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Dinoflagellates of the genus Gambierdiscus have been associated with ciguatera, the most common non-bacterial fish-related intoxication in the world. Many studies report the presence of potentially toxic Gambierdiscus species along the Atlantic coasts including G. australes, G. silvae and G. excentricus. Estimates of their toxicity, as determined by bio-assays, vary substantially, both between species and strains of the same species. Therefore, there is a need for additional knowledge on the metabolite production of Gambierdiscus species and their variation to better understand species differences. Using liquid chromatography coupled to mass spectrometry, toxin and metabolomic profiles of five species of Gambierdiscus found in the Atlantic Ocean were reported. In addition, a molecular network was constructed aiming at annotating the metabolomes. Results demonstrated that G. excentricus could be discriminated from the other species based solely on the presence of MTX4 and sulfo-gambierones and that the variation in toxin content for a single strain could be up to a factor of two due to different culture conditions between laboratories. While untargeted analyses highlighted a higher variability at the metabolome level, signal correction was applied and supervised multivariate statistics performed on the untargeted data set permitted the selection of 567 features potentially useful as biomarkers for the distinction of G. excentricus, G. caribaeus, G. carolinianus, G. silvae and G. belizeanus. Further studies will be required to validate the use of these biomarkers in discriminating Gambierdiscus species. The study also provided an overview about 17 compound classes present in Gambierdiscus, however, significant improvements in annotation are still required to reach a more comprehensive knowledge of Gambierdiscus’ metabolome

Dinoflagellates of the genus Gambierdiscus have been associated with ciguatera, the most common non-bacterial fish-related intoxication in the world. Many studies report the presence of potentially toxic Gambierdiscus species along the Atlantic coasts including G. australes, G. silvae and G. excentricus. Estimates of their toxicity, as determined by bio-assays, vary substantially, both between species and strains of the same species. Therefore, there is a need for additional knowledge on the metabolite production of Gambierdiscus species and their variation to better understand species differences. Using liquid chromatography coupled to mass spectrometry, toxin and metabolomic profiles of five species of Gambierdiscus found in the Atlantic Ocean were reported. In addition, a molecular network was constructed aiming at annotating the metabolomes. Results demonstrated that G. excentricus could be discriminated from the other species based solely on the presence of MTX4 and sulfo-gambierones and that the variation in toxin content for a single strain could be up to a factor of two due to different culture conditions between laboratories. While untargeted analyses highlighted a higher variability at the metabolome level, signal correction was applied and supervised multivariate statistics performed on the untargeted data set permitted the selection of 567 features potentially useful as biomarkers for the distinction of G. excentricus, G. caribaeus, G. carolinianus, G. silvae and G. belizeanus. Further studies will be required to validate the use of these biomarkers in discriminating Gambierdiscus species. The study also provided an overview about 17 compound classes present in Gambierdiscus, however, significant improvements in annotation are still required to reach a more comprehensive knowledge of Gambierdiscus’ metabolome.

Plant metabolite profiling reveals the diversity of secondary or specialized metabolites in the plant kingdom withits hundreds of thousands of species. Specialized plant metabolites constitute a vast class of chemicals posingsignificant challenges in analytical chemistry. In order to be of maximum scientific relevance, reports dealingwith these compounds and their source species must be transparent, make use of standards and reference ma-terials, and be based on correctly and traceably identified plant material. Essential aspects in qualitative plantmetabolite profiling include: (i) critical review of previous literature and a reasoned sampling strategy; (ii)transparent plant sampling with wild material documented by vouchers in public herbaria and, optimally, seedbanks; (iii) if possible, inclusion of generally available reference plant material; (iv) transparent, documentedstate-of-the art chemical analysis, ideally including chemical reference standards; (v) testing for artefacts duringpreparative extraction and isolation, using gentle analytical methods; (vi) careful chemical data interpretation,avoiding over- and misinterpretation and taking into account phytochemical complexity when assigning iden-tification confidence levels, and (vii) taking all previous scientific knowledge into account in reporting thescientific data. From the current stage of the phytochemical literature, selected comments and suggestions aregiven. In the past, proposed revisions of botanical taxonomy were sometimes based on metabolite profiles, butthis approach (“chemosystematics” or “chemotaxonomy”) is outdated due to the advent of DNA sequence-basedphylogenies. In contrast, systematic comparisons of plant metabolite profiles in a known phylogenetic frameworkremain relevant. This approach, known as chemophenetics, allows characterizing species and clades based ontheir array of specialized metabolites, aids in deducing the evolution of biosynthetic pathways and coevolution,and can serve, Plant metabolite profiling reveals the diversity of secondary or specialized metabolites in the plant kingdom with its hundreds of thousands of species. Specialized plant metabolites constitute a vast class of chemicals posing significant challenges in analytical chemistry. In order to be of maximum scientific relevance, reports dealing with these compounds and their source species must be transparent, make use of standards and reference materials, and be based on correctly and traceably identified plant material. Essential aspects in qualitative plant metabolite profiling include: (i) critical review of previous literature and a reasoned sampling strategy; (ii) transparent plant sampling with wild material documented by vouchers in public herbaria and, optimally, seed banks; (iii) if possible, inclusion of generally available reference plant material; (iv) transparent, documented state-of-the art chemical analysis, ideally including chemical reference standards; (v) testing for artefacts during preparative extraction and isolation, using gentle analytical methods; (vi) careful chemical data interpretation, avoiding over- and misinterpretation and taking into account phytochemical complexity when assigning identification confidence levels, and (vii) taking all previous scientific knowledge into account in reporting the scientific data. From the current stage of the phytochemical literature, selected comments and suggestions are given. In the past, proposed revisions of botanical taxonomy were sometimes based on metabolite profiles, but this approach (“chemosystematics” or “chemotaxonomy”) is outdated due to the advent of DNA sequence-based phylogenies. In contrast, systematic comparisons of plant metabolite profiles in a known phylogenetic framework remain relevant. This approach, known as chemophenetics, allows characterizing species and clades based on their array of specialized metabolites, aids in deducing the evolution of biosynthetic pathways and coevolutio

Plant metabolite profiling reveals the diversity of secondary or specialized metabolites in the plant kingdom withits hundreds of thousands of species. Specialized plant metabolites constitute a vast class of chemicals posingsignificant challenges in analytical chemistry. In order to be of maximum scientific relevance, reports dealingwith these compounds and their source species must be transparent, make use of standards and reference ma-terials, and be based on correctly and traceably identified plant material. Essential aspects in qualitative plantmetabolite profiling include: (i) critical review of previous literature and a reasoned sampling strategy; (ii)transparent plant sampling with wild material documented by vouchers in public herbaria and, optimally, seedbanks; (iii) if possible, inclusion of generally available reference plant material; (iv) transparent, documentedstate-of-the art chemical analysis, ideally including chemical reference standards; (v) testing for artefacts duringpreparative extraction and isolation, using gentle analytical methods; (vi) careful chemical data interpretation,avoiding over- and misinterpretation and taking into account phytochemical complexity when assigning iden-tification confidence levels, and (vii) taking all previous scientific knowledge into account in reporting thescientific data. From the current stage of the phytochemical literature, selected comments and suggestions aregiven. In the past, proposed revisions of botanical taxonomy were sometimes based on metabolite profiles, butthis approach (“chemosystematics” or “chemotaxonomy”) is outdated due to the advent of DNA sequence-basedphylogenies. In contrast, systematic comparisons of plant metabolite profiles in a known phylogenetic frameworkremain relevant. This approach, known as chemophenetics, allows characterizing species and clades based ontheir array of specialized metabolites, aids in deducing the evolution of biosynthetic pathways and coevolution,and can serve, Plant metabolite profiling reveals the diversity of secondary or specialized metabolites in the plant kingdom with its hundreds of thousands of species. Specialized plant metabolites constitute a vast class of chemicals posing significant challenges in analytical chemistry. In order to be of maximum scientific relevance, reports dealing with these compounds and their source species must be transparent, make use of standards and reference materials, and be based on correctly and traceably identified plant material. Essential aspects in qualitative plant metabolite profiling include: (i) critical review of previous literature and a reasoned sampling strategy; (ii) transparent plant sampling with wild material documented by vouchers in public herbaria and, optimally, seed banks; (iii) if possible, inclusion of generally available reference plant material; (iv) transparent, documented state-of-the art chemical analysis, ideally including chemical reference standards; (v) testing for artefacts during preparative extraction and isolation, using gentle analytical methods; (vi) careful chemical data interpretation, avoiding over- and misinterpretation and taking into account phytochemical complexity when assigning identification confidence levels, and (vii) taking all previous scientific knowledge into account in reporting the scientific data. From the current stage of the phytochemical literature, selected comments and suggestions are given. In the past, proposed revisions of botanical taxonomy were sometimes based on metabolite profiles, but this approach (“chemosystematics” or “chemotaxonomy”) is outdated due to the advent of DNA sequence-based phylogenies. In contrast, systematic comparisons of plant metabolite profiles in a known phylogenetic framework remain relevant. This approach, known as chemophenetics, allows characterizing species and clades based on their array of specialized metabolites, aids in deducing the evolution of biosynthetic pathways and coevolutio

Chemotaxonomy presents various challenges that need to be overcome in order to obtain valid and reliable results. Individual genetic and environmental variations can give a false picture and lead to wrong conclusions. Applying a holistic approach, based on multivariate data analysis, these challenges can be overcome. Thus, a metabolomics approach has to be optimized depending on the subject of research. We used 1H NMR-based metabolomics as a potential chemotaxonomic tool on the selected Euphorbia species growing wild in Serbia. Principal components analysis (PCA), soft independent modeling by class analogy (SIMCA) and Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) were used to analyze obtained NMR data in order to reveal chemotaxonomic biomarkers. The standard protocol for plant metabolomics was optimized aiming to extract more specific metabolites, which are characteristic for the Euphorbia genus. The obtained models were validated, which revealed that variables unique for each species were associated with certain classes of molecules according to literature data. In E. salicifolia, acacetin-7-O-glycoside (not found before in the species) was detected, and the structure of the aglycone part was solved based on 2D NMR data. In the presented paper, we have shown that metabolomics can be successfully used in Euphorbia chemotaxonomy.

Different fungal species belonging to the Colletotrichum genus cause anthracnose disease in a range of major crops, resulting in huge economic losses worldwide. Typical symptoms include dark, sunken lesions on leaves, stems, or fruits. Colletotrichum spp. have synthesized, in vitro, a number of biologically active and structurally unusual metabolites that are involved in their host’s infection process. In this study, we applied a one strain many compounds (OSMAC) approach, integrated with targeted and non-targeted metabolomics profiling, to shed light on the secondary phytotoxic metabolite panels produced by pathogenic isolates of Colletotrichum truncatum and Colletotrichum trifolii. The phytotoxicity of the fungal crude extracts was also assessed on their primary hosts and related legumes, and the results correlated with the metabolite profile that arose from the different cultural conditions. To the best of our knowledge, this is the first time that the OSMAC strategy integrated with metabolomics approaches has been applied to Colletotrichum species involved in legume diseases.

