Antioxidative compounds from leaves of Tahongai (<Emphasis Type="Italic">Klienhovia hospita</Emphasis>)

In our effort to find antioxidant agent, we focused on Tahongai (Kleinhovia hospita) which have been used traditionally in Indonesia as medicinal herbal to cure liver disease. Based on the biologically guided fractionation using DPPH radical scavenging assay, eleutherol and kaempferol 3-O-β-d-glucoside was isolated from the leaves of K. hospita. Kaempferol 3-O-β-d-glucoside (1) and eleutherol (2) scavenged the radical with IC₅₀ of 71.4 and 491.8 μM, respectively. In addition, both of the compounds did not exhibit cytotoxicity on HepG2 cells.     

Antioxidative activity and anti-inflammatory effects of diarylheptanoids isolated from <Emphasis Type="Italic">Alnus hirsuta</Emphasis>

Two diarylheptanoids, 1,7-bis-(3,4-dihydroxyphenyl)-heptane-3-one-5-O-β-d-xylopyranoside (I) and 1,7-bis-(3,4-dihydroxyphenyl)-heptane-5-O-β-d-xylopyranoside (II), were isolated from the bark of Alnus hirsuta. Compounds I and II exhibited strong antioxidative activity against 1,1-diphenyl-2-picrylhydrazyl radicals, with IC₅₀ values of 8.36 ± 0.54 and 8.67 ± 1.46 μM, respectively. In addition, we demonstrated that compounds I and II inhibited the production of nitric oxide and reactive oxygen species and the expression of proinflammatory molecules such as inducible nitric oxide synthase and cyclooxygenase-2 in lipopolysaccharide-induced macrophages. According to our results, the two diarylheptanoids isolated from the bark of A. hirsuta exhibited significant antioxidative activity and anti-inflammatory effects and may be useful in the pharmaceutical industry for alleviating oxidative stress.     

A glucosylceramide with antimicrobial activity from the edible mushroom <Emphasis Type="Italic">Pleurotus citrinopileatus</Emphasis>

The golden oyster mushroom Pleurotus citrinopileatus is a popular edible mushroom with multifunctional biological activities, but there are a limited number of previous studies on its chemical composition. This is the first report of the isolation of glucosylceramide with antimicrobial activity from the fruiting body of this mushroom. This compound was identified as 1-O-β-d-glucopyranosyl-(2S,3R,4E,8E)-2-[(2R)-2-hydroxyhexadecanoylamino]-9-methyl-4,8-octadecadiene-1,3-diol. The IC₅₀ value of this compound for the growth of Escherichia coli and Staphylococcus aureus was 275.1 μM (200 μg/ml) and 323.2 μM (235 μg/ml), respectively.     

Insecticidal activity of synthetic antioxidants,natural phytochemicals,and essential oils against an <Emphasis Type="Italic">Aspergillus</Emphasis> section <Emphasis Type="Italic">Flavi</Emphasis> vector (<Emphasis Type="Italic">Oryzaephilus surinamensis</Emphasis> L.) in microcosm

The purpose of this study is to investigate the insecticidal activity of two benzoic acids 2(3)-tert-butyl-4 hydroxyanisole (BHA) and 2,6-di(tert-butyl)-p-cresol (BHT); two phenolic acids 3-phenyl-2-propenoic acid (CA) and trans-4-hydroxy-3-methoxycinnamic acid (FA); and two essential oils of Eugenia caryophyllata (clove tree) and Thymus vulgaris (thyme) against Oryzaephilus surinamensis (L.) vector carrier of aflatoxigenic fungi on stored peanut. The weight loss of peanut, susceptibility of insects, Aspergillus isolation frequency from insects and peanut, and aflatoxin B₁ analyses from peanut were determined. BHA, BHT, and BHA/BHT mixture were highly effective against O. surinamensis, these chemical agents give 100% mortality at the doses assayed. Essential oil of thyme at 2,000 and 3,000 ppm were highly effective against O. surinamensis, these concentrations gave 100% mortality. No Aspergillus section Flavi contamination was observed in dead insects collected from peanut treated with BHA/BHT mixture. No differences were observed in the fresh weight of pods peanut among treatments with and without chemical agents. All samples of treated peanut pods showed complete inhibition of this toxin after 120 days of storage. Our results indicate that these substances could be evaluated further for the control of pest vectors of aflatoxigenic fungi of stored peanut.     

