1H NMR metabolic phenotyping of Dipterocarpus alatus as a novel tool for age and growth determination.

Dipterocarpus alatus belongs to Family Dipterocarpaceae that can be commonly found in Southeast Asian countries. It is a perennial plant with oval-shaped leaves and oleoresin-rich wood. It has been considered as a multipurpose plant since all parts can be practically utilized. One of the major problems for utilizing Dipterocarpus alatus is the difficulty knowing the exact age as this kind of plant is ready for multipurpose use after 20 years of age. At present, the most commonly used method for determining age of Dipterocarpus alatus is the annual ring estimation. However, this conventional method is unable to provide the high precision and accuracy of age determination due to its limitation including blurry annual rings caused by enriched oleoresin in the wood. The current study aimed to investigate the differences of 1H -NMR spectroscopy-based metabolic profiles from bark and leaf of Dipterocarpus alatus at different ages including 2, 7, 15 and 25 years. Our findings demonstrated that there is a total of 56 metabolites shared between bark and leaf. It is noticeable that bark at different ages exhibited the strongest variation and sugar or sugar derivatives that were found in higher concentrations in bark compared with those in leaf. We found that decreasing levels of certain metabolites including tagatose, 1'kestose and 2'-fucosyllactose exhibited the promising patterns. In conclusion, panel metabolites involved in the sucrose biosynthesis can precisely determine the age and growth of Dipterocarpus alatus.

Treatment of urea manufacturing facility effluent by Hopea odorata and Khaya ivorensis.

Phytoremediation is an environmentally friendly and sustainable alternative for treatment of nitrogen-enriched wastewaters. In this study, Ta-khian (Hopea odorata) and Lagos mahogany (Khaya ivorensis), two tropical timber plants, were investigated for their performances in treatment of urea manufacturing factory effluent with high nitrogen (N) content. Plant seedlings received four concentrations of N (190, 240, 290 and 340 mg/L N) in laboratory-scale constructed wetlands every 4 days for a duration of 8 weeks. The solution volumes supplied to each container, amount of N recovered by plants and plant growth characteristics were measured throughout the experiment. Results showed that Ta-khian plants were highly effective at reducing N concentration and volume of water. A maximum of 63.05% N recovery was obtained by Ta-khian plants grown in 290 mg/L N, which was assimilated in the chlorophyll molecule structure and shoot biomass. Significant positive correlations have been shown between N recovery percentages and plant growth parameters. Ta-Khian plants can be applied as suitable phytoremediators for mitigating N pollution in water sources.

Hopeahainol A binds reversibly at the acetylcholinesterase (AChE) peripheral site and inhibits enzyme activity with a novel higher order concentration dependence.

Natural product inhibitors of AChE are of interest both because they offer promise as inexpensive drugs for symptomatic relief in Alzheimer's disease and because they may provide insights into the structural features of the AChE catalytic site. Hopeahainol A is an uncharged polyphenol AChE inhibitor from the stem bark of Hopea hainanensis with a constrained, partially dearomatized bicyclic core. Molecular modeling indicates that hopeahainol A binds at the entrance of the long but narrow AChE active site gorge because it is too bulky to be accommodated within the gorge without severe distortion of the gorge as depicted in AChE crystal structures. We conducted inhibitor competition experiments in which AChE inhibition was measured with hopeahainol A together with either edrophonium (which binds at the base of the gorge) or thioflavin T (which binds to the peripheral or P-site near the gorge mouth). The results agreed with the molecular modeling and indicated that hopeahainol A at lower concentrations (<200 µM) bound only to the P-site, as hopeahainol A and thioflavin T were unable to form a ternary complex with AChE while hopeahainol A and edrophonium did form a ternary complex with essentially no competition between them. Inhibition increased to a striking extent at higher concentrations of hopeahainol A, with plots analogous to classic Dixon plots showing a dependence on hopeahainol A concentrations to the third- or fourth order. The inhibition at higher hopeahainol A concentrations was completely reversed on dilution and blocked by bound edrophonium. We hypothesize that bound hopeahainol A induces conformational changes in the AChE active site that allow binding of additional hopeahainol A molecules, a phenomenon that would be unprecedented for a reversible inhibitor that apparently forms no covalent bonds with AChE.

Oligostilbenoids from the heartwood of N.†Heimii: role of non-covalent association in their biogenesis.

Four new oligostilbenes, including one dimer and three tetramers of resveratrol, that is, heimiols B-E (1-4) were isolated from the heartwood of Neobalanocarpus heimii (Dipterocarpaceae), together with thirteen known resveratrol oligomers (5-17). Examination of the structural diversity of the isolated oligostilbenes led to hypothesis of their biogenetic origin through a small number of versatile chemical pathways. These hypotheses are strongly supported by computational calculations (based on the density functional theory, DFT) that were performed to rationalize conformational re-arrangements and thus provide insights into the mechanism of oligostilbenoid biosynthesis. Non-covalent complexes are believed to drive the regio- and stereoselectivity of the oligomerization reactions.

High CO2 concentration increases relative leaf carbon gain under dynamic light in Dipterocarpus sublamellatus seedlings in a tropical rain forest, Malaysia.

