Freshwater algae chemotaxonomy by high-performance liquid chromatographic(HPLC)analysis

The study of community composition of algae is essential for understanding the structure and dynamics of the aquatic ecosystem and for evaluating the eutrophic level of the water body.A high-performance liquid chromatographic(HPLC)method based on a reversephase C_(18) nonpolar column was developed for the main algal taxa,which includes cyanophytes,bacillariophytes,euglenophytes,dinophytes,and chlorophytes.Based on the elution order using HPLC,19 pigments were identified,and they were chlorophyllide a,19′-butanoyloxyfucoxanthin,chlorophyll c_(1)+c_(2),phephorbides a,peridinin,methyl-chlorophyllide a,fucoxanthin,neoxanthin,violaxanthin,myxoxanthophyll,diadinoxanthin,diatoxanthin,lutein,zeaxanthin,chlorophyll b allomer,chlorophyll b,chlorophyll a allomer,chlorophyll a,andβ,β-carotene.A comparison study of cell microscopic counts and accessory pigment analysis indicated that HPLC analysis could be a useful tool for monitoring phytoplankton communities and their abundance.

Review of the El Soplao Amber Outcrop,Early Cretaceous of Cantabria,Spain

El Soplao outcrop, an Early Cretaceous amber deposit recently discovered in northern Spain （Cantabria）, has been shown to be the largest site of amber with arthropod inclusions that has been found in Spain so far. Relevant data provided herein for biogeochemistry of the amber, palynology, taphonomy and arthropod bioinclusions complement those previously published. This set of data suggests at least two botanical sources for the amber of El Soplao deposit. The first （type A amber） strongly supports a source related to Cheirolepidiaceae, and the second （type B amber） shows non-specific conifer biomarkers. Comparison of molecular composition of type A amber with Frenelopsis leaves （Cheirolepidiaceae） strongly suggests a biochemical affinity and a common botanical origin. A preliminary palynological study indicates a regional high taxonomical diversity, mainly of pteridophyte spores and gymnosperm pollen grains. According to the preliminary palynological data, the region was inhabited by conifer forests adapted to a dry season under a subtropical climate. The abundant charcoalified wood associated with the amber in the same beds is evidence of paleofires that most likely promoted both the resin production and an intensive erosion of the litter, and subsequent great accumulation of amber plus plant cuticles. In addition, for the first time in the fossil record, charcoalified plant fibers as bioinclnsions in amber are reported. Other relevant taphonomic data are the exceptional presence of serpulids and bryozoans on the surfaces of some amber pieces indicating both a long exposure on marine or brackish-water and a mixed assemblage of amber. Lastly, new findings of insect bioiuclusions, some of them uncommon in the fossil record or showing remarkable adaptations, are reported. In conclusion, a documented scenario for the origin of the El Soplao amber outcrop is provided.

Furoquinoline Alkaloids and Methoxyflavones from the Stem Bark of Melicope madagascariensis(Baker)T.G.Hartley

Melicope madagascariensis(Rutaceae)is an endemic plant species of Madagascar that was first classified as a member of the genus Euodia J.R.&G.Forst(Rutaceae)under the scientific name Euodia madagascariensis Baker.Based on morphological characteristics,Thomas Gordon Hartley taxonomically revised E.madagascariensis Baker to be M.madagascariensis(Baker)T.G.Hartley.Chemotaxonomical studies have long been used to help the identification and confirmation of taxonomical classification of plant species and botanicals.Aiming to find more evidences to support the taxonomical revision performed on E.madagascariensis,we carried out phytochemical investigation of two samples of the plant.Fractionation of the ethanol extracts prepared from two stem bark samples of M.madagascariensis(Baker)T.G.Hartley led to the isolation of seven known furoquinoline alkaloids 1–7 and two known methoxyflavones 8 and 9.The presence of furoquinoline alkaloids and methoxyflavones in the title species is in agreement with its taxonomic transfer from Euodia to Melicope.Antiprotozoal evaluation of the isolated compounds showed that 6-methoxy-7-hydroxydictamnine(heliparvifoline,3)showed weak antimalarial activity(IC_(50)=35μM)against the chloroquine-resistant strain Dd2 of Plasmodium falciparum.Skimmianine(4)displayed moderate cytotoxicity with IC_(50)value of 1.5 lM against HT-29 colon cancer cell line whereas 3,5-dihydroxy-30,40,7-trimethoxyflavone(9)was weakly active in the same assay(IC_(50)=13.9μM).