Chemotaxonomy presents various challenges that need to be overcome in order to obtain valid and reliable results. Individual genetic and environmental variations can give a false picture and lead to wrong conclusions. Applying a holistic approach, based on multivariate data analysis, these challenges can be overcome. Thus, a metabolomics approach has to be optimized depending on the subject of research. We used 1H NMR-based metabolomics as a potential chemotaxonomic tool on the selected Euphorbia species growing wild in Serbia. Principal components analysis (PCA), soft independent modeling by class analogy (SIMCA) and Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) were used to analyze obtained NMR data in order to reveal chemotaxonomic biomarkers. The standard protocol for plant metabolomics was optimized aiming to extract more specific metabolites, which are characteristic for the Euphorbia genus. The obtained models were validated, which revealed that variables unique for each species were associated with certain classes of molecules according to literature data. In E. salicifolia, acacetin-7-O-glycoside (not found before in the species) was detected, and the structure of the aglycone part was solved based on 2D NMR data. In the presented paper, we have shown that metabolomics can be successfully used in Euphorbia chemotaxonomy.

Chemotaxonomy presents various challenges that need to be overcome in order to obtain valid and reliable results. Individual genetic and environmental variations can give a false picture and lead to wrong conclusions. Applying a holistic approach, based on multivariate data analysis, these challenges can be overcome. Thus, a metabolomics approach has to be optimized depending on the subject of research. We used 1H NMR-based metabolomics as a potential chemotaxonomic tool on the selected Euphorbia species growing wild in Serbia. Principal components analysis (PCA), soft independent modeling by class analogy (SIMCA) and Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) were used to analyze obtained NMR data in order to reveal chemotaxonomic biomarkers. The standard protocol for plant metabolomics was optimized aiming to extract more specific metabolites, which are characteristic for the Euphorbia genus. The obtained models were validated, which revealed that variables unique for each species were associated with certain classes of molecules according to literature data. In E. salicifolia, acacetin-7-O-glycoside (not found before in the species) was detected, and the structure of the aglycone part was solved based on 2D NMR data. In the presented paper, we have shown that metabolomics can be successfully used in Euphorbia chemotaxonomy.

Species misidentification in the field of natural products is an acknowledged problem. These errors are especially widespread in sponge studies, albeit rarely assessed and documented. As a case study, we aim to revisit reports of isomalabaricane triterpenes, isolated from four demosponge genera: Jaspis, Geodia, Stelletta and Rhabdastrella. From a total of 44 articles (1981-2022), 27 unique vouchers were listed, 21 of which were accessed and re-examined here: 11 (52.4%) of these were misidentified. Overall, 65.9% of the studies published an incorrect species name: previously identified Jaspis and Stelletta species were all in fact Rhabdastrella globostellata. We conclude that isomalabaricane triterpenes were isolated from only two Rhabdastrella species and possibly one Geodia species. In addition to shedding a new light on the distribution of isomalabaricane triterpenes, this study is an opportunity to highlight the crucial importance of vouchers in natural product studies. Doing so, we discuss the impact of species misidentification and poor accessibility of vouchers in the field of sponge natural products. We advocate for stricter voucher guidelines in natural product journals and propose a common protocol of good practice, in the hope of reducing misidentifications in sponge studies, ensure reproducibility of studies, and facilitate follow-up work on the original material.

Species misidentification in the field of natural products is an acknowledged problem. These errors are especially widespread in sponge studies, albeit rarely assessed and documented. As a case study, we aim to revisit reports of isomalabaricane triterpenes, isolated from four demosponge genera: Jaspis, Geodia, Stelletta and Rhabdastrella. From a total of 44 articles (1981-2022), 27 unique vouchers were listed, 21 of which were accessed and re-examined here: 11 (52.4%) of these were misidentified. Overall, 65.9% of the studies published an incorrect species name: previously identified Jaspis and Stelletta species were all in fact Rhabdastrella globostellata. We conclude that isomalabaricane triterpenes were isolated from only two Rhabdastrella species and possibly one Geodia species. In addition to shedding a new light on the distribution of isomalabaricane triterpenes, this study is an opportunity to highlight the crucial importance of vouchers in natural product studies. Doing so, we discuss the impact of species misidentification and poor accessibility of vouchers in the field of sponge natural products. We advocate for stricter voucher guidelines in natural product journals and propose a common protocol of good practice, in the hope of reducing misidentifications in sponge studies, ensure reproducibility of studies, and facilitate follow-up work on the original material.

Species misidentification in the field of natural products is an acknowledged problem. These errors are especially widespread in sponge studies, albeit rarely assessed and documented. As a case study, we aim to revisit reports of isomalabaricane triterpenes, isolated from four demosponge genera: Jaspis, Geodia, Stelletta and Rhabdastrella. From a total of 44 articles (1981-2022), 27 unique vouchers were listed, 21 of which were accessed and re-examined here: 11 (52.4%) of these were misidentified. Overall, 65.9% of the studies published an incorrect species name: previously identified Jaspis and Stelletta species were all in fact Rhabdastrella globostellata. We conclude that isomalabaricane triterpenes were isolated from only two Rhabdastrella species and possibly one Geodia species. In addition to shedding a new light on the distribution of isomalabaricane triterpenes, this study is an opportunity to highlight the crucial importance of vouchers in natural product studies. Doing so, we discuss the impact of species misidentification and poor accessibility of vouchers in the field of sponge natural products. We advocate for stricter voucher guidelines in natural product journals and propose a common protocol of good practice, in the hope of reducing misidentifications in sponge studies, ensure reproducibility of studies, and facilitate follow-up work on the original material.

The chemical composition of recent and fossil organic-walled dinoflagellate cyst walls and its diversity is poorly understood and analyses on single microscopic specimens are rare. A series of infrared spectroscopic experiments resulted in the proposition of a standardized attenuated total reflection micro-Fourier transform infrared-based method that allows the collection of robust data sets consisting of spectra from individual dinocysts. These data sets are largely devoid of nonchemical artifacts inherent to other infrared spectrochemical methods, which have typically been used to study similar specimens in the past. The influence of sample preparation, specimen morphology and size and spectral data processing steps is also assessed within this methodological framework. As a result, several guidelines are proposed which facilitate the collection and qualitative interpretation of highly reproducible and repeatable spectrochemical data. These, in turn, pave the way for a systematic exploration of dinocyst chemistry and its assessment as a chemotaxonomical tool or proxy.

Species misidentification in the field of natural products is an acknowledged problem. These errors are especially widespread in sponge studies, albeit rarely assessed and documented. As a case study, we aim to revisit reports of isomalabaricane triterpenes, isolated from four demosponge genera: Jaspis, Geodia, Stelletta and Rhabdastrella. From a total of 44 articles (1981-2022), 27 unique vouchers were listed, 21 of which were accessed and re-examined here: 11 (52.4%) of these were misidentified. Overall, 65.9% of the studies published an incorrect species name: previously identified Jaspis and Stelletta species were all in fact Rhabdastrella globostellata. We conclude that isomalabaricane triterpenes were isolated from only two Rhabdastrella species and possibly one Geodia species. In addition to shedding a new light on the distribution of isomalabaricane triterpenes, this study is an opportunity to highlight the crucial importance of vouchers in natural product studies. Doing so, we discuss the impact of species misidentification and poor accessibility of vouchers in the field of sponge natural products. We advocate for stricter voucher guidelines in natural product journals and propose a common protocol of good practice, in the hope of reducing misidentifications in sponge studies, ensure reproducibility of studies, and facilitate follow-up work on the original material.

The chemical composition of recent and fossil organic-walled dinoflagellate cyst walls and its diversity is poorly understood and analyses on single microscopic specimens are rare. A series of infrared spectroscopic experiments resulted in the proposition of a standardized attenuated total reflection micro-Fourier transform infrared-based method that allows the collection of robust data sets consisting of spectra from individual dinocysts. These data sets are largely devoid of nonchemical artifacts inherent to other infrared spectrochemical methods, which have typically been used to study similar specimens in the past. The influence of sample preparation, specimen morphology and size and spectral data processing steps is also assessed within this methodological framework. As a result, several guidelines are proposed which facilitate the collection and qualitative interpretation of highly reproducible and repeatable spectrochemical data. These, in turn, pave the way for a systematic exploration of dinocyst chemistry and its assessment as a chemotaxonomical tool or proxy.

Tribe Eucalypteae, commonly known as the Australian eucalypts, is an economical, pharmacological, chemically, and historically important group consisting of over 850 species. These angiosperms are the number one source of hardwood globally with over 200,000 km2 of plantation dedicated to E. globulus. E. grandis and E. robusta, they possess cultural and ethnomedicinal significance, rooted deep in the heritage and culture of the Australian indigenous populations and have acted as sources of biologically active new small molecules in the modern era. The relationship between these plants has been the source of scientific debate and intrigue since the early 1900s, coming to a head in 1995 when the genera Angophora and Corymbia were delineated from the large genus Eucalyptus. The rearrangement of the relative taxonomic rank of these taxa sparked intensive research into the Australian eucalypts, with 86.7 % of the 36,264 publications containing Eucalyptus, Corymbia and Angophora being published since the 1995 taxonomic revision. Despite this large quantity of research, the delineation of these genera remains blurred due to morphological similarities, and ambiguous results from chemotaxonomic studies. Due to this, we have performed the largest chemical investigation of tribe Eucalypteae to date, utilizing a mixture of traditional natural product isolation and discovery methodologies accompanied by modern chemoinformatic analysis, statistical and machine learning investigation of the NMR assignments of published formyl phloroglucinol and [beta]-triketones, and the novel application of non-uniformly sampled HMBC as a chemotaxonomic probe to qualitatively identify the presence of key moieties of chemotaxonomic significance. [...], Thesis (PhD Doctorate), Doctor of Philosophy (PhD), School of Environment and Sc, Science, Environment, Engineering and Technology, Full Text

The presence of Narcissus munnozii-garmendiae in Extremadura is commented. Bulb and leaf extracts are analysed by GC-MS for alkaloid identification and alkaloid profile determination. The analysis is used as a chemotaxonomic tool to compare the population cited near Alburquerque in the province of Badajoz and the classical population in the province of Ciudad Real., Se comenta la presencia de Narcissus munnozii-garmendiae en Extremadura. Se analizan extractos de bulbos y hojas mediante GC-MS para la identificación de alcaloides y determinación del perfil alcaloídico. El análisis es usado como una herramienta quimiotaxonómica para comparar la población citada cerca de Alburquerque en la provincia de Badajoz y la población clásica de la provincia de Ciudad Real.

Species misidentification in the field of natural products is an acknowledged problem. These errors are especially widespread in sponge studies, albeit rarely assessed and documented. As a case study, we aim to revisit reports of isomalabaricane triterpenes, isolated from four demosponge genera: Jaspis, Geodia, Stelletta and Rhabdastrella. From a total of 44 articles (1981-2022), 27 unique vouchers were listed, 21 of which were accessed and re-examined here: 11 (52.4%) of these were misidentified. Overall, 65.9% of the studies published an incorrect species name: previously identified Jaspis and Stelletta species were all in fact Rhabdastrella globostellata. We conclude that isomalabaricane triterpenes were isolated from only two Rhabdastrella species and possibly one Geodia species. In addition to shedding a new light on the distribution of isomalabaricane triterpenes, this study is an opportunity to highlight the crucial importance of vouchers in natural product studies. Doing so, we discuss the impact of species misidentification and poor accessibility of vouchers in the field of sponge natural products. We advocate for stricter voucher guidelines in natural product journals and propose a common protocol of good practice, in the hope of reducing misidentifications in sponge studies, ensure reproducibility of studies, and facilitate follow-up work on the original material.