Cultivation of <Emphasis Type="Italic">Pleurotus eryngii</Emphasis> on umbrella plant (<Emphasis Type="Italic">Cyperus alternifolius</Emphasis>) substrate

Mycelial growth and mushroom yields of three strains of Pleurotus eryngii produced on wheat bran-supplemented umbrella plant (Cyperus alternifolius) substrate were assessed using surface brightness, bromophenol blue color reactions, ergosterol and glucosamine contents, and water potential as indicators of strain performance. Mycelial growth was 31%–46% greater, depending on strain, on the umbrella plant substrate compared with the mushroom industry standard sugi (Cryptomeria japonica) substrate. Mushroom yields on the first flush were 20%–23% higher, depending on strain, on the plastic bottle-contained umbrella plant substrate. However, yields on the second break were lower from the umbrella plant substrate. Because many growers in Japan only harvest one flush, production of P. eryngii on umbrella plant substrate may offer commercial producers an alternative basal ingredient to diminishing supplies of sugi sawdust.Part of this report was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002.     

Chemical changes in terpenes of sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) wood during steam drying in kiln at high temperature

Sawdusts of sugi (Cryptomeria japonica) wood prepared before and after steam drying at 120°C in a kiln were extracted with n-hexane and ethyl acetate to give n-hexane extracts and ethyl acetate extracts. From gas chromatography-mass spectrometry analysis of the ethyl acetate extracts from woods before and after steam drying, the components of 4-epi-cubebol, cubebol, and 2,7(14),10-bis-abolatrien-1-ol-4-one, which existed in the raw sugi wood, were proved to disappear in the steam-dried wood. These components were also absent in the ethyl acetate extract of the steam-condensed solution of waste steam from the kiln outlet. When these three components were treated with 0.2% (v/v) acetic acid solution at 120°C, δ-cadinene was produced as a major product from both 4-epi-cubebol and cubebol by dehydration and cleavage of the cyclopropane ring, and cryptomerone from 2,7(14),10-bisabolatrien-1-ol-4-one by hydration. The chemical changes of the three components presumably occur during steam drying of the sugi wood. This study was presented in part at the 85th Spring Meeting of the Chemical Society of Japan, Kanagawa, Japan, March 26–29, 2005     

Inhibitory effects of <Emphasis Type="SmallCaps">l</Emphasis>-pipecolic acid from the edible mushroom, <Emphasis Type="Italic">Sarcodon aspratus</Emphasis>, on angiotensin I-converting enzyme

The aqueous extract of the edible mushroom Sarcodon aspratus showed inhibitory effects against angiotensin I-converting enzyme (ACE). l-Pipecolic acid (l-piperidine-2-carboxylic acid) was isolated from a hot-water extract in a 0.02% yield as an active principle. The mode of inhibition of l-pipecolic acid was found to be competitive, whereas its d-isomer showed no significant inhibitory effects against ACE, suggesting that the configuration of the carboxyl group in the molecule plays an important role in the enzyme inhibition.     

Effects of stilbenes from bark of <Emphasis Type="Italic">Picea glehnii</Emphasis> (Sieb. et Zucc) and their related compounds against feeding behaviour of <Emphasis Type="Italic">Reticulitermes speratus</Emphasis> (Kolbe)

The effects of stilbene glucosides and related compounds on termite feeding behavior were investigated using paper disc methods against the subterranean termite Reticulitermes speratus. The stilbene-rich fraction and isorhapontin (3-methoxy-3,4, 5-trihydroxystilbene-3--d-glucoside) from bark extracts of Picea glehnii showed avoidance by termites in choice tests. In the no-choice tests using compounds purified from the stilbene-rich fraction, the largest feeding deterrent effect was observed for piceid (3,4,5-trihydroxystilbene-3--d-glucoside), followed by isorhapontin, and astringin (3,3,4,5-tetrahydroxystilbene-3--d-glucoside), at the concentrations from 0.63 to 2.5µmol/disc. No change in activity was observed at retentions of more than 5.0µmol/disc. When the activities of isorhapontin and its aglycone derivative (isorhapontigenin: 3-methoxy-3,4,5-trihydroxystilbene) were compared with that of taxifolin (3,3,4,5,7-pentahydroxyflavanone) in the no-choice test, the stilbenes exhibited a larger antifeedant potential. Methylation of isorhapontigenin increased its termiticidal activity.Part of this study was presented at the 32nd Annual Meeting of the International Research Group on Wood Preservation, 2001     

