frog metamorphosis, and oak and maple litter reduced green frog tadpole mass. Our results suggest that attention to local forest composition, as well as the timing of pesticide application might help ameliorate the harmful effects of pesticides observed in freshwater systems.

Probability of foliar injury for Acer sp. based on foliar fluoride concentrations.

Fluoride is considered one of the most phytotoxic elements to plants, and indicative fluoride injury has been associated over a wide range of foliar fluoride concentrations. The aim of this study was to determine the probability of indicative foliar fluoride injury based on Acer sp. foliar fluoride concentrations using a logistic regression model. Foliage from Acer nedundo, Acer saccharinum, Acer saccharum and Acer platanoides was collected along a distance gradient from three separate brick manufacturing facilities in southern Ontario as part of a long-term monitoring programme between 1995 and 2014. Hydrogen fluoride is the major emission source associated with the manufacturing facilities resulting with highly elevated foliar fluoride close to the facilities and decreasing with distance. Consistent with other studies, indicative fluoride injury was observed over a wide range of foliar concentrations (9.9-480.0 µg F- g-1). The logistic regression model was statistically significant for the Acer sp. group, A. negundo and A. saccharinum; consequently, A. negundo being the most sensitive species among the group. In addition, A. saccharum and A. platanoides were not statistically significant within the model. We are unaware of published foliar fluoride values for Acer sp. within Canada, and this research provides policy maker and scientist with probabilities of indicative foliar injury for common urban Acer sp. trees that can help guide decisions about emissions controls. Further research should focus on mechanisms driving indicative fluoride injury over wide ranging foliar fluoride concentrations and help determine foliar fluoride thresholds for damage.

Analysis of the embryo proteome of sycamore (Acer pseudoplatanus L.) seeds reveals a distinct class of proteins regulating dormancy release.

Acer pseudoplatanus seeds are characterized by a deep physiological embryo dormancy that requires a few weeks of cold stratification in order to promote germination. Understanding the function of proteins and their related metabolic pathways, in conjunction with the plant hormones implicated in the breaking of seed dormancy, would expand our knowledge pertaining to this process. In this study, a proteomic approach was used to analyze the changes occurring in seeds in response to cold stratification, which leads to dormancy release. In addition, the involvement of abscisic (ABA) and gibberellic acids (GA) was also examined. Fifty-three proteins showing significant changes were identified by mass spectrometry. An effect of ABA on protein variation was observed at the beginning of stratification, while the influence of GA on protein abundance was observed during the middle phase of stratification. The majority of proteins associated with dormancy breaking in the presence of only water, and also ABA or GA, were classified as being involved in metabolism and genetic information processing. For metabolic-related proteins, the effect of ABA on protein abundance was stimulatory for half of the proteins and inhibitory for half of the proteins. On the other hand, the effect on genetic information processing related proteins was stimulatory. GA was found to upregulate both metabolic-related and genetic information processing-related proteins. While seed dormancy breaking depends on proteins involved in a variety of processes, proteins associated with methionine metabolism (adenosine kinase, methionine synthase) and glycine-rich RNA binding proteins appear to be of particular importance.

Light compensation points in shade-grown seedlings of deciduous broadleaf tree species with different successional traits raised under elevated CO2.

We measured leaf photosynthetic traits in shade-grown seedlings of four tree species native to northern Japan, raised under an elevated CO2 condition, to investigate the effects of elevated CO2 on shade tolerance of deciduous broadleaf tree species with different successional traits. We considered Betula platyphylla var. japonica and Betula maximowicziana as pioneer species, Quercus mongolica var. crispula as a mid-successional species, and Acer mono as a climax species. The plants were grown under shade conditions (10% of full sunlight) in a CO2 -regulated phytotron. Light compensation points (LCPs) decreased in all tree species when grown under elevated CO2 (720 µmol·mol(-1) ), which were accompanied by higher apparent quantum yields but no photosynthetic down-regulation. LCPs in Q. mongolica and A. mono grown under elevated CO2 were lower than those in the two pioneer birch species. The LCP in Q. mongolica seedlings was not different from that of A. mono in each CO2 treatment. However, lower dark respiration rates were observed in A. mono than in Q. mongolica, suggesting higher shade tolerance in A. mono as a climax species in relation to carbon loss at night. Thus, elevated CO2 may have enhanced shade tolerance by lowering LCPs in all species, but the ranking of shade tolerance related to successional traits did not change among species under elevated CO2 , i.e. the highest shade tolerance was observed in the climax species (A. mono), followed by a gap-dependent species (Q. mongolica), while lower shade tolerance was observed in the pioneer species (B. platyphylla and B. maximowicziana).