Fungi are rich in complexes of cryptic species that need a combination of different approaches to be delimited, including genomic information. Beauveria (Cordycipitaceae, Hypocreales) is a well-known genus of entomopathogenic fungi, used as a biocontrol agent. In this study we present a polyphasic taxonomy regarding two widely distributed complexes of Beauveria: B. asiatica and B. bassiana s.lat. Some of the genetic groups as previously detected within both taxa were either confirmed or fused using population genomics. High levels of divergence were found between two clades in B. asiatica and among three clades in B. bassiana, supporting their subdivision as distinct species. Morphological examination focusing on the width and the length of phialides and conidia showed no difference among the clades within B. bassiana while conidial length was significantly different among clades within B. asiatica. The secondary metabolite profiles obtained by liquid chromatography-mass spectrometry (LC-MS) allowed a distinction between B. asiatica and B. bassiana, but not between the clades therein. Based on these genomic, morphological, chemical data, we proposed a clade of B. asiatica as a new species, named B. thailandica, and two clades of B. bassiana to respectively represent B. namnaoensis and B. neobassiana spp. nov. Such closely related but divergent species with different host ranges have potential to elucidate the evolution of host specificity, with potential biocontrol application.

Premise of research. Geochemical fingerprinting of fossil plants is a relatively new research field complementing morphological analyses and providing information for paleoenvironmental interpretations. Ginkgoales contains a single extant species but was diverse through the Mesozoic and is an excellent target for biochemical analyses. Methodology. Cuticles derived from fresh and fallen autumn leaves of extant Ginkgo biloba and seven fossil gink- goalean leaf taxa, one seed fern taxon, and two taxa with bennettitalean affinity were analyzed by infrared (IR) microspec- troscopy at the D7 beamline in the MAX IV synchrotron laboratory, Sweden. The fossil material derives from Triassic and Jurassic successions of Greenland. Spectral data sets were compared and evaluated by hierarchical cluster analysis (HCA) and principal component analysis performed on vector-normalized, first-derivative IR absorption spectra. Pivotal results. The IR absorption spectra of the fossil leaves all reveal signatures that clearly indicate the pres- ence of organic compounds. Spectra of the extant G. biloba leaves reveal the presence of aliphatic chains, aromatic and ester carbonyl functional groups from polymer cutin and other waxy compounds, and polysaccharides. Inter- estingly, both the extant autumn leaves and the fossil specimens reveal the presence of carboxyl/ketone molecules, suggesting that chemical alterations during the initial stages of decomposition are preserved through fossilization. Two major subclusters were identified through HCA of the fossil spectra. Conclusions. Consistent chemical IR signatures, specific for each fossil taxon are present in cuticles, and suf- ficient molecular content is preserved in key regions to reflect the plants’ original chemical signatures. The alter- ations of the organic compounds are initiated as soon as the leaves are shed, with loss of proteins and increased ester and carboxyl/ketone compound production in the fallen leaves. We further show th, Also funded by a Utrecht NetworkYoung Researchers grant to M. Pucetaite

Premise of research. Geochemical fingerprinting of fossil plants is a relatively new research field complementing morphological analyses and providing information for paleoenvironmental interpretations. Ginkgoales contains a single extant species but was diverse through the Mesozoic and is an excellent target for biochemical analyses. Methodology. Cuticles derived from fresh and fallen autumn leaves of extant Ginkgo biloba and seven fossil gink- goalean leaf taxa, one seed fern taxon, and two taxa with bennettitalean affinity were analyzed by infrared (IR) microspec- troscopy at the D7 beamline in the MAX IV synchrotron laboratory, Sweden. The fossil material derives from Triassic and Jurassic successions of Greenland. Spectral data sets were compared and evaluated by hierarchical cluster analysis (HCA) and principal component analysis performed on vector-normalized, first-derivative IR absorption spectra. Pivotal results. The IR absorption spectra of the fossil leaves all reveal signatures that clearly indicate the pres- ence of organic compounds. Spectra of the extant G. biloba leaves reveal the presence of aliphatic chains, aromatic and ester carbonyl functional groups from polymer cutin and other waxy compounds, and polysaccharides. Inter- estingly, both the extant autumn leaves and the fossil specimens reveal the presence of carboxyl/ketone molecules, suggesting that chemical alterations during the initial stages of decomposition are preserved through fossilization. Two major subclusters were identified through HCA of the fossil spectra. Conclusions. Consistent chemical IR signatures, specific for each fossil taxon are present in cuticles, and suf- ficient molecular content is preserved in key regions to reflect the plants’ original chemical signatures. The alter- ations of the organic compounds are initiated as soon as the leaves are shed, with loss of proteins and increased ester and carboxyl/ketone compound production in the fallen leaves. We further show th, Also funded by a Utrecht NetworkYoung Researchers grant to M. Pucetaite

Premise of research. Geochemical fingerprinting of fossil plants is a relatively new research field complementing morphological analyses and providing information for paleoenvironmental interpretations. Ginkgoales contains a single extant species but was diverse through the Mesozoic and is an excellent target for biochemical analyses. Methodology. Cuticles derived from fresh and fallen autumn leaves of extant Ginkgo biloba and seven fossil ginkgoalean leaf taxa, one seed fern taxon, and two taxa with bennettitalean affinity were analyzed by infrared (IR) microspectroscopy at the D7 beamline in the MAX IV synchrotron laboratory, Sweden. The fossil material derives from Triassic and Jurassic successions of Greenland. Spectral data sets were compared and evaluated by hierarchical cluster analysis (HCA) and principal component analysis performed on vector-normalized, first-derivative IR absorption spectra. Pivotal results. The IR absorption spectra of the fossil leaves all reveal signatures that clearly indicate the presence of organic compounds. Spectra of the extant G. biloba leaves reveal the presence of aliphatic chains, aromatic and ester carbonyl functional groups from polymer cutin and other waxy compounds, and polysaccharides. Interestingly, both the extant autumn leaves and the fossil specimens reveal the presence of carboxyl/ketone molecules, suggesting that chemical alterations during the initial stages of decomposition are preserved through fossilization. Two major subclusters were identified through HCA of the fossil spectra. Conclusions. Consistent chemical IR signatures, specific for each fossil taxon are present in cuticles, and sufficient molecular content is preserved in key regions to reflect the plants' original chemical signatures. The alterations of the organic compounds are initiated as soon as the leaves are shed, with loss of proteins and increased ester and carboxyl/ketone compound production in the fallen leaves. We further show that the group

Fungi are rich in complexes of cryptic species that need a combination of different approaches to be delimited, including genomic information. Beauveria (Cordycipitaceae, Hypocreales) is a well-known genus of entomopathogenic fungi, used as a biocontrol agent. In this study we present a polyphasic taxonomy regarding two widely distributed complexes of Beauveria: B. asiatica and B. bassiana s.lat. Some of the genetic groups as previously detected within both taxa were either confirmed or fused using population genomics. High levels of divergence were found between two clades in B. asiatica and among three clades in B. bassiana, supporting their subdivision as distinct species. Morphological examination focusing on the width and the length of phialides and conidia showed no difference among the clades within B. bassiana while conidial length was significantly different among clades within B. asiatica. The secondary metabolite profiles obtained by liquid chromatography-mass spectrometry (LC-MS) allowed a distinction between B. asiatica and B. bassiana, but not between the clades therein. Based on these genomic, morphological, chemical data, we proposed a clade of B. asiatica as a new species, named B. thailandica, and two clades of B. bassiana to respectively represent B. namnaoensis and B. neobassiana spp. nov. Such closely related but divergent species with different host ranges have potential to elucidate the evolution of host specificity, with potential biocontrol application.

The class Alphaproteobacteria is comprised of a diverse assemblage of Gram-negative bacteria that includes organisms of varying morphologies, physiologies and habitat preferences many of which are of clinical and ecological importance. Alphaproteobacteria classification has proved to be difficult, not least when taxonomic decisions rested heavily on a limited number of phenotypic features and interpretation of poorly resolved 16S rRNA gene trees. Despite progress in recent years regarding the classification of bacteria assigned to the class, there remains a need to further clarify taxonomic relationships. Here, draft genome sequences of a collection of genomes of more than 1000 Alphaproteobacteria and outgroup type strains were used to infer phylogenetic trees from genome-scale data using the principles drawn from phylogenetic systematics. The majority of taxa were found to be monophyletic but several orders, families and genera, including taxa recognized as problematic long ago but also quite recent taxa, as well as a few species were shown to be in need of revision. According proposals are made for the recognition of new orders, families and genera, as well as the transfer of a variety of species to other genera and of a variety of genera to other families. In addition, emended descriptions are given for many species mainly involving information on DNA G+C content and (approximate) genome size, both of which are confirmed as valuable taxonomic markers. Similarly, analysis of the gene content was shown to provide valuable taxonomic insights in the class. Significant incongruities between 16S rRNA gene and whole genome trees were not found in the class. The incongruities that became obvious when comparing the results of the present study with existing classifications appeared to be caused mainly by insufficiently resolved 16S rRNA gene trees or incomplete taxon sampling. Another probable cause of misclassifications in the past is the partially low overall fit of phe

Aspergillus burnettii is a new species belonging to the A. alliaceus clade in Aspergillus subgenus Circumdati section Flavi isolated from peanut-growing properties in southern Queensland, Australia. A. burnettii is a fast-growing, floccose fungus with distinctive brown conidia and is a talented producer of biomass-degrading enzymes and secondary metabolites. Chemical profiling of A. burnettii revealed the metabolites ochratoxin A, kotanins, isokotanins, asperlicin E, anominine and paspalinine, which are common to subgenus Circumdati, together with burnettiene A, burnettramic acids, burnettides, and high levels of 14α-hydroxypaspalinine and hirsutide. The genome of A. burnettii was sequenced and an annotated draft genome is presented. A. burnettii is rich in secondary metabolite biosynthetic gene clusters, containing 51 polyketide synthases, 28 non-ribosomal peptide synthetases and 19 genes related to terpene biosynthesis. Functional annotation of digestive enzymes of A. burnettii and A. alliaceus revealed overlapping carbon utilisation profiles, consistent with a close phylogenetic relationship.