Neuropharmacological activities of phytoncide released from <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

Forest-air bathing and walking (shinrin-yoku) is beneficial to human heath. In this study the phytoncide (volatile compounds) released from Cryptomeria japonica plantation forest was characterized by using solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS). The main volatile compounds were α-pinene (19.35%), β-myrcene (16.98%), d-limonene (15.21%), and γ-muurolene (7.42%). Furthermore, the neuropharmacological activity of the essential oils from leaves of C. japonica (ECJ) was evaluated by several animal behavior tests. ECJ could prolong the sleeping phase of ICR (imprinting control region) mice in the pentobarbital-induced sleeping time model. Furthermore, both EJC and one of its monoterpenes, d-limonene, possessed potent anxiolytic and analgesic activities based on the results obtained from elevated plus maze and writhing tests. The volatile compounds released from C. japonica provide relaxing and stress-relieving effects on mice, and further study on the effect of phytoncide on humans is worthwhile.     

Antioxidant properties and structural analysis of phenolic glucosides from bark of <Emphasis Type="Italic">Populus ussuriensis</Emphasis> Kom.

The methanol (MeOH) extract of Populus ussuriensis Kom. bark was analyzed for antioxidant assessing by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging potential. Among fractions using several solvents, the ethyl acetate (EtOAc) soluble fraction, which showed strong antioxidant activity (IC₅₀ 2.02 ± 0.01 μg/ml), was further purified by Thin layer chromatography (TLC) guided Sephadex LH-20 column chromatography. Three known phenolic glucosides, picein (I), salicortin (II), grandidentatin (III), and that of a new, 2-hydroxycyclohexyl-4′-O-p-coumaroyl-β-d-glucopyranoside (isograndidentatin A), were isolated and their structures were elucidated on the basis of physiochemical and spectroscopic methods. This was the first report of the chemical composition of P. ussuriensis bark. Phenolic glucosides III and IV exhibited strong antioxidant activities, with IC₅₀ values 6.73 ± 0.01 and 6.69 ± 0.01 μM, respectively, comparable to the control (α-tocopherol, IC₅₀ 6.80 ± 0.01 μM). P. ussuriensis bark EtOAc-soluble fraction and compounds III and IV could be used as biopreservatives in food applications as well as for cosmetic and medicinal preparations, to alleviate oxidative stress.     

At5g54160 gene encodes <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> 5-hydroxyconiferaldehyde <Emphasis Type="Italic">O</Emphasis>-methyltransferase

The function of an Arabidopsis thaliana gene, At5g54160 annotated as a caffeic acid O-methyltransferase CAOMT gene was characterized. The recombinant enzyme of this gene (AtOMT1) catalyzed the O-methylation of phenylpropanoid and flavonoid substrates. The specificity constants (k _cat/K _m) for 5-hydroxyconiferaldehyde (5-HCAld) and quercetin were both 0.11 μM⁻¹·min⁻¹. On the other hand, lignins of At5g54160-knockout Arabidopsis mutants lacked syringyl units. In addition, we showed that the gene silencing also resulted in significant accumulation of caffeyl alcohol (CaAlc). These results strongly suggested that At5g54160 gene is involved in syringyl lignin synthesis for the methylation of both 5-hydroxyconiferaldehyde and 3,4-dihydroxyphenyl compound(s). Part of this work was presented at the Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry, March 24–27, 2007     

Spectrophotometric assay of a wood preservative, <Emphasis Type="Italic">N,N</Emphasis>-didecyl-<Emphasis Type="Italic">N</Emphasis>-methylpoly(oxyethyl) ammonium propionate (DMPAP), in aqueous solution

