New formylated phloroglucinol compounds from Eucalyptus globulus foliage

Two new acylphloroglucinols were isolated from the leaves of Eucalyptus globulus Labill and identified as macrocarpals P (1) and Q (2). Structural elucidations were carried out using conventional 1D and 2D NMR and mass spectrometry together with complementary techniques (UV and IR). Macrocarpal Q was a diastereoisomer of macrocarpal E (3), configuration of which was not precised. Simultaneous isolation of macrocarpals E and Q allowed to determine the configurations of both compounds. The diformylphloroglucinol (4) was also isolated as well as already known compounds grandinol, macrocarpals D, I, L, N, O and am-1.     

Identification of quantitative trait loci influencing foliar concentrations of terpenes and formylated phloroglucinol compounds in Eucalyptus nitens

Leaves of eucalypt species contain a variety of plant secondary metabolites, including terpenoids and formylated phloroglucinol compounds (FPCs). Both terpene and FPC concentrations are quantitative traits that can show large variation within a population and have been shown to be heritable. The molecular genetic basis of this variation is currently unknown. Progeny from a field trial of a three-generation mapping pedigree of Eucalyptus nitens were assayed for terpenes and FPCs. Quantitative trait loci (QTL) analyses were conducted using a map constructed from 296 markers to locate regions of the genome influencing foliar concentrations of these plant secondary compounds. A large number of significant QTL for 14 traits were located across nine linkage groups, with significant clustering of QTL on linkage groups 7, 8 and 9. As expected, QTL for biosynthetically related compounds commonly colocated, but QTL for unrelated monterpenes and FPCs also mapped closely together. Colocation of these QTL with mapped candidate genes from the various biosynthetic pathways, and subsequent use of these genes in association mapping, will assist in determining the causes of variation in plant secondary metabolites in eucalypts.     

Population divergence in the ontogenetic trajectories of foliar terpenes of a Eucalyptus species

Background and Aims The development of plant secondary metabolites during early life stages can have significant ecological and evolutionary implications for plant–herbivore interactions. Foliar terpenes influence a broad range of ecological interactions, including plant defence, and their expression may be influenced by ontogenetic and genetic factors. This study investigates the role of these factors in the expression of foliar terpene compounds in Eucalyptus globulus seedlings.Methods Seedlings were sourced from ten families each from three genetically distinct populations, representing relatively high and low chemical resistance to mammalian herbivory. Cotyledon-stage seedlings and consecutive leaf pairs of true leaves were harvested separately across an 8-month period, and analysed for eight monoterpene compounds and six sesquiterpene compounds.Key Results Foliar terpenes showed a series of dynamic changes with ontogenetic trajectories differing between populations and families, as well as between and within the two major terpene classes. Sesquiterpenes changed rapidly through ontogeny and expressed opposing trajectories between compounds, but showed consistency in pattern between populations. Conversely, changed expression in monoterpene trajectories was population- and compound-specific.Conclusions The results suggest that adaptive opportunities exist for changing levels of terpene content through ontogeny, and evolution may exploit the ontogenetic patterns of change in these compounds to create a diverse ontogenetic chemical mosaic with which to defend the plant. It is hypothesized that the observed genetically based patterns in terpene ontogenetic trajectories reflect multiple changes in the regulation of genes throughout different terpene biosynthetic pathways.     

Antiplasmodial phloroglucinol derivatives from Syncarpia glomulifera

Bioassay guided fractionation of a MeOH extract of the stem bark of Syncarpia glomulifera (Myrtaceae) led to the isolation of the two new phloroglucinol derivatives (±)-rhodomyrtosone F (1) and (±)-calliviminone C (2), the three known triterpenes, betulinic acid (3), ursolic acid-3-acetate (4), and ursolic acid (5), and 1-(2,4,6-trihydroxyphenyl)-1-hexanone (6). Compound 1 exhibited strong antiplasmodial activity, while compounds 2–4 were moderately active and 5 and 6 were inactive in this assay. The structures of 1 and 2 were elucidated based on analyses of their mass spectrometric data, 1D and 2D NMR spectra, and comparison with related compounds.     