Mycological investigation of various foods (mainly cowpea, groundnut, maize, rice, sorghum) and agricultural soils from two states in north-central Nigeria (Nasarawa and Niger), was conducted in order to understand the role of filamentous fungi in food contamination and public health. A total of 839 fungal isolates were recovered from 84% of the 250 food and all 30 soil samples. Preliminary identifications were made, based on macro- and micromorphological characters. Representative strains (n = 121) were studied in detail using morphology and DNA sequencing, involving genera/species-specific markers, while extrolite profiles using LC-MS/MS were obtained for a selection of strains. The representative strains grouped in seven genera (Aspergillus, Fusarium, Macrophomina, Meyerozyma, Neocosmospora, Neotestudina and Phoma). Amongst the 21 species that were isolated during this study was one novel species belonging to the Fusarium fujikuroi species complex, F. madaense sp. nov., obtained from groundnut and sorghum in Nasarawa state. The examined strains produced diverse extrolites, including several uncommon compounds: averantinmethylether in A. aflatoxiformans; aspergillimide in A. flavus; heptelidic acid in A. austwickii; desoxypaxillin, kotanin A and paspalitrems (A and B) in A. aflatoxiformans, A. austwickii and A. cerealis; aurasperon C, dimethylsulochrin, fellutanine A, methylorsellinic acid, nigragillin and pyrophen in A. brunneoviolaceus; cyclosporins (A, B, C and H) in A. niger; methylorsellinic acid, pyrophen and secalonic acid in A. piperis; aspulvinone E, fonsecin, kojic acid, kotanin A, malformin C, pyranonigrin and pyrophen in A. vadensis; and all compounds in F. madaense sp. nov., Meyerozyma, Neocosmospora and Neotestudina. This study provides snapshot data for prediction of food contamination and fungal biodiversity exploitation.

The class Alphaproteobacteria is comprised of a diverse assemblage of Gram-negative bacteria that includes organisms of varying morphologies, physiologies and habitat preferences many of which are of clinical and ecological importance. Alphaproteobacteria classification has proved to be difficult, not least when taxonomic decisions rested heavily on a limited number of phenotypic features and interpretation of poorly resolved 16S rRNA gene trees. Despite progress in recent years regarding the classification of bacteria assigned to the class, there remains a need to further clarify taxonomic relationships. Here, draft genome sequences of a collection of genomes of more than 1000 Alphaproteobacteria and outgroup type strains were used to infer phylogenetic trees from genome-scale data using the principles drawn from phylogenetic systematics. The majority of taxa were found to be monophyletic but several orders, families and genera, including taxa recognized as problematic long ago but also quite recent taxa, as well as a few species were shown to be in need of revision. According proposals are made for the recognition of new orders, families and genera, as well as the transfer of a variety of species to other genera and of a variety of genera to other families. In addition, emended descriptions are given for many species mainly involving information on DNA G+C content and (approximate) genome size, both of which are confirmed as valuable taxonomic markers. Similarly, analysis of the gene content was shown to provide valuable taxonomic insights in the class. Significant incongruities between 16S rRNA gene and whole genome trees were not found in the class. The incongruities that became obvious when comparing the results of the present study with existing classifications appeared to be caused mainly by insufficiently resolved 16S rRNA gene trees or incomplete taxon sampling. Another probable cause of misclassifications in the past is the partially low overall fit of phe

Mycological investigation of various foods (mainly cowpea, groundnut, maize, rice, sorghum) and agricultural soils from two states in north-central Nigeria (Nasarawa and Niger), was conducted in order to understand the role of filamentous fungi in food contamination and public health. A total of 839 fungal isolates were recovered from 84% of the 250 food and all 30 soil samples. Preliminary identifications were made, based on macro- and micromorphological characters. Representative strains (n = 121) were studied in detail using morphology and DNA sequencing, involving genera/species-specific markers, while extrolite profiles using LC-MS/MS were obtained for a selection of strains. The representative strains grouped in seven genera (Aspergillus, Fusarium, Macrophomina, Meyerozyma, Neocosmospora, Neotestudina and Phoma). Amongst the 21 species that were isolated during this study was one novel species belonging to the Fusarium fujikuroi species complex, F. madaense sp. nov., obtained from groundnut and sorghum in Nasarawa state. The examined strains produced diverse extrolites, including several uncommon compounds: averantinmethylether in A. aflatoxiformans; aspergillimide in A. flavus; heptelidic acid in A. austwickii; desoxypaxillin, kotanin A and paspalitrems (A and B) in A. aflatoxiformans, A. austwickii and A. cerealis; aurasperon C, dimethylsulochrin, fellutanine A, methylorsellinic acid, nigragillin and pyrophen in A. brunneoviolaceus; cyclosporins (A, B, C and H) in A. niger; methylorsellinic acid, pyrophen and secalonic acid in A. piperis; aspulvinone E, fonsecin, kojic acid, kotanin A, malformin C, pyranonigrin and pyrophen in A. vadensis; and all compounds in F. madaense sp. nov., Meyerozyma, Neocosmospora and Neotestudina. This study provides snapshot data for prediction of food contamination and fungal biodiversity exploitation.

Metabolomika je nova naučna multidisciplinarna oblast koja obuhvata različiteaspekte biologije, hemije i matematike. Koristi moderne spektroskopske ihromatografske tehnike (NMR, IC, UV, MS, GC, LC) i multivarijantnu statističkuanalizu podataka sa ciljem da se izmere kvantitativno i kvalitativno svi metaboliti uispitivanom organizmu i na taj način dobije jasna metabolomička slika pod ispitivanimuslovima. Primena metabolomike nije ograničena samo na biljni svet, već se možeprimeniti na sve žive organizme, pa tako i na ljude. Posebno je značajna mogućnostprimene u medicini kao dijagnostičke metode ili u farmakognoziji za otkrivanje novihlekovitih sirovina. U ovoj disertaciji primena metabolomike na endemski biljni rodAmphoricarpos Vis. imala je dva cilja: (a) sticanje novih saznanja o mogućnostiprimene dobijenih rezultata u diferencijaciji balkanskih vrsta ovog roda i (b) razvojmetode za brzo otkrivanje novih farmakološki aktivnih sastojaka u biljnom materijalu.Vrste roda Amphoricarpos Vis. rasprostranjene su na Balkanskom poluostrvu.Anatoliji i Kavkazu. Da bi se dobilo više uvida u taksonomski status balkanskih taksonaovog roda, koji nije do kraja raščišćen, primenjena je metabolomička studija. Po Blečićui Mayeru1 postoje tri taksona: dve vrste A. neumayeri Vis. i A. autariatus Blečić etMayer, koja se deli na dve podvrste - ssp. autariatus i ssp. bertisceus Blečić et Mayer. Sdruge strane, Webb2 priznaje samo jednu vrstu - A. neumayeri Vis., koja sadrži dvepodvrste - ssp. neumayeri i ssp. murbeckii Bošnjak. U novije vreme, na osnovuispitivanja genoma vrsta ovog roda predložena je samo jedna vrsta, А. neumayerianus(Vis.) Greuter.3 U ovom radu analizirana su sva tri taksona (prema Blečiću i Mayeru), auzorci su sakupljeni na različitim lokalitetima u Crnoj Gori u fazi cvetanja. Primenjenaje 1H NMR spektroskopija kombinovana sa multivarijantnom analizom podataka.Pokazano je da je OPLS-DA (engl. Orthogonal Partial Least Squares-DiscriminantAnalysis) najefikasniji metod za d, Metabolomics is a new scientific multidisciplinary field that includes variousaspects of biology, chemistry and mathematics. It uses modern spectroscopic andchromatographic techniques (NMR, IC, UV, MS, GC, LC) and multivariate statisticaldata analysis to measure quantitatively and qualitatively all of the metabolites in the testorganism and thereby obtain a clear metabolic profile under test conditions. The use ofmetabolomics is not limited to the plant world, but can be applied to all livingorganisms as well as humans. A particularly important possibility is the application inmedicine as a diagnostic method or in pharmacognosy for the discovery of new naturalsources of drugs. In this thesis, the application of metabolomics to the endemic plantgenus Amphoricarpos Vis. was applied with two objectives: (a) acquiring newknowledge of the systematization of the genus, and (b) developing a method for rapiddetection of new pharmacologically active ingredients in plant material.Genus Amphoricarpos Vis. is an endemic of the western part of the BalkanPeninsula. In order to gain more insight into the taxonomic status of the genus that isstill ambiguous, a metabolomic study was applied. Blečić and Mayer1 proposed threetaxa: two species A. neumayeri Vis. and A. autariatus Blečić et Mayer, the latter dividedinto two subspecies, ssp. autariatus i ssp. bertisceus Blečič et Mayer. On the other hand,Webb2 recognizeed only one species, A. neumayeri Vis. divided into two subspecies,ssp. neumayeri i ssp. murbeckii Bošnjak. Recently, on the basis of a genomeexamination of the taxa from the West Balkans, only one species was proposed, A.neumayerianus (Vis.) Greuter.3In this dissertation all three taxa (according to Blečić and Mayer)1 were analysed. Thesamples were collected at different locations in Montenegro at the stage of flowering.1H NMR spectroscopy combined with multivariate data analysis was applied.Orthogonal Partial Least Squares-Discriminant Analysis (OPLS-DA) has been shown to

Metabolomika je nova naučna multidisciplinarna oblast koja obuhvata različiteaspekte biologije, hemije i matematike. Koristi moderne spektroskopske ihromatografske tehnike (NMR, IC, UV, MS, GC, LC) i multivarijantnu statističkuanalizu podataka sa ciljem da se izmere kvantitativno i kvalitativno svi metaboliti uispitivanom organizmu i na taj način dobije jasna metabolomička slika pod ispitivanimuslovima. Primena metabolomike nije ograničena samo na biljni svet, već se možeprimeniti na sve žive organizme, pa tako i na ljude. Posebno je značajna mogućnostprimene u medicini kao dijagnostičke metode ili u farmakognoziji za otkrivanje novihlekovitih sirovina. U ovoj disertaciji primena metabolomike na endemski biljni rodAmphoricarpos Vis. imala je dva cilja: (a) sticanje novih saznanja o mogućnostiprimene dobijenih rezultata u diferencijaciji balkanskih vrsta ovog roda i (b) razvojmetode za brzo otkrivanje novih farmakološki aktivnih sastojaka u biljnom materijalu.Vrste roda Amphoricarpos Vis. rasprostranjene su na Balkanskom poluostrvu.Anatoliji i Kavkazu. Da bi se dobilo više uvida u taksonomski status balkanskih taksonaovog roda, koji nije do kraja raščišćen, primenjena je metabolomička studija. Po Blečićui Mayeru1 postoje tri taksona: dve vrste A. neumayeri Vis. i A. autariatus Blečić etMayer, koja se deli na dve podvrste - ssp. autariatus i ssp. bertisceus Blečić et Mayer. Sdruge strane, Webb2 priznaje samo jednu vrstu - A. neumayeri Vis., koja sadrži dvepodvrste - ssp. neumayeri i ssp. murbeckii Bošnjak. U novije vreme, na osnovuispitivanja genoma vrsta ovog roda predložena je samo jedna vrsta, А. neumayerianus(Vis.) Greuter.3 U ovom radu analizirana su sva tri taksona (prema Blečiću i Mayeru), auzorci su sakupljeni na različitim lokalitetima u Crnoj Gori u fazi cvetanja. Primenjenaje 1H NMR spektroskopija kombinovana sa multivarijantnom analizom podataka.Pokazano je da je OPLS-DA (engl. Orthogonal Partial Least Squares-DiscriminantAnalysis) najefikasniji metod za d, Metabolomics is a new scientific multidisciplinary field that includes variousaspects of biology, chemistry and mathematics. It uses modern spectroscopic andchromatographic techniques (NMR, IC, UV, MS, GC, LC) and multivariate statisticaldata analysis to measure quantitatively and qualitatively all of the metabolites in the testorganism and thereby obtain a clear metabolic profile under test conditions. The use ofmetabolomics is not limited to the plant world, but can be applied to all livingorganisms as well as humans. A particularly important possibility is the application inmedicine as a diagnostic method or in pharmacognosy for the discovery of new naturalsources of drugs. In this thesis, the application of metabolomics to the endemic plantgenus Amphoricarpos Vis. was applied with two objectives: (a) acquiring newknowledge of the systematization of the genus, and (b) developing a method for rapiddetection of new pharmacologically active ingredients in plant material.Genus Amphoricarpos Vis. is an endemic of the western part of the BalkanPeninsula. In order to gain more insight into the taxonomic status of the genus that isstill ambiguous, a metabolomic study was applied. Blečić and Mayer1 proposed threetaxa: two species A. neumayeri Vis. and A. autariatus Blečić et Mayer, the latter dividedinto two subspecies, ssp. autariatus i ssp. bertisceus Blečič et Mayer. On the other hand,Webb2 recognizeed only one species, A. neumayeri Vis. divided into two subspecies,ssp. neumayeri i ssp. murbeckii Bošnjak. Recently, on the basis of a genomeexamination of the taxa from the West Balkans, only one species was proposed, A.neumayerianus (Vis.) Greuter.3In this dissertation all three taxa (according to Blečić and Mayer)1 were analysed. Thesamples were collected at different locations in Montenegro at the stage of flowering.1H NMR spectroscopy combined with multivariate data analysis was applied.Orthogonal Partial Least Squares-Discriminant Analysis (OPLS-DA) has been shown to