Electrochemical Identification of Yulania spp. by Fingerprinting of Leaves Using Glassy Carbon Electrode

In this communication,we used electrochemical sensor for recording the electrochemical profiles of eleven species of Yulania spp.from leaf extract.Two solvents and two buffer conditions were used for electrochemical fingerprints collection.Their electrochemical fingerprints can be converted to different patterns and consequently for species recognition.The results indicate the pattern recognition is much convenient than that of the recognition of species directly using voltammetric signal.The current information in electrochemical fingerprinting represents the type and amount of electrochemically active molecules,which linked to the genetic differences among the plants.Therefore,the electrochemical fingerprints were applied for further phylogenetic study.The phylogenetic tree deduced from voltametric curves is divided into three main groups.The first clade contains Y.denudate,Liriodendron chinense,Y.cylindrica,Y.biondii,Y.sprengeri.The second clade contains Y.zenii,Y.liliiflora,Y.kobus,and Y.amoena.The third clade contains Y.×soulangeana,Manglietia fordiana and Y.sinostellata.In addition,Y.salicifolia is not in these main clades.The results demonstrate that electrochemical fingerprinting can be used as a com-plementary tool in the study of phylogenetics.

Beauvericin production by the Lepidoptera pathogenic fungus Isaria tenuipes: Analysis of natural specimens, synnemata from cultivation, and mycelia from liquid-media fermentation

Beauvericin was analyzed in three forms of the Lepidoptra pathogenic fungus Isaria tenuipes(4 isolates):(a)natural specimen,(b)cultivated synnemata on rice media,and(c)mycelia from fermentation in liquid media.Beauvericin was detected in very low amounts in all tested natural specimens.Synnemata on rice contained much higher concentrations of beauvericin than the corresponding natural materials,although the concentrations were lower than mycelia from liquid fermentation.The results casted a caution that beauvericin concentration should be carefully checked,as a possible toxic constituent,upon mass production of a selected strain of Isaria tenuipes for health food purposes.

Chemotaxonomic Study Based on Flavonoids as Taxonomic Markers in the Roots of the Selected Species Belonged to Family Solanaceae, Sudan

This work is a taxonomic study on flavonoids in the roots of selected species belonged to family Solanaceae. These species considered as: Lycopersicum esculentum, Solanum melongena and Solanum tuberosum. These species distributed in different localities in Sudan. The selected members have nutritive, medicinal and economic importance, extra of that, the present study included botanical and chemical studies. The collected species have been updated due to nomenclature and synonymy. The geographical distribution of the selected members has been indicated. The chemical studies included identification of the flavonoid compounds using Gas Chromatography Mass Spectrophotometer (GC-MS). Eighty-three flavonoid compounds were detected in the roots of family Solanaceae. The highest number (32) was detected in the roots of Solanum tuberosum. A single flavonoid compound was restricted only to the roots of Solanum melongena. This was: (cyclohexanol-5-methyl-2-(1methylene), {1R-1.alpha, 2beta, 5.alpha}). Four taxonomic markers were identified for the roots of Lycopersicum esculentum. Ten taxonomic markers were identified in the roots of Solanum tuberosum.

Essential Oils from Different Parts of the Sea Buckthorn <i>Hippophae rhamnoides</i>L.

The study investigated essential oils of berries and green biomass of the sea buckthorn at various growth stages. Plant material of H. rhamnoídes ssp. mongolica was collected in the experimental fields of the Institute of Cytology and Genetics SB RAS (Siberia, Russia). Volatile compounds and essential oils were detected. Green spring shoots without leaves, first-year seedlings without leaves and berries were analyzed. In all studied exemplars, 103 volatile compounds were defined among which considerable proportion wasn’t found in a sea-buckthorn earlier. Phenylpropanoids prevailed in essential oils composition from seedlings. Alkanes and phenylpropanoids prevailed in shoots. Berries contained mostly carboxylic acids and their esters. These data can be applied to a fingerprint of various cultivars of a sea-buckthorn with relevance to food science and technology.

Recent Advance in Chemical and Biological Studies on Cimicifugeae Pharmaceutical Resources

Cimicifugeae is one of the rich sources for various active components and the health promoting and therapeutic values of the components have been corroborated by long-term use in folk medicine and traditional Chinese medicine.Increasing interest in Cimicifugeae pharmaceutical resources has led to the further discoveries of triterpenoid saponins, phenolic compounds, chromones, and many other compounds in various species of Cimicifugeae, and to the investigations on their chemotaxonomy, molecular phylogeny, and bioactivities. Based on our pharmacophylogenetic studies, the progress in phytochemistry, chemotaxonomy, molecular biology, and phylogeny of Cimicifugeae had been summarized since 2007, especially Cimicifuga L. ex Wernisch. and Actaea L., and their relevance to therapeutic efficacy. An exhaustive literature survey is used to characterize the global scientific effort in the phytochemical and biological studies of Cimicifugeae. More triterpenoid saponins have been found in various species, among which the cimigenol type(type A) is predominant. The versatile bioactivities of saponins and extracts,as well as those of phenolics and other ingredients, were summarized and discussed. The morphology-based five-genus classification of Cimicifugeae is not supported by molecular phylogeny. Molecular phylogeny based on nuclear and chloroplast DNA sequences tends to merge Cimicifuga Wernisch., Souliea Franch., and Actaea L. into a single genus. It is indispensable to integrate the emerging technologies into Cimicifugeae research for both the sustainable utilization of Cimicifugeae pharmaceutical resources and finding novel compounds with potential clinical utility and less adverse effects. Systems biology and omics technologies would play an increasingly important role in booming pharmaceutical research involving bioactive compounds of Cimicifugeae.

