Alkaloids and other constituents of Zanthoxylum williamsii,Z. monophyllum and Z. fagara

Zanthoxylum williamsii (Rutaceae) was found to contain (+)-asaranin, (+)-sesamin, esculetin dimethyl ether, nitidine, chelerythrine, magnoflorine, laurifoline, skimmianine and edulinine. The quaternary alkaloid fraction of Z. monophyllum contained berberine, magnoflorine, chelerythrine and a 1,2,9,10-substituted dihydroxydimethoxy-N,N-dimethylaporphinium salt. Leaves of Z. fagara were found to contain synephrine. Leaves of each species were examined for the presence of bishordeninyl terpene alkaloids, but none was found. Some chemotaxonomic relationships among Zanthoxylum species are discussed.     

The chemosystematics of the genus Zanthoxylum L. (Rutaceae) in Kenya

Background and aimThere is currently a significant taxonomic confusion in the genus Zanthoxylum which has a wide application in Kenyan herbal medicine. The significance of proper identification/classification of medicinal plants is key in connecting the ethnobotanical uses with the existing chemical and biological knowledge. This study aims at solving the challenges encountered in the classification of Zanthoxylum species in Kenya using metabolomics as a tool for chemosystematics.MethodsFive Zanthoxylum species (Z. chalybeum, Z. gilletii, Z. holtzianum, Z. paracanthum and Z. usambarense) were analyzed using metabolomics approach where LC-MS analysis was used to establish the chemical profiles of each species. Hierarchical cluster analysis was used to show the relationship among the Zanthoxylum spp studied.ResultsBased on phytochemical production, the five Zanthoxylum species segregated into two clusters; cluster one composed of Z. gilletii Z. usambarense and Z. paracanthum while Z. chalybeum and Z. holtzianum constituted cluster two. The chemical markers (discriminants) were identified as; laudanine, kalambroside A, kalambroside B, gamma-sanshool, ochotensine, cis-N-methylcanadine, 18-hydroxylinoleic acid, dihydrochelerythrine, N-feruloylglycine, p-coumaric acid ethyl ester, dihydroberberine and one unknown compound.ConclusionsChemometric analysis of relatedness of Zanthoxylum group of plants conformed to the current classical taxonomy of Zanthoxylum species in Kenya.     

Target enrichment improves phylogenetic resolution in the genus Zanthoxylum (Rutaceae) and indicates both incomplete lineage sorting and hybridization events

Background and Aims Zanthoxylum is the only pantropical genus within Rutaceae, with a few species native to temperate eastern Asia and North America. Efforts using Sanger sequencing failed to resolve the backbone phylogeny of Zanthoxylum. In this study, we employed target-enrichment high-throughput sequencing to improve resolution. Gene trees were examined for concordance and sectional classifications of Zanthoxylum were evaluated. Off-target reads were investigated to identify putative single-copy markers for bait refinement, and low-copy markers for evidence of putative hybridization events.MethodsA custom bait set targeting 354 genes, with a median of 321 bp, was designed for Zanthoxylum and applied to 44 Zanthoxylum species and one Tetradium species as the outgroup. Illumina reads were processed via the HybPhyloMaker pipeline. Phylogenetic inferences were conducted using coalescent and maximum likelihood methods based on concatenated datasets. Concordance was assessed using quartet sampling. Additional phylogenetic analyses were performed on putative single and low-copy genes extracted from off-target reads.Key ResultsFour major clades are supported within Zanthoxylum: the African clade, the Z. asiaticum clade, the Asian–Pacific–Australian clade and the American–eastern Asian clade. While overall support has improved, regions of conflict are similar to those previously observed. Gene tree discordances indicate a hybridization event in the ancestor of the Hawaiian lineage, and incomplete lineage sorting in the American backbone. Off-target putative single-copy genes largely confirm on-target results, and putative low-copy genes provide additional evidence for hybridization in the Hawaiian lineage. Only two of the five sections of Zanthoxylum are resolved as monophyletic.ConclusionsTarget enrichment is suitable for assessing phylogenetic relationships in Zanthoxylum. Our phylogenetic analyses reveal that current sectional classifications need revision. Quartet tree concordance indicates several instances of reticulate evolution. Off-target reads are proven useful to identify additional phylogenetically informative regions for bait refinement or gene tree based approaches.     

