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.     

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.     

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 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.     

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.     

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.     

Pollen Morphology of Hydrocotyloideae and Saniculoideae (Apiaceae) in China

Pollen grains of 30 species and 2 varieties from China, belonging to 5 genera (out of 36 species in 5 genera) of Hydrocotyloideae and Saniculoideae, were examined with light microscope and scanning electron microscope. 1.Six types of the pollen shape are recognized in Hydrocotyloideae and Saniculoideae from China, and their evolutionary trend is from rhomboidal→subspheroidal→ellipsoidal→subrectangular→superrectangular to equatorially-constricted. 2.Pollen grains of Hydrocotyloideae are mostly ellipsoidal with P / E ratio 1.07-1.6, the polar axis 22.5-46μm long, the equatorial axis 13.75-27.5μm long, and with the size index 21.1-31.8.They are angulapeturate in the great majority, and the exine sculpture is recticulate under SEM. Those of Saniculoideae are mostly superrectangular or rectangular, with P/E ratio 1.3-2.1, the polar axis 35-65μm long, the equatorial axis 17.5-42.5μm long, and with the size index 28.2-49.7. They are planaperturate in the great majority and the exine sculpture is striate-reticulate or cerebro-reticulate under SEM. Therefore, the latter might be more advanced than the former. 3.Pollen grains of Dickinsia Franch., endemic to China, are ellipsoidal,with P/E ratio 1.5,the polar axis 31.25-37.5μm long, the equatorial axis 20-25μm, and with the size index 27.81. They are angulaperturate, and the exine sculpture is cerebroid-reticulate under SEM. The genus is greatly similar to the members of Hydrocotyloideae in pollen morphology, which supports the treatment of the genus asa member of Hydrocotyloideae by Handel-Mazzetti (1933).     

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.     

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。     

中国伞形科变豆菜亚科的果实解剖特征及其系统学意义

对中国伞形科变豆菜亚科 (Saniculoideae) 2属 (变豆菜属SaniculaL .和刺芹属EryngiumL .) 13种植物的果实形态结构进行了比较解剖学观察 ,从果实横切面形状及其合生面宽度、外果皮和中果皮外突所呈皮刺和鳞片或瘤状体的形态及皮刺中木化厚壁细胞组织的分布、外果皮细胞形状及其角质层厚度、中果皮细胞层数和结晶体类型及分布、色素块沉积、伴生分泌管和油管的大小及分布等方面对上述各属种进行了观察和分析 ,归纳出各属的果实解剖特征 ,探讨了属间演化水平 ,认为刺芹属果实较变豆菜属果实演化程度高。结合已有的研究结果 ,讨论了该亚科与伞形科天胡荽亚科 (Hydrocotyloideae)间的区别及其演化关系。     

THE USE OF PROTEIN-SEROLOGICAL CHARACTERS IN THE SYSTEMATICS OF THE FAMILY OLEACEAE

Serological analyses of soluble seed proteins of 12 representative taxa of the family Oleaceae by the techniques of Ouchterlony, presaturation, and immunoelectrophoresis (IEP) yielded complementary taxonomic information. Ouchterlony reactions differentiated among protein extracts of three species, and the combined serological techniques permitted the detection of protein differences of respective taxa of the two subfamilies Jasminoideae and Oleoideae. IEP enabled the separation of the 12 taxa investigated and the distribution into the two subfamilies, by the differential electrophoretic positions of precipitin arcs, which were consistent with members of each of the two subfamilies. Presaturation data, when analyzed by two cluster analysis computer programs, were taxonomically significant. One program, which calculated amalgamation distances from the presaturation data, clustered the 12 taxa into subgroups which corresponded to tribes, and within two groups, which corresponded to two subfamilies. A monothetic clustering program provided theoretical information on evolution of taxa within the family Oleaceae based on serological correspondences obtained from the presaturation data; the subfamily Jasminoideae was found phylogenetically primitive and the subfamily Oleoideae was advanced. Additionally, IEP data supported those theories suggesting that taxa of the Oleoideae evolved from taxa of the Jasminoideae. The groupings of taxa and different information obtained from the cluster analyses of data and all serological techniques reinforced each other, as well as contemporary taxonomic and phylogenetic treatments of taxa of the family Oleaceae. This research demonstrated the taxonomic value of protein-serological data, particularly as applied to the taxonomy of the Oleaceae.     

