Phytochemistry (v.85, #C)

Medicinal property, phytochemistry and pharmacology of several Jatropha species (Euphorbiaceae): A review by Carla W. Sabandar; Norizan Ahmat; Faridahanim Mohd Jaafar; I. Sahidin (7-29).
Species from Jatropha are used in medicinal folklore and known as a purgative. Crude extracts and compounds such as cyclic peptide alkaloids, diterpenes with various skeletons and other compounds from this genus have been reported for cytotoxicity, tumor-promoting, antimicrobial, antiprotozoal, anticoagulant, immunomodulating, anti-inflammatory, antioxidant, protoscolicidal, insecticidal, molluscicidal, inhibition AChE and toxicity activities.Display Omitted► The medicinal property of several Jatropha species are reviewed. ► The phytochemistry of mostly alkaloid cyclic peptides and diterpenoids is presented. ► The biological activity of crude extracts and pure compounds is succinctly discussed. ► The relationship of medicinal property, biological activity and phytochemistry is also described.The genus Jatropha (Euphorbiaceae) comprises of about 170 species of woody trees, shrubs, subshrubs or herbs in the seasonally dry tropics of the Old and the New World. They are used in medicinal folklore to cure various diseases of 80% of the human population in Africa, Asia and Latin America. Species from this genus have been popular to cure stomachache, toothache, swelling, inflammation, leprosy, dysentery, dyscrasia, vertigo, anemia, diabetis, as well as to treat HIV and tumor, opthalmia, ringworm, ulcers, malaria, skin diseases, bronchitis, asthma and as an aphrodisiac. They are also employed as ornamental plants and energy crops. Cyclic peptides alkaloids, diterpenes and miscellaneous compounds have been reported from this genus. Extracts and pure compounds of plants from this genus are reported for cytotoxicity, tumor-promoting, antimicrobial, antiprotozoal, anticoagulant, immunomodulating, anti-inflammatory, antioxidant, protoscolicidal, insecticidal, molluscicidal, inhibition AChE and toxicity activities.
Keywords: Euphorbiaceae; Jatropha; Medicinal properties; Phytochemical; Pharmacology;

Characterization of Arabidopsis serine:glyoxylate aminotransferase, AGT1, as an asparagine aminotransferase by Qianyi Zhang; Jamie Lee; Sudhakar Pandurangan; Matthew Clarke; Agnieszka Pajak; Frédéric Marsolais (30-35).
Recombinant serine:glyoxylate aminotransferase, AGT1, from Arabidopsis, preferred asparagine over serine or alanine as substrate. AGT1 transcripts were induced by asparagine in roots suggesting a function in asparagine metabolism.Display Omitted► Recombinant AGT1 prefers asparagine over serine or alanine as substrate. ► AGT1 transcripts were induced by asparagine in roots. ► The results suggest that AGT1 is involved in asparagine metabolism.Asparagine (Asn) is a major form of nitrogen transported to sink tissues. Results from a previous study have shown that an Arabidopsis mutant lacking asparaginase activity develops relatively normally, highlighting a possible compensation by other types of asparagine metabolic enzymes. Prior studies with barley and tobacco mutants have associated Asn aminotransferase activity with the photorespiratory enzyme, serine (Ser):glyoxylate aminotransferase. This enzyme is encoded by AGT1 in Arabidopsis thaliana. Recombinant N-terminal His-tagged AGT1 purified from Escherichia coli was characterized with Ser, alanine (Ala) and Asn as amino acid donors and glyoxylate, pyruvate and hydroxypyruvate as organic acid acceptors. The V max of AGT1 with Asn was higher than with Ser or Ala by ca. 5- to 20-fold. As a result, the catalytic efficiency (V max/K m) was slightly higher with Asn than with the two other amino acids. In the roots of 10-day-old seedlings treated for 2 h with 20 mM Asn, the AGT1 transcript levels were raised by 2-fold. During this treatment, the concentration of Asn in root was raised by ca. 5-fold. These results suggest that AGT1 is involved in Asn metabolism in Arabidopsis.
Keywords: Arabidopsis thaliana; Cruciferae; Serine:glyoxylate aminotransferase; AGT1; Asparagine; Substrate preference; Transcriptional regulation;

Characterization of the plastidial geraniol synthase from Madagascar periwinkle which initiates the monoterpenoid branch of the alkaloid pathway in internal phloem associated parenchyma by Andrew J. Simkin; Karel Miettinen; Patricia Claudel; Vincent Burlat; Grégory Guirimand; Vincent Courdavault; Nicolas Papon; Sophie Meyer; Stéphanie Godet; Benoit St-Pierre; Nathalie Giglioli-Guivarc’h; Marc J.C. Fischer; Johan Memelink; Marc Clastre (36-43).
The Catharanthus roseus geraniol synthase is localized in plastids and transcripts are detected in internal phloem associated parenchyma (IPAP) cells in young developing leaves.Display Omitted► The CrGES cDNA from Catharanthus roseus was cloned and characterized. ► CrGES displays geraniol synthase activity. ► The CrGES gene is up-regulated by methyl jasmonate. ► The protein is localized in plastid stroma and stromules. ► CrGES transcripts are expressed in internal phloem associated parenchyma.Madagascar periwinkle (Catharanthus roseus [L.] G. Don, Apocynaceae) produces monoterpene indole alkaloids (MIAs), secondary metabolites of high interest due to their therapeutic value. A key step in the biosynthesis is the generation of geraniol from geranyl diphosphate (GPP) in the monoterpenoid branch of the MIA pathway. Here we report on the cloning and functional characterization of C. roseus geraniol synthase (CrGES). The full-length CrGES was over-expressed in Escherichia coli and the purified recombinant protein catalyzed the conversion of GPP into geraniol with a K m value of 58.5 μM for GPP. In vivo CrGES activity was evaluated by heterologous expression in a Saccharomyces cerevisiae strain mutated in the farnesyl diphosphate synthase gene. Analysis of culture extracts by gas chromatography-mass spectrometry confirmed the excretion of geraniol into the growth medium. Transient transformation of C. roseus cells with a Yellow Fluorescent Protein-fusion construct revealed that CrGES is localized in plastid stroma and stromules. In aerial plant organs, RNA in situ hybridization showed specific labeling of CrGES transcripts in the internal phloem associated parenchyma as observed for other characterized genes involved in the early steps of MIA biosynthesis. Finally, when cultures of Catharanthus cells were treated with the alkaloid-inducing hormone methyl jasmonate, an increase in CrGES transcript levels was observed. This observation coupled with the tissue-specific expression and the subcellular compartmentalization support the idea that CrGES initiates the monoterpenoid branch of the MIA biosynthetic pathway.
Keywords: Catharanthus roseus; Geraniol synthase; Alkaloid; Internal phloem associated parenchyma;

