Alkaloidal content of argentine Argemone

The alkaloid content of different Argentine Argemone has been determined. Two varieties of A. subfusiformis subsp. subfusiformis Ownb. and A. subfusiformis subsp. subinermis Ownb. yielded a similar ratio and content of the following alkaloids: protopine, allocrytopine, berberine, sanguinarine, and chelerythrine. The A. subfusiformis taxa showed a markedly high sanguinarine content in roots as opposed to aerial parts. A. polyanthemos (Fedde) Ownb. showed a different ratio between alkaloids but a qualitative similar result. N-Norchelerythrine was isolated from A. polyanthemos. The chemotaxonomic value of the alkaloid analyses is discussed.     

A chemotaxonomic study of free flavone aglycones from some Iberian Thymus species

Sixteen free flavone aglycones exuded onto the leaf surfaces have been analysed in twenty-three taxa of genus Thymus growing in the Iberian Peninsula. A chemotaxonomic study of these compounds has been carried out. Correlations with morphological classifications and the usefulness of excreted flavonoids as taxonomic markers are discussed.     

The use of an oligonucleotide that hybridizes to DNA sequences interspersed in the genome of members of the genusPhytophthora as an adjunct to morphological criteria in species identification

An oligonucleotide (primer), designed from a conserved region of the multi-allelicb locus of the basidiomycete fungusUstilago maydis, generated reproducible PCR fingerprints inPhytophthora species. The primer hybridized in a species-specific manner to nucleotide sequences interspersed in the genome of the closely related members ofPhytophthora taxonomic group IV. We recommend the use of this PCR procedure as an alternative method for resolving the close taxonomic affinity of some members of the genusPhytophthora.     

Investigation of serological determinants from single storage plant proteins

The main storage proteins of the angiosperms provide valuable characters for taxonomic research. By comparing the reactivity of one specific protein in different taxa (mainly Ranunculaceae). determinants or determinant groups can be discriminated. Their number is a function of the number of the reference or anti-systems and of the number of taxa compared. These serological determinants prove a posteriori to be good characters and most useful in taxonomy. Their systematic range extends from one genus to the whole of the angiosperms. The resolving power is best between genera within a family. Using such a single protein system, it is possible to co-ordinate the determinants of the taxa investigated and to map their distribution; this is an advantage over the multi-protein systems commonly used. By serological comparison, it should be possible to establish the homology of the main storage globulins throughout the angiosperms.     

Alkaloids and triterpenes from the african toddalioideae

Stem and root barks of Araliopsis tabouensis, Diphasia klaineana and Teclea verdoorniana have been investigated. A. tabouensis yielded flindissol (I), (+)-N-methylplatydesminium ion (III) and a second furoquinoline alkaloid: D. klaineana yielded lupeol (IX). evoxanthine (XI), arborinine (X) and skimmianine (V). T. verdoorniana gave lupeol (IX) and evoxanthine (XI). A further alkaloid, designated DK/1, was isolated from both the Diphasia and Teclea species. The value, as chemotaxonomic indicators, of the alkaloids and other constituents known to occur in six African genera of the Toddalioideae has been examined and possible affinities with other rutaceous taxa are suggested.     

The potential of AFLPs in biosystematics: A first application inSolanum taxonomy(Solanaceae)

Using the AFLP technique highly informative DNA fingerprints were generated from 19 taxa ofSolanum sect.Petota (potatoes) and three taxa ofSolanum sect.Lycopersicum (tomatoes). Both phenetic and cladistic analyses were conducted from the individual genotypic level to the species level. An AFLP fingerprint, using a combination of suitable AFLP primers, generated 12 to 71 scorable fragments per genotype which was sufficient for taxonomic interpretation. The classifications based on the molecular markers were generally in agreement with current taxonomic opinions. Unexpectedly,S. microdontum was associated with ser.Megistacroloba rather than with ser.Tuberosa, andS. demissum (ser.Demissa) and species of ser.Acaulia appeared closely affiliated. AFLP is an efficient and reliable technique to generate biosystematic data and therefore a promising tool for evolutionary studies.     