Metabolomika je nova naučna multidisciplinarna oblast koja obuhvata različite aspekte biologije, hemije i matematike. Koristi moderne spektroskopske i hromatografske tehnike (NMR, IC, UV, MS, GC, LC) i multivarijantnu statističku analizu podataka sa ciljem da se izmere kvantitativno i kvalitativno svi metaboliti u ispitivanom organizmu i na taj način dobije jasna metabolomička slika pod ispitivanim uslovima. Primena metabolomike nije ograničena samo na biljni svet, već se može primeniti na sve žive organizme, pa tako i na ljude. Posebno je značajna mogućnost primene u medicini kao dijagnostičke metode ili u farmakognoziji za otkrivanje novih lekovitih sirovina. U ovoj disertaciji primena metabolomike na endemski biljni rod Amphoricarpos Vis. imala je dva cilja: (a) sticanje novih saznanja o mogućnosti primene dobijenih rezultata u diferencijaciji balkanskih vrsta ovog roda i (b) razvoj metode za brzo otkrivanje novih farmakološki aktivnih sastojaka u biljnom materijalu. Vrste roda Amphoricarpos Vis. rasprostranjene su na Balkanskom poluostrvu. Anatoliji i Kavkazu. Da bi se dobilo više uvida u taksonomski status balkanskih taksona ovog roda, koji nije do kraja raščišćen, primenjena je metabolomička studija. Po Blečiću i Mayeru1 postoje tri taksona: dve vrste A. neumayeri Vis. i A. autariatus Blečić et Mayer, koja se deli na dve podvrste - ssp. autariatus i ssp. bertisceus Blečić et Mayer. S druge strane, Webb2 priznaje samo jednu vrstu - A. neumayeri Vis., koja sadrži dve podvrste - ssp. neumayeri i ssp. murbeckii Bošnjak. U novije vreme, na osnovu ispitivanja genoma vrsta ovog roda predložena je samo jedna vrsta, А. neumayerianus (Vis.) Greuter.3 U ovom radu analizirana su sva tri taksona (prema Blečiću i Mayeru), a uzorci su sakupljeni na različitim lokalitetima u Crnoj Gori u fazi cvetanja. Primenjena je 1H NMR spektroskopija kombinovana sa multivarijantnom analizom podataka. Pokazano je da je OPLS-DA (engl. Orthogonal Partial Least Squares-Discriminant Analysis), Metabolomics is a new scientific multidisciplinary field that includes various aspects of biology, chemistry and mathematics. It uses modern spectroscopic and chromatographic techniques (NMR, IC, UV, MS, GC, LC) and multivariate statistical data analysis to measure quantitatively and qualitatively all of the metabolites in the test organism and thereby obtain a clear metabolic profile under test conditions. The use of metabolomics is not limited to the plant world, but can be applied to all living organisms as well as humans. A particularly important possibility is the application in medicine as a diagnostic method or in pharmacognosy for the discovery of new natural sources of drugs. In this thesis, the application of metabolomics to the endemic plant genus Amphoricarpos Vis. was applied with two objectives: (a) acquiring new knowledge of the systematization of the genus, and (b) developing a method for rapid detection of new pharmacologically active ingredients in plant material. Genus Amphoricarpos Vis. is an endemic of the western part of the Balkan Peninsula. In order to gain more insight into the taxonomic status of the genus that is still ambiguous, a metabolomic study was applied. Blečić and Mayer1 proposed three taxa: two species A. neumayeri Vis. and A. autariatus Blečić et Mayer, the latter divided into two subspecies, ssp. autariatus i ssp. bertisceus Blečič et Mayer. On the other hand, Webb2 recognizeed only one species, A. neumayeri Vis. divided into two subspecies, ssp. neumayeri i ssp. murbeckii Bošnjak. Recently, on the basis of a genome examination of the taxa from the West Balkans, only one species was proposed, A. neumayerianus (Vis.) Greuter.3 In this dissertation all three taxa (according to Blečić and Mayer)1 were analysed. The samples were collected at different locations in Montenegro at the stage of flowering. 1H NMR spectroscopy combined with multivariate data analysis was applied. Orthogonal Partial Least Squares-Discriminant Analysis (OP

Fossil resin (amber) has been recently reported as common, but small, sedimentary components throughout thelower Upper Cretaceous (Cenomanian; 99–94 Ma) strata of the Tupuangi Formation, Chatham Islands, easternZealandia. From these deposits, resin has also been identified and obtained from well-preserved, coalified specimensof the conifer fossil Protodammara reimatamorioriMays and Cantrill, 2018. Here, we employed attenuatedtotal reflectance Fourier-transform infrared spectroscopy (ATR-FTIR) to both dispersed and in situ amber specimens.These resulted in very similar chemical signatures, indicating that these fossils are likely from the same orclosely-related botanical sources. The FTIR data are typical of a conifer source within the ‘cupressaceous resins’category of Tappert et al. (2011). Carbon-13 nuclear magnetic resonance spectroscopy (13C NMR) facilitatedthe probable identification of these ambers as ‘Class Ib' (sensu Anderson et al. 1992). Based on these spectraldata sets, the likely botanical sources of the amber were either Araucariaceae or Cupressaceae; both of these coniferfamilies were common and widespread in the Southern Hemisphere during the Cretaceous. However, themorphology and anatomy of P. reimatamoriori support an affinity to the latter family, thus indicating that the Cretaceousamber of the Chatham Islands was generally produced by members of the Cupressaceae. Comparing theFTIR data to the published spectra of modern resins, we also identify a band ratio which may aid in distinguishingbetween the FTIR spectra of Araucariaceae and Cupressaceae, and outline the limitations to this approach. A highconcentration of ester bonds in Chatham amber specimens, which exceeds typical Cupressaceae resins, is probablycaused by taphonomic alteration via thermal maturation. The source of thermal alteration was likely preburialwildfires,conditions forwhich P. reimatamoriori was adapted to as part of its life cycle. A comparison of ambersof the Chatham Islands wi, Fieldwork and research supported by National Geographic Society (grant 9761-15) awarded to C.M.; additional financial support provided by the Paleontological Society and Monash University.

Fossil resin (amber) has been recently reported as common, but small, sedimentary components throughout thelower Upper Cretaceous (Cenomanian; 99–94 Ma) strata of the Tupuangi Formation, Chatham Islands, easternZealandia. From these deposits, resin has also been identified and obtained from well-preserved, coalified specimensof the conifer fossil Protodammara reimatamorioriMays and Cantrill, 2018. Here, we employed attenuatedtotal reflectance Fourier-transform infrared spectroscopy (ATR-FTIR) to both dispersed and in situ amber specimens.These resulted in very similar chemical signatures, indicating that these fossils are likely from the same orclosely-related botanical sources. The FTIR data are typical of a conifer source within the ‘cupressaceous resins’category of Tappert et al. (2011). Carbon-13 nuclear magnetic resonance spectroscopy (13C NMR) facilitatedthe probable identification of these ambers as ‘Class Ib' (sensu Anderson et al. 1992). Based on these spectraldata sets, the likely botanical sources of the amber were either Araucariaceae or Cupressaceae; both of these coniferfamilies were common and widespread in the Southern Hemisphere during the Cretaceous. However, themorphology and anatomy of P. reimatamoriori support an affinity to the latter family, thus indicating that the Cretaceousamber of the Chatham Islands was generally produced by members of the Cupressaceae. Comparing theFTIR data to the published spectra of modern resins, we also identify a band ratio which may aid in distinguishingbetween the FTIR spectra of Araucariaceae and Cupressaceae, and outline the limitations to this approach. A highconcentration of ester bonds in Chatham amber specimens, which exceeds typical Cupressaceae resins, is probablycaused by taphonomic alteration via thermal maturation. The source of thermal alteration was likely preburialwildfires,conditions forwhich P. reimatamoriori was adapted to as part of its life cycle. A comparison of ambersof the Chatham Islands wi, Fieldwork and research supported by National Geographic Society (grant 9761-15) awarded to C.M.; additional financial support provided by the Paleontological Society and Monash University.

Volatile terpenoid resins represent a diverse group of plant defense chemicals involved in defense against herbivory, abiotic stress, and communication. However, their composition in tropical forests remains poorly characterized. As a part of tree identification, the 'smell' of damaged trunks is widely used, but is highly subjective. Here, we analyzed trunk volatile monoterpene emissions from 15 species of the genus Protium in the central Amazon. By normalizing the abundances of 28 monoterpenes, 9 monoterpene 'fingerprint' patterns emerged, characterized by a distinct dominant monoterpene. While 4 of the 'fingerprint' patterns were composed of multiple species, 5 were composed of a single species. Moreover, among individuals of the same species, 6 species had a single 'fingerprint' pattern, while 9 species had two or more 'fingerprint' patterns among individuals. A comparison of 'fingerprints' between 2015 and 2017 from 15 individuals generally showed excellent agreement, demonstrating a strong dependence on species identity, but not time of collection. The results are consistent with a previous study that found multiple divergent copies of monoterpene synthase enzymes in Protium. We conclude that the monoterpene 'fingerprint' database has important implications for constraining Protium species identification and phylogenetic relationships and enhancing understanding of physiological and ecological functions of resins and their potential commercial applications.

