Pollen morphology of the tribe Cynoglosseae of Boraginoideae (Boraginaceae) in China

Pollen morphology of 16 Chinese species representing 7 genera in the tribe Cynoglosseae of Boraginoideae(Boraginaceae) was examined under LM and SEM, and 5 species under TEM. Pollen grains are cocoon-shaped, rarely subprolate, prolate or ovoid, very small, 7～ 15.7μm× 3.5 ~ 13.9 μm in size, P/E = 1.6～2.02; 3-colporate apertures alternate with 3-pseudocolpi, with equatorial endocingulus except those in Bothriospermum; exine surface is usually smooth, with or without perforations in two poles, rarely with tuberculate ornamentation; exine is rather thin and includes ectexine and endexine, while ectexine consists of imperforate tectum, columellae and foot-layer. The tribe Cynoglosseae has many common characters of pollen grains, but there are some differences among genera. A key to the genera is given based on pollen morphology. Bothriospermum, without endocingulus, may be a primitive genus in this tribe, and its pollen morphology is more similar to that of Eritrichieae than to Cynoglosseae, thus it seems more reasonable to put it into Eritrichieae. Solenanthus may be the most advanced genus in the tribe Cynoglosseae for its pollen grains of sub-isopolar and ovoid shape.     

Phylogenetic relationships within Pappophoreae s.l. (Poaceae: Chloridoideae): Additional evidence based on ITS and trnL-F sequence data

Historically, Pappophoreae included the genera Cottea, Enneapogon, Kaokochloa, Pappophorum and Schmidtia. Some authors consider this tribe as a well-supported monophyletic group; while other evidences reveals Pappophoreae as polyphyletic, with Pappophorum separated from the rest of the tribe. When the latter happens, it can form a clade with Tridens flavus. Molecular phylogenetic analyses of the subfamily Chloridoideae have included few species of Pappophoreae; therefore, further research involving more representatives of this tribe is needed. With the aim of providing new evidence to help clarify the phylogenetic position of Pappophorum and its relationships with other genera of the tribe and the subfamily Chloridoideae, eight new sequences of ITS and trnL-F regions of Pappophoreae species were generated. These sequences were analyzed together with other available sequence data obtained from GenBank, using maximum parsimony and Bayesian inference, for individual (trnL-F or ITS) or combined trnL-F/ITS data sets. All analyses reveal that Pappophoreae is polyphyletic, with Pappophorum separated from the rest of the tribe forming a well-supported clade sister to Tridens flavus.     

Generic relationships among the baccate-fruited Amaryllidaceae (tribe Haemantheae) inferred from plastid and nuclear non-coding DNA sequences

Using sequences from the plastid trnL-F region and nrDNA ITS, we investigated the phylogeny of the fleshy-fruited African tribe Haemantheae of the Amaryllidaceae across 19 species representing all genera of the tribe. ITS and a combined matrix produce the most resolute and well-supported tree with parsimony analysis. Two main clades are resolved, one comprising the monophyletic rhizomatous genera Clivia and Cryptostephanus, and a larger clade that unites Haemanthus and Scadoxus as sister genera to an Apodolirion/Gethyllis subclade. One of four included Gethyllis species, G. lanuginosa, resolves as sister to Apodolirion with ITS. Relationships among the Clivia species are not in agreement with a previous published phylogeny. Biogeographic analysis using the divergence/vicariance method roots the tribe in Eastern South Africa, with several subsequent dispersals to the winter rainfall Western Cape region. Chromosomal change from an ancestral 2n=22 (characteristic of Clivia) is associated with each main clade. Reduction in number has occurred in all but Cryptostephanus, which has 2n=24 chromosomes. Increasing the sampling across all of the species in the tribe will allow a more detailed understanding of the biogeographic patterns inherent in the parsimony topology, which undoubtedly reflect Quaternary climatic changes in Southern Africa.     

