Chemotypification of Astrantia major L. (Apiaceae): Essential‐Oil and Lignan Profiles of Fruits

The fruit essential oils of two populations of Astrantia major L. (Apiaceae, subfamily Saniculoideae) were analyzed in detail by GC and GC/MS analyses. Seventy‐six constituents identified accounted for 92.7–94.0% of the oils. The two oils differed significantly: the wild‐growing population from Serbia contained zingiberene (47.9%), β‐bisabolene (9.7%), and β‐sesquiphellandrene (7.9%), while the one from Poland (botanical gardens) was sesquiterpene‐poor with the major contributors oleic acid (38.6%), nonacosane (15.4%), and linoleic acid (5.1%). Motivated by the unresolved taxonomical relations between the Saniculoideae and Apioideae subfamilies, we performed multivariate statistical analyses on the compositional data of these A. major samples, and additional 14 Saniculoideae and 31 Apioideae taxa. This allowed us to assess the chemotaxonomical usefulness of such chemical data in differentiating taxa from these two Apiaceae subfamilies and to corroborate the existence of at least two A. major chemotypes. Diethyl ether extracts of the two samples of A. major fruits yielded seven diaryltetrahydrofurofurano lignans. Except for eudesmin that has been found for the first time in a Saniculoideae taxon, all other lignans (magnolin, epimagnolins A and B, epieudesmin, yangambin, and epiyangambin) are new for the entire plant family Apiaceae. The lignan profiles also supported the existence of two separate A. major chemotypes.     

Molecular systematics of Apiaceae subfamily Apioideae: phylogenetic analyses of nuclear ribosomal DNA internal transcribed spacer and plastid RPO C1 intron sequences

Evolutionary relationships among representatives of Apiaceae (Umbelliferae) subfamily Apioideae have been inferred from phylogenetic analyses of nuclear ribosomal DNA internal transcribed spacer (ITS 1 and ITS 2) and plastid rpoC1 intron sequences. High levels of nucleotide sequence variation preclude the use of the ITS region for examining relationships across subfamilial boundaries in Apiaceae, whereas the rpoC1 intron is more suitably conserved for family-wide phylogenetic study but is too conserved for examining relationships among closely related taxa. In total, 126 ITS sequences from subfamily Apioideae and 100 rpoC1 intron sequences from Apiaceae (all three subfamilies) and outgroups Araliaceae and Pittosporaceae were examined. Phylogenies estimated using parsimony, neighbor-joining, and maximum likelihood methods reveal that: (1) Apiaceae subfamily Apioideae is monophyletic and is sister group to Apiaceae subfamily Saniculoideae; (2) Apiaceae subfamily Hydrocotyloideae is not monophyletic, with some members strongly allied to Araliaceae and others to Apioideae + Saniculoideae; and (3) Apiaceae subfamily Apioideae comprises several well-supported subclades, but none of these coincide with previously recognized tribal divisions based largely on morphological and anatomical characters of the fruit. Four major clades in Apioideae are provisionally recognized and provide the framework for future lower level phylogenetic analyses. A putative secondary structure model of the Daucus carota (carrot) rpoC1 group II intron is presented. Of its six major structural domains, domains II and III are the most, and domains V and VI the least, variable.     

A phylogeny of the flowering plant family Apiaceae based on chloroplast DNA rpl16 and rpoC1 intron sequences: towards a suprageneric classification of subfamily Apioideae

The higher level relationships within Apiaceae (Umbelliferae) subfamily Apioideae are controversial, with no widely acceptable modern classification available. Comparative sequencing of the intron in chloroplast ribosomal protein gene rpl16 was carried out in order to examine evolutionary relationships among 119 species (99 genera) of subfamily Apioideae and 28 species from Apiaceae subfamilies Saniculoideae and Hydrocotyloideae, and putatively allied families Araliaceae and Pittosporaceae. Phylogenetic analyses of these intron sequences alone, or in conjunction with plastid rpoC1 intron sequences for a subset of the taxa, using maximum parsimony and neighbor-joining methods, reveal a pattern of relationships within Apioideae consistent with previously published chloroplast DNA and nuclear ribosomal DNA ITS based phylogenies. Based on consensus of relationship, seven major lineages within the subfamily are recognized at the tribal level. These are referred to as tribes Heteromorpheae M. F. Watson & S. R. Downie Trib. Nov., Bupleureae Spreng. (1820), Oenantheae Dumort. (1827), Pleurospermeae M. F. Watson & S. R. Downie Trib. Nov., Smyrnieae Spreng. (1820), Aciphylleae M. F. Watson & S. R. Downie Trib. Nov., and Scandiceae Spreng. (1820). Scandiceae comprises subtribes Daucinae Dumort. (1827), Scandicinae Tausch (1834), and Torilidinae Dumort. (1827). Rpl16 intron sequences provide valuable characters for inferring high-level relationships within Apiaceae but, like the rpoC1 intron, are insufficient to resolve relationships among closely related taxa.     