Nitrogen-containing non-steroidal secondary metabolites of Solanum, Cyphomandra, Lycianthes and Margaranthus

Of the 49 species of Solanum studied, cuscohygrine has been detected in 25, solamine and related amines in 17 and solamine-derived amides in 16. Five species of Cyphomandra examined all contained both amines and amides. From roots of Margaranthus solanaceus cuscohygrine has been isolated which probably occurs, too, in roots of Lycianthes rantonnettii. The distribution of these compounds throughout the taxa could be of chemotaxonomic value.     

Phylogenetic relationships among cultivated <Emphasis Type="Italic">Zanthoxylum</Emphasis> species in China based on cpDNA markers

We here present a molecular phylogenetic analysis of cultivated Zanthoxylum species which have a long history of cultivation both for economic and for chemical values in China. Three cpDNA markers, including matK, rbcL, and trnL-F, were sequenced, with the goals of untangling phylogenetic relationships and inferring biogeographic origin and patterns of distribution among Zanthoxylum species. Based on three cpDNA markers, 19 haplotypes with 64 polymorphic sites in Zanthoxylum provenances were identified in our study. A low genetic differentiation (G _ST ?=?0.271, N _ST ?=?0.373) was observed within Zanthoxylum provenances. Based on phylogenetic tree and haplotype network, all 19 haplotypes were grouped into six clusters. Our results also supported the hypothesis that the so-called “Green Huajiao” belongs to the species Zanthoxylum armatum rather than Zanthoxylum schinifolium. The results also revealed that haplotypes of two cultivated species, Zanthoxylum bungeanum and Z. armatum, most probably diverged during the Late Miocene. Ancestral area reconstruction indicated that cultivated Zanthoxylum species experienced multiple long-distance dispersal events and several vicariance events and the ancestors of Zanthoxylum first colonized Yunnan and Guizhou provinces (D). Accordingly, the current disjunct distribution of Z. bungeanum and Z. armatum may represent long-distance dispersal of ancestors popularly named “Dahongpao” and “Qinghuajiao,” respectively. It is concluded that cpDNA markers may provide a new conceptual and practical opportunity to evaluate genetic diversity and to identify local cultivars of Zanthoxylum, making it a valuable source to include into potential breeding programs.     

A hydroxygermacrene and other constituents from Pseudobrickellia brasiliensis

Pseudobrickellia brasiliensis afforded, in addition to known compounds, a new germacradiene derivative and a hydroxy-α-amyrin.     

Phylogeny and molecular evolution of the rbcL gene of St genome in Elymus sensu lato (Poaceae: Triticeae)

Analysis of the patterns and levels of diversity in duplicate gene not only traces evolutionary history of polyploids, but also provides insight into how the evolutionary process differs between lineages and between homoeologous loci within lineages. Elymus sensu lato is a group of allopolyploid species, which share a common St genome and with the different combinations of H, Y, P, and W genomes. To estimate the evolutionary process of the rbcL gene in species of Elymus s. l. and its putative dioploid relatives, 74 sequences were obtained from 21 species of Elymus s. l. together with 24 diploid taxa representing 19 basic genomes in Triticeae. Phylogeny and sequence diversity pattern analysis suggested that (1) species of Pseudoroegneria (Nevski) Á. Löve might serve as the maternal donor of the species of Elymus s. l; (2) differentiation of St genome were shown in the species of Elymus s. l. following polyploidy event; (3) divergences within the species might associate with geographic diversity and morphological variability; (4) differences in the levels and patterns of nucleotide diversity of the rbcL gene implied that the St genome lineages in the species of Elymus s. l. have differently evolutionary potentials.     

The Biochemical of Tetradium,Euodia and Melicope and their significance in the Rutaceae

From the stem and root barks of Tetradium glabrifolium (Rutaceae) benzophenanthridine alkaloids have been isolated together with furoquinoline alkaloids, coumarins and limonoids. The occurrence of the l-benzyltetrahydroisoquinolinederived benzophenanthridines adds Tetradium to the small group of rutaceous genera (Fagaropsis, Phellodendron, Toddalia, Zanthoxylum) known to produce alkaloids of this type. These findings support Hartley's decision to reassign taxa from Euodia (sensu Engler) into the three genera Tetradium, Euodia s. s. and Melicope and his contention that Tetradium is closely allied to Zanthoxylum and Phellodendron, with which it shares several unusual chemical characters. Hartley's re-alignment of Tetradium, which cuts across Engler's sub-families Rutoideae and Toddalioideae, is therefore sustained by the known distribution of secondary metabolites, whilst available information suggests a wide disparity between the chemical profiles of Tetradium and Melicope. The chemosystematic significance of the addition of Tetradium to the 1-benzyltetrahydroisoquinoline producing genera and the potential of these taxa to have acted as a starting point for biochemical evolution within the Rutaceae is discussed.     