Tomato transformed with ZAT12 gene for drought stress tolerance were obtained. BcZAT12 transformed lines ZT1 and ZT5 were drought tolerant having potential in tomato yield under drought conditions.Display Omitted► Focuses on transformation of tomato with ZAT12 gene that plays a role in abiotic stress tolerance. ► Six successful transformants (ZT1–ZT6) of tomato cv. H-86 harbouring ZAT12 gene resulted. ► Lowered EL and improved RWC in transformed tomato plants were significant. ► Decreased ROS with elevated CAT activity were noted in transformed tomato plants. ► Tolerant tomato lines ZT1 and ZT5 have potential in improved tomato yield under drought stress.Efficient genetic transformation of cotyledonary explants of tomato (Solanum lycopersicum, cv. H-86, Kashi vishesh) was obtained. Disarmed Agrobacterium tumifaciens strain GV 3101 was used in conjugation with binary vector pBinAR containing a construct consisting of the coding sequence of the BcZAT12 gene under the regulatory control of the stress inducible Bclea1a promoter. ZAT12 encodes a C2H2 zinc finger protein which confers multiple abiotic stress tolerance to plants. Integration of ZAT12 gene into nuclear genome of individual kanamycin resistant transformed To tomato lines was confirmed by Southern blot hybridization with segregation analysis of T1 plants showing Mendelian inheritance of the transgene. Expression of ZAT12 in drought-stressed transformed tomato lines was verified in T2 generation plants using RT-PCR. Of the six transformed tomato lines (ZT1–ZT6) the transformants ZT1 and ZT5 showed maximum expression of BcZAT12 gene transcripts when exposed to 7 days drought stress. Analysis of relative water content (RWC), electrolyte leakage (EL), chlorophyll colour index (CCI), H2O2 level and catalase activity suggested that tomato BcZAT12 transformants ZT1 and ZT5 have significantly increased levels of drought tolerance. These results suggest that BcZAT12 transformed tomato cv. H-86 has real potential for molecular breeding programs aimed at augmenting yield of tomato in regions affected with drought stress.
Keywords: Solanum lycopersicum; Tomato; Drought; Transformation; ZAT12;

Cell wall compositional modifications of Miscanthus ecotypes in response to cold acclimation by Jean-Marc Domon; Laëtitia Baldwin; Sébastien Acket; Elodie Caudeville; Stéphanie Arnoult; Hélène Zub; Françoise Gillet; Isabelle Lejeune-Hénaut; Maryse Brancourt-Hulmel; Jérôme Pelloux; Catherine Rayon (51-61).
The cell wall composition of three Miscanthus clones with contrasted frost tolerance has been determined during cold acclimation. The plants were harvested at the juvenile stage and cell wall composition of each clone during cold acclimation was compared to the one of non cold-treated plants.Display Omitted► PAL activity was increased in the frost-tolerant Miscanthus clones under cold stress. ► CAD activity under cold stress was largest in the frost-tolerant clones. ► During cold acclimation, an increase in β-glucan was observed in all genotypes. ► GAX content changed in the frost tolerant clones under the ambient conditions. Miscanthus, a potential energy crop grass, can be damaged by late frost when shoots emerge too early in the spring and during the first winter after planting. The effects of cold acclimation on cell wall composition were investigated in a frost-sensitive clone of Miscanthus x giganteus compared to frost-tolerant clone, Miscanthus sinensis August Feder, and an intermediate frost-tolerant clone, M. sinensis Goliath. Cellulose and lignin contents were higher in M. x giganteus than in the M. sinensis genotypes. In ambient temperature controls, each clone displayed different glucuronoarabinoxylan (GAX) contents and degree of arabinose substitution on the xylan backbone. During cold acclimation, an increase in (1 → 3),(1 → 4)-β-d-glucan content was observed in all genotypes. Uronic acid level increased in the frost sensitive genotype but decreased in the frost tolerant genotypes in response to cold. In all clones, major changes in cell wall composition were observed with modifications in phenylalanine ammonia-lyase (PAL) and cinnamyl alcohol dehydrogenase (CAD) activities in both non- and cold-acclimated experiments. A large increase in CAD activity under cold stress was displayed in each clone, but it was largest in the frost-tolerant clone, M. sinensis August Feder. The marked increase in PAL activity observed in the frost-tolerant clones under cold acclimation, suggests a reorientation of the products towards the phenylpropanoid pathway or aromatic synthesis. How changes in cell wall physical properties can impact frost tolerance is discussed.
Keywords: Miscanthus; Poales; Cell wall; Cold stress; Glucuronoarabinoxylan; (1 → 3),(1 → 4)-β-d-glucan; Cellulose; Uronic acid; Lignin; PAL; CAD;