Although considerable progress has been made in recent years regarding the classification of bacteria assigned to the phylum Bacteroidetes, there remains a need to further clarify taxonomic relationships within a diverse assemblage that includes organisms of clinical, piscicultural, and ecological importance. Bacteroidetes classification has proved to be difficult, not least when taxonomic decisions rested heavily on interpretation of poorly resolved 16S rRNA gene trees and a limited number of phenotypic features. Here, draft genome sequences of a greatly enlarged collection of genomes of more than 1,000 Bacteroidetes and outgroup type strains were used to infer phylogenetic trees from genome-scale data using the principles drawn from phylogenetic systematics. The majority of taxa were found to be monophyletic but several orders, families and genera, including taxa proposed long ago such as Bacteroides, Cytophaga, and Flavobacterium but also quite recent taxa, as well as a few species were shown to be in need of revision. According proposals are made for the recognition of new orders, families and genera, as well as the transfer of a variety of species to other genera. In addition, emended descriptions are given for many species mainly involving information on DNA G+C content and (approximate) genome size, both of which can be considered valuable taxonomic markers. We detected many incongruities when comparing the results of the present study with existing classifications, which appear to be caused by insufficiently resolved 16S rRNA gene trees or incomplete taxon sampling. The few significant incongruities found between 16S rRNA gene and whole genome trees underline the pitfalls inherent in phylogenies based upon single gene sequences and the impediment in using ordinary bootstrapping in phylogenomic studies, particularly when combined with too narrow gene selections. While a significant degree of phylogenetic conservation was detected in all phenotypic characters

Although considerable progress has been made in recent years regarding the classification of bacteria assigned to the phylum Bacteroidetes, there remains a need to further clarify taxonomic relationships within a diverse assemblage that includes organisms of clinical, piscicultural, and ecological importance. Bacteroidetes classification has proved to be difficult, not least when taxonomic decisions rested heavily on interpretation of poorly resolved 16S rRNA gene trees and a limited number of phenotypic features. Here, draft genome sequences of a greatly enlarged collection of genomes of more than 1,000 Bacteroidetes and outgroup type strains were used to infer phylogenetic trees from genome-scale data using the principles drawn from phylogenetic systematics. The majority of taxa were found to be monophyletic but several orders, families and genera, including taxa proposed long ago such as Bacteroides, Cytophaga, and Flavobacterium but also quite recent taxa, as well as a few species were shown to be in need of revision. According proposals are made for the recognition of new orders, families and genera, as well as the transfer of a variety of species to other genera. In addition, emended descriptions are given for many species mainly involving information on DNA G+C content and (approximate) genome size, both of which can be considered valuable taxonomic markers. We detected many incongruities when comparing the results of the present study with existing classifications, which appear to be caused by insufficiently resolved 16S rRNA gene trees or incomplete taxon sampling. The few significant incongruities found between 16S rRNA gene and whole genome trees underline the pitfalls inherent in phylogenies based upon single gene sequences and the impediment in using ordinary bootstrapping in phylogenomic studies, particularly when combined with too narrow gene selections. While a significant degree of phylogenetic conservation was detected in all phenotypic characters

Volatile terpenoid resins represent a diverse group of plant defense chemicals involved in defense against herbivory, abiotic stress, and communication. However, their composition in tropical forests remains poorly characterized. As a part of tree identification, the 'smell' of damaged trunks is widely used, but is highly subjective. Here, we analyzed trunk volatile monoterpene emissions from 15 species of the genus Protium in the central Amazon. By normalizing the abundances of 28 monoterpenes, 9 monoterpene 'fingerprint' patterns emerged, characterized by a distinct dominant monoterpene. While 4 of the 'fingerprint' patterns were composed of multiple species, 5 were composed of a single species. Moreover, among individuals of the same species, 6 species had a single 'fingerprint' pattern, while 9 species had two or more 'fingerprint' patterns among individuals. A comparison of 'fingerprints' between 2015 and 2017 from 15 individuals generally showed excellent agreement, demonstrating a strong dependence on species identity, but not time of collection. The results are consistent with a previous study that found multiple divergent copies of monoterpene synthase enzymes in Protium. We conclude that the monoterpene 'fingerprint' database has important implications for constraining Protium species identification and phylogenetic relationships and enhancing understanding of physiological and ecological functions of resins and their potential commercial applications.

The relationships between the structure of the phytoplankton community and the bio-optical properties of surface waters were studied during the TransPEGASO cruise along a transect across the Atlantic Ocean that covered seven biogeographical provinces, from the Alborán Sea (SW Mediterranean) to the Patagonian Shelf. We characterized the composition of the phytoplankton community by means of high-performance liquid chromatography and CHEMTAX pigment analyses applied to whole water and two filtration size classes (<3 and ≥3 µm), flow cytometric determinations and microscopic observations. Additionally, the study was complemented by measurements of the absorption of particulate matter and coloured dissolved organic matter (CDOM). The size class distribution of the chlorophyll a (Chl a) obtained from the size-fractionated filtration (SFF) was compared with that resulting from the diagnostic pigment algorithms (VU) developed by Vidussi et al. (2001) and Uitz et al. (2006), and the total Chl a–based expressions (HI) of Hirata et al. (2011). The seven provinces crossed by the transect could be divided into an oligotrophic group with Chl a<0.25 mg m–3 comprising the tropical and subtropical Atlantic (including the Canary Current Coastal Province), and a eutrophic group (Chl a >0.5 mg m–3) with a single Mediterranean (MEDI) sample and those from the southwestern Atlantic Shelf (SWAS). According to CHEMTAX, the most important taxa in the tropical and subtropical Atlantic were Prochlorococcus, haptophytes and Synechoccoccus, while the MEDI and SWAS were dominated by diatoms and haptophytes. Both the VU and HI algorithms, which are based on pigment composition or Chl a concentration, predicted for SWAS a high proportion of nano- and microphytoplankton, while the SFF indicated dominance of the <3 µm size class. In addition, the CHEMTAX results indicated a high average diatom contribution in this province. However, at several SWAS stations with relatively high values of diatom Chl

[EN] The relationships between the structure of the phytoplankton community and the bio-optical properties of surface waters were studied during the TransPEGASO cruise along a transect across the Atlantic Ocean that covered seven biogeographical provinces, from the Alborán Sea (SW Mediterranean) to the Patagonian Shelf. We characterized the composition of the phytoplankton community by means of high-performance liquid chromatography and CHEMTAX pigment analyses applied to whole water and two filtration size classes (<3 and ≥3 µm), flow cytometric determinations and microscopic observations. Additionally, the study was complemented by measurements of the absorption of particulate matter and coloured dissolved organic matter (CDOM). The size class distribution of the chlorophyll a (Chl a) obtained from the size-fractionated filtration (SFF) was compared with that resulting from the diagnostic pigment algorithms (VU) developed by Vidussi et al. (2001) and Uitz et al. (2006), and the total Chl a–based expressions (HI) of Hirata et al. (2011). The seven provinces crossed by the transect could be divided into an oligotrophic group with Chl a<0.25 mg m–3 comprising the tropical and subtropical Atlantic (including the Canary Current Coastal Province), and a eutrophic group (Chl a >0.5 mg m–3) with a single Mediterranean (MEDI) sample and those from the southwestern Atlantic Shelf (SWAS). According to CHEMTAX, the most important taxa in the tropical and subtropical Atlantic were Prochlorococcus, haptophytes and Synechoccoccus, while the MEDI and SWAS were dominated by diatoms and haptophytes. Both the VU and HI algorithms, which are based on pigment composition or Chl a concentration, predicted for SWAS a high proportion of nano- and microphytoplankton, while the SFF indicated dominance of the <3 µm size class. In addition, the CHEMTAX results indicated a high average diatom contribution in this province. However, at several SWAS stations with relatively high values of diato, [ES] Durante la campaña TransPEGASO, realizada a lo largo de un transecto a través del Océano Atlántico que cubrió siete provincias biogeográficas, desde el mar de Alborán (Mediterráneo SO) hasta la Plataforma Patagónica, se estudiaron las relaciones entre la estructura de la comunidad fitoplanctónica y las propiedades bio-ópticas del agua. La composición del fitoplancton en muestras de agua entera y de dos fracciones de tamaño (<3 y ≥3 µm) obtenidas por filtración se caracterizó por medio de análisis de pigmentos por HPLC (de high-performance liquid chromatographyy), seguido de la aplicación del algoritmo CHEMTAX. Además, se llevaron a cabo determinaciones de citometría de flujo y observaciones microscópicas, y el estudio se complementó con mediciones de absorción de material particulado y materia orgánica disuelta coloreada (CDOM, de coloured dissolved organic matter). La distribución de la clorofila a (Chl a) entre las diversas clases de tamaño obtenidas mediante filtración fraccionada (SFF, de size-fractionated filtration) se comparó con las distribuciones derivadas de los algoritmos desarrollados por Vidussi et al. (2001) y Uitz et al. (2006) (VU), y por Hirata et al. (2011) (HI). Las siete provincias atravesadas por el –3 transecto podían clasificarse en un grupo oligotrófico, con Chl a <0.25 mg m , que comprende el Atlántico tropical y subtropical –3 (incluida la provincia costera de Canarias) y un grupo eutrófico (Chl a >0.5 mg m ) con una sola muestra mediterránea (MEDI) y las de la plataforma patagónica, en el sudoeste del Atlántico (SWAS). Según CHEMTAX, los taxones más importantes en el Atlántico tropical y subtropical fueron Prochlorococcus, haptofitos y Synechoccoccus, mientras que las provincias MEDI y SWAS estuvieron dominadas por diatomeas y haptofitos. Tanto los algoritmos VU como los HI, que se basan en la composición de pigmentos o en la concentración de Chl a, predijeron para SWAS una alta proporción de nano y microfitoplancton, mientras que la, Por otra parte, los resultados de CHEMTAX indicaron que, en promedio, la contribución de las diatomeas era importante en esta provincia. Sin embargo, en varias estaciones de SWAS para las que CHEMTAX estimaba una elevada contribución de diatomeas, las observaciones microscópicas encontraron solamente escasas concentraciones de células de diatomeas de tamaño nano- o microplanctónico. Esta discrepancia parece deberse a la presencia, confirmada por microscopía electrónica de barrido, de pequeñas células (<3 µm) de la diatomea Minidiscus sp. y de Parmales (un grupo que comparte la composición pigmentaria con las diatomeas). Estos hallazgos advierten contra una asignación rutinaria de los pigmentos de las diatomeas a la clase de tamaño de microplancton. La absorción total (sin contar la propia del agua) en la columna de agua estuvo dominada por CDOM. En promedio, la contribución de la absorción de fitoplancton para las diferentes provincias osciló de 19.3% en MEDI a 45.7% en SWAS y 47% en la provincia del Atlántico Tropical 2 –1 Occidental (WTRA). La absorción del fitoplancton por unidad de Chl a [aph*(443), m mg ] fue menor en MEDI y SWAS que en las provincias oligotróficas. aph*(443) se correlacionó negativamente con el primer componente derivado de un análisis de los componentes principales basado en la concentración de los pigmentos más importantes y no se correlacionó con indicadores de la estructura de tamaños de la comunidad fitoplanctónica tales como la proporción de Chl a en la clase <3 µm o un índice de tamaño (SI) derivado de la distribución de clases de tamaño obtenida mediante el algoritmo VU. Estas observaciones indican que la variabilidad observada en aph*(443) se relacionaba principalmente con diferencias en la composición pigmentaria y posiblemente también con procesos de fotoaclimatación del fitoplancton, y que cualquier efecto de empaquetamiento debido al tamaño de las células quedaba probablemente enmascarado por otros factores. Este último resultado