Morphology and biogeography of Apiaceae subfamily Saniculoideae as inferred by phylogenetic analysis of molecular data

The phylogenetic placements of several African endemic genera at the base of Apiaceae subfamilies Saniculoideae and Apioideae have revolutionized ideas of relationships that affect hypotheses of character evolution and biogeography. Using an explicit phylogeny of subfamily Saniculoideae, we reconstructed the evolutionary history of phenotypic characters traditionally important in classification, identified those characters most useful in supporting relationships, and inferred historical biogeography. The 23 characters examined include those of life history, vegetative morphology, inflorescences, and fruit morphology and anatomy. These characters were optimized over trees derived from maximum parsimony analysis of chloroplast DNA trnQ-trnK sequences from 94 accessions of Apiaceae. The results revealed that many of these characters have undergone considerable modification and that traditional assumptions regarding character-state polarity are often incorrect. Infrasubfamilial relationships inferred by molecular data are supported by one to five morphological characters. However, none of these morphological characters support the monophyly of subfamilies Saniculoideae or Apioideae, the clade of Petagnaea, Eryngium and Sanicula, or the sister-group relationship between Eryngium and Sanicula . Southern African origins of Saniculoideae and of its tribes Steganotaenieae and Saniculeae are supported based on dispersal-vicariance analysis.     

A SEROLOGICAL COMPARISON OF UMBELLIFERAE SUBFAMILIES

Serological techniques were employed to compare the three subfamilies Hydrocotyloideae, Saniculoideae, and Apioideae of the Umbelliferae. Data obtained from turbidimetric and double diffusion analyses of seed proteins from thirteen genera demonstrated three distinct serological groupings. These groupings correspond to the three subfamilies. The serological data indicated that one grouping (Apioideae) was more similar to Saniculoideae than to Hydrocotyloideae.     

Clarification of the relationship beteen Apiaceae and Araliaceae based on matK and rbcL sequence data

Apiaceae and Araliaceae (Apiales) represent a particularly troublesome example of the difficulty in understanding evolutionary relationships between tropical-temperate family pairs. Previous studies based on rbcL sequence data provided insights at higher levels, but were unable to resolve fully the family-pair relationship. In this study, sequence data from a more rapidly evolving gene, matK, was employed to provide greater resolution. In Apiales, matK sequences evolve an average of about two times faster than rbcL sequences. Results of phylogenetic analysis of matK sequences were first compared to those obtained previously from rbcL data; the two data sets were then combined and analyzed together. Molecular analyses confirm the polyphyly of apiaceous subfamily Hydrocotyloideae and suggest that some members of this subfamily are more closely related to Araliaceae than to other Apiaceae. The remainder of Apiaceae forms a monophyletic group with well-defined subclades corresponding to subfamilies Apioideae and Saniculoideae. Both the matK and the combined rbcL-matK analyses suggest that most Araliaceae form a monophyletic group, including all araliads sampled except Delarbrea and Mackinlaya. The unusual combination of morphological characters found in these two genera and the distribution of matK and rbcL indels suggest that these taxa may be the remnants of an ancient group of pro-araliads that gave rise to both Apiaceae and Araliaceae. Molecular data indicate that the evolutionary history of the two families is more complex than simple derivation of Apiaceae from within Araliaceae. Rather, the present study suggests that there are two well-defined "families," both of which may have been derived from a lineage (or lineages) or pro-araliads that may still have extant taxa.     