The anthocyanin compositions of Schizanthus grahamii and Schizanthus hookeri flowers are qualitatively and quantitatively different, thus contributing to the different pollination syndromes that they show.Display Omitted► Total anthocyanin content is much higher in Schizanthus grahamii than in Schizanthus hookeri. ► Delphinidin 3-O-glucoside is the major anthocyanin in S. grahamii red flowers (hummingbird). ► Petanin is the major anthocyanin in S. hookeri bluish-pink flowers (bee). ► Common pigments in both species can be indicative of a common ancestor. ► Synthesis of simpler red pigments by S. grahamii is related to specialisation.Three different pollination syndromes (bee, hummingbird and moth-pollination syndromes) have been described for the different species of the genus Schizanthus Ruiz & Pav. (Solanaceae). Two closely related species from a phylogenetic point of view, Schizanthus grahamii and Schizanthus hookeri, show hummingbird and bee-pollination syndromes, respectively. One of the traits used to assign these pollination syndromes is the colour of the petals, which is red in S. grahamii and bluish-pink in S. hookeri. The objective of this work was to establish the differences in the anthocyanin composition of these two Schizanthus species that contribute to the different pollination syndrome. Identification of the pigments was carried out from the chromatographic and spectral data supplied by the HPLC-DAD-MS n analyses of the samples. Alkaline and acid hydrolyses were also performed in the extracts and in some isolated compounds in order to confirm the identities. First difference between these two species was found in the total anthocyanin content, which was about 13-fold higher in S. grahamii than in S. hookeri. Furthermore, the major peak was also different in both cases, thus explaining quantitative and qualitative colour differences between species. Delphinidin 3-O-rutinoside represented ca. 72% of the total area in S. grahamii and petanin (Petunidin 3-O-p-coumaroylrutinoside-5-O-glucoside) accounted for almost 62% of the total area in S. hookeri. The presence of the p-coumaroylrutinosyl moiety in the petanin makes the intramolecular copigmentation possible, thus conferring the bluish-pink hue to the petals of S. hookeri. Delphinidin 3-O-rutinoside is in turn responsible for the red colour exhibited by the petals of S. grahamii and along with other floral traits, responsible for the attraction of hummingbirds to the plant. Pigments structurally related to petanin, which have been often detected in other genus of Solanaceae, were detected in both species and in similar contents, supporting results from previous studies that pointed to a bee-pollinated common ancestor for both species that further specialised acquiring traits attractive to hummingbirds and among them, red colour of the petals which is mainly supplied by delphinidin 3-O-rutinoside.
Keywords: Schizanthus grahamii; Schizanthus hookeri; Solanaceae; HPLC-DAD-MS n ; Anthocyanins; Hummingbird and bee-pollination syndromes;

Guatemalan potato moth Tecia solanivora distinguish odour profiles from qualitatively different potatoes Solanum tuberosum L. by Miriam Frida Karlsson; Göran Birgersson; Peter Witzgall; Jonas Duus Stevens Lekfeldt; P.A. Nimal Punyasiri; Marie Bengtsson (72-81).
Volatile compound mediated oviposition and larval survival of Tecia solanivora on healthy vs. stressed tubers. This supports the preference performance hypothesis for insect herbivores.Display Omitted► Oviposition preference supports insect herbivores preference performance hypothesis. ► Potato metabolites affect larval survival. ► Oviposition avoidance by Tecia solanivora towards stressed potato tubers. ► Larval survival is negatively correlated with α-solanine and α-chaconine. ► Qualitatively different potatoes emit different volatile chemical composition.Guatemalan potato moth, Tecia solanivora, lay eggs in the soil nearby potato Solanum spp. and larvae feed on the tubers. We investigated the oviposition behaviour of T. solanivora females and the survival of larval offspring on healthy vs. stressed, i.e. light exposed and/or damaged potato tubers. In choice tests, females laid significantly more eggs in response to potato odour of healthy tubers and female oviposition preference correlated with higher larval survival. Survival of larvae was negatively correlated with the tuber content of the steroid glycoalkaloids α-solanine and α-chaconine: healthy potatoes contained lower amounts than stressed tubers, ranging from 25 to 500 μg g−1 and from 30 to 600 μg g−1, respectively. Analysis of volatile compounds emitted by potato tubers revealed that stressed tubers could clearly be distinguished from healthy tubers by the composition of their volatile profiles. Compounds that contributed to this difference were e.g. decanal, nonanal, isopropyl myristate, phenylacetaldehyde, benzothiazole, heptadecane, octadecane, myristicin, E,E-α-farnesene and verbenone. Oviposition assays, when female moths were not in contact with the tubers, clearly demonstrated that volatiles guide the females to lay fewer eggs on stressed tubers that are of inferior quality for the larvae. We propose that volatiles, such as sesquiterpenes and aldehydes, mediate oviposition behaviour and are correlated with biosynthetically related, non-volatile compounds, such as steroidal glycoalkaloids, which influence larval survival. We conclude that the oviposition response and larval survival of T. solanivora on healthy vs. stressed tubers supports the preference performance hypothesis for insect herbivores.
Keywords: Solanum tuberosum; Solanum; Gelechiidae; Preference performance; Oviposition; SGAs; Phenols; α-Solanine; α-Chaconine;