Genetic structure of cultivated <Emphasis Type="Italic">Zanthoxylum</Emphasis> species investigated with SSR markers

Zanthoxylum is an economically and ecologically important genus of the Rutaceae family, of which Z. bungeanum and Z. armatum have a long history of cultivation in China. However, how the natural processes such as selection and drift and agriculture practices have influenced the genetic variation of cultivated Zanthoxylum species during long-term domestication remains elusive. Herein, we determined the population genetic structure of current widely cultivated Zanthoxylum species, Z. bungeanum and Z. armatum. Microsatellite markers revealed a high level of genetic variation and significant genetic differentiation for both species despite Z. bungeanum showed higher genetic diversity than Z. armatum. AMOVA indicated that most of the genetic variation exists within individuals rather than among provenances for both species. Population structure analyses generated three distinct groups within the entire accessions. All Z. bungeanum accessions were distinguished into two major geographic groups, north and south groups, with Qinling Mountains as the main geographic barrier to gene flow while a significant genetic differentiation was observed between cultivated and wild Z. armatum accessions. Mantel test of Z. bungeanum displayed a significant correlation between genetic and geographic distances within each inferred group but no correlation between genetic and geographic distance was observed when comparing genetic and geographic distances focusing only on pairwise of north vs. south provenances, ruling out the hypothesis that gene flow between north and south provenances followed an isolation-by-distance model. Our research provided a fundamental genetic profile that will improve the conservation and responsible exploitation of the extant germplasm of Zanthoxylum.     

The genus Laetiporus (Basidiomycota, Polyporales) in East Asia

Relationships among East Asian, North American and European Laetiporus sulphureus s. lat., a cosmopolitan brown rot species complex, were assessed with phylogenetic analyses and incompatibility tests. Three East Asian taxa, Laetiporus cremeiporus sp. nov., Laetiporus montanus and Laetiporus versisporus, are described and illustrated as well as compared with related taxa from Southeast Asia, North America and Europe. Phylogenetic analyses showed that L. cremeiporus and L. versisporus are clearly distinct species among Laetiporus taxa. The three conifer inhabiting species, Laetiporus conifericola, Laetiporus huroniensis and L. montanus, are closely related to each other. The European population of L. montanus exhibits two sequence variants of the EF1α: one is the same as observed in L. sulphureus in Europe and the other is that observed in East Asian population of L. montanus. A key to the known species of Laetiporus in the northern hemisphere is provided.     

Elaeodendrol and elaeodendradiol,new nor-triterpenes from Elaeodendron glaucum

The isolation of six known and two new D:A-friedooleananes is reported from the bark of Elaeodendron glaucum. The structures of the new nor-triterpenes, elaeodendrol and elaeodendradiol, were established respectively as 17β- hydroxy-28-norfriedelan-3-one and l7β,25-dihydroxy-28-norfriedelan-3-one by a study of the methyl chemical shifts in their ¹H NMR spectra.     

The influence of environmental instability on the reproductive strategy of the critically endangered Acıgöl killifish (Aphanius transgrediens)

To investigate the variability in the reproductive tactics of Acıgöl killifish (Aphanius transgrediens), one of the most seriously threatened freshwater teleost species, this study compared its reproductive ecology in two contrasting habitats that differ substantially in terms of stability of environmental parameters, particularly salinity regime (stable vs. unstable). Fish were sampled monthly from October 2013 to September 2014 with the aim of testing whether the reproductive life-history response of fish to stable and unstable conditions differed. The reproductive effort (gonad weight) of both sexes did not differ significantly between the two habitats, but females in the unstable habitat had significantly lower fecundity and larger eggs. The relationship between fecundity and fish size was stronger in the stable habitat, whereas the relationship was quite variable and uncertain in the unstable habitat. Fish born in the unstable habitat reached their first maturity at a smaller size than those in the stable habitat. The gonado-somatic index and the duration of hydrated eggs showed that reproduction continued from February to May in both habitats; nonetheless, a second spawning event occurred during July and August in the unstable habitat, which included the reproductive contributions of YOY individuals and older generations. This study’s results suggest that A. transgrediens employs varying reproductive strategies against environmental instability in its restricted unique range. This may have further implications for the ways in which habitat-specific conservation methods are used.     