A spectrophotometric assay based on the color reaction between N,N-didecyl-N-methyl-poly(oxyethyl) ammonium propionate (DMPAP) and 4-[4-(dipropylamino) phenylazo]-benzenesulfonic acid (propyl orange) was used to determine DMPAP concentrations as a wood preservative. The assay was carried out using a propyl orange solution at pH 2.9. The visible absorption spectrum of propyl orange showed an absorption maximum at 510 nm, which decreased linearly with increasing DMPAP concentration from 0 to 10 ppm. To apply this assay method to determine DMPAP retention in treated wood, the influence of wood extractives on the assay was investigated. Extractives from Japanese cedar, hinoki cypress, and Japanese larch were found to increase apparent DMPAP concentration. However, it was also found that measuring visible absorption at 477 nm prevented overestimation and gave precise values. This assay can be a viable alternative to the current methods for the determination of DMPAP concentrations.     

Physiological effects of touching hinoki cypress (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>)

This study aimed to clarify the physiological effects of touching hinoki cypress (hereinafter called “hinoki”) (Chamaecyparis obtusa) wood with the palm of the right hand on brain activity and autonomic nervous activity. Twenty-two female university students (mean age 21.1?±?0.2 years) participated in the study. As an indicator of brain activity, oxyhemoglobin (oxy-Hb) concentrations were measured in the left/right prefrontal cortex using near-infrared time-resolved spectroscopy. Heart rate variability (HRV) was used as an indicator of autonomic nervous activity. The high-frequency (HF) component of HRV, which reflected parasympathetic nervous activity, and the low-frequency (LF)/HF ratio, which reflected sympathetic nervous activity, were measured. Plate (300?×?300 mm) of uncoated hinoki was used as tactile stimulation. Marble was used as a control material. After sitting at rest with their eyes closed, the participants touched the materials for 90 s with their eyes still closed. The results showed that tactile stimulation with hinoki significantly decreased oxy-Hb concentration in the left prefrontal cortex and increased ln(HF) reflected parasympathetic nervous activity compared with marble. In conclusion, our study showed that touching hinoki wood with the palm calms prefrontal cortex activity and increases parasympathetic nervous activity, thereby inducing physiological relaxation.     

Successful <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Populus tomentosa</Emphasis> with apple <Emphasis Type="Italic">SPDS</Emphasis> gene

The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase （SPDS） genes derived from an apple by an Agrobacterium-mediated method. Four transgenic clones were confu＇med by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR.     

New piezoelectric moduli of wood: <Emphasis Type="Italic">d</Emphasis><Subscript>31</Subscript> and <Emphasis Type="Italic">d</Emphasis><Subscript>32</Subscript>

This article reports the piezoelectric moduli of wood d ₃₁, d ₃₂, and d ₃₆. The piezoelectric moduli of wood d ₃₁ and d ₃₂ have not been previously reported, although there has been much research on the d ₁₄ and d ₂₅ moduli of wood. The moduli d ₃₁, d ₃₂, and d ₃₆ were measured carefully because their absolute values were considerably smaller than those of d ₁₄ and d ₂₅. For Softwoods, d ₃₆ values were mostly negative, whereas the values for hardwoods had either positive or negative values. The other moduli, d ₃₁ and d ₃₂, were a mixture of positive and negative values in softwoods and hardwoods. The existence of d ₃₁ and d ₃₂ suggests the presence of an electrical polarity of the cellulose crystal in the fiber direction of the wood. The polarities of d ₃₁ and d ₃₂ became clear from wood in the outer part of the trunk, where the crystallinity of cellulose is large and the alignment of the crystals becomes parallel to the fiber direction.     