红花变豆菜叶绿体基因组结构及同属种间关系研究

红花变豆菜（Sanicula rubriflora F. Schmidt）是有药用价值的植物,全株干燥后与其他药用同属植物易混淆,种间关系存在争议,通过高通量测序技术对红花变豆菜叶绿体基因组测序,利用生物信息学方法对测序数据进行拼接、注释,首次报道红花变豆菜叶绿体基因组结构及特点,利用叶绿体基因组数据,提供种间分类新证据,并且分析相关类群的进化关系。S. rubriflora叶绿体基因组序列的长度为155 721 bp,其中包括一个85 981 bp的大单拷贝区（large single copy,LSC）和一个17 060 bp的小单拷贝区（small single-copy region,SSC）,它们被两个26 340 bp的反向重复区（inverted repeat sequence,IRs）隔开。红花变豆菜叶绿体基因组GC含量为38.20%,包含129个基因,其中84个蛋白质编码基因,37个tRNA基因和8个rRNA基因。红花变豆菜叶绿体基因组结构具有高度保守性,其中编码基因共有51 907个密码子,最多编码5 095个亮氨酸,最少编码689个色氨酸,简单重复序列分析共发现32个位点,大多数是单碱基重复的A/T类型。叶绿体基因组聚类结果支持天胡荽亚科（Hydrocotyloideae）是伞形科（Umbelliferae）内比较原始的类群;变豆菜亚科（Saniculoideae）和芹亚科（Apioideae）为姊妹类群,是伞形科较进化的类群;变豆菜属植物是一个相对自然的类群;红花变豆菜与黄花变豆菜（S. flavovirens）为近缘姊妹种,但是两者形态和地理分布差异较大。该研究结果为变豆菜属属下种间鉴定及其种间演化奠定基础。     

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.     

A study of the systematics of cyprinid fishes by two-dimensional gel electrophoresis

This study was carried out to shed light on confused subfamilial groupings in the Cyprinidae from the biochemical viewpoint at the molecular level, specifically by using two-dimensional gel electrophoresis of liver proteins. Six pairs of cypriniform fishes, which are different from one another at familial, subfamilial, generic, specific, subspecific, and individual levels, were compared. The genetic distances between pairs of fishes increased as taxonomic ranks of the pairs became higher, confirming the reliable usefulness of this technique. Four species representing the different subfamilies, Cyprininae, Gobioninae, Acheilognathinae, and Leuciscinae, were compared to give new insight into relationships at the subfamilial level. Cyprinus carpio (Cyprininae) and Pseudogobio esocinus esocinus (Gobioninae) gave the smallest genetic distance and the largest values were obtained between either one of the above species and Acheilognathus melanogaster (Acheilognathinae), suggesting that the former two subfamilies compose the most closely related group that is in turn distantly related to Acheilognathinae. Tribolodon hakonensis (Leuciscinae) had almost equal genetic distances to the three other species.     

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.     

Nucleolar organizing region in theApiaceae (Umbelliferae)

Data available for the nucleolus organizing region in the familyApiaceae are reviewed. An attempt has been made to establish the exact number of this region in various subfamilies and tribes through studies on the karyotype and nucleolus. Most of the taxa have a single nucleolar chromosome per haploid complement. The location of the nucleolar organizing region (NOR) on the chromosome varies. Members ofHydrocotyloideae differ drastically from those of subfam.Saniculoideae andApioideae, with respect to the location of NOR. Despite wide geographical distribution, varied ecological preferences and differences in morphology, anatomy and cytology, Umbellifers have attained stability in the number and location of NORs. Characters of NOR offer scope for utilization in understanding phylogenetic relationships at higher levels of taxonomic hierarchy.Studies on the Nucleolus and Nucleolar Chromosomes in Angiosperms XII.     