Research progress in the phytochemistry and biology of Ilex pharmaceutical resources

Ilex is a botanical source for various health-promoting and pharmaceutically active compounds that have been used in traditional Chinese medicine and food for thousands of years.Increasing interest in Ilex pharmaceutical and food resources has led to additional discoveries of terpenoids,saponins,polyphenols(especially flavonoids),glycosides,and many other compounds in various Ilex species,and to investigation of their chemotaxonomy,molecular phylogeny and pharmacol-ogy.In continuation of our studies on Ilex pharmacology and phylogeny,we review the phytochemistry,chemotaxonomy,molecular biology and phylogeny of Ilex species and their relevance to health-promotion and therapeutic efficacy.The similarity and dissimilarity between Ilex paraguariensis,the source plant of mate tea,and the source plants of large-leaved K udingcha(e.g.,Ilex kudingcha and Ilex latifolia)are discussed.It is essential to utilize emerging technologies in non-Camellia tea studies to promote the sustainable utilization of Ilex resources and the identification and development of novel compounds with potential health and clinical utility.Systems biology and "-omics"technologies will play an increasingly important role in pharmaceutical and food research on the bioactive compounds of Ilex species.

Molecular phylogenetic analyses redefine seven major clades and reveal 22 new generic clades in the fungal family Boletaceae

Mushrooms in the basidiomycete family Boletaceae are ecologically and economically very important.However,due to the morphological complexity and the limited phylogenetic information on the various species and genera of this fungal family,our understanding of its systematics and evolution remains rudimentary.In this study,DNA sequences of four genes(nrLSU,tef1-α,rpb1,and rpb2)were newly obtained from ca.200 representative specimens of Boletaceae.Our phylogenetic analyses revealed seven major clades at the subfamily level,namely Austroboletoideae,Boletoideae,Chalciporoideae,Leccinoideae,Xerocomoideae,Zangioideae,and the Pulveroboletus Group.In addition,59 genus-level clades were identified,of which 22 were uncovered for the first time.These 22 clades were mainly placed in Boletoideae and the Pulveroboletus Group.The results further indicated that the characters frequently used in the morphology-based taxonomy of Boletaceae,such as basidiospore ornamentation,the form of the basidioma,and the stuffed pores each had multiple origins within the family,suggesting that the use of such features for high-level classification of Boletaceae should be de-emphasized and combined with other characters.

Lenormandins A—G,new azaphilones from Hypoxylon lenormandii and Hypoxylon jaklitschii sp.nov.,recognised by chemotaxonomic data

Seven new azaphilone pigments named lenormandins A–G were isolated from stromata(fruiting bodies)of the xylariaceous fungus,Hypoxylon lenormandii using preparative High Performance Liquid Chromatography(HPLC)and their structures were elucidated by NMR spectroscopy,high resolution mass spectrometry and other spectral methods.Their occurrence in herbarium specimens(including various type materials collected in the 19th and early 20th century)and several fresh collections was studied by analytical HPLC with diode array and mass spectrometric detection(HPLC-DAD/MS),revealing that this group of pigments is specific for Hypoxylon lenormandii from various geographic regions and otherwise only occurs in closely related taxa,i.e.H.sublenormandii and an undescribed fungus from Sri Lanka,for which we propose the name H.jaklitschii.The status of the new species was established by using molecular phylogenetic data based on ITS and partial beta-tubulin sequences,and by detailed morphological studies.

New Hypoxylon species from Martinique and new evidence on the molecular phylogeny of Hypoxylon based on ITS rDNA and β-tubulin data

Three new species of Hypoxylon(Xylariaceae)collected from Martinique in the French Caribbean are recognised by new combinations of morphological characters.Their status as undescribed taxa was supported by secondary metabolite profiling based on High performance liquid chromatography with diode array and mass spectrometric detection(HPLC/DAD-MS)as well as by comparison of ITS and partialß-tubulin DNA sequences with related taxa.In the course of this study,the teleomorph of Nodulisporium griseobrunneum was found,and this species could be transferred to Hypoxylon.Moreover,several names in Hypoxylon are epitypified by selecting recently collected specimens from the same geographic areas as the holotypes came from.Despite the fact that our study used the hitherto most extensive taxon sampling,the phylogenetic analyses inferred from ITS andß-tubulin sequences remain contradictory to each other,and neither genealogy was found fully in agreement with phenotype-derived traits.We conclude that the right gene(or multi-gene genealogies)to reflect the phylogeny and evolution of Hypoxylon still remains to be found.For the time being,we recommend that the application of polyphasic taxonomic concepts should be continued in taxonomic studies of Hypoxylon.