Phylogeny and genetic diversity of D-genome species of Aegilops and Triticum (Triticeae, Poaceae) from Iran based on microsatellites, ITS, and trnL-F

Cereal species of the grass tribe Triticeae are economically important and provide staple food for large parts of the human population. The Fertile Crescent of Southwest Asia harbors high genetic and morphological diversity of these species. In this study, we analyzed genetic diversity and phylogenetic relationships among D genome-bearing species of the wheat relatives of the genus Aegilops from Iran and adjacent areas using allelic diversity at 25 nuclear microsatellite loci, nuclear rDNA ITS, and chloroplast trnL-F sequences. Our analyses revealed high microsatellite diversity in Aegilops tauschii and the D genomes of Triticum aestivum and Ae. ventricosa, low genetic diversity in Ae. cylindrica, two different Ae. tauschii gene pools, and a close relationship among Ae. crassa, Ae. juvenalis, and Ae. vavilovii. In the latter species group, cloned sequences revealed high diversity at the ITS region, while in most other polyploids, homogenization of the ITS region towards one parental type seems to have taken place. The chloroplast genealogy of the trnL-F haplotypes showed close relationships within the D genome Aegilops species and T. aestivum, the presence of shared haplotypes in up to three species, and up to three different haplotypes within single species, and indicates chloroplast capture from an unidentified species in Ae. markgrafii. The ITS phylogeny revealed Triticum as monophyletic and Aegilops as monophyletic when Amblyopyrum muticum is included.     

Molecular phylogeny and systematics of the tribe Chorisporeae (Brassicaceae)

Sequence data from nuclear (ITS) and chloroplast (trnL-F) regions for 89 accessions representing 56 out of 64 species from all five genera of the tribe Chorisporeae (plus Dontostemon tibeticus) have been studied to test the monophyly of the tribe and its component genera, clarify its boundaries, and elucidate its phylogenetic position in the family. Both data sets showed strong support for the monophyly of the Chorisporeae as currently delimited, though the position of its tentative member D. tibeticus was not resolved by ITS. Parrya and Pseudoclausia are poly- and paraphyletic with regard to each other, and Chorispora is either polyphyletic or at least paraphyletic (comprising Diptychocarpus) within a weakly supported monophyletic clade. The incongruence in branching pattern among the markers was most likely caused by hybridization and possibly influenced by incomplete lineage sorting. The present results suggest uniting Pseudoclausia, Clausia podlechii, and Achoriphragma with Parrya and transferring P. beketovii and P. saposhnikovii to Leiospora (Euclidieae). We also obtained support for splitting Chorispora into two geographically defined groups, one of which is closer to Diptychocarpus. Both data sets revealed a close relationship of the Chorisporeae to Dontostemoneae, while ITS also indicated affinity to Hesperideae. Therefore, the position of Chorisporeae needs further verification.     

Tribes of Boraginoideae (Boraginaceae) and placement of Antiphytum, Echiochilon, Ogastemma and Sericostoma: A phylogenetic analysis based on atpB plastid DNA sequence data

 The genera Antiphytum, Echiochilon, Ogastemma and Sericostoma, which have been difficult to place within the tribes of the subfamily Boraginoideae (Boraginaceae), are analysed using plastid atpB sequence data. A selection of Boraginaceae genera was used to obtain a framework for the phylogenetic position of Antiphytum, Echiochilon, Ogastemma and Sericostoma. Sericostoma is found to belong within Echiochilon. The new tribe Echiochileae, Boragineae and Lithospermeae are monophyletic but the tribes Eritrichieae and Cynoglosseae are paraphyletic. The biogeography of Echiochileae (Echiochilon and Ogastemma from Africa and western Asia, and Antiphytum from America) is discussed. Received November 27, 2001; accepted March 20, 2002 Published online: November 14, 2002 ? Springer-Verlag 2002 Addresses of the authors: Elisabeth L?ngstr?m (e-mail: elisabeth.langstrom@ebc.uu.se), Department of Systematic Botany, Evolutionary Biology Centre, Uppsala University, Norbyv?gen 18D, SE-752 36 Uppsala, Sweden. Mark W. Chase, Jodrell Laboratory, Royal Botanic Gardens, Kew, TW9 3DS, UK.     

Phylogenetic analysis of Lappula Moench (Boraginaceae) based on molecular and morphological data

Lappula Moench includes ca. 70 species and exhibits a wide range of nutlet variation. Currently, the evolutionary relationships among species of Lappula have not been examined; therefore, to elucidate phylogenetic relationships and morphological evolution within Lappula and related genera, we conducted phylogenetic analyses with matrices that include 48 species as well as four DNA regions (ITS, trnL-trnF, rpS16 and psbA-trnH) and 18 morphological characters. These matrices were analyzed using maximum parsimony, maximum likelihood, and Bayesian inference methods. Analyses of the combined molecular and morphological data result in a phylogeny that is better resolved than that based solely on molecular sequence data. Phylogenetic results suggest that the current infrageneric classification of Lappula, at least at the subsectional and series level, is artificial. The evolutionary patterns of 18 morphological characters are investigated in a phylogenetic context. In Lappula, nutlet homomorphism and small corollas are resolved as ancestral, while nutlet heteromorphism and larger corollas are derived.     