Geographical and environment-related variations of essential oils in isolated populations of Thymus richardii Pers. in the Mediterranean basin

Composition of essential oils of different populations of Thymus richardii grex of six localities from Bosnia-Herzegovina (Konjic, Borci), Spain (Majorca, Ibiza, Valencia) and Italy (Marettimo, Sicily) were determined by GC/FID and GC/MS. The main constituents in most of the samples were aromatic monoterpenes corresponding to non-phenolic cyclic compounds (p-cymene, γ-terpinene). The highest monoterpene concentrations were found in the Bosnian samples (70%), and the lowest in samples from the Balearic Islands (<30%; Ibiza and Majorca). Sesquiterpenes were the major component (average > 50%) in samples from Majorca with β-bisabolene (>40%) being the principal constituent. Discriminant analysis (LDA) shows the differentiation of two chemotypes: A (phenol chemotype), with p-cymene and γ-terpinene as characteristic compounds and B, with β-bisabolene and carvacrol, as major and significative compounds. The occurrence of the chemotypes was related to summer positive precipitation and to deep of soils.     

Volatile secondary metabolites of Micromeria dalmatica Benth. (Lamiaceae): biosynthetical and chemotaxonomical aspects

Analysis by GC and GC/MS of the essential oil obtained from above-ground parts of Micromeria dalmatica Benth. allowed the identification of 116 components, comprising 93.6% of the total oil composition. The major compounds are 3-oxygenated p-menthane monoterpenes and were identified as pulegone (29.6%), menthone (11.7%), and piperitenone (10.8%). The chemical composition of this and additional 30 oils obtained from selected Micromeria Benth. taxa were compared by using multivariate statistical analysis (agglomerative hierarchical cluster analysis and principal component analysis (PCA)). The results of statistical analyses, as well as the domination of different concurrent p-menthane-skeleton-type monoterpene biosynthetical sub-branches in the compared M. dalmatica samples, implied the occurrence of at least two different chemotypes of the mentioned species.     

CHROMOSOME NUMBERS IN UMBELLIFERAE. IV.

Chromosome numbers are reported for 167 collections representing 100 taxa of Umbelliferae. More than four-fifths of the counts apply to members of subfamilies Hydrocotyloideae (29) and. Saniculoideae (50); the remaining 21 belong to Apioideae. Chromosome numbers of plants belonging to 68 taxa are published here for the first time; chromosome numbers are verified for 23 taxa; and chromosome numbers differing from those published previously are reported in nine instances. No chromosome counts have previously been reported for 19 of the genera included. Polyploidy has been established for Azorella, Mulinum, Coaxana, Enantiophylla, and Tiozimia.     

Essential oil composition of two variants of Prostanthera lasianthos Labill. from Australia

The essential oils from the leaves of two variants of Prostanthera lasianthos Labill. have been analysed by GC and GC/MS. The different samples studied showed two chemotypes, the rheophytic variant, chemotype 1,8-cineole and β-pinene and the smooth-leaved variant with the chemotype linalool, linalyl acetate and β-selinene. The percentage composition of these compounds were 57.3–66.0%, 9.2–10.2%, 13.8–24.6%, 13.8–19.1% and 7.8–14.2%, respectively. One of the samples (P.l.n2) showed intermediate values so it could be a hybrid although it was morphologically similar to smooth-leaved variant. According to our chemical results and previous morphological studies we think that both variants could be recognised as distinct taxa level (subspecies or species) although further genetic research should be done to confirm this hypothesis.     

A molecular phylogenetic study of southern African Apiaceae

It has been suggested that southern Africa is the origin of the predominantly herbaceous Apiaceae subfamily Apioideae and that the woody habit is plesiomorphic. We expand previous molecular phylogenetic analyses of the family by considering all but three of the approximately 38 genera native to southern Africa, including all genera whose members, save one, have a woody habit. Representatives of five other genera are included because they may be closely related to these southern African taxa. Chloroplast DNA rps16 intron and/or nuclear rDNA ITS sequences for 154 accessions are analyzed using maximum parsimony, Bayesian, and maximum likelihood methods. Within Apioideae, two major clades hitherto unrecognized in the subfamily are inferred. The monogeneric Lichtensteinia clade is sister group to all other members of the subfamily, whereas the Annesorhiza clade (Annesorhiza, Chamarea, and Itasina) plus Molopospermum (and Astydamia in the ITS trees) are the successive sister group to all Apioideae except Lichtensteinia. Tribe Heteromorpheae is expanded to include Pseudocarum, "Oreofraga" ined., and five genera endemic to Madagascar. The southern African origin of subfamily Apioideae is corroborated (with subsequent migration northward into Eurasia along two dispersal routes), and the positions of the herbaceous Lichtensteinia and Annesorhiza clades within the subfamily suggest, surprisingly, that its ancestor was herbaceous, not woody.     