Chemical and principal-component analyses of the essential oils of Apioideae taxa (Apiaceae) from central Balkan

The volatile constituents of the essential oils of 23 taxa belonging to the Apioideae subfamily were studied in detail. The investigated taxa were Pimpinella serbica (Vis.) Bentham & Hooker, Libanotis montana Cr., Cnidium silaifolium (Jacq.) Simk. ssp. orientale (Boiss.) Tutin, Bupleurum praealtum L., B. sibthorpianum S. S. var. diversifolium (Roch.) Hay, Aegopodium podagraria L., Torilis anthriscus (L.) Gmel., Orlaya grandiflora (L.) Hoffm., Laserpitium siler L., Laser trilobum (L.) Brokh., Chaerophyllum aureum L., C. hirsutum L., C. temulum L., Pastinaca sativa L., P. hirsuta Pancic., Tordylium maximum L., Physospermum cornubiense (L.) DC., Peucedanum alsaticum L., P. oreoselinum (L.) Moench, P. cervaria (L.) Cuss., P. austriacum (Jacq.) Koch, P. longifolium W. et K., and P. officinale L. All of these species grow wild in the central part of the Balkan Peninsula. The essential oils were found to be complex mixtures of various compounds, more than 100 constituents being in each taxon, with contributions of main products never exceeding 25% of the total content. Sesquiterpene hydrocarbons were found to be the main group of constituents of all taxa, except for Peucedanum species, where monoterpene hydrocarbons were identified as the main components. The chemotaxonomic value of the essential-oil composition is discussed according to results of principal-component analysis (PCA). The essential-oil composition mainly reflects current taxonomic relationships between the investigated taxa.     

Comparison of ISSR,IRAP and REMAP markers for assessing genetic diversity in different species of <Emphasis Type="Italic">Brassica</Emphasis> sp.

Molecular markers provide facilities in order to study genetic diversity and relationship among genotypes. In this study, genetic diversity among 35 genotype of Brassica sp. (belonging B. napus, B. juncea, B. rapa, B. nigra) were determined using 13 ISSR, 3 IRAP markers and 18 REMAP (primer combinations of ISSR and retrotransposon primer). The percentage of polymorphism for ISSR, IRAP and REMAP was 96.38, 94 and 96%, respectively. By comparison between markers, ISSRs indicated the highest expected heterozygosity (He) and Shannon’s information index (I) with value of 0.34 and 0.51, respectively, while REMAP marker had by far the highest number of polymorphic bands (340) and marker index (7.1) among all fragments scored over all markers. In pattern of clustering based on Bayesian methods, K = 8 was resulted for combined data clustering that was more organized clustering for genotypes compared to others. This research suggests the combined data of ISSR, IRAP and REMAP markers are most reliable than each solely marker whilst have been clustered genotypes in their taxonomic classification of Brassica without any mixture. Principle coordinate analysis (PCoA) separated 35 genotypes in four groups which all of genotypes were clustered correctly based on their taxonomic classification. The findings of this study provide the valuable insight into the Brassica species relationships in terms of similarity among genotypes which can be helpful in breeding programs, and also demonstrate that retrotransposon markers are legible for genetic diversity and next genetic analysis in Brassica genus.     

Genetic relationship and genetic diversity among Typha taxa from East Asia based on AFLP markers

The genetic relationship and diversity among four Typha taxa in East Asia were evaluated using amplified fragment length polymorphism (AFLP) markers. Three AFLP selective primer combinations generated a total of 707 amplification products, of which 704 (99.6%) were polymorphic. The unweighted pair-group method with arithmetic mean (UPGMA) dendrogram and principal component analysis (PCA) plot confirmed the taxonomic status of four separate species. East Asian Typha taxa separated into two groups: the first with Typha angustifolia and the second with T. orientalis, T. laxmanni, and T. latifolia with a high bootstrap value for UPGMA (93%) and a low first score for PCA (25%). The two clusters corresponded with two sections based on the bracteoles in the female flower: section Bracteolatae and section Ebracteolatae. T. angustifolia showed the highest genetic diversity among the four Typha taxa (percentage of polymorphic loci [PPL] = 71%, H_o = 0.157), whereas T. latifolia had the lowest genetic diversity (PPL = 40%, H_o = 0.117). Genetic diversity was related to the presence of the gap between male and female inflorescences. A positive correlation between genetic distance and geographic distance was clearly found in the two species with continuous inflorescences (T. latifolia and T. orientalis). This positive correlation was not observed in the other species with discontinuous spikes (T. angustifolia and T. laxmanni).     