Variation in the chemical composition of cone volatiles within the African cycad genus Encephalartos by Terence N. Suinyuy; John S. Donaldson; Steven D. Johnson (82-91).
Chemical diversity of cone volatiles in some Encephalartos species.Display Omitted► Diverse volatiles characterise Encephalartos odours with similarities among species. ► There is interspecific variation in odours of 19 Encephalartos taxa. ► The compounds (3E)-1,3-octadiene and (3E,5Z)-1,3,5-octatriene occur in most species. ► Variations in odour profile of Encephalartos taxa suggest adaptive response. ► Geography, phylogeny nor pollination can fully explain the interspecific variations.Volatiles play a key role in attraction of pollinators to cycad cones, but the extent to which volatile chemistry varies among cycad species is still poorly documented. Volatile composition of male and female cones of nineteen African cycad species (Encephalartos; Zamiaceae) was analysed using headspace technique and gas chromatography–mass spectrometry (GC–MS). A total of 152 compounds were identified among the species included in this study, the most common of which were monoterpenes, nitrogen-containing compounds and unsaturated hydrocarbons. Male and female cones emitted similar volatile compounds which varied in relative amounts with two unsaturated hydrocarbons (3E)-1,3-octadiene and (3E,5Z)-1,3,5-octatriene present in the volatile profile of most species. In a multivariate analysis of volatile profiles using non-metric multidimensional scaling (NMDS), a number of species clusters were identified according to shared emission of unsaturated hydrocarbons, pyrazines, benzenoids, aldehydes, alkanes and terpenoids. In comparison, terpenoids are common in Zamia and dominant in Macrozamia species (both in the family Zamiaceae) while benzenoids, esters, and alcohols are dominant in Cycas (Cycadaceae) and in Stangeria (Stangeriaceae). It is likely that volatile variation among Encephalartos species reflects both phylogeny and adaptations to specific beetle pollinators.
Keywords: Zamiaceae; Gas chromatography–mass spectrometry; Phylogenetic signal; Pollination system; Vegetation type; Interspecific variation;

Pavettamine, the toxic agent of gousiekte, is produced in the highest concentration in the youngest leaves. Most pavettamine was found to occur in a conjugated form. The bacterial endosymbiont does not produce pavettamine in in vitro culture.Display Omitted► Fadogia homblei, a plant known to cause gousiekte in ruminants contains pavettamine in their leaves. ► Most pavettamine was found to be conjugated in Fadogia homblei. ► The endosymbiont of Fadogia homblei revealed no in vitro production of pavettamine. ► We report a mass spectrometry method to determine and quantify pavettamine.Plant poisoning of livestock is responsible for considerable economic losses in southern Africa. Six plant species of the Rubiaceae family are known to cause gousiekte, a cardiac syndrome of ruminants induced by ingestion of the toxic compound pavettamine. Progress in understanding the etiology of this disease is largely hampered by the variable toxicity of the plants and the absence of a quantification method for pavettamine. The pavettamine concentration in leaf samples of Fadogia homblei, a known gousiekte causing plant, was analyzed by mass-spectrometry. In the most apical leaf pair, the highest concentration of pavettamine was detected. Distal leaves contained progressively less pavettamine. Besides a significant amount of free pavettamine, most pavettamine was found to occur in a conjugated form. To which molecules the pavettamine is conjugated remains unknown as is the function of conjugated pavettamine in the development of gousiekte. All know gousiekte-causing plants contain symbiotic bacteria in their leaves; it was hypothesized that these bacteria might be involved in the production of pavettamine. However, analysis of in vitro cultures of the F. homblei endosymbiont revealed no production of pavettamine. Pavettamine is therefore not produced by the bacteria alone. It is either the product of the interaction with the plant or solely produced by the host.
Keywords: Fadogia homblei; Rubiaceae; Burkholderia; Endosymbiont; Mass-spectrometry; Pavettamine; Gousiekte;

Chemical taxonomy of red-flowered wild Camellia species based on floral anthocyanins by Jian-Bin Li; Fumio Hashimoto; Keiichi Shimizu; Yusuke Sakata (99-106).
The evolution of anthocyanin biosynthesis of the red-flowered Camellia species occurred from 3,5-di-O-glucosylation and 3-O-sambubiosylation (primitive trait) to 3-O-glycosylation (advanced trait) and from p-coumaroylation to non-esterification. Our data support the notion that the Xinan district in southwestern China is the site/center of origin and that the Huanan district might be considered as the secondary site/center of origin of the red-flowered Camellia species.Display Omitted► The Xinan district in China is the presumed site/center of origin of red-flowered Camellia species. ► The Huanan district might be considered as the secondary site/center of origin. ► Camellia saluenensis and Camellia reticulata are the most primitive species. ► The process of biosynthesis evolved from: 3,5-di-O-glucosylation to 3-O-glycosylation; 3-O-sambubiosylation to 3-O-glycosylationl; and p-coumaroylation to non-esterification.This study uses anthocyanins in the red flowers of section Camellia as taxonomic markers to investigate the phenetic relationships among 33 wild species from China, Taiwan, and Japan. The 25 anthocyanins from section Camellia produced 38 pigment patterns that serve as phenetic markers. Principal Component Analysis (PCA) indicated that the attachment of one or two glucoses to the cyanidin-core structure at the 3- or the 3- and 5-positions, respectively, was the most influential pattern against the first factor, Z 1. In addition, two alternative pigment patterns, acylated or non-acylated, and the structural isomerism (cis- or trans-) of the p-coumaroyl group were relatively significant patterns. Ward’s minimum-variance cluster analysis (WMVCA) produced a dendrogram that consisted of two sub-clusters. One sub-cluster (A) was constructed by species that have mainly two types of anthocyanins: 3,5-di-O-β-glucosides (Camellia saluenensis) and sambubioside of cyanidin (Camellia reticulata). The other sub-cluster (B) was made up of the 3-O-β-glucosides of cyanidin (Camellia japonica) and delphinidin (Camellia hongkongensis), with a higher proportion of the 3-O-β-galactosides (Camellia mairei and Camellia boreali-yunnanica). The former group showed a higher proportion of acylation, over 63%, but with the exception of Camellia azalea. The latter group showed less than 52% acylation, but with the exception of C. hongkongensis and C. boreali-yunnanica. PCA and WMVCA indicated that the greater the amount of di-O-glycosides and acylation, the more primitive anthocyanin traits the species possess. Based on these results, in conjunction with geographical and literary information, the data suggest that the Xinan district is the site/center of origin for the red-flowered Camellia species of which both C. saluenensis and C. reticulata have the most primitive anthocyanin traits.
Keywords: Camellia spp.; Theaceae; Chemical taxonomy; Floral pigment; Lathyloside; Principal component analysis; Phylogeny; Anthocyanin; Sambubioside;