Chemical constituents from the leaves of Zanthoxylum nitidum (Roxb.) DC

Twelve compounds were isolated from the ethyl acetate soluble portion of the 70% hydroalcoholic extract from the leaves of Zanthoxylum nitidum (Roxb.) DC. All of the compounds were determined on the basis of UV, IR, NMR and MS and comparison with literature. Among them, compounds 4, 7, 10 and 12 were first acquired from Rutaceae, compound 2 was isolated from the genus of Zanthoxylum for the first time and compounds 1, 3, 6, 9, 11 were isolated from Z. nitidum for the first time. Our present study exhibited the taxonomic relationships between Z. nitidum and other species in the same genus; those between Rutaceae and other families. Furthermore, we have summarized the chemotaxonomic significance of these compounds in Rutaceae and Zanthoxylum.     

Ovarian dynamics in Heliconius butterflies: Correlations among daily oviposition rates,egg weights,and quantitative aspects of oögenesis

Both oviposition and ovarian morphology were studied in individual butterflies of the neotropical genus Heliconius, which as adults ingest amino acids from pollen, live up to 6 months, and have continuous oögenesis. Among female Heliconius charitonius, daily oviposition correlates directly with the total number of oöcytes developing in the ovaries. The calculated time required for complete growth of one oöcyte is, however, reasonably constant among individuals of a species with widely varying oviposition rates. Thus, within a species, butterflies laying more eggs per day are not necessarily maturing each at a faster rate, but are processing more oöcytes simultaneously in their ovaries. A further correlation between oviposition rate and adult size suggests that in H. charitonius both ovarian capacity and daily egg production are determined ultimately by extent and/or quality of larval nutrition. Among other Heliconius species, those producing larger eggs generally take longer to make them, but also may develop more oöcytes at the same time in their ovaries. Finally, maximal volume attained by the cap of seven nurse cells associated with each growing oöcyte appears constant within a species and among species is directly proportional to average mature egg weight.     

Eudesmanolides,guaianolides, germacranolides and elemanolides from Zinnia species

The investigation of six Zinnia species afforded in addition to already known compounds two eudesmanolides, five guaianolides, three germacranolides, five elemanolides and a dihydroxygeranylgeraniol. The structures were elucidated by spectroscopic methods and some chemical transformations. The chemotaxonomic situation is discussed briefly.     

Genetic structure in aquatic bladderworts: clonal propagation and hybrid perpetuation

• Background and Aims The free-floating aquatic bladderwort Utricularia australis f. australis is a sterile F₁ hybrid of U. australis f. tenuicaulis and U. macrorhiza. However, co-existence of the hybrids and parental species has not been observed. In the present study, the following questions are addressed. (a) Does the capacity of the two parental species to reproduce sexually contribute to higher genotypic diversity than that of sterile F₁ hybrid? (b) Are there any populations where two parental species and their hybrid co-exist? (c) If not, where and how do hybrids originate?• Methods The presence and absence of Utricularia was thoroughly investigated in two regions in Japan. An amplified fragment length polymorphism (AFLP) analysis was conducted for 397 individuals collected from all populations (33 in total) where Utricularia was observed.• Key Results The mean number of genotypes per population (G) and genotypic diversity (D) were extremely low irrespective of the capacity to reproduce sexually: G was 1·1–1·2 and D was 0·02–0·04. The hybrid rarely co-existed with either parental species, and the co-existence of two parental species was not observed. Several AFLP bands observed in the hybrid are absent in both parental genotypes, and parent and hybrid genotypes in the same region do not show greater genetic similarity than those in distant regions.• Conclusions The capacity to reproduce sexually in parental species plays no role in increasing genotypic diversity within populations. The observed genotypes of the hybrid could not have originated from hybridization between the extant parental genotypes within the study regions. Considering the distribution ranges of three investigated taxa, it is clear that the hybrid originated in the past, and hybrid populations have been maintained exclusively by clonal propagation, which may be ensured by both hybrid vigor and long-distance dispersal of clonal offspring.     