Chronic drought stress reduced but not protected Shantung maple (Acer truncatum Bunge) from adverse effects of ozone (O3) on growth and physiology in the suburb of Beijing, China.

A two-year experiment exposing Acer truncatum Bunge seedlings to elevated ozone (O3) concentrations above ambient air (AO) and drought stress (DS) was carried out using open-top chambers (OTCs) in a suburb of Beijing in north China in 2012-2013. The results suggested that AO and DS had both significantly reduced leaf mass area (LMA), stomatal conductance (Gs), light saturated photosynthetic rate (Asat) as well as above and below ground biomass at the end of the experiment. It appeared that while drought stress mitigated the expression of foliar injury, LMA, leaf photosynthetic pigments, height growth and basal diameter, due to limited carbon fixation, the O3 - induced reductions in Asat, Gs and total biomass were enhanced 23.7%. 15.5% and 8.1% respectively. These data suggest that when the whole plant was considered that drought under the conditions of this experiment did not protect the Shantung maple seedlings from the effects of O3.

Effects of CO2 on Acer negundo pollen fertility, protein content, allergenic properties, and carbohydrates.

Atmospheric gaseous pollutants can induce qualitative and quantitative changes in airborne pollen characteristics. In this work, it was investigated the effects of carbon dioxide (CO2) on Acer negundo pollen fertility, protein content, allergenic properties, and carbohydrates. Pollen was collected directly from the anthers and in vitro exposed to three CO2 levels (500, 1000, and 3000 ppm) for 6 and 24 h in an environmental chamber. Pollen fertility was determined using viability and germination assays, total soluble protein was determined with Coomassie Protein Assay Reagent, and the antigenic and allergenic properties were investigated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunological techniques using patients' sera. Also, pollen fructose, sucrose, and glucose values were determined. Carbon dioxide exposure affected negatively pollen fertility, total soluble protein content, and fructose content. The patient sera revealed increased IgE reactivity to proteins of A. negundo pollen exposed to increasing levels of the pollutant. No changes were detected in the SDS-PAGE protein profiles and in sucrose and glucose levels. Our results indicate that increase in atmospheric CO2 concentrations can have a negative influence of some features of A. negundo airborne pollen that can influence the reproductive processes as well as respiratory pollen allergies in the future.

Elevated carbon dioxide and ozone alter productivity and ecosystem carbon content in northern temperate forests.

Three young northern temperate forest communities in the north-central United States were exposed to factorial combinations of elevated carbon dioxide (CO2 ) and tropospheric ozone (O3 ) for 11 years. Here, we report results from an extensive sampling of plant biomass and soil conducted at the conclusion of the experiment that enabled us to estimate ecosystem carbon (C) content and cumulative net primary productivity (NPP). Elevated CO2 enhanced ecosystem C content by 11%, whereas elevated O3 decreased ecosystem C content by 9%. There was little variation in treatment effects on C content across communities and no meaningful interactions between CO2 and O3 . Treatment effects on ecosystem C content resulted primarily from changes in the near-surface mineral soil and tree C, particularly differences in woody tissues. Excluding the mineral soil, cumulative NPP was a strong predictor of ecosystem C content (r(2) = 0.96). Elevated CO2 enhanced cumulative NPP by 39%, a consequence of a 28% increase in canopy nitrogen (N) content (g N m(-2) ) and a 28% increase in N productivity (NPP/canopy N). In contrast, elevated O3 lowered NPP by 10% because of a 21% decrease in canopy N, but did not impact N productivity. Consequently, as the marginal impact of canopy N on NPP (∆NPP/∆N) decreased through time with further canopy development, the O3 effect on NPP dissipated. Within the mineral soil, there was less C in the top 0.1 m of soil under elevated O3 and less soil C from 0.1 to 0.2 m in depth under elevated CO2 . Overall, these results suggest that elevated CO2 may create a sustained increase in NPP, whereas the long-term effect of elevated O3 on NPP will be smaller than expected. However, changes in soil C are not well-understood and limit our ability to predict changes in ecosystem C content.

Calcium and aluminum impacts on sugar maple physiology in a northern hardwood forest.