Glycosylceramides from marine green microalga Tetraselmis sp. by Atsushi Arakaki; Daisuke Iwama; Yue Liang; Nagisa Murakami; Masaharu Ishikura; Tsuyoshi Tanaka; Tadashi Matsunaga (107-114).
Twelve representative strains of marine eukaryotic and prokaryotic microalgae were screened for glycosylceramide producers using TLC and ESI-MS/MS analyses. Glycosylceramides were only isolated from the marine microalgae Tetraselmis sp. Their structures were determined by a combination of one- and two-dimensional NMR techniques, and mass spectrometry.Display Omitted► A screening method for glycosylceramides was established using TLC and ESI-MS/MS. ► Glycosylceramides were isolated from marine microalgae Tetraselmis sp. strain NKG400013. ► The two lipids consist of a botanical sphingoid base and fungal fatty acyl chains. ► This is the first indication of the presence of glycosylceramide in Prasinophyceae.Glycosylceramides are ubiquitous and important components of the plasma membrane in most eukaryotic cells and a few bacteria. They play significant roles in a variety of cellular functions. Their molecular structures are well recognized in animals, higher plants, and fungi, but are poorly characterized in lower plants. In this study, a high glycosylceramide-producing microalgal strain Tetraselmis sp. NKG 400013 was found. TLC and MS analyses established the presence of glycosylceramides, GT1 and GT2, in this strain. Their chemical structures were determined by NMR spectroscopy and GC/MS, and were identified as glycosylceramides consisting of the typical botanical sphingoid base ([4E, 8E]-sphinga-4, 8-dienine) and 2-hydroxy-Δ3-unsaturated fatty acyl chains, respectively. To our knowledge, the occurrence of glycosylceramides in microalga of the class Prasinophyceae was previously unknown.
Keywords: Tetraselmis sp.; Prasinophyceae; Glucosylceramides; Sphingolipids; Δ3-unsaturatation;

Endosymbiotic heterocystous cyanobacteria synthesize different heterocyst glycolipids than free-living heterocystous cyanobacteria by Stefan Schouten; Tracy A. Villareal; Ellen C. Hopmans; Anchelique Mets; Kathleen M. Swanson; Jaap S. Sinninghe Damsté (115-121).
We report the glycolipid composition of several marine free-living heterocystous cyanobacteria as well as the heterocystous endosymbiont Richelia intracellularis found in the diatoms Hemiaulus hauckii and Hemiaulus membranaceus.Display Omitted► Glycolipids rarely investigated for marine heterocystous cyanobacteria. ► Glycolipids of selected marine cyanobacteria were similar to that of freshwater representatives. ► Heterocystous endosymbionts found in diatoms synthesize different glycolipids. ► These glycolipids contain C5 sugars and longer chain lengths for the aglycone moiety.The heterocysts of limnetic nitrogen-fixing filamentous cyanobacteria contain unique glycolipids in their cell wall that create the distinctive gas impermeability of the heterocyst cell wall as well as serve as biomarker lipids for these microbes. It has been assumed that marine free-living and endosymbiotic cyanobacteria synthesize the same glycolipids although they have not been investigated in any detail. Here we report the glycolipid composition of several marine free-living heterocystous cyanobacteria as well as the heterocystous endosymbiont Richelia intracellularis found in the biogeochemically important diatoms Hemiaulus hauckii and Hemiaulus membranaceus. In the marine cyanobacteria Nostoc muscorum and Calothrix sp., we detected the same glycolipids as found in freshwater representatives of these genera. However, we did not detect these glycolipids in the HemiaulusRichelia association. Instead, we identified glycolipids which comprised a C5 sugar, ribose, rather than the C6 sugars normally encountered in glycolipids of free-living cyanobacteria. In addition, the glycolipids had slightly longer chain lengths (C30 and C32 versus C26 and C28) in the aglycone moiety. The different glycolipid composition of the marine endosymbotic heterocystous cyanobacteria compared to their free-living counterparts may be an adaptation to the high intracellular O2 concentrations within their host. These glycolipids may provide unique tracers for the presence of these microbes in marine environments and permit exploration of the evolutionary origins of these symbioses.
Keywords: Heterocystous cyanobacteria; Hemiaulus; Richelia intracellularis; Glycolipids;

Sesquiterpenoids from Curcuma wenyujin with anti-influenza viral activities by Jian-Yong Dong; Xi-Yan Ma; Xiao-Qin Cai; Peng-Cheng Yan; Lei Yue; Chen Lin; Wei-Wei Shao (122-128).
Five sesquiterpenoids, together with seven known analogs, were isolated from the rhizomes of Curcuma wenyujin. Several compounds showed significant in vitro antiviral activity against the influenza virus A.Display Omitted► Twelve sesquiterpenoids (five hitherto unknown) were isolated from the rhizomes of Curcuma wenyujin. ► Their structures were elucidated using NMR spectroscopy and X-ray crystallography. ► Several compounds showed significant in vitro antiviral activity against influenza virus A.Five sesquiterpenoids, 1α,8α-epidioxy-4α-hydroxy- 5αH-guai-7(11),9-dien- 12,8-olide. (1), 8,9-seco-4β-hydroxy-1α,5βH-7(11)-guaen-8,10-olide (2), 8α-hydroxy-1α, 4β,7βH-guai-10(15)-en- 5β,8β-endoxide(3), 7β,8α-dihydroxy-1α,4αH-guai-10(15)-en-5β,8β-endoxide(4) and 7-hydroxy-5(10),6,8-cadinatriene-4-one(5), together with seven known analogs were isolated from the rhizomes of Curcuma wenyujin. Their structures and relative configurations were determined on the basis of spectroscopic methods including 2D NMR techniques, and the structures of 1 and 2 were confirmed by single-crystal X-ray diffraction experiment. Compounds 1–10 and 12 showed significant in vitro antiviral activity against the influenza virus A with IC50 values ranged from 6.80 to 39.97 μM, and SI values ranged from 6.35 to 37.25.
Keywords: Curcuma wenyujin; Zingiberaceae; Sesquiterpenoid; Guaiane; Seco-guaiane; Cadinane; Germacrane; Eudesmane; Anti-influenza viral A activity;