A spectral theory for Wright’s inbreeding coefficients and related quantities

Wright’s inbreeding coefficient, F_ST, is a fundamental measure in population genetics. Assuming a predefined population subdivision, this statistic is classically used to evaluate population structure at a given genomic locus. With large numbers of loci, unsupervised approaches such as principal component analysis (PCA) have, however, become prominent in recent analyses of population structure. In this study, we describe the relationships between Wright’s inbreeding coefficients and PCA for a model of K discrete populations. Our theory provides an equivalent definition of F_ST based on the decomposition of the genotype matrix into between and within-population matrices. The average value of Wright’s F_ST over all loci included in the genotype matrix can be obtained from the PCA of the between-population matrix. Assuming that a separation condition is fulfilled and for reasonably large data sets, this value of F_ST approximates the proportion of genetic variation explained by the first (K − 1) principal components accurately. The new definition of F_ST is useful for computing inbreeding coefficients from surrogate genotypes, for example, obtained after correction of experimental artifacts or after removing adaptive genetic variation associated with environmental variables. The relationships between inbreeding coefficients and the spectrum of the genotype matrix not only allow interpretations of PCA results in terms of population genetic concepts but extend those concepts to population genetic analyses accounting for temporal, geographical and environmental contexts.     

6β-[2-Methylbutanoyloxy]tropan-3α-ol,a new alkaloid from Datura ceratocaula structure and biosynthesis

6β-[2-methylbutanoyloxy]tropan-3α-ol, a new alkaloid, has been isolated from the aerial parts of Datura ceratocaula (Solanaceae). Its chemotaxonomic significance within the genus is discussed. Biosynthetically, the 2-methylbutanoyl moiety is derived from L-isoleucine.     

The chemotaxonomic value of the diglucoside anthrone homonataloside B in the genus Aloe

The chemotaxonomic value of the only known diglucoside anthrone from Aloe, homonataloside B, is discussed. This compound has been detected in only 14 species in a chemotaxonomic survey of 380 Aloe species. The homonataloside B-accumulating species are only found in Africa with none of the Malagasy endemics producing this compound. A summary of the morphological variation is presented together with the taxonomic distribution of this unique anthrone. The representatives of this distinct chemotype occupy disparate positions in the largely artificial hierarchy of the present classification system. Species included in this chemotype provide chemical evidence of hybridisation in Aloe.     

Contrasting patterns of niche partitioning between two anaerobic terminal oxidizers of organic matter

Understanding the ecological principles underlying the structure and function of microbial communities remains an important goal for microbial ecology. We examined two biogeochemically important taxa, the sulfate-reducing bacterial genus, Desulfobulbus, and the methanogenic archaeal genus, Methanosaeta, to compare and contrast niche partitioning by these two taxa that are ecologically linked as anaerobic terminal oxidizers of organic material. An observational approach utilizing functional gene pyrosequencing was combined with a community-based reciprocal incubation experiment and characterization of a novel Desulfobulbus isolate. To analyze the pyrosequencing data, we constructed a data analysis pipeline, which we validated with several control data sets. For both taxa, particular genotypes were clearly associated with certain portions of an estuarine gradient, consistent with habitat or niche partitioning. Methanosaeta genotypes were generally divided between those found almost exclusively in the marine habitat (∼30% of operational taxonomic units (OTUs)), and those which were ubiquitously distributed across all or most of the estuary (∼70% of OTUs). In contrast to this relatively monotonic distribution, for Desulfobulbus, there were many more genotypes, and their distributions represented a wide range of inferred niche widths from specialist genotypes found only at a single site, to ubiquitous or generalist genotypes found in all 10 sites examined along the full estuarine gradient. Incubation experiments clearly showed that, for both taxa, communities from opposite ends of the estuary did not come to resemble one another, regardless of the chemical environment. Growth of a Desulfobulbus isolated into pure culture indicated that the potential niche of this organism is significantly larger than the realized niche. We concluded that niche partitioning can be an important force structuring microbial populations, with biotic and abiotic components having very different effects depending on the physiology and ecology of each taxon.     