Anti-inflammatory phloroglucinol derivatives from Hypericum empetrifolium

Phytochemical investigation of Hypericum empetrifolium Willd. (Clusiaceae), a species native to Greece and Turkey has led to the bioassay-guided identification of two acylphloroglucinol derivatives with potent in vitro anti-inflammatory activity. Using NMR spectroscopy and mass spectrometry, the acylphloroglucinol derivatives were characterized as 3-geranyl-1-(2'-methylpropanoyl)phloroglucinol (1) and 3-geranyl-1-(2'-methylbutanoyl)phloroglucinol (2). Hypotheses are proposed regarding the biosynthetic origin of these and similar acylphloroglucinols from related Hypericum species. Compounds 1 and 2 were evaluated for in vitro inhibitory activity against COX-1, COX-2 and 5-LOX catalyzed LTB(4) formation. Compound 1 displayed good activity (IC(50) values: 6.0, 29.9, and 2.2 μM, respectively) in all three assays. Compound 2 showed good activity (IC(50) value: 5.8 μM) against LTB(4) formation and moderate activity (IC(50) value: 26.2 μM) against COX-1.     

Eucalyptus foliar chemistry explains selective feeding by koalas

The koala is the quintessential specialist herbivore, feeding almost exclusively on Eucalyptus foliage. Consequently, the limitations imposed on the koala's diet by plant defences indicate the extent to which evolutionary adaptations allow mammalian herbivores to circumvent such defences. We tested whether a recently discovered group of plant secondary metabolites, the formylated phloroglucinol compounds (FPCs), deters koalas from feeding on some eucalypt foliage. We found that captive koalas ate less foliage in a single night from trees with high FPC concentrations. Individual trees also differ in the types of FPC they possess, but for a given eucalypt species, most FPCs were similarly effective deterrents. Two closely related and sympatric eucalypt species could be clearly separated by the amounts that koalas ate from each; however, this difference could not be explained by total FPC concentrations alone. We suggest, that in this case, the presence of a distinct type of FPC deters koala herbivory on the less palatable species, and may have facilitated the evolutionary divergence of these species. We conclude that plant defences probably play an important role in determining the distribution and abundance of koalas.     

Further phloroglucinol derivatives in the fruits of Mallotus japonicus

Two new rottlerin-like phloroglucinol derivatives were detected from the fruits of Mallotus japonicus and identified as 3-(3,3-dimethylallyl)-5-(3-acetyl-2,4-dihydroxy-5-methyl-6-methoxybenzyl)-phlorobutyrophenone and -phloroisobutyrophenone by spectral studies. 2,6-Dihydroxy-3-methyl-4-methoxyacetophenone was also isolated     

Relationships between growth and foliar nutrient concentrations in Eucalyptus deglupta

Summary Relationships beween nutrient concentrations and the growth of the fast growing tropical eucalypt E. deglupta were examined at two sites in Papua New Guinea. At the Gogol Valley site a predominantly linear relationship was found between growth and foliar N over the range 0.68–2.04 per cent N. At the Kerevat site the relationship between growth and foliar nutrients was less strongly developed (foliar N varied from 1.77 to 3.36 per cent). The combined data could be described by a second degree polynomial. Height=11.05N – 2.03N² – 3.52P – 4.46.This accounted for 72 per cent of the variation in height. Based on this relationship a tentative critical foliar N concentration (corresponding with 90 per cent of maximum height) of 2.1 per cent is proposed. re]19760128     

Tomoeones A-H, cytotoxic phloroglucinol derivatives from Hypericum ascyron

Phloroglucinol derivatives tomoeones A-H (1-8) and three known compounds were isolated from leaves of Hypericum ascyron. Their structures were established based on spectroscopic analyses. They are all acylphloroglucinol derivatives possessing a spiro skeleton with geminal isoprenyl groups and a monoterpene moiety, and they are stereoisomers to each other at C-4 and C-13. They appear to be a class of phloroglucinol derivatives. Cytotoxicities of the isolated phloroglucinol derivatives against human tumor cell lines, including multidrug-resistant (MDR) cancer cell lines, were evaluated. Tomoeone F (6) demonstrated significant cytotoxicity against KB cells with an IC50 value of 6.2 microM. Compound 6 was also cytotoxic against MDR cancer cell lines (KB-C2 and K562/Adr), which was more potent than doxorubicin.     