From two species of Sutera (S. foetida and S. cordata) (Scrophulariaceae tribe Limoselleae) were isolated three known secoiridoid glucosides (12–14) as well as four iridoid congeners (8–11), all biosynthetically derived from iridodial glucoside (and/or deoxyloganic acid). In addition, two previously unknown compounds were found, namely a terpenoid glucoside lactone (suterolide, 21) and the phenylethanoid glycoside 2′′′′-O-acetyl-angoroside A (19) as well as verbascoside, echinacoside and tubuloside A(15–17, respectively). Two other species, Jamesbrittenia dissecta and Lyperia antirrhinoides, previously considered to belong to the same genus (Sutera) were shown to be members of two different genera, respectively. Significantly, these two species contained iridoids derived from 8-epi-iridodial (and 8-epideoxyloganic acid), namely aucubin (2), melittoside (3) and acetylharpagide (4). In addition we investigated Melanospermum transvaalense, Lyperia tristis and Microdon dubius likewise from Limoselleae and all of these contained iridoid glucosides from the 8-epi-pathway. Thus, secoiridoid distribution confirms the DNA-based circumscription of Sutera and its sister-group relationship with Manulea. In addition, the results show that the clade including these two genera has a biosynthetic pathway to iridoids fundamentally different from the rest of the tribe and from the whole family Scrophulariaceae.

Harmful blooms formed by planktonic microalgae (HABs) in both freshwater and coastal waters regularly lead to severe mortalities of fish and invertebrates causing substantial economic losses of marine products worldwide. The mixotrophic haptophyte Prymnesium parvum is one of the most important microalgae associated with fish kills. Here 26 strains of P. parvum with a wide geographical distribution were screened for the production of prymnesins, the suspected causative allelochemical toxins. All investigated strains produced prymnesins, indicating that the toxins play an important role for the organism. The prymnesins can be classified into three types based on the length of the carbon backbone of the compound and each algal strain produced only one of these types. Biogeographical mapping of the prymnesin distribution indicated a global distribution of each type. In addition, phylogenetic analyses based on internal transcribed spacer (ITS) sequences revealed monophyletic origin of all prymnesin types and clades could therefore be defined based on the toxic compound. It might be that evolution of new species within the P. parvum species complex is driven by changes in toxin type or that they are a result of it. Such a correlation between chemotype and phylotype has never been documented before for a harmful microalga. Chemotaxonomy and ITS-type classification may thus be used to further delimit the P. parvum species complex.

Цель статьи – провести ревизию всех доступных образцов лишайников группы «Xanthoparmelia pulla», установить их таксономическую принадлежность, изучить распространение и экологию установленных видов в пределах Беларуси. = The aim of the research was to revise all available specimens of lichen group «Xanthoparmelia pulla», to state their taxonomy, distribution and ecology in Belarus.

Ovim istraživanjem obuhvaćene su dve populacije Alnus glutinosa (L.) Gaertn. (Prilički kiseljak i Rimski most) i dve populacije Alnus incana (L.) Moench (Golijska reka i Sastavci) uzorkovane na planini Goliji. Osnovni ciljevi bili su: 1) ispitivanje varijabilnosti i odnosa populacija unutar i između vrsta u odnosu na koncentraciju diarilheptanoida u kori prirodnih populacija A. glutinosa i A. incana; 2) ispitivanje sezonske varijabilnosti diarilheptanoida u kori vrsta A. glutinosa i A. incana; 3) procena hemotaksonomskog značaja diarilheptanoida u ispitivanim vrstama; 4) procena mogućnosti primene diarilheptanoida u detekciji spontanih hibrida između A. glutinosa i A. incana; 5) ispitivanje varijabilnosti oblika, veličine i nervature lista u populacijama A. glutinosa i A. incana; 6) procena spontane hibridizacije između ove dve vrste na osnovu morfologije lista. Koncentracija najzastupljenijih diarilheptanoida (hirsutanonol-5-O-β-D-glukopiranozida, oregonina, hirsutanonola, alnuzida A, platifilozida i rubranozida A) u etanolnim ekstraktima kore ispitivanih populacija određena je pomoću UHPLC-MS/MS. Međupopulaciona varijabilnost koncentracije diarilheptanoida ispitana je pomoću parametarskih (ANOVA) i neparametarskih (Kruskal-Wallis) testova, analize glavne komponente (PCA) i diskriminantne analize (DA). Prvi uvid u kvantitativnu distribuciju odabranih diarilheptanoida kod A. glutinosa i A. incana je procenjen na osnovu analize populacija Rimski most i Sastavci. DA je pokazala 100% diskriminacije između tri grupe individua koje je izdvojila PCA, na osnovu čega je postavljena hipoteza da je jedna grupa od četiri individue hibridogenog porekla. Diarilheptanoidi su se pokazali dobrim hemotaksonomskim markerima na interspecijskom nivou i utvrđena je njihova mogućnost primene u detekciji spontanih hibrida..., In this study, two populations of Alnus glutinosa (L.) Gaertn. (Prilički kiseljak and Rimski most) and two populations of Alnus incana (L.) Moench (Golijska reka and Sastavci) were sampled on mountain Golija with the aims to investigate: 1) variability and relations of populations within and among species based on the concentration of diarylheptanoids in the bark of natural populations of A. glutinosa and A. incana; 2) seasonal variability of diarylheptanoids in the bark of A. glutinosa and A. incana; 3) the reliability of diarylheptanoids as chemotaxonomic markers in the studied species; 4) the reliability of diarylheptanoids for the detection of spontaneous hybrids between A. glutinosa and A. incana; 5) variability of leaf shape, size and venation in the populations of A. glutinosa and A. incana; 6) spontaneous hybridization between the studied species based on leaf morphology. Six diarylheptanoids (hirsutanonol-5-O-β-D-glucopyranoside, oregonin, hirsutanonol, alnuside A, platyphylloside and rubranoside A) were quantified in bark ethanol extracts of four Alnus spp. populations by UHPLC-MS/MS. Interpopulation variability of diarylheptanoid concentration was investigated with ANOVA, Kruskal-Wallis test, principal component analysis (PCA) and discriminant analysis (DA). Quantitative distribution of selected diarylheptanoids in the bark of A. glutinosa and A. incana was first analyzed in populations Rimski most and Sastavci, respectively. DA completely discriminates between three groups of individuals that were implied by PCA. Based on the results, a hypothesis is proposed that the group of four individuals is of hybridogenic origin. Diarylheptanoids proved to be good chemotaxonomic markers at the interspecific level and may also reveal hybrid species...

Amber is a complex, organic polymer that offers unparalleled utility as a preservation medium, providing insights into past organisms and environments. However, under specific circumstances, this information can be compromised through alteration of the amber structure. Understanding the degradation of amber in the geosphere could improve prospecting techniques and maximise the quality and validity of chemical information from altered samples. This study analysed 114 amber samples retrieved from two new Eocene Australian deposits at Strahan, Tasmania and Anglesea, Victoria using a combination of attenuated total reflectance Fourier transform infrared and solid-state 13C cross-polarised magic angle spinning nuclear magnetic resonance spectroscopy. Results identified both Class Ib polylabdanoid and Class II cadinene-based amber. The presence of Class II amber in Australia suggests one of two possibilities: (1) a local Dipterocarpaceae source, the primary producer of Class II resins, despite the absence of this family from the Australian Eocene fossil record; or (2) a local, unidentified botanical source of cadinene-based amber. A third alternative, that Class II amber was transported to Australia from Southeast Asia via ocean currents, is rejected. Taphonomic analysis revealed four mechanisms of alteration prevalent in amber across the two study regions, with evidence of oxidation and metal carboxylate formation. Both the nature and extent of these alterations were found to vary significantly between classes I and II, suggesting that amber class may play a defining role in determining the chemical pathways by which amber degrades. Of note was the high proportion of amber that exhibited no significant chemical changes despite extensive visible alteration features, suggesting the integrity of palaeobiological and palaeoenvironmental information in these samples may be preserved.

Amber is a complex, organic polymer that offers unparalleled utility as a preservation medium, providing insights into past organisms and environments. However, under specific circumstances, this information can be compromised through alteration of the amber structure. Understanding the degradation of amber in the geosphere could improve prospecting techniques and maximise the quality and validity of chemical information from altered samples. This study analysed 114 amber samples retrieved from two new Eocene Australian deposits at Strahan, Tasmania and Anglesea, Victoria using a combination of attenuated total reflectance Fourier transform infrared and solid-state 13C cross-polarised magic angle spinning nuclear magnetic resonance spectroscopy. Results identified both Class Ib polylabdanoid and Class II cadinene-based amber. The presence of Class II amber in Australia suggests one of two possibilities: (1) a local Dipterocarpaceae source, the primary producer of Class II resins, despite the absence of this family from the Australian Eocene fossil record; or (2) a local, unidentified botanical source of cadinene-based amber. A third alternative, that Class II amber was transported to Australia from Southeast Asia via ocean currents, is rejected. Taphonomic analysis revealed four mechanisms of alteration prevalent in amber across the two study regions, with evidence of oxidation and metal carboxylate formation. Both the nature and extent of these alterations were found to vary significantly between classes I and II, suggesting that amber class may play a defining role in determining the chemical pathways by which amber degrades. Of note was the high proportion of amber that exhibited no significant chemical changes despite extensive visible alteration features, suggesting the integrity of palaeobiological and palaeoenvironmental information in these samples may be preserved.

The application of phylogenetic taxonomic procedures led to improvements in the classification of bacteria assigned to the phylum Actinobacteria but even so there remains a need to further clarify relationships within a taxon that encompasses organisms of agricultural, biotechnological, clinical, and ecological importance. Classification of the morphologically diverse bacteria belonging to this large phylum based on a limited number of features has proved to be difficult, not least when taxonomic decisions rested heavily on interpretation of poorly resolved 16S rRNA gene trees. Here, draft genome sequences of a large collection of actinobacterial type strains were used to infer phylogenetic trees from genome-scale data using principles drawn from phylogenetic systematics. The majority of taxa were found to be monophyletic but several orders, families, and genera, as well as many species and a few subspecies were shown to be in need of revision leading to proposals for the recognition of 2 orders, 10 families, and 17 genera, as well as the transfer of over 100 species to other genera. In addition, emended descriptions are given for many species mainly involving the addition of data on genome size and DNA G+C content, the former can be considered to be a valuable taxonomic marker in actinobacterial systematics. Many of the incongruities detected when the results of the present study were compared with existing classifications had been recognized from 16S rRNA gene trees though whole-genome phylogenies proved to be much better resolved. The few significant incongruities found between 16S/23S rRNA and whole genome trees underline the pitfalls inherent in phylogenies based upon single gene sequences. Similarly good congruence was found between the discontinuous distribution of phenotypic properties and taxa delineated in the phylogenetic trees though diverse non-monophyletic taxa appeared to be based on the use of plesiomorphic character states as diagnostic features.