Phylogenetic and chemotaxonomic resolution of the genus Annulohypoxylon(Xylariaceae)including four new species

This study deals with an extensive evaluation focusing on phylogenetic and chemotaxonomic infrageneric relationships of the genus Annulohypoxylon(Xylariaceae,Ascomycota),whose species are ubiquitously associated with seed plants as endophytes and saprotrophs in all forested areas of the world.Using evidence from phylogenetic,chemotaxonomic and morphological data,various varieties within the genus are raised to species level,leading to the new combinations for A.areolatum(:A.bovei var.microsporum),A.macrosporum(:A.leptascum var.macrosporum),and A.microdiscum(:A.moriforme var.microdiscum).Annulohypoxylon substygium nom.nov.is applied for A.stygium var.annulatumand the four new tropical and subtropical species A.massivum,A.violaceopigmentum,A.viridistratum and A.yungensis are introduced.Furthermore,A.leucadendri is excluded from the genus as its morphological characters disagree with the generic concept,the recently erected A.palmicola is synonymized with A.leptascum and A.austrobahiense has been reassigned to the genus Hypoxylon.In addition,the key taxa A.annulatum and A.truncatum have been reinvestigated and epitypes have been defined.For the first time,a dichotomous key to the genus is provided.A phylogenetic b-tubulin tree in conjunction with stromatal HPLC profiles clearly shows that Annulohypoxylon comprises two distinct lineages.The A.cohaerens/multiforme group might eventually warrant segregation into a new genus as further molecular data become available.

Chemical Constituents from Leaves of Oplopanax horridus

Objective To study the chemical constituents from the leaves of Oplopanax horridus. Methods The chemical constituents were isolated and purified by column chromatography on silica gel and Sephadex LH-20 gel columns, 1H-NMR and 13C-NMR were applied for the identification of chemical structure. Results Ten compounds were isolated and identified as dammara-20,24-dien-3β-ol acetate（1）, phytol（2）, 16 Z,19Z-pentacosadienoic acid（3）, β-sitosterol（4）,（3S,8S）-falcarindiol（5）, maltol（6）, acankoreagenin（7）, daucosterol（8）, stigmasterol-3-O-β-D-glucopyranoside（9）, and acankoreoside A（10）. Conclusion Compounds 1-3, 6, and 10 are isolated from this plant for the first time. Compounds 1-3 and 6 are isolated from the plants in genus Oplopanax Miq. for the first time. Moreover, Compounds 1, 3, and 6 are isolated from the plants in the family of Araliaceae for the first time.

Identification of fungal fossils and novel azaphilone pigments in ancient carbonised specimens of Hypoxylon fragiforme from forest soils of Chatillon-sur-Seine(Burgundy)

Fungal stromata were recently discovered in association with charcoal and burnt soil aggregates during an archaeological survey in the Chaˆtillon-sur-Seine forest massif.The wood and soil in the samples were dated to the medieval period(between 738 and 1411 AD).Light microscopy and scanning electron microscopy revealed that a few of the stromatal fragments still contained ascospores.Their macromorphological characters were described and secondary metabolite profiles were generated using high performance liquid chromatography with diode array and mass spectrometric detection(HPLC–DAD/MS).The combination of these two data lines then allowed species identification.Most of the fragments were assigned to Hypoxylon fragiforme,the type species of the Hypoxylaceae(Xylariales).Two further species whose stromata grew on the fossil charcoal could be tentatively identified as Jackrogersella cohaerens and(more tentatively)as Hypoxylon vogesiacum.These three species are still commonly encountered in the forests of Central Europe today.Furthermore,the HPLC-HRMS data of H.fragiforme suggested the presence of unknown azaphilone dimers and of further new pigments.These archaeological compounds were compared to fresh stromata of H.fragiforme collected in Germany and subjected to the same analytical protocol.While the major components in both samples were identified as the known mitorubrin type azaphilones and orsellinic acid,the chemical structures of seven novel complex azaphilone pigments,for which we propose the trivial names rutilins C-D and fragirubrins A-E,were elucidated using spectral methods(NMR and CD spectroscopy,high resolution mass spectrometry).It appears that these pigments had indeed persisted for millennia in the fossil stromata.