Forests of northeastern North America have been exposed to anthropogenic acidic inputs for decades, resulting in altered cation relations and disruptions to associated physiological processes in multiple tree species, including sugar maple (Acer saccharum Marsh.). In the current study, the impacts of calcium (Ca) and aluminum (Al) additions on mature sugar maple physiology were evaluated at the Hubbard Brook Experimental Forest (Thornton, NH, USA) to assess remediation (Ca addition) or exacerbation (Al addition) of current acidified conditions. Fine root cation concentrations and membrane integrity, carbon (C) allocation, foliar cation concentrations and antioxidant activity, foliar response to a spring freezing event and reproductive ability (flowering, seed quantity, filled seed and seed germination) were evaluated for dominant sugar maple trees in a replicated plot study. Root damage and foliar antioxidant activity were highest in Al-treated trees, while growth-associated C, foliar re-flush following a spring frost and reproductive ability were highest in Ca-treated trees. In general, we found that trees on Ca-treated plots preferentially used C resources for growth and reproductive processes, whereas Al-treated trees devoted C to defense-based processes. Similarities between Al-treated and control trees were observed for foliar cation concentrations, C partitioning and seed production, suggesting that sugar maples growing in native forests may be more stressed than previously perceived. Our experiment suggests that disruption of the balance of Ca and Al in sugar maples by acid deposition continues to be an important driver of tree health.

Foraging strategies in trees of different root morphology: the role of root lifespan.

Resource exploitation of patches is influenced not simply by the rate of root production in the patches but also by the lifespan of the roots inhabiting the patches. We examined the effect of sustained localized nitrogen (N) fertilization on root lifespan in four tree species that varied widely in root morphology and presumed foraging strategy. The study was conducted in a 12-year-old common garden in central Pennsylvania using a combination of data from minirhizotron and root in-growth cores. The two fine-root tree species, Acer negundo L. and Populus tremuloides Michx., exhibited significant increases in root lifespan with local N fertilization; no significant responses were observed in the two coarse-root tree species, Sassafras albidum Nutt. and Liriodendron tulipifera L. Across species, coarse-root tree species had longer median root lifespan than fine-root tree species. Localized N fertilization did not significantly increase the N concentration or the respiration of the roots growing in the N-rich patch. Our results suggest that some plant species appear to regulate the lifespan of different portions of their root system to improve resource acquisition while other species do not. Our results are discussed in the context of different strategies of foraging of nutrient patches in species of different root morphology.

Effects of elevated [CO2] and low soil moisture on the physiological responses of Mountain Maple (Acer spicatum L.) seedlings to light.

Global climate change is expected to affect how plants respond to their physical and biological environments. In this study, we examined the effects of elevated CO2 ([CO2]) and low soil moisture on the physiological responses of mountain maple (Acer spicatum L.) seedlings to light availability. The seedlings were grown at ambient (392 µmol mol(-1)) and elevated (784 µmol mol(-1)) [CO2], low and high soil moisture (M) regimes, at high light (100%) and low light (30%) in the greenhouse for one growing season. We measured net photosynthesis (A), stomatal conductance (g s), instantaneous water use efficiency (IWUE), maximum rate of carboxylation (V cmax), rate of photosynthetic electron transport (J), triose phosphate utilization (TPU)), leaf respiration (R d), light compensation point (LCP) and mid-day shoot water potential (Ψx). A and g s did not show significant responses to light treatment in seedlings grown at low soil moisture treatment, but the high light significantly decreased the C i/C a in those seedlings. IWUE was significantly higher in the elevated compared with the ambient [CO2], and the effect was greater at high than the low light treatment. LCP did not respond to the soil moisture treatments when seedlings were grown in high light under both [CO2]. The low soil moisture significantly reduced Ψx but had no significant effect on the responses of other physiological traits to light or [CO2]. These results suggest that as the atmospheric [CO2] rises, the physiological performance of mountain maple seedlings in high light environments may be enhanced, particularly when soil moisture conditions are favourable.

The within-season and between-tree distribution of imidacloprid trunk-injected into Acer platanoides (Sapindales: Sapindaceae).