Spatial distributions of foliar nitrogen and phosphorus in crowns of Eucalyptus grandis

Summary Eucalyptus grandis trees were grown in plantations with and without added fertiliser to examine the effects of plant nutrition on photosynthesis and growth. Leaves were sampled from known locations within canopies of selected trees and leaf N and P concentrations were measured. Contour maps of N and P distributions were then produced for crowns of trees aged between 6 and 16 months. Gas exchange measurements on sample leaves were used to estimate parameters of a model of C₃ photosynthesis as a function of leaf N and P contentrations. Linear relationships were obtained between model parameters and leaf N concentration, but P appeared to be present in excess, since no correlation was found with P contentration. Photosynthetic light response curves were calculated for model leaves with differing N concentrations. The curves show that optimal concentrations of N in leaves depend on mean levels of irradiance during growth.     

Diagnosis of nitrogen deficiency and toxicity of Eucalyptus globulus seedlings by foliar analysis

The relationship between shoot growth and foliar nitrogen (N) in E. globulus seedlings was studied in the glasshouse to determine standard values for N deficiency and toxicity diagnosis. Seedlings were grown for 9 weeks in yellow sand, at 10 rates of N, applied as ammonium sulphate, calcium nitrate or ammonium nitrate. Shoot dry weight (DW) increased linearly with N rate for all forms of N in the deficiency range. Seedlings continued to respond to higher rates of ammonium and ammonium nitrate than to nitrate. Maximum shoot DW for nitrate fed plants and ammonium nitrate fed plants were 51% and 84% respectively of ammonium fed plants. Total N concentration in the youngest fully expanded leaf (YFEL) ranged from 1.0% to 3.3% in deficient and adequate plants. The critical N concentration for deficiency diagnosis (corresponding to 90% maximum yield) in the YFEL, determined from these growth response curves averaged over all N forms, was 2.6% N. For ammonium nitrate fed plants, total N concentration in the YFEL for the severely deficient, deficient, adequate, and toxic ranges were <1.4%, 1.4–2.5%, 2.6–3.5%, > 4.3%. High total N concentrations were associated with growth depression and toxicity symptoms, which differed with N form. For nitrate fed plants, a total N concentration above 3.3% in the YFEL was associated with severe growth depression, and leaf tip necrosis. The adequate concentration range for ammonium nitrate was similar to values found on a field trial with 7 month old E. globulus trees grown on an exforest site.     

Variation in foliar nutrients in Eucalyptus trees in eastern and Western Australia

Abstract Levels of nitrogen, phosphorus and potassium were measured for the foliage of two co-dominant eucalypts at each of two sites, one in eastern Australia and the other in Western Australia. In eastern Australia, foliage was sampled in the canopy and subcanopy for narrow-leaved ironbark Eucalyptus crebra and grey box E. mollucana and in Western Australia, for jarrah E. marginata and marri E. calophylla. The Western Australian trees were also sampled for ‘young’ and ‘old’ leaves. Both eucalypts in eastern Australia had greater nitrogen and phosphorus levels, but lower potassium, than E. marginata or E. calophylla. Eucalyptus calophylla foliage had greater levels of all three nutrients than E. marginata foliage as did E. crebra relative to E. mollucana. At both sites, foliar nutrient levels were greater in the canopy than subcanopy foliage, and, at least in Western Australia, the younger leaves had greater nutrient levels than the older leaves. The observed differences in foliar nutrient levels are consistent with observed trends in the abundance and diversity of foliage arthropods and the use of the trees as foraging substrates by birds.     

The molecular basis of quantitative variation in foliar secondary metabolites in Eucalyptus globulus

Eucalyptus is characterized by high foliar concentrations of plant secondary metabolites with marked qualitative and quantitative variation within a single species. Secondary metabolites in eucalypts are important mediators of a diverse community of herbivores. We used a candidate gene approach to investigate genetic associations between 195 single nucleotide polymorphisms (SNPs) from 24 candidate genes and 33 traits related to secondary metabolites in the Tasmanian Blue Gum (Eucalyptus globulus). We discovered 37 significant associations (false discovery rate (FDR) Q < 0.05) across 11 candidate genes and 19 traits. The effects of SNPs on phenotypic variation were within the expected range (0.018 < r(2) < 0.061) for forest trees. Whereas most marker effects were nonadditive, two alleles from two consecutive genes in the methylerythritol phosphate pathway (MEP) showed additive effects. This study successfully links allelic variants to ecologically important phenotypes which can have a large impact on the entire community. It is one of very few studies to identify the genetic variants of a foundation tree that influences ecosystem function.     