Phylogenetic status of Vavilovia formosa (Fabaceae-Fabeae) based on nrDNA ITS and cpDNA sequences

The phylogenetic status of the monotypic genus Vavilovia was studied using nrDNA ITS and cpDNA trnL-F and trnS-G regions. The results from the analysis of each dataset and the combined dataset, revealed that Vavilovia is closely related to Pisum, forming a group that is sister to Lathyrus. The molecular data and some morphological and biological characteristics strongly indicate that Vavilovia should be subsumed under Pisum, as Pisum formosum.     

Molecular phylogeny of Plumbaginaceae with emphasis on Acantholimon Boiss. based on nuclear and plastid DNA sequences in Iran

The present study represents phylogenetic analyses of Plumbaginaceae with emphasis on Acantholimon from Iran using nrDNA ITS and plastid trnY-T sequences. The analyses support the monophyly and the close relationship of Limonium, Armeria and Psylliostachys. This is the first report of the close relationship between Acantholimon and Cephalorhizum. The data for the position of Cephalorhizum is unclear. The Shimodaira–Hasegawa test of nrDNA ITS and the combined datasets indicated that Acantholimon and Cephalorhizum are distinct genera. The molecular data revealed that the traditionally recognized multi-specific Acantholimon sections (Acantholimon, Acmostegia, Glumaria, Microstegia, Staticopsis and Tragacanthina) are not monophyletic. Their members are intermixed with each other and scattered across the Acantholimon clade, but the smaller sections including Platystegia and Pterostegia, each comprising two species, are monophyletic.     

Hybridization between the threatened plant, <Emphasis Type="Italic">Lespedeza leptostachya</Emphasis> Englem. and its co-occurring congener <Emphasis Type="Italic">Lespedeza capitata</Emphasis> Michx.: morphological and molecular evidence

Natural hybridization is common in the genus Lespedeza. No hybrids between Lespedeza leptostachya Englem. and Lespedeza capitata Michx. are formally recognized in any of the current floras, however observations in the field suggest that hybridization might occur in many of their shared habitats. Putative hybrids were compared to L. leptostachya and L. capitata using morphological measurements and screened for the presence of species-specific trnL-F gene region (cpDNA) and the ITS gene region (nrDNA). A discriminate analysis of 10 morphological measurements identified the hybrids as intermediate to both parents with two PCA axes explaining 99% of the variation between taxa. The presence of hybrids was confirmed by genetic markers with individuals morphologically identified as hybrids having cpDNA trnL-F genotypes identical to L. leptostachya and the ITS (nrDNA) phenotypes in most cases contain the ITS genotype of both parents, however, some putative hybrid individuals contained the ITS genotype of only one parents. Those individuals with L. leptostachya ITS and trnL-F could be a case of misclassification, but the presence of both L. capitata ITS genotypes and L. leptostachya trnL-F genotypes suggest segregation has occurred, which may result from either selfing or backcrossing.     

Phylogeny of Crocus (Iridaceae) based on one chloroplast and two nuclear loci: Ancient hybridization and chromosome number evolution

Crocus consists of about 100 species distributed from western Europe and northern Africa to western China, with the center of diversity on the Balkan Peninsula and in Asia Minor. Our study focuses on clarifying phylogenetic relationships and chromosome number evolution within the genus using sequences of the chloroplast trnL-F region, the nuclear ribosomal DNA internal transcribed spacer (ITS) region, and a part of the nuclear single-copy gene pCOSAt103.In a combined dataset of ITS and trnL-F sequences, 115 individuals representing 110 taxa from both subgenera and all sections and series of Crocus were analyzed with Bayesian phylogenetic inference. For pCOSAt103 79 individuals representing 74 Crocus taxa were included, and for the majority of them PCR amplicons were cloned and up to eight clones per individual were sequenced to detect allopolyploidization events. Romulea species were included as outgroup in both analyses. Characteristics of seed surface structures were evaluated by scanning electron microscopy.Phylogenetic analysis of ITS/trnL-F data resulted in a monophyletic genus Crocus, probably monophyletic sections Crocus and Nudiscapus, and inferred monophyly for eight of the 15 series of the genus. The C. biflorus aggregate, thought to be consisting of closely related subspecies, was found to be polyphyletic, the taxa occurring within three major clades in the phylogenetic tree. Cloning of pCOSAt103 resulted in the detection of homoeologous copies in about one third of the taxa of section Nudiscapus, indicating an allotetraploid origin of this section. Reconstruction of chromosome number evolution along the phylogenetic tree using a probabilistic and a parsimony approach arrived at partly contradictory results. Both analyses agreed however on the occurrence of multiple polyploidization and dysploidy events. B chromosomes evolved at least five times independently within the genus, preferentially in clades characterized by karyotype changes.     