Heartwood extractives from the Amazonian trees <Emphasis Type="Italic">Dipteryx odorata,Hymenaea courbaril</Emphasis>, and <Emphasis Type="Italic">Astronium lecointei</Emphasis> and their antioxidant activities

Heartwood extracts from Amazonian trees cumaru-ferro (Dipteryx odorata), jatoba (Hymenaea courbaril), and guarita (Astronium lecointei) exhibit antioxidant activities comparable with that of α-tocopherol, a well-known antioxidant. This article reports the characterization of the antioxidant compounds in the extracts of the three heartwoods. Silica gel column chromatography of the cumaru-ferro EtOAc extract yielded (−)-(3R)-7,2′,3′-trihydroxy-4′-methoxyisoflavan and (+)-(3R)-8,2′,3′-trihydroxy-7,4′-dimethoxyisoflavan. Silica gel column chromatography followed by preparative high-performance liquid chromatography of the jatoba EtOAc extract yielded (−)-fisetinidol and (+)-trans-taxifolin. Chemical structures were assigned using electron-ionization mass spectrometry, ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy including nuclear Overhauser effect spectroscopy (NOESY), as well as optical rotation and circular dichroism. Gas chromatography-mass spectrometry demonstrated that the isolated compounds were predominant in the EtOAc extracts. In the guarita EtOAc extract, catechin and gallic acid were identified by comparing their retention times and mass fragmentation patterns with those of authentic samples. Antioxidant activity determined by the 1,1-diphenyl-2-picrylhydrazyl assay demonstrated that all these compounds had activities comparable with that of α-tocopherol. Part of this report was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, August 2007     

Bioremediation of CCA-treated wood by brown-rot fungi <Emphasis Type="Italic">Fomitopsis palustris</Emphasis>, <Emphasis Type="Italic">Coniophora puteana</Emphasis>, and <Emphasis Type="Italic">Laetiporus sulphureus</Emphasis>

This study evaluated oxalic acid accumulation and bioremediation of chromated copper arsenate (CCA)-treated wood by three brown-rot fungi Fomitopsis palustris, Coniophora puteana, and Laetiporus sulphureus. The fungi were first cultivated in a fermentation broth to accumulate oxalic acid. Bioremediation of CCA-treated wood was then carried out by leaching of heavy metals with oxalic acid over a 10-day fermentation period. Higher amounts of oxalic acid were produced by F. palustris and L. sulphureus compared with C. puteana. After 10-day fermentation, oxalic acid accumulation reached 4.2 g/l and 3.2 g/l for these fungi, respectively. Fomitopsis palustris and L. sulphureus exposed to CCA-treated sawdust for 10 days showed a decrease in arsenic of 100% and 85%, respectively; however, C. puteana remediation removed only 18% arsenic from CCA-treated sawdust. Likewise, chromium removal in F. palustris and L. sulphureus remediation processes was higher than those for C. puteana. This was attributed to low oxalic acid accumulation. These results suggest that F. palustris and L. sulphureus remediation processes can remove inorganic metal compounds via oxalic acid production by increasing the acidity of the substrate and increasing the solubility of the metals.An erratum to this article can be found at     

Response of <Emphasis Type="Italic">Eucalyptus camaldulensis</Emphasis> and <Emphasis Type="Italic">Acacia mangium</Emphasis> kraft pulp in different ECF bleaching options

In this study, the response of Eucalyptus camaldulensis and Acacia mangium kraft pulp to different options of ECF bleaching was investigated. Oxygen-delignified pulps of both species having similar kappa number (c.a. 12) were used. Both the pulps were prebleached using conventional ClO₂ stage (D₀), combined use of ClO₂ and O₃ (DZ), hot acid treatment before D₀ (A_hotD₀) and hot acid treatment before (DZ) i.e., A_hot(DZ), hot ClO₂ treatment (D_HT) and A_hot followed by D_HT (A_hotD_HT). All prebleached pulps were finally bleached to 90%+ ISO brightness using D₁ option. The overall benefits of other options were not considerable compared to D₀ and (DZ). From the ClO₂ consumption point of view, for a certain species, the option A_hot(DZ) was the best. Instead of D₁ in final bleaching, pressurized hydrogen peroxide (PO) was also investigated in the case of D₀ and (DZ) prebleached pulps. (PO)-bleached pulp produced higher brightness stability but lower viscosity compared to D₁.     