Leaf-Associated Bacterial and Fungal Taxa Shifts in Response to Larvae of the Tree Hole Mosquito, <Emphasis Type="Italic">Ochlerotatus triseriatus</Emphasis>

Larvae of the eastern tree hole mosquito, Ochlerotatus triseriatus (Say), and related container-breeding species are known to feed upon substrate-associated microorganisms. Although the importance of these microbial resources to larval growth has been established, almost nothing is known about the taxonomic composition and dynamics of these critical microbial food sources. We examined bacterial and fungal community compositional changes on oak leaves tethered in natural tree hole habitats of O. triseriatus. We eliminated larvae experimentally in a subset of the tree holes and examined 16S rDNA gene sequences for bacteria and ergosterol concentrations and 18S rRNA gene sequences for fungi collected from leaf material subsamples. Leaf ergosterol content varied significantly with time, but not treatment. Principal component analysis (PCA) was used to compare microbial taxonomic patterns found in leaves incubated with or without larvae present, and we found that larval presence affected both bacterial and fungal groups, either from loosely attached or strongly adherent categories. Bacterial communities generally grouped more tightly when larvae were present, and class level taxa proportions changed when larvae were present, suggesting selection by larval feeding or activities for particular taxa such as members of the Bacteroidetes, Alphaproteobacteria, and Betaproteobacteria classes. Fungal taxa composite scores also separated along PC axes related to the presence of larvae and indicated larval feeding effects on several higher taxonomic groups, including Saccharomycetes, Dothideomycetes, and Chytridiomycota. These results support the hypothesis that larval mosquito feeding and activities altered microbial communities associated with substrate surfaces, potentially leading to decreased food value of the resource and affecting decomposition of particulate matter in the system.     

Comparison of essential oils and genetic relationship of Origanum × intercedens to its parental taxa in the island of Crete

Plants of the taxa Origanum onites and Origanum vulgare were allowed to cross-pollinate under natural open field conditions and to produce a mixed population of putative hybrid Origanum × intercedens and parent genotypes. Randomly collected plants were classified as putative hybrids or parent genotypes by inspection of their inflorescence. They were then subjected to analysis of their essential oil composition and were fingerprinted by randomly amplified polymorphic DNA (RAPD) fragments. DNA primers identifying the genotype as well as showing the distance of a particular putative hybrid plant from the parent genotypes were found. Alone or in combination with the essential oil composition they can be used as reliable tools for the genetic identification of the two parental taxa and the putative hybrid plants in natural populations.     

Metabolic fingerprinting of Leontopodium species (Asteraceae) by means of ¹H NMR and HPLC-ESI-MS

The genus Leontopodium, mainly distributed in Central and Eastern Asia, consists of ca. 34-58 different species. The European Leontopodium alpinum, commonly known as Edelweiss, has a long tradition in folk medicine. Recent research has resulted in the identification of prior unknown secondary metabolites, some of them with interesting biological activities. Despite this, nearly nothing is known about the Asian species of the genus. In this study, we applied proton nuclear magnetic resonance (1H NMR) spectroscopy and liquid chromatography-mass spectrometry (LC-MS) metabolic fingerprinting to reveal insights into the metabolic patterns of 11 different Leontopodium species, and to conclude on their taxonomic relationship. Principal component analysis (PCA) of 1H NMR fingerprints revealed two species groups. Discriminators for these groups were identified as fatty acids and sucrose for group A, and ent-kaurenoic acid and derivatives thereof for group B. Five diterpenes together with one sesquiterpene were isolated from Leontopodium franchetii roots; the compounds were described for the first time for L. franchetii: ent-kaur-16-en-19-oic acid, methyl-15α-angeloyloxy-ent-kaur-16-en-19-oate, methyl-ent-kaur-16-en-19-oate, 8-acetoxymodhephene, 19-acetoxy-ent-kaur-16-ene, methyl-15β-angeloyloxy-16,17-epoxy-ent-kauran-19-oate. In addition, differences in the metabolic profile between collected and cultivated species could be observed using a partial least squares-discriminant analysis (PLS-DA). PCA of the LC-MS fingerprints revealed three groups. Discriminating signals were compared to literature data and identified as two bisabolane derivatives responsible for discrimination of group A and C, and one ent-kaurenoic acid derivative, discriminating group B. A taxonomic relationship between a previously unidentified species and L. franchetii and Leontopodium sinense could be determined by comparing NMR fingerprints. This finding supports recent molecular data. Furthermore, Leontopodium dedekensii and L. sinense, two closely related species in terms of morphology and DNA-fingerprints, could be distinguished clearly using 1H NMR and LC-MS metabolic fingerprinting.     