Norway maple trees, Acer platanoides L. (Sapindales: Sapindaceae), that were trunk-injected with imidacloprid as part of an Asian longhorned beetle eradication program, were used to study the temporal and between-tree distribution of imidacloprid in twigs from June through September. The effect of injection time during spring on imidacloprid residues across the summer season and the distribution of imidacloprid in twig bark versus twig xylem were also investigated. Overall, we observed a significant decline in imidacloprid concentrations within each plant part sampled across the study period, although the 19 trees used in the study varied greatly in the pattern of imidacloprid residues over time. The concentration of imidacloprid in twig bark per dry mass was approximately two times higher than that of the twig xylem (means +/- SD of 1.21 +/- 2.16 ppm vs. 0.63 +/- 1.08 ppm imidacloprid, respectively). The majority (> 50%) of whole twig, twig bark and twig xylem samples from injected trees contained < 1 ppm imidacloprid and 37% of twig samples contained 0 ppm. Maximum values were 9 ppm for whole twigs from trunk-injected trees, 12 ppm imidacloprid for twig bark, and 5 ppm for twig xylem. Leaves, sampled only in September, had much higher imidacloprid residues than twigs collected at the same time; the majority (53%) of leaf samples contained > 5 ppm imidacloprid, with a maximum of 49 ppm. The concentrations ofimidacloprid in whole twigs, twig bark, and twig xylem were highly correlated, and levels in leaves were correlated with imidacloprid levels in whole twigs.

Cyclic diarylheptanoids as inhibitors of NO production from Acer nikoense.

We prepared a series of acerogenins A and B derivatives as inhibitors of nitric oxide (NO) production in vitro. Our results suggested that an ester group at a hydroxyl at C-2 improved inhibitory effects without cytotoxicity. A benzoyl ester derivative of acerogenin C showed the most potent inhibitory activity of NO production from lipopolysaccharide-activated macrophages.

Leaf shape responds to temperature but not CO2 in Acer rubrum.

The degree of leaf dissection and the presence of leaf teeth, along with tooth size and abundance, inversely correlate with mean annual temperature (MAT) across many plant communities. These relationships form the core of several methods for reconstructing MAT from fossils, yet the direct selection of temperature on tooth morphology has not been demonstrated experimentally. It is also not known if atmospheric CO(2) concentration affects leaf shape, limiting confidence in ancient climate reconstructions because CO(2) has varied widely on geologic timescales. Here I report the results of growing Acer rubrum (red maple) in growth cabinets at contrasting temperature and CO(2) conditions. The CO(2) treatment imparted no significant differences in leaf size and shape, while plants grown at cooler temperatures tended to have more teeth and more highly dissected leaves. These results provide direct evidence for the selection of temperature on leaf shape in one species, and support a key link in many leaf-climate methods. More broadly, these results increase confidence for using leaf shape in fossils to reconstruct paleoclimate.

Metal resistance in populations of red maple (Acer rubrum L.) and white birch (Betula papyrifera Marsh.) from a metal-contaminated region and neighbouring non-contaminated regions.

Metal resistance in populations of Acer rubrum and Betula papyrifera in the industrially contaminated region of Sudbury, Ontario, was compared with resistance in populations from neighbouring uncontaminated regions. In two one-season experiments, seedlings were grown outdoors on contaminated (mainly Cu, Ni) and uncontaminated substrates. Sudbury populations of both species responded less to contamination than populations from uncontaminated regions. In A. rubrum this difference was small. For both species, Sudbury plants were smaller when grown on uncontaminated substrate. B. papyrifera from Sudbury grew better on contaminated substrate than the other populations. There is indication of variation in metal resistance within the populations from the non-contaminated regions. The data shows that trees may develop adaptive resistance to heavy metals, but the low degree of resistance indicates that the development of such resistances are slower than observed for herbaceous species with shorter generation times.

Maplexins, new α-glucosidase inhibitors from red maple (Acer rubrum) stems.

Thirteen gallic acid derivatives including five new gallotannins, named maplexins A-E, were isolated from red maple (Acer rubrum) stems. The compounds were identified by spectral analyses. The maplexins varied in number and location of galloyl groups attached to 1,5-anhydro-d-glucitol. The isolates were evaluated for α-glucosidase inhibitory and antioxidant activities. Maplexin E, the first compound identified with three galloyl groups linked to three different positions of 1,5-anhydro-d-glucitol, was 20 fold more potent than the α-glucosidase inhibitory drug, Acarbose (IC(50)=8 vs 160 µM). Structure-activity related studies suggested that both number and position of galloyls attached to 1,5-anhydro-d-glucitol were important for α-glucosidase inhibition.