Isolation and characterization of sesquiterpenes from Arecophila saccharicola YMJ96022401 with NO production inhibitory activity by Lin-Wen Lee; Guei-Jane Wang; Mei-Hsiang Lin; Yu-Min Ju; Yen-Wen Lin; Fang-Yu Chen; Tzong-Huei Lee (129-136).
The monocot-associated fungus, Arecophila saccharicola, was fermented in a malt extract medium. From the ethyl acetate extract of the fermented broth of this fungus, drimane-type sesquiterpenes 16 and an eremophilane-type sesquiterpene 7 were isolated together with two known compounds 8 and 9. Their structures were established by spectroscopic analyses. The inhibitory effects of 19 on the nitric oxide (NO) production in lipopolysaccharide-activated murine macrophage RAW264.7 cells were also evaluated.Display Omitted► The monocot-associated fungus Arecophila saccharicola was cultured. ► Nine chemical entities were isolated from the fermented broth and their structures were elucidated. ► Their inhibitory activities on NO production in LPS-activated RAW264.7 cells were evaluated.Sesquiterpenes, arecoic acids A–F and arecolactone, were isolated from the ethyl acetate extracts of the fermented broth of Arecophila saccharicola YMJ96022401 along with two known analogues 1,7α,10α-trihydroxyeremophil-11(13)-en-12,8-olide and 1,10α,13-trihydroxyeremophil-7(11)-en-12,8-olide. Their structures were elucidated on the basis of spectroscopic data analyses. The inhibitory effects of all of these compounds on nitric oxide (NO) production in lipopolysaccharide (LPS, 200 μg/mL)-activated murine macrophage RAW264.7 cells were also evaluated. Among these compounds, 1,7α,10α-trihydroxyeremophil-11(13)-en-12,8-olide significantly inhibited NO production without any cytotoxicity, and its average maximum inhibition (E max) at 100 μM and median inhibitory concentration (IC50) were 85.7% ± 0.8% and 16.5 ± 1.0 μM, respectively. Arecolactone was the most potent, with the E max at 12.5 μM and IC50 being 94.7% ± 0.8% and 1.32 ± 0.1 μM, respectively, but displayed cytotoxicity at considerable higher concentrations than 25 μM. Analyses of Western blotting indicated that arecolactone (0.8–12.5 μM) inhibited induction of inducible NO synthase (iNOS) by LPS, which involved suppression of NF-κB activation and the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun NH(2)-terminal kinase (JNK) and p38 mitogen-activated protein kinases (MAPKs) in activated RAW 264.7 cells. In addition, arecolactone concentration-dependently prevented the vascular hyporeactivity to phenylephrine induced by LPS (300 ng/mL) through iNOS pathway in isolated rat thoracic aortic rings. These results indicated that both of these naturally occurring iNOS inhibitors may provide a rationale for the potential anti-inflammatory effect of A. saccharicola YMJ96022401.
Keywords: Arecophila saccharicola; Ascomycete; Nitric oxide; Sesquiterpene; Anti-inflammation;

Gastroprotective effect and structure of a rhamnogalacturonan from Acmella oleracea by Adamara M. Nascimento; Lauro M. de Souza; Cristiane H. Baggio; Maria Fernanda de P. Werner; Daniele Maria-Ferreira; Luisa M. da Silva; Guilherme L. Sassaki; Philip A.J. Gorin; Marcello Iacomini; Thales R. Cipriani (137-142).
A methyl esterified rhamnogalacturonan was isolated from the plant Acmella oleracea. Its structure was elucidated by methylation analysis and NMR spectroscopy. It inhibited ethanol-induced gastric ulcers in rats with an ED50 of 1.5 mg/kg.Display Omitted► A methyl esterified rhamnogalacturonan was isolated and characterized from Acmella oleracea. ► The rhamnogalacturonan is partially substituted by side-chains of type II arabinogalactans. ► The polysaccharide reduced ethanol-induced gastric lesions in rats with an ED50 of 1.5 mg/kg. ► Rhamnogalacturonan of A. oleracea may be a gastroprotective agent.The plant Acmella oleracea (L.) R.K.Jansen (Asteraceae), locally known as jambu, is widely used in Legal Amazon in local dishes and in folk medicine. A polysaccharide (SC) was isolated from this plant, following aqueous extraction, which contained uronic acid, galactose, arabinose, rhamnose, and glucose in a 15:2:1:1:0.5 molar ratio and had a M w 226,000 g/mol. Methylation analysis and NMR spectroscopy indicated that SC is a rhamnogalacturonan composed of a long chain of →4)-6-OMe-α-d-GalpA-(1→, interspersed with some α-l-Rhap residues, partly substituted by side-chains of type II arabinogalactans. SC significantly inhibited ethanol-induced gastric ulcers in rats with an ED50 of 1.5 mg/kg, indicating that SC acts as gastroprotective agent.
Keywords: Acmella oleracea; Asteraceae; Rhamnogalacturonan; Anti-ulcer activity;