Petiolins A-C, phloroglucinol derivatives from Hypericum pseudopetiolatum var. kiusianum

Two new phloroglucinol derivatives possessing chromane skeleton, petiolins A (1) and B (2), and a new phloroglucinol derivative containing a dihydrofuran ring, petiolin C (3), were isolated from aerial parts of Hypericum pseudopetiolatum var. kiusianum. The gross structures of 1-3 were elucidated by spectroscopic data, and the relative stereochemistry of 3 was elucidated by NOESY data. Petiolins A-C (1-3) showed modest cytotoxicity, while petiolin C (3) exhibited antifungal activity.     

Effects of grandinol and related phloroglucinol derivatives on transpiration and stomatal closure

Grandinol, an inhibitor of seed germination and photosynthesis in Eucalyptus sp., inhibits transpiration and stomatal opening. The acylphloroglucinol structure in grandinol seemed to be essential for these activities. Enhancement of activity was achieved by the introduction of a formyl group into the molecule. Therefore, structural requirements for these activities were very similar to that for the inhibition of seed germination and photosynthesis. Other grandinol-related compounds having two electron-withdrawing groups on the phloroglucinol nuclei were also active in these assays.     

The resistance to waterlogging of three Eucalyptus species

Summary Seedlings of Ecalyptus grandis W. Hill ex Maiden were found to be more resistant to waterlogging than those of Eucalyptus robusta Sm. and seedlings of both species were more resistant than those of Eucalyptus saligna Sm. Resistance depended on the stress avoidance mechanism of adventitious root formation. The ethylene-releasing growth substance 2-chloroethylphosphonic acid (ethephon) applied to non-flooded plants was used as a tool to study the role of ethylene in the response of plants to waterlogging. Plant height and total dry weight decreased linearly with plant ethylene content. There was no difference between the species in their response to ethephon.     

Distribution of the ability for citrate utilization amongst Clostridia

44 species of the genus Clostridium were examined with regard to their ability to grow on citrate. In addition to Clostridium sphenoides, a known citrate utilizer, the following species were found to utilize citrate: C. sporosphaeroides, C. symbiosum, C. rectum, C. indolis, C. subterminale and C. sporogenes. The major products formed from citrate were acetate, ethanol and carbon dioxide (not measured). Minor products were butyrate, butanol, acetone and acetoin.The enzyme citrate lyase was detectable in cell extracts of C. sporosphaeroides and C. symbiosum using an optical assay. Evidence for the presence of this enzyme in the other species was obtained in immunological experiments and in experiments with [1,5-¹⁴C]citrate.     

Metabolic responses of Eucalyptus species to different temperature regimes

Species and hybrids of Eucalyptus are the world's most widely planted hardwood trees. They are cultivated across a wide range of latitudes and therefore environmental conditions. In this context, comprehensive metabolomics approaches have been used to assess how different temperature regimes may affect the metabolism of three species of Eucalyptus, E. dunnii, E. grandis and E. pellita.Young plants were grown for 53 d in the greenhouse and then transferred to growth chambers at 10°C, 20°C or30°C for another 7 d. In all three species the leaf chlorophyll content was positively correlated to temperature, and in E.pellita the highest temperature also resulted in a significant increase in stem biomass. Comprehensive metabolomics was performed using untargeted gas chromatography mass spectrometry(GC-MS) and liquid chromatography(LC)-MS.This approach enabled the comparison of the relativeabundance of 88 polar primary metabolites from GC-MS and625 semi-polar secondary metabolites from LC-MS. Using principal components analysis, a major effect of temperature was observed in each species which was larger than that resulting from the genetic background. Compounds mostly affected by temperature treatment were subsequently selected using partial least squares discriminant analysis and were further identified. These putative annotations indicated that soluble sugars and several polyphenols, including tannins, triterpenes and alkaloids were mostly influenced.