Major lineages within Apiaceae subfamily Apioideae: a comparison of chloroplast restriction site and DNA sequence data

Traditional sources of taxonomic characters in the large and taxonomically complex subfamily Apioideae (Apiaceae) have been confounding and no classification system of the subfamily has been widely accepted. A restriction site analysis of the chloroplast genome from 78 representatives of Apioideae and related groups provided a data matrix of 990 variable characters (750 of which were potentially parsimony-informative). A comparison of these data to that of three recent DNA sequencing studies of Apioideae (based on ITS, rpoCl intron, and matK sequences) shows that the restriction site analysis provides 2.6–3.6 times more variable characters for a comparable group of taxa. Moreover, levels of divergence appear to be well suited to studies at the subfamilial and tribal levels of Apiaceae. Cladistic and phenetic analyses of the restriction site data yielded trees that are visually congruent to those derived from the other recent molecular studies. On the basis of these comparisons, six lineages and one paraphyletic grade are provisionally recognized as informal groups. These groups can serve as the starting point for future, more intensive studies of the subfamily.     

伞形科柴胡属部分物种的果实特征及系统学意义

对21个代表种果实形态与解剖特征观察发现：（1）中国柴胡属（Bupleurum L.）果实以芹亚科（Apioideae）原始的两侧压扁型为主,在外形上可分为圆柱形、矩圆形和卵形3类;（2）在柴胡属中果皮细胞内没有发现广泛存在于芹亚科基部类群变豆菜亚科（Saniculoideae）和天胡荽亚科（Hydrocotyloideae）的晶体后含物;（3）从果实的横切面看,可将柴胡属划分为棱槽单油管型和棱槽多油管型两大类。结合前人对伞形科其它类群果实形态解剖特征的研究及近年来分子系统学的证据,认为：（1）中国柴胡属植物的多油管类型为本属较进化类群;（2）柴胡属可能是芹亚科中连接基部类群与其它类群的过渡环节或进化旁支;（3）果实形态与解剖特征可作为探讨国产柴胡属种内和种间关系的辅助性状,并给出了分种检索表。     

Occurrence of 1,5-dimethylcyclodeca-1,5,7-triene (Pregeijerene) in Pimpinella species and chemosystematic implications

The essential oils from fresh roots of 66 species, representing 39 general of the Apiaceae (Umbelliferae), subfamily Apioideae, were surveyed for pregeijerene. Of all plants examined, only the 11 species of the genus Pimpinella were shown to contain pregeijerene. According to its extremely restricted occurrence in the Apiaceae, its possible taxonomic value with regard to the definition of the genus Pimpinella is discussed.     

The phylogenetic significance of the carpophore in Apiaceae

Background and aims

Fruit structural characters have traditionally been important in the taxonomy of the family Apiaceae. Previous investigations using a limited number of taxa have shown that the carpophore may be especially useful in helping to circumscribe subfamily Azorelloideae. The present study examines, for the first time, carpophore structure in 92 species from 43 genera, representing all subfamilies of Apiaceae, and including all genera assigned to subfamily Azorelloideae. Phylogenetic interpretations are made for the first time, using all available information, and a standard terminology is proposed to describe the various character states found in carpophores.

Methods

Carpophore structure was studied in detail using light microscopy.