Bisabololoxide derivatives from Artemisia persica, and determination of their absolute configurations by ECD by Fahimeh Moradi-Afrapoli; Samad Nejad Ebrahimi; Martin Smiesko; Melanie Raith; Stefanie Zimmermann; Farsad Nadjafi; Reto Brun; Matthias Hamburger (143-152).
Absolute configurations of bisabololoxides 15 were achieved by comparison of experimental ECD spectra with simulated data for possible stereoisomers, using time dependent density function theory.Display Omitted► A series of bisabololoxide diesters were identified by HPLC-based activity profiling. ► The compounds possess in vitro antiplasmodial activity. ► Absolute configuration was established by ECD.Five antiplasmodial bisabololoxide sesquiterpene diesters were isolated from an EtOAc extract of the aerial parts of Artemisia persica following an HPLC-time-based activity profiling of the extract. Structure elucidation was achieved by 1D and 2D NMR experiments. Relative configurations of cyclohexenone/cyclohexene and tetrahydropyran moieties of 15 were established on the basis of 3 J H–H coupling constants and NOE difference spectra. Stereochemical correlation of the two rings, and assignment of absolute configuration of 15 were achieved by comparison of experimental ECD spectra with simulated ECD data for possible stereoisomers, by using time dependent density function theory (TDDFT). Bisaboloids 14 exhibited in vitro antimalarial activity against Plasmodium falciparum, with IC50 values ranging from 2.8 to 20.1 μM, and selectivity indices (SI) in L-6 cells of 3.7–11.9.
Keywords: Artemisia persica; Plasmodium falciparum; TDDFT; Absolute configurations; Bisabololoxide;

Biphenyl-type neolignans from Magnolia officinalis and their anti-inflammatory activities by Wen-Lung Kuo; Ching-Yi Chung; Tsong-Long Hwang; Jih-Jung Chen (153-160).
Five biphenyl-type neolignans, 5-allyl-5′-(1″-hydroxyallyloxy)biphenyl-2,2′-diol (1), 5,5′-diallyl-2′-(allyloxy)biphenyl-2-ol (2), 5,5′-diallyl-2′-(3-methylbut-2-enyloxy)biphenyl-2-ol (3), (E)-5-allyl-3′-(prop-1-enyl)biphenyl-2,4′-diol (4), and 4-allyl-2-(2′-methylbenzofuran-5′-yl)phenol (5) were isolated from stem bark of Magnolia officinalis, together with twelve known compounds. This paper describes the structural elucidation of 15 and their anti-inflammatory activities.Display Omitted► Five compounds were isolated from Magnolia officinalis. ► Several isolated compounds inhibited fMLP-induced O2 •−generation (IC50  ⩽ 10.7 μM). ► Several inhibited fMLP-induced elastase release (IC50  ⩽ 8.76 μM). ► Two are potential candidates for treatment of inflammatory diseases.Reactive oxygen species (ROS) and granule proteases produced by human neutrophils contribute to the pathogenesis of inflammatory diseases. The MeOH extract of the stem bark of Magnolia officinalis showed potent inhibitory effects on superoxide anion generation and elastase release by human neutrophils in response to formyl-L-methionyl-L-leucyl-L-phenylalanine/cytochalasin B (fMLP/CB). Five biphenyl-type neolignan derivatives, 5-allyl-5′-(1″-hydroxyallyloxy)biphenyl-2,2′-diol, 5,5′-diallyl-2′-(allyloxy)biphenyl-2-ol, 5,5′-diallyl-2′-(3-methylbut-2-enyloxy)biphenyl-2-ol, (E)-5-allyl-3′-(prop-1-enyl)biphenyl-2,4′-diol, and 4-allyl-2-(2′-methylbenzofuran-5′-yl)phenol, have been isolated from the stem bark of M. officinalis, together with 12 known compounds. Several exhibited inhibition (IC50 values ⩽ 10.7 μM) of superoxide anion generation by human neutrophils in response to fMLP/CB. Others inhibited fMLP/CB-induced elastase release with IC50 values ⩽ 8.76 μM.
Keywords: Magnolia officinalis; Magnoliaceae; Structure elucidation; Biphenyl-type neolignan; Anti-inflammatory activity;

Friedolanostanes and xanthones from the twigs of Garcinia hombroniana by Saranyoo Klaiklay; Yaowapa Sukpondma; Vatcharin Rukachaisirikul; Souwalak Phongpaichit (161-166).
17,14-Friedolanostanes (garcihombronanes K–L, 12), and the xanthones (garcihombronones A–D, 36), together with 14 known compounds, were isolated from Garcinia hombroniana. Their antibacterial activity against methicillin-resistant Staphylococcus aureus and S. aureus was evaluated.Display Omitted► Two 17,14-friedolanostanes and four xanthones were isolated from Garcinia hombroniana. ► Xanthones were isolated for the first time from this plant. ► Their antibacterial activity was evaluated.The twigs of Garcinia hombroniana yielded six compounds: two 17,14-friedolanostanes (garcihombronanes K–L, 12) and four xanthones (garcihombronones A–D, 36) together with 14 known compounds including four friedolanostanes, one lanostane, six xanthones, two benzoic acid derivatives and one biflavonoid. Their structures were elucidated by analysis of spectroscopic data and comparison of the NMR data with those reported previously. Their antibacterial activity against methicillin-resistant Staphylococcus aureus and S. aureus was evaluated.
Keywords: Garcinia hombroniana; Guttiferae; Friedolanostane; Lanostane; Xanthone; Antibacterial;