Revision and tribal placement of the Argentinean genus Parodiodoxa (Brassicaceae)

Parodiodoxa is a monotypic genus of Brassicaceae endemic to northwestern Argentina. It is poorly known and until now remained the only South American genus of the family that had not been assigned to a particular tribe. Sequence data from the nuclear ribosomal ITS region and the chloroplast trnL intron/trnL-F spacer region were used in this study to determine the systematic position of Parodiodoxa. For this purpose, taxa were sampled both at the tribal and generic levels. Results from tribal-level sampling support the inclusion of Parodiodoxa in the tribe Thelypodieae, whereas those at the generic level reveal a relationship to Weberbauera (W. rosulans and W. herzogii). Topologies within the Thelypodieae were poorly resolved, in agreement with previous studies. Morphological characteristics of Parodiodoxa are also discussed in relation to other genera of the tribe.     

Phylogenetic relationships in tribe Spiraeeae (Rosaceae) inferred from nucleotide sequence data

Tribe Spiraeeae has generally been defined to include Aruncus, Kelseya, Luetkea, Pentactina, Petrophyton, Sibiraea, and Spiraea. Recent phylogenetic analyses have supported inclusion of Holodiscus in this group. Spiraea, with 50-80 species distributed throughout the north temperate regions of the world, is by far the largest and most widespread genus in the tribe; the remaining genera have one to several species each. Phylogenetic analyses of nuclear ITS and chloroplast trnL-trnF nucleotide sequences for 33 species representing seven of the aforementioned genera plus Xerospiraea divided the tribe into two well supported clades, one including Aruncus, Luetkea, Holodiscus, and Xerospiraea, the second including the other genera. Within Spiraea, none of the three sections recognized by Rehder based on inflorescence morphology is supported as monophyletic. Our analyses suggest a western North American origin for the tribe, with several biogeographic events involving vicariance or dispersal between the Old and New Worlds having occurred within this group.     

A Preliminary Molecular Phylogeny of the Rhynchosporeae (Cyperaceae)

The tribe Rhynchosporeae comprises the genera Rhynchospora and Pleurostachys and has never been studied using molecular techniques. The objective of this study was to use an analysis of trnL-F sequences to evaluate the hypothesized taxonomic divisions within the Rhynchosporeae including the monophyly of the genera and the soundness of the subgenera and sections. A total of 44 ingroup species were studied, 41 of Rhynchospora representing 22 of Kükenthal’s 28 sections, and three of Pleurostachys. Five outgroup species from other genera were also included. The cladistic analysis of 50 trnL intron and trnL-F intergenic spacer sequences resulted in 16 trees and a strict consensus tree. The Rhynchosporeae form two well-supported primary clades with several well-supported smaller clades, many of which agree with previously hypothesized sections. Pleurostachys is embedded within the second primary clade.     

Molecular phylogenetic analysis of key <Emphasis Type="Italic">Jatropha</Emphasis> species inferred from nrDNA ITS and chloroplast (<Emphasis Type="Italic">trn</Emphasis>L-F and <Emphasis Type="Italic">rbc</Emphasis>L) sequences