Ovim istraživanjem obuhvaćene su dve populacije Alnus glutinosa (L.) Gaertn. (Prilički kiseljak i Rimski most) i dve populacije Alnus incana (L.) Moench (Golijska reka i Sastavci) uzorkovane na planini Goliji. Osnovni ciljevi bili su: 1) ispitivanje varijabilnosti i odnosa populacija unutar i između vrsta u odnosu na koncentraciju diarilheptanoida u kori prirodnih populacija A. glutinosa i A. incana; 2) ispitivanje sezonske varijabilnosti diarilheptanoida u kori vrsta A. glutinosa i A. incana; 3) procena hemotaksonomskog značaja diarilheptanoida u ispitivanim vrstama; 4) procena mogućnosti primene diarilheptanoida u detekciji spontanih hibrida između A. glutinosa i A. incana; 5) ispitivanje varijabilnosti oblika, veličine i nervature lista u populacijama A. glutinosa i A. incana; 6) procena spontane hibridizacije između ove dve vrste na osnovu morfologije lista. Koncentracija najzastupljenijih diarilheptanoida (hirsutanonol-5-O-β-D-glukopiranozida, oregonina, hirsutanonola, alnuzida A, platifilozida i rubranozida A) u etanolnim ekstraktima kore ispitivanih populacija određena je pomoću UHPLC-MS/MS. Međupopulaciona varijabilnost koncentracije diarilheptanoida ispitana je pomoću parametarskih (ANOVA) i neparametarskih (Kruskal-Wallis) testova, analize glavne komponente (PCA) i diskriminantne analize (DA). Prvi uvid u kvantitativnu distribuciju odabranih diarilheptanoida kod A. glutinosa i A. incana je procenjen na osnovu analize populacija Rimski most i Sastavci. DA je pokazala 100% diskriminacije između tri grupe individua koje je izdvojila PCA, na osnovu čega je postavljena hipoteza da je jedna grupa od četiri individue hibridogenog porekla. Diarilheptanoidi su se pokazali dobrim hemotaksonomskim markerima na interspecijskom nivou i utvrđena je njihova mogućnost primene u detekciji spontanih hibrida. U narednoj vegetacijskoj sezoni, ispitana je sezonska, unutarpopulaciona, međupopulaciona i interspecijska varijabilnost koncentracije diarilheptanoida u dve geografski, In this study, two populations of Alnus glutinosa (L.) Gaertn. (Prilički kiseljak and Rimski most) and two populations of Alnus incana (L.) Moench (Golijska reka and Sastavci) were sampled on mountain Golija with the aims to investigate: 1) variability and relations of populations within and among species based on the concentration of diarylheptanoids in the bark of natural populations of A. glutinosa and A. incana; 2) seasonal variability of diarylheptanoids in the bark of A. glutinosa and A. incana; 3) the reliability of diarylheptanoids as chemotaxonomic markers in the studied species; 4) the reliability of diarylheptanoids for the detection of spontaneous hybrids between A. glutinosa and A. incana; 5) variability of leaf shape, size and venation in the populations of A. glutinosa and A. incana; 6) spontaneous hybridization between the studied species based on leaf morphology. Six diarylheptanoids (hirsutanonol-5-O-β-D-glucopyranoside, oregonin, hirsutanonol, alnuside A, platyphylloside and rubranoside A) were quantified in bark ethanol extracts of four Alnus spp. populations by UHPLC-MS/MS. Interpopulation variability of diarylheptanoid concentration was investigated with ANOVA, Kruskal-Wallis test, principal component analysis (PCA) and discriminant analysis (DA). Quantitative distribution of selected diarylheptanoids in the bark of A. glutinosa and A. incana was first analyzed in populations Rimski most and Sastavci. DA completely discriminates three groups of individuals that were implied by PCA. Based on the results, a hypothesis is proposed that the group of four individuals is of hybridogenic origin. Diarylheptanoids proved to be good chemotaxonomic markers at the interspecific level and may also reveal hybrid species. In the next vegetation season, seasonal, within population, among population and interspecies variability of diarylheptanoid concentration were investigated in two geographically close (Rimski most and Sastavci), and two relatively distant p

Ovim istraživanjem obuhvaćene su dve populacije Alnus glutinosa (L.) Gaertn. (Prilički kiseljak i Rimski most) i dve populacije Alnus incana (L.) Moench (Golijska reka i Sastavci) uzorkovane na planini Goliji. Osnovni ciljevi bili su: 1) ispitivanje varijabilnosti i odnosa populacija unutar i između vrsta u odnosu na koncentraciju diarilheptanoida u kori prirodnih populacija A. glutinosa i A. incana; 2) ispitivanje sezonske varijabilnosti diarilheptanoida u kori vrsta A. glutinosa i A. incana; 3) procena hemotaksonomskog značaja diarilheptanoida u ispitivanim vrstama; 4) procena mogućnosti primene diarilheptanoida u detekciji spontanih hibrida između A. glutinosa i A. incana; 5) ispitivanje varijabilnosti oblika, veličine i nervature lista u populacijama A. glutinosa i A. incana; 6) procena spontane hibridizacije između ove dve vrste na osnovu morfologije lista. Koncentracija najzastupljenijih diarilheptanoida (hirsutanonol-5-O-β-D-glukopiranozida, oregonina, hirsutanonola, alnuzida A, platifilozida i rubranozida A) u etanolnim ekstraktima kore ispitivanih populacija određena je pomoću UHPLC-MS/MS. Međupopulaciona varijabilnost koncentracije diarilheptanoida ispitana je pomoću parametarskih (ANOVA) i neparametarskih (Kruskal-Wallis) testova, analize glavne komponente (PCA) i diskriminantne analize (DA). Prvi uvid u kvantitativnu distribuciju odabranih diarilheptanoida kod A. glutinosa i A. incana je procenjen na osnovu analize populacija Rimski most i Sastavci. DA je pokazala 100% diskriminacije između tri grupe individua koje je izdvojila PCA, na osnovu čega je postavljena hipoteza da je jedna grupa od četiri individue hibridogenog porekla. Diarilheptanoidi su se pokazali dobrim hemotaksonomskim markerima na interspecijskom nivou i utvrđena je njihova mogućnost primene u detekciji spontanih hibrida..., In this study, two populations of Alnus glutinosa (L.) Gaertn. (Prilički kiseljak and Rimski most) and two populations of Alnus incana (L.) Moench (Golijska reka and Sastavci) were sampled on mountain Golija with the aims to investigate: 1) variability and relations of populations within and among species based on the concentration of diarylheptanoids in the bark of natural populations of A. glutinosa and A. incana; 2) seasonal variability of diarylheptanoids in the bark of A. glutinosa and A. incana; 3) the reliability of diarylheptanoids as chemotaxonomic markers in the studied species; 4) the reliability of diarylheptanoids for the detection of spontaneous hybrids between A. glutinosa and A. incana; 5) variability of leaf shape, size and venation in the populations of A. glutinosa and A. incana; 6) spontaneous hybridization between the studied species based on leaf morphology. Six diarylheptanoids (hirsutanonol-5-O-β-D-glucopyranoside, oregonin, hirsutanonol, alnuside A, platyphylloside and rubranoside A) were quantified in bark ethanol extracts of four Alnus spp. populations by UHPLC-MS/MS. Interpopulation variability of diarylheptanoid concentration was investigated with ANOVA, Kruskal-Wallis test, principal component analysis (PCA) and discriminant analysis (DA). Quantitative distribution of selected diarylheptanoids in the bark of A. glutinosa and A. incana was first analyzed in populations Rimski most and Sastavci, respectively. DA completely discriminates between three groups of individuals that were implied by PCA. Based on the results, a hypothesis is proposed that the group of four individuals is of hybridogenic origin. Diarylheptanoids proved to be good chemotaxonomic markers at the interspecific level and may also reveal hybrid species...

The application of phylogenetic taxonomic procedures led to improvements in the classification of bacteria assigned to the phylum Actinobacteria but even so there remains a need to further clarify relationships within a taxon that encompasses organisms of agricultural, biotechnological, clinical, and ecological importance. Classification of the morphologically diverse bacteria belonging to this large phylum based on a limited number of features has proved to be difficult, not least when taxonomic decisions rested heavily on interpretation of poorly resolved 16S rRNA gene trees. Here, draft genome sequences of a large collection of actinobacterial type strains were used to infer phylogenetic trees from genome-scale data using principles drawn from phylogenetic systematics. The majority of taxa were found to be monophyletic but several orders, families, and genera, as well as many species and a few subspecies were shown to be in need of revision leading to proposals for the recognition of 2 orders, 10 families, and 17 genera, as well as the transfer of over 100 species to other genera. In addition, emended descriptions are given for many species mainly involving the addition of data on genome size and DNA G+C content, the former can be considered to be a valuable taxonomic marker in actinobacterial systematics. Many of the incongruities detected when the results of the present study were compared with existing classifications had been recognized from 16S rRNA gene trees though whole-genome phylogenies proved to be much better resolved. The few significant incongruities found between 16S/23S rRNA and whole genome trees underline the pitfalls inherent in phylogenies based upon single gene sequences. Similarly good congruence was found between the discontinuous distribution of phenotypic properties and taxa delineated in the phylogenetic trees though diverse non-monophyletic taxa appeared to be based on the use of plesiomorphic character states as diagnostic features.

This project was designed to investigate the effects of carbon dioxide (CO2) levels on the growth and pigment ratios (chemotaxonomy) of freshwater algal species typical to the south Florida surface waters. Green algae, diatoms, and cyanobacteria were cultured under 400 or 800 ppm CO3 in air for several weeks. Growth monitoring used a cell counter, hemocytometer, and chlorophyll fluorescence. Pigments were analyzed using HPLC-PDA. Experiments with certified CO2 concentrations (400, 600, 800, 1200 ppm) were conducted with helium degassed ultrapure water and each of three culture media. Theoretical and experimental pH values with water matched exactly. However, each culture media proved to exhibit significant buffer capacity. Cell growth monitoring was problematic except for the cyanobacterium Microcystis aeruginosa. That species responded to increased CO2 (800 ppm) with increased growth rates as predicted. The other species gave erratic results mainly due to difficulties in obtaining valid consistent cell counts., 2018, Includes bibliography., Degree granted: Thesis (M.S.)--Florida Atlantic University, 2018., Collection: FAU Electronic Theses and Dissertations Collection

Morphology and phylogeny has been used to distinguish members of the plant pathogenic fungal genus Stemphylium. A third method for distinguishing species is by chemotaxonomy. The main goal of the present study was to investigate the chemical potential of Stemphylium via HPLC-UV-MS analysis, while also exploring the potential of chemotaxonomy as a robust identification method for Stemphylium. Several species were found to have species-specific metabolites, while other species were distinguishable by a broader metabolic profile rather than specific metabolites. Many previously described metabolites were found to be important for distinguishing species, while some unknown metabolites were also found to have important roles in distinguishing species of Stemphylium. This study is the first of its kind to investigate the chemical potential of Stemphylium across the whole genus.