Triterpenoid glycosides from the stems of Gordonia kwangsiensis by Hui-Zheng Fu; Chuang-Jun Li; Jing-Zhi Yang; Xiao-Guang Chen; Dong-Ming Zhang (167-174).
Eleven oleanane-type triterpenoid glycosides, named, gordonsaponins A−K (111), were isolated from the stems of Gordonia kwangsiensis. Their cytotoxic activities were tested.Display Omitted► Eleven oleanane-type triterpenoid glycosides were isolated from the stems of Gordonia kwangsiensis. ► All were evaluated for their cytotoxic activities. ► Only one showed activity against all tested cell lines, with IC50 values ranging from 0.1 to 2.41 μM.Eleven oleanane-type triterpenoid glycosides, named gordonsaponins A−K, were isolated from the stems of Gordonia kwangsiensis. Their structures were elucidated by spectroscopic and chemical methods. The cytotoxic activities of all eleven were evaluated against five human tumor cell lines (HCT-8, Bel-7402, BGC-823, A549, and A2780), with only one having activity against all tested cell lines, with IC50 values ranging from 0.1 to 2.41 μM.
Keywords: Gordonia kwangsiensis; Theaceae; Triterpenoid glycosides; Gordonsaponins A−K; Cytotoxic activity;

Chemical constituents from Brucea javanica by Shi-Hui Dong; Jia Liu; Ying-Zi Ge; Lei Dong; Cheng-Hui Xu; Jian Ding; Jian-Min Yue (175-184).
Fourteen triterpenoids, four known quassinoids and seven known lignans were isolated from Brucea javanica. Their structures were elucidated by spectroscopic analysis. The structure of a previously reported triterpenoid was revised as its C-21 epimer. The cytotoxicities of triterpenoids and quassinoids against two human tumor cell lines were evaluated.Display Omitted► Report of 14 triterpenoids, four known quassinoids, and seven known lignans. ► Isolation of triterpenes in the twigs, and quassinoids and lignans only in the seeds. ► Revision of the structure of a previously reported triterpenoid as its C-21 epimer.Fourteen apotirucallane-type triterpenoids, named brujavanones A–N, were isolated from the twigs of Brucea javanica, along with four known quassinoids and seven known lignans from the seeds of B. javanica. Their structures were elucidated on the basis of extensive spectroscopic data analysis. The structure of a previously reported triterpenoid, bruceajavanin C, was revised as its C-21 epimer. The cytotoxic activities of triterpenoids and quassinoids against two human tumor cell lines, HL-60 and A-549, were evaluated, but all the compounds were inactive (IC50  > 10 μM).
Keywords: Brucea javanica; Simaroubaceae; Triterpenoids; Quassinoids; Lignans; Brujavanones A–N; Bruceajavanin C;

Characterization of chlorinated valepotriates from Valeriana jatamansi by Sheng Lin; Zhong-Xiao Zhang; Tao Chen; Ji Ye; Wei-Xing Dai; Lei Shan; Juan Su; Yun-Heng Shen; Hui-Liang Li; Run-Hui Liu; Xi-ke Xu; Hui Wang; Wei-dong Zhang (185-193).
Fifteen previously unreported and six known chlorinated valepotriates were isolated from Valeriana jatamansi. Most compounds showed moderate cytotoxicity against A 549, PC-3M, HCT-8 and Bel 7402 cell lines with IC50 values of 0.89–9.76 μM.Display Omitted► Target isolation was performed by dereplication principle. ► Isolation of 19 chlorinated valepotriates. ► A part of the isolates exhibited moderate cytotoxicity against four tumor cells.HPLC–PDA–MS and TLC analysis were used to look for minor cytotoxic chlorinated valepotriates from whole plants of Valeriana jatamansi (syn. Valeriana wallichii DC.). This resulted in isolation of 15 chlorinated valepotriates, designated as chlorovaltrates A-O, together with six known analogues, (1S,3R,5R,7S,8S,9S)-3,8-epoxy-1,5-dihydroxyvalechlorine, volvaltrate B, chlorovaltrate, rupesin B, (1S,3R,5R,7S,8S,9S)-3,8-epoxy-1-O-ethyl-5-hydroxyvalechlorine, and (1R,3R,5R,7S,8S,9S)-3,8-epoxy-1-O-ethyl-5-hydroxyvalechlorine. Their structures were elucidated by spectroscopic methods including homo- and heteronuclear two-dimensional NMR experiments. Chlorovaltrates K-N, chlorovaltrate and rupesin B showed moderate cytotoxicity against lung adenocarcinoma (A 549), metastatic prostate cancer (PC-3M), colon cancer (HCT-8) and hepatoma (Bel 7402) cell lines with IC50 values of 0.89–9.76 μM.
Keywords: Valeriana jatamansi; Valerianaceae; Chlorinated valepotriates; Dereplication; Cytotoxicity;

Macroline–sarpagine and macroline–pleiocarpamine bisindole alkaloids from Alstonia angustifolia by Shin-Jowl Tan; Kuan-Hon Lim; G. Subramaniam; Toh-Seok Kam (194-202).
Nine binsindole alkaloids, comprising four belonging to the macroline–sarpagine group, and five belonging to the macroline–pleiocarpamine group, were isolated from the stem-bark extracts of Alstonia angustifolia.Display Omitted► Nine bisindole alkaloids were isolated from stem-bark extract of Alstonia angustifolia. ► Four are macroline–sarpagine bisindoles. ► Five are macroline–pleiocarpamine bisindoles.Nine bisindole alkaloids, comprising four belonging to the macroline–sarpagine group, and five belonging to the macroline–pleiocarpamine group, were isolated from the stem-bark extracts of Alstonia angustifolia (Apocynacea). Their structures were established using NMR and MS analyses.
Keywords: Alstonia angustifolia; Apocynaceae; Alkaloids; Bisindoles; NMR;