Leaf-level responses to light in two co-occurring <Emphasis Type="Italic">Quercus</Emphasis> (<Emphasis Type="Italic">Quercus ilex</Emphasis> and <Emphasis Type="Italic">Quercus suber</Emphasis>): leaf structure,chemical composition and photosynthesis

We studied morphological, biochemical and physiological leaf acclimation to incident Photon-Photosynthetic-Flux-Density (PPFD) in Quercus ilex (holm oak) and Quercus suber (cork oak) at Mediterranean evergreen oak woodlands of southern Portugal. Specific leaf area (SLA) decreased exponentially with increasing PPFD in both species. Q. ilex had lower SLA values than Q. suber. Leaf nitrogen, cellulose and lignin concentration (leaf area-based) scaled positively with PPFD. Maximum rate of carboxylation (V_cmax), capacity for maximum photosynthetic electron transport (J_max), rate of triose-P utilization (V_TPU) and the rate of nonphotorespiratory light respiration (Rd) were also positively correlated with PPFD in both Quercus species, when expressed in leaf area but not on leaf mass basis. Q suber showed to have higher photosynthetic potential (V_cmax, J_max^m and V_TPU^m) and a higher nitrogen efficient nitrogen use than Q.ilex. Leaf chlorophyll concentration increased with decreasing PPFD, improving apparent quantum use efficiency (Φ) in both Quercus species. We concluded that, in Q.ilex and Q.suber, leaf structural plasticity is a stronger determinant for leaf acclimation to PPFD than biochemical and physiological plasticity.     

Gas exchange responses of <Emphasis Type="Italic">Eucalyptus</Emphasis>, <Emphasis Type="Italic">C. africana</Emphasis> and <Emphasis Type="Italic">G. robusta</Emphasis> to varying soil moisture content in semi-arid (Thika) Kenya

A dramatic decline in forest cover in eastern Africa along with a growing population means that timber and poles for building and fuelwood are in short supply. To overcome this shortage, the region is increasingly turning to eucalyptus. But eucalyptus raises environmental concerns of its own. Fears that it will deplete water supply, affect wildlife and reduce associated crop yields have caused many countries in the region to discourage farmers from planting this exotic. This paper is part of a series of investigations on the growth and water use efficiency of faster growing eucalyptus hybrids, which was introduced from South Africa to Kenya. The hypothesis is that the new hybrids are more efficient in using water and more suitable for the semi-arid tropics than existing eucalyptus and two popular agroforestry species. Gas exchange characteristics of juvenile Eucalyptus grandis (W. Hill ex Maiden), two eucalyptus hybrids (E. grandis × Eucalyptus camaldulensis Dehnh.), Grevillea robusta (A. Cunn) and Cordia africana (Lam) was studied under field and pot conditions using an infrared gas analyzer was used to measure photosynthetic active radiation (PAR), net photosynthetic rate (A), stomatal conductance (g _s) and transpiration rate (E) at CO₂ concentrations of 360 μmol mol⁻¹ and ambient humidity and temperature. A, E and g _s varied between species, being highest in eucalyptus hybrid GC 15 (24.6 μmol m⁻² s⁻¹) compared to eucalyptus hybrid GC 584 (21.0 μmol m⁻² s⁻¹), E. grandis (19.2 μmol m⁻² s⁻¹), C. africana (17.7 μmol m⁻² s⁻¹) and G. robusta (11.1 μmol m⁻² s⁻¹). C. africana exhibited high E values (7.0 mmol m⁻² s⁻¹) at optimal soil moisture contents than G. robusta (3.9 mmol m⁻² s⁻¹) and eucalyptus (5.3 mmol m⁻² s⁻¹) in field experiment and G. robusta (3.2 mmol m⁻² s⁻¹) and eucalyptus (4.2 mmol m⁻² s⁻¹) in pot-grown trees. At very low soil moisture content, extremely small g _s values were recorded in GC 15 and E. grandis (8 mmol m⁻² s⁻¹) and G. robusta (14 mmol m⁻² s⁻¹) compared to GC 584 (46.9 mmol m⁻² s⁻¹) and C. africana (90.0 mmol m⁻² s⁻¹) indicating strong stomatal control by the species. Instantaneous water use efficiency ranged between 3 and 5 μmol mmol⁻¹ and generally decreased with decline in soil moisture in pot-grown trees but increased with declining soil moisture in field-grown trees.