Strong plastid degradation is consistent within section Chondrophyllae,the most speciose lineage of Gentiana

Recovering phylogenetic relationships in lineages experiencing intense diversification has always been a persistent challenge in evolutionary studies, including in Gentiana section Chondrophyllae sensu lato (s.l.). Indeed, this subcosmopolitan taxon encompasses more than 180 mostly annual species distributed around the world. We sequenced and assembled 22 new plastomes representing 21 species in section Chondrophyllae s.l. In addition to previously released plastome data, our study includes all main lineages within the section. We reconstructed their phylogenetic relationships based on protein‐coding genes and recombinant DNA (rDNA) cistron sequences, and then investigated plastome structural evolution as well as divergence time. Despite an admittedly humble species cover overall, we recovered a well‐supported phylogenetic tree based on plastome data, and found significant discordance between phylogenetic relationships and taxonomic treatments. Our results show that G. capitata and G. leucomelaena diverged early within the section, which is then further divided into two clades. The divergence time estimation showed that section Chondrophyllae s.l. evolved in the second half of the Oligocene. We found that section Chondrophyllae s.l. had the smallest average plastome size (128 KB) in tribe Gentianeae (Gentianaceae), with frequent gene and sequence losses such as the ndh complex and its flanking regions. In addition, we detected both expansion and contraction of the inverted repeat (IR) regions. Our study suggests that plastome degradation parallels the diversification of this group, and illustrates the strong discordance between phylogenetic relationships and taxonomic treatments, which now need to be carefully revised.     

Gynoecium and perianth inZanthoxylum s.l. (Rutaceae)

The question whether the uniseriate perianth ofZanthoxylum L. s. str. is homologous with the calyx or the corolla of taxa included inFagara, or of an independent origin, has been controversial for a long time, but the arguments mostly have remained theoretical. The present investigation of floral structures indicates that there are two different types of uniseriate perianth inZanthoxylum s. str. Therefore, this taxon does not represent a natural group and should be united withFagara asZanthoxylum s.l. The infrageneric taxonomy of this genus is still very ambiguous. It is shown that differences in indumentum, number of sepals and petals (5-4-3) resp. perianth segments (4–9), stamens (3–6), and free carpels (1–5) are of systematic relevance. Particularly important but so far neglected is carpel shape, where an acrostylous and an anacrostylous-basistylous type can be recognized. Stigmata of 2 or more carpels mostly fuse to form a compitum. 4–5-merous flowers with calyx and corolla, and acrostylous carpels are considered as plesiomorphic character states in the genus. On the basis of ± corresponding morphological and phytochemical progressions a working hypothesis about the relationships withinZanthoxylum s.l. is presented in graphical form (Fig. 9).Adapted from a lecture held at the 10th Symposion on Morphology, Anatomy, and Systematics in Göttingen, February 1991.     

Vitellin and vitellogenin concentrations during oögenesis in the first gonotrophic cycle of the house fly,Musca domestica

The house fly, Musca domestica, contains at least two native vitellin and two vitellogenin proteins. Both vitellins appear to have an identical vitellogenin partner. The major native vitellin has a mol. wt of 281 K Daltons, and the major native vitellogenin has a mol. wt of 283 K Daltons. These proteins are composed of three subunits with mol. wt of 48, 45 and 40 K Daltons. The relationship of the subunits to the native proteins is not known.Haemolymph vitellogenin levels are cyclical during oögenesis with no detectable amounts in previtellogenic flies and low levels in postvitellogenic flies. The highest level of vitellogenin, 10.5 μg/μl, occurred in flies with stage-7 ovaries. The vitellogenin levels during oögenesis fit a parabolic curve and the fat body vitellogenin content during oögenesis showed this same pattern.Uptake of vitellogenin into the ovary during each stage of oögenesis also fit a parabolic curve and produced a high linear correlation with haemolymph vitellogenin levels. The greatest uptake was 37 μg/stage and occurred during stage 6.     

ω-ethyl, ω-ethenyl and ω-ethynyl-α-alkylidene-γ-lactones from Clinostemon mahuba

The trunk wood of Clinostemon mahuba contains eight (3R)-2-alkylidene-3-hydroxy-4-methylenebutanolides, seven (3R,4S)-2-alkylidene-3-hydroxy-4-methylbutanolides and seven (3S,4S)-2-alkylidene-3-hydroxy-4-methylbutanolides distinguished by the alkylidene side chains with respect to their E- or Z-geometry, ethenyl, ethynyl or ethyl terminals and lengths (C₁₆ or C₁₈).