Wood and bark anatomy of Steganotaenia and Polemanniopsis (tribe Steganotaenieae,Apiaceae) with notes on phylogenetic implications

The wood and bark structure of the distinctive southern African genera Polemanniopsis (including the newly described species P. namibensis) and Steganotaenia have been described. To allow for comparisons with the traditional subfamily Saniculoideae, a shrubby species of Eryngium from the Juan Fernández Islands was also studied. Polemanniopsis and Steganotaenia were recently considered as two closely related genera forming a new tribe Steganotaenieae of subfamily Saniculoideae (Apiaceae), whereas Eryngium is commonly recognized as a member of Saniculoideae. Eryngium differs significantly from the other two genera in the smaller size of intervessel pits, sclerification and radial dilatation in collapsed secondary phloem, the absence of crystals in the phelloderm cells and the occurrence of druse crystals in secondary phloem ray cells. Steganotaenia and Polemanniopsis share features, including the presence of marginal axial parenchyma, the occurrence of radial secretory canals in secondary xylem, dilatation of the secondary phloem by axial parenchyma stretching, cortical periderm initiation and the presence of chambered phelloderm cells containing druse crystals. These characters (especially the occurrence of chambered crystalliferous cells in phelloderm, which has not yet been reported for Apiaceae) support both the monophyly and the isolated position of the Steganotaeniae. No reliable synapomorphic features could be found to support a relationship with Saniculoideae. Steganotaenia is remarkable in the presence of axial secretory canals in the phelloderm: these structures have not yet been found in the periderm of any member of Apiales. Our results do not provide any support for the suggestion that the woody habit in the three genera examined was derived from herbaceous ancestors secondarily. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 163 , 55–59.     

Phylogenetic relationships among the Braconidae (Hymenoptera: Ichneumonoidea) inferred from partial 16S rDNA, 28S rDNA D2, 18S rDNA gene sequences and morphological characters

Phylogenetic relationships among the Braconidae were examined using homologous 16S rDNA, 28S rDNA D2 region, and 18S rDNA gene sequences and morphological data using both PAUP* 4.0 and MRBAYES 3.0B4 from 88 in-group taxa representing 35 subfamilies. The monophyletic nature of almost all subfamilies, of which multiple representatives are present in this study, is well-supported except for two subfamilies, Cenocoelinae and Neoneurinae that should probably be treated as tribal rank taxa in the subfamily Euphorinae. The topology of the trees generated in the present study supported the existence of three large generally accepted lineage or groupings of subfamilies: two main entirely endoparasitic lineages of this family, referred to as the "helconoid complex" and the "microgastroid complex," and the third "the cyclostome." The Aphidiinae was recovered as a member of the non-cyclostomes, probably a sister group of Euphorinae or Euphorinae-complex. The basal position of the microgastroid complex among the non-cyclostomes has been found in all our analyses. The cyclostomes were resolved as a monophyletic group in all analyses if two putatively misplaced groups (Mesostoa and Aspilodemon) were excluded from them. Certain well-supported relationships evident in this family from the previous analyses were recovered, such as a sister-group relationships of Alysiinae+Opiinae, of Braconinae+Doryctinae, and a close relationship between Macrocentrinae, Xiphozelinae, Homolobinae, and Charmontinae. The relationships of "Ichneutinae + ((Adeliinae + Cheloninae) + (Miracinae + (Cardiochilinae + Microgastrinae)))" was confirmed within the microgastroid complex. The position of Acampsohelconinae, Blacinae, and Trachypetinae is problematic.