Key Results

Carpophores, when present, may be categorized into two main groups (B and C) based mainly on the arrangement of the vascular bundles in transverse section, and further divided into six sub-types according to the length of the carpophore (short in B1 and C1) and whether they are entire (B1–B3 and C1) or bifurcate (B4 and C2). Free carpophores are absent in subfamily Mackinlayoideae, and in tribes Lichtensteinieae and Phlyctidocarpeae, which have two opposite vascular bundles (Group A). Entire carpophores with one or two vascular bundles, or bifurcate carpophores with lateral vascular bundles (arranged side by side within the commissural plane), are the main types characterizing Azorelloideae. The short, hygroscopic carpophores found in Choritaenia are unique in Apiaceae and provide additional evidence for the exclusion of this genus from Azorelloideae. Carpophore type C2 is typical for most Apioideae sensu lato (exceptions are, for example, Arctopus and Alepidea, which have type B2).

Conclusions

A single carpophore and ventral vascular bundles not forming free carpophores are proposed to be the ancestral conditions in Apiaceae, while bifurcate carpophores with opposite vascular bundles are the derived state, present in most Apioideae. Secondary reductions seem to have occurred in several unrelated lineages in all major groups, e.g. many Azorelloideae, several protoapioids (including nearly all members of the tribe Saniculeae) and 29 euapioid genera (e.g. some Oenantheae).     

Using RAD seq for reconstructing phylogenies of highly diverged taxa: A test using the tribe Scandiceae (Apiaceae)

The angiosperm Apiaceae tribe Scandiceae includes four major clades—subtribes Daucinae, Ferulinae, Torilidinae, and Scandicinae—that originated ca. 20 Mya. Although all four subtribes are highly supported in molecular analyses, and morphological data indicate a sister relationship between Daucinae and Torilidinae, their branching order has not been resolved using standard Sanger multilocus data. Therefore, in this study, we test the utility of genomic RAD seq data in resolving deep phylogenetic relationships (up to 20 Mya) in Apiaceae subfamily Apioideae, with special emphasis on tribe Scandiceae using 12 representative species. We used two bioinformatic pipelines, pyRAD and RADIS (based on STACKS), to assemble RAD seq data and we tested the influence of various combinations of parameters on the robustness of the inferred tree topologies. Although different data processing approaches produced alignments with various amounts of missing data, they converged to two well‐supported topologies, irrespective of the phylogenetic method applied. Highly supported trees showed Scandicinae as sister to all other clades and indicated that Daucinae and Torilidinae are sister groups, thus confirming the relationship inferred from morphology. We conclude that the RAD seq method can be successfully used to resolve deep relationships formed 20 Mya within Apiaceae. We provide recommendations for parameter settings in RADIS and pyRAD for the analysis of taxa that have accumulated considerable genomic divergence.     

Polyphyletic origin in <Emphasis Type="Italic">Pimpinella</Emphasis> (Apiaceae): evidence in Western Europe

The genus Pimpinella L. comprises about 150 species, being one of the largest genera within the family Apiaceae (subfamily Apioideae). Previous molecular phylogenetic studies have shown that Pimpinella is a taxonomically complex group. In this study, evolutionary relationships among representatives from Western Europe have been inferred from phylogenetic analyses of nuclear ribosomal DNA internal transcribed spacer (ITS 1 and ITS 2) and plastid sequences (trnL intron and the trnL-F spacer), with a representative sampling included (168 accessions in the ITS analysis, representing 158 species; and 42 accessions in the cpDNA analysis representing 35 taxa of Pimpinella and closely related species). All analyses resolved that Pimpinella is a non-monophyletic group, and Pimpinella’s taxa that grow in Western Europe are part of phylogenetically independent groups that correspond to three different tribes of the subfamily Apioideae: Pimpinelleae (core group), Pyramidoptereae and Smyrnieae.     

Chemical composition of hips essential oils of some Rosa L. species

The chemical composition of the hips essential oils of 9 taxa of Rosa L. was analyzed and compared using the standardized analytical GC and GC/MS methods. The volatile hips oil compositions for these species are presented. All oil samples were dominated by following components: vitispiran (isomer), a-E-acaridial, dodecanoic acid, hexadecanoic acid, docosane (C22), beta-ionone, 6-methyl-5-hepten-2-one, 2-heptanone, heptanal, myristic acid and linolic acid. Statistical analyses of 97 GC peaks of these oils were used to distinguish compositional patterns. There appeared to be correlation between the essential oil patterns and the classification within Rosa L. Cluster analysis of the composition of main components clearly showed two groups, one constituted by R. rugosa Thunb. from the Cinnamomea section, and the other constituted by the remaining taxa from the Caninae section.