Understory plants in tropical forests often experience a low-light environment combined with high CO2 concentration. We hypothesized that the high CO2 concentration may compensate for leaf carbon loss caused by the low light, through increasing light-use efficiency of both steady-state and dynamic photosynthetic properties. To test the hypothesis, we examined CO2 gas exchange in response to an artificial lightfleck in Dipterocarpus sublamellatus Foxw. seedlings under contrasting CO2 conditions: 350 and 700 µmol CO2 mol(-1) air in a tropical rain forest, Pasoh, Malaysia. Total photosynthetic carbon gain from the lightfleck was about double when subjected to the high CO2 when compared with the low CO2 concentration. The increase of light-use efficiency in dynamic photosynthesis contributed 7% of the increased carbon gain, most of which was due to reduction of photosynthetic induction to light increase under the high CO2. The light compensation point of photosynthesis decreased by 58% and the apparent quantum yield increased by 26% at the high CO2 compared with those at the low CO2. The study suggests that high CO2 increases photosynthetic light-use efficiency under both steady-state and fluctuating light conditions, which should be considered in assessing the leaf carbon gain of understory plants in low-light environments.

ROS production and lipid catabolism in desiccating Shorea robusta seeds during aging.

Reactive oxygen species (ROS) and lipid peroxidation products appear to correlate strongly with the desiccation induced loss of viability in recalcitrant sal seeds. The 100% germination in fresh sal seeds declined with dehydration under natural storage conditions (26 ± 1 °C, relative humidity 52 ± 2%). Seeds became non-viable within 8 days. Desiccation induced disturbances in the metabolic activity of seeds resulted in generation of enormous amounts of ROS that are responsible for cellular damage and viability loss. Oxidative stress in the dehydrating aging sal seeds was further aggravated by inducing lipid peroxidation as the amounts of free fatty acid, conjugated diene, lipid hydroperoxide and secondary free radicals; malondialdehyde and 4-hydroxy-2-nonenal, were also promoted. In addition, significant rise in lipid degrading enzymes; lipase (EC 3.1.1.3) and lipoxygenase (LOX, EC 1.13.11.12) were detected in dehydrating sal seeds. Our results indicated multiple pathways (ROS, lipid peroxidation & lipase and LOX) that operate in the dehydrating recalcitrant sal seeds finally contributing to loss of viability.

Vertical variation in leaf gas exchange parameters for a Southeast Asian tropical rainforest in Peninsular Malaysia.

Vertical variation in leaf gas exchange characteristics of trees grown in a lowland dipterocarp forest in Peninsular Malaysia was investigated. Maximum net photosynthetic rate, stomatal conductance, and electron transport rate of leaves at the upper canopy, lower canopy, and forest floor were studied in situ with saturated condition photosynthetic photon flux density. The dark respiration rate of leaves at the various heights was also studied. Relationships among gas exchange characteristics, and also with nitrogen content per unit leaf area and leaf dry matter per area were clearly detected, forming general equations representing the vertical profile of several important parameters related to gas exchange. Numerical analysis revealed that the vertical distribution of gas exchange parameters was well determined showing both larger carbon gain for the whole canopy and at the same time positive carbon gain for the leaves of the lowest layer. For correct estimation of gas exchange at both leaf and canopy scales using multi-layer models, it is essential to consider the vertical distribution of gas exchange parameters with proper scaling coefficients.

Patchy stomatal behavior during midday depression of leaf CO2 exchange in tropical trees.

We investigated effects of heterogeneous stomatal behavior on diurnal patterns of leaf gas exchange in 10 tree species. Observations were made in middle and upper canopy layers of potted tropical rainforest trees in a nursery at the Forest Research Institute Malaysia. Measurements were taken from 29 January to 3 February 2010. We measured in situ diurnal changes in net photosynthetic rate and stomatal conductance in three leaves of each species under natural light. In both top-canopy and sub-canopy species, midday depression of net assimilation rate occurred in late morning. Numerical analysis showed that patchy bimodal stomatal behavior occurred only during midday depression, suggesting that the distribution pattern of stomatal apertures (either uniform or non-uniform stomatal behavior) varies flexibly within single days. Direct observation of stomatal aperture using Suzuki's Universal Micro-Printing (SUMP) method demonstrated midday patchy stomatal closure that fits a bimodal pattern in Shorea leprosula Miq., Shorea macrantha Brandis. and Dipterocarpus tempehes V.Sl. Inhibition of net assimilation rate and stomatal conductance appears to be a response to changes in vapor pressure deficit (VPD). Variable stomatal closure with increasing VPD is a mechanism used by a range of species to prevent excess water loss from leaves through evapotranspiration (viz., inhibition of midday leaf gas exchange). Bimodal stomatal closure may occur among adjacent stomata within a single patch, rather than among patches on a single leaf. Our results suggest the occurrence of patches at several scales within single leaves. Further analysis should consider variable spatial scales in heterogeneous stomatal behavior between and within patches and within single leaves.

Total synthesis of hopeahainol A and hopeanol.