Geographic and temporal distributions of four genotypes found in Erysiphe gracilis var. gracilis,a powdery mildew of evergreen oaks (Erysiphales)

A survey of 402 samples of Erysiphe gracilis var. gracilis on evergreen oaks collected from a wide area of western Japan showed that they were divided into four distinct genotypes each forming a separate clade with high bootstrap support, which were referred to as E. hiratae (genotype I), E. uncinuloides (genotype II), E. gracilis s. str. (genotype III), and E. pseudogracilis (genotype IV) in a separate taxonomic treatment. However, there are no clear differences in geographic distributions among these four genotypes. Quercus myrsinifolia was only infected by genotype II and Q. salicina only by genotype IV, whereas Q. glauca was infected by all four. These results strongly suggest an association between host species and speciation of these genotypes. A further 312 samples of Q. glauca with E. gracilis s. lat. colonies were collected from four locations in the Mie University campus to investigate frequency of genotypes I and II every month from May 2015 to January 2016. No temporal isolation was found in genotype frequencies. These genotypes frequently co-existed on a single leaf surface, especially at the locations disturbed by human activities. Two oak powdery mildews, E. gracilis s. lat. and Cystotheca wrightii, produced conidia only one month a year and their life cycle differed from most other powdery mildew species. This suggests that these oak mildews developed their unique life cycles to synchronize with the life cycle of evergreen oaks.     

Genotypic and Phenotypic Diversity within Species of Purple Nonsulfur Bacteria Isolated from Aquatic Sediments

To assess the extent of genotypic and phenotypic diversity within species of purple nonsulfur bacteria found in aquatic sediments, a total of 128 strains were directly isolated from agar plates that had been inoculated with sediment samples from Haren and De Biesbosch in The Netherlands. All isolates were initially characterized by BOX-PCR genomic DNA fingerprinting, and 60 distinct genotypes were identified. Analyses of 16S rRNA gene sequences of representatives of each genotype showed that five and eight different phylotypes of purple nonsulfur bacteria were obtained from the Haren and De Biesbosch sites, respectively. At the Haren site, 80.5% of the clones were Rhodopseudomonas palustris, whereas Rhodoferax fermentans and Rhodopseudomonas palustris were numerically dominant at the De Biesbosch site and constituted 45.9 and 34.4% of the isolates obtained, respectively. BOX-PCR genomic fingerprints showed that there was a high level of genotypic diversity within each of these species. The genomic fingerprints of Rhodopseudomonas palustris isolates were significantly different for isolates from the two sampling sites, suggesting that certain strains may be endemic to each sampling site. Not all Rhodopseudomonas palustris isolates could degrade benzoate, a feature that has previously been thought to be characteristic of the species. There were differences in the BOX-PCR genomic fingerprints and restriction fragment length polymorphisms of benzoate-coenzyme A ligase genes and form I and form II ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) genes between benzoate-degrading and non-benzoate-degrading genotypes. The ability to distinguish these two Rhodopseudomonas palustris groups based on multiple genetic differences may reflect an incipient speciation event resulting from adaptive evolution to local environmental conditions.     

Chemotaxonomic characterization and chemical similarity of tribes/genera of the Solanoideae subfamily (Solanaceae) based on occurrence of withanolides

Withanolides are well established as chemotaxonomic markers in the Solanaceae. A large number of withanolides have been isolated from the subfamily Solanoideae (Solanaceae). A database of the occurrences of withanolides in Solanoideae was compiled to help characterize the chemical profile and verify the chemical similarity of the tribes/genera of this subfamily. The study revealed the importance of this group of compounds as chemotaxonomic markers, especially for Physaleae, Deprea, and Jaborosa, ratifying the importance of chemotaxonomy as a tool for elucidation of taxonomic problems, such as undefined relationships of genera that are not yet included in tribes of Solanoideae.     

The rDNA Internal Transcribed Spacer Region as a Taxonomic Marker for Nematodes

The ITS region from a wide taxonomic range of nematodes, including secernentean and adenophorean taxa, and free-living, entomopathogenic, and plant-parasitic species, was evaluated as a taxonomic marker. Size of the amplified product aided in the initial determination of group membership, and also suggested groups that may require taxonomic reevaluation. Congeneric species often displayed identically sized ITS regions, but genera such as Pratylenchus and Tylenchorhynchus had species with large differences in size. ITS heterogeneity in individuals and populations was identified in several nematode taxa. PCR-RFLP of ITS1 is advocated as a method of taxonomic analysis in genera such as Helicotylenchus that contain numerous species with few diagnostic morphological characteristics.