Similar, but different: Possessing almost the same but not identical structures, the recently discovered natural products hopeahainol A (1) and hopeanol (2) exhibit important but differing biological properties (see structures). Their first total synthesis has now been achieved through a series of novel cascade reactions and skeletal rearrangements.

Sustained diurnal photosynthetic depression in uppermost-canopy leaves of four dipterocarp species in the rainy and dry seasons: does photorespiration play a role in photoprotection?

Diurnal and seasonal changes in gas exchange and chlorophyll fluorescence of the uppermost-canopy leaves of four evergreen dipterocarp species were measured on clear days. The trees, that were growing in a plantation stand in southern Yunnan, China, had canopy heights ranging from 17 to 22 m. In the rainy season, Dipterocarpus retusus Bl. had higher photosynthetic capacity (A(max)) than Hopea hainanensis Merr. et Chun, Parashorea chinensis Wang Hsie and Vatica xishuangbannaensis G.D. Tao et J.H. Zhang (17.7 versus 13.9, 11.8 and 7.7 micromol m(-2) s(-1), respectively). In the dry season, A(max) in all species decreased by 52-64%, apparent quantum yield and dark respiration rate decreased in three species, and light saturation point decreased in two species. During the diurnal courses, all species exhibited sustained photosynthetic depression from midmorning onward in both seasons. The trees were able to regulate light energy allocation dynamically between photochemistry and heat dissipation during the day, with reduced actual photochemistry and increased heat dissipation in the dry season. Photorespiration played an important role in photoprotection in all species in both seasons, as indicated by a continuous increase in photorespiration rate in the morning toward midday and a high proportion of electron flow (about 30-65% of total electron flow) allocated to oxygenation for most of the day. None of the species suffered irreversible photoinhibition, even in the dry season. The sustained photosynthetic depression in the uppermost-canopy leaves of these species could be a protective response to prevent excessive water loss and consequent catastrophic leaf hydraulic dysfunction.

Midday depression of leaf CO2 exchange within the crown of Dipterocarpus sublamellatus in a lowland dipterocarp forest in Peninsular Malaysia.

We observed diurnal and seasonal patterns of leaf-scale gas exchange within the crown of a Dipterocarpus sublamellatus Foxw. tree growing in a lowland dipterocarp forest at Pasoh, Peninsular Malaysia. Observations were carried out nine times over 6 years, from September 2002 to December 2007. Observation periods included both wet and mild-dry periods, and natural and saturated photosynthetic photon flux density (PPFD) light conditions. In situ measurements of the diurnal change in net photosynthetic rate and in stomatal conductance were carried out on canopy leaves of a 40-m-tall D. sublamellatus tree, which was accessed from a canopy corridor. A diurnal change in electron transport rate was observed under saturated PPFD conditions. The maximum net assimilation rate was approximately 10 micromol m(-2) s(-1). There was a clear inhibition of the net assimilation rate coupled with stomatal closure after late morning and this inhibition occurred year-round. Although the electron transport rate decreased alongside this inhibition, it sometimes followed on. Numerical analysis showed that the main factor in the inhibition of the net assimilation rate was patchy bimodal stomatal closure, which occurred in both mild-dry and wet periods. The midday depression occurred year-round, though there are fluctuations in soil moisture during the mild-dry and wet periods. The magnitude of the inhibition was not related to soil water content but was related to vapor pressure deficit (VPD): that is, whether the days were sunny and hot or cloudy and cool. On cloudy, cool days in the wet period, the net photosynthesis was only moderately inhibited, but it still decreased in the afternoon and was coupled with patchy stomatal closure, even in quite moderate VPD, leaf temperature and PPFD conditions. Our results suggest that patchy stomatal closure signaled by the increase in VPD, in transpiration and by circadian rhythms, was the key factor in constraining midday leaf gas exchange of the D. sublamellatus canopy leaves.

Changes of leaf morphological, anatomical structure and carbon isotope ratio with the height of the Wangtian tree (Parashorea chinensis) in Xishuangbanna, China.

Leaf morphological and anatomical structure and carbon isotope ratio (delta13C) change with increasing tree height. To determine how tree height affects leaf characteristics, we measured the leaf area, specific leaf mass (ratio of leaf mass to leaf area [LMA]), thickness of the total leaf, cuticle, epidermis, palisade and sponge mesophyll, stomata traits and delta13C at different heights of Parashorea chinensis with methods of light and scanning electron microscopy (SEM) and isotope-ratio mass spectrometry. The correlation and stepwise regression between tree height and leaf structure traits were carried out with SPSS software. The results showed that leaf structures and delta13C differed significantly along the tree height gradient. The leaf area, thickness of sponge mesophyll and size of stomata decreased with increasing height, whereas the thickness of lamina, palisade mesophyll, epidermis, and cuticle, ratios of palisade to spongy thickness, density of stomata and vascular bundles, LMA and delta13C increased with tree height. Tree height showed a significant relationship with all leaf indices and the most significant relationship was with epidermis thickness, leaf area, cuticle thickness, delta13C. The delta13C value showed a significantly positive relationship with LMA (R = 0.934). Our results supported the hypothesis that the leaf structures exhibited more xeromorphic characteristics with the increasing gradient of tree height.