The genus Jatropha (Euphorbiaceae) contains species that are of significant economic and ornamental value. However, Jatropha breeding material is rather limited due to incomplete information regarding phylogenetic relationships among germplasm resources. Phylogenetic analyses were performed based on the internal transcribed spacer of nuclear ribosomal DNA (nrDNA ITS), two chloroplast regions (trnL-F and rbcL), and the combined (ITS+trnL-F+rbcL) dataset among twenty-five specimens representing six key Jatropha species. Phylogenetic relationships of Jatropha were well resolved between subgenus Curcas and subgenus Jatropha, and demonstrated the intermediate position of section Polymorphae among sections of both subgenera. Jatropha curcas and J. integerrima demonstrated a close phylogenetic relationship. The molecular data agreed with the morphological classification that recognized J. multifida and J. podagrica in sec. Peltatae. The distinct intraspecific divergence that occurred in J. curcas could be attributed to restricted gene flow caused by geographical isolation and different ecological conditions. Phylograms produced with trnL-F and rbcL sequence data suggested slow rates of sequence divergence among Jatropha spp., while the ITS gene tree had good resolution suggesting high genetic variation of ITS among Jatropha species.     

Phylogeny of the Southeast Asian endemic genus Neocinnamomum H. Liu (Lauraceae)

A phylogenetic analysis of Neocinnamomum H. Liu and related genera was conducted using psbA–trnH, trnK cpDNA regions, and the ITS nrDNA segment. Neocinnamomum was confirmed to be monophyletic, and an evolutionary series of inflorescence development within the genus was recognized. The compound thyrse seen in N. caudatum is reduced to the few- to many-flowered condensed inflorescences with a poorly defined branching system seen in most species and ultimately to the 1-flowered inflorescence seen in N. atjehense. Consensus network analysis (CNA) suggested that long-branch attraction is responsible for the observed close relationship between Neocinnamomum and Cassytha L. in a combined analysis of the complete data. In contrast, the sister relationship of Neocinnamomum and Caryodaphnopsis seen in the Bayesian analyses of the partial combined matrix was supported by CNA and is also supported by morphology and wood and bark anatomy. The close similarity of the compound thyrse of less derived Neocinnamomum species to the thyrsoid inflorescences of some Caryodaphnopsis species is also seen as strong support for their affinity.     

Phylogeny and biogeography of the African genus Virectaria Bremek. (Sabiceeae s.l., Ixoroideae, Rubiaceae)

The phylogenetic relationships of the tropical African genus Virectaria with its associated genera within the tribe Sabiceeae s.l. (Ixoroideae and Rubiaceae) were inferred from the combined analysis of nuclear ITS and chloroplast rpoC1 and trnT-F nucleotide sequence data. Phylogenetic relationships within Virectaria were investigated using combined analyses of ETS (nrDNA), ITS, rpoC1 and trnT-F sequence data. The present analyses further show that Hekistocarpa is sister to the Tamridaea–Virectaria–Sabicea clade, Tamridaea and Virectaria are sister genera, and Sabicea s.l. is sister to the Tamridaea–Virectaria clade. Our results strongly support the monophyly of Virectaria and the sister-group relationships between V. multiflora and V. herbacoursi, V. angustifolia and V. procumbens, and V. major and V. belingana. Our analyses indicate a tropical African origin for Sabiceeae s.l., a long isolated evolution for Tamridaea and a wide range of dispersal of Virectaria species in the Lower-Guinean, Upper-Guinean and Congolian regions, without a clearly defined direction of migration.     

Differentiation of the endemic Greek genus <Emphasis Type="Italic">Hymenonema</Emphasis> and its relatives of subtribe Scolyminae (Compositae,Cichorieae) based on a multilocus species tree reconstruction

Hymenonema (Compositae, tribe Cichorieae) together with the genera Catananche, Gundelia, and Scolymus forms the subtribe Scolyminae. It is endemic to Greece and consists of two species, Hymenonema laconicum and Hymenonema graecum, which occur in the south Peloponnisos and central Aegean area, respectively. The present contribution aims at a phylogenetic reconstruction of evolutionary relationships among the 12 species of the subtribe, focusing on the temporal and spatial framework for its evolution. The phylogenetic relationships among the members of Scolyminae were inferred from molecular data based on the multi-copy region of the nrDNA internal transcribed spacers ITS1 and ITS2, two intergenic spacers of the cpDNA (trnL-trnF, rpl32-trnL), and one single-copy nuclear region (D10). The gene trees were reconstructed using Bayesian phylogenetic methods. All gene trees support the monophyly of Hymenonema and the sister-group relationship with the genus Scolymus. The further sister-group relationship of this group (Hymenonema–Scolymus) with Catananche is also supported by nrDNA and cpDNA analyses. Finally, a species tree (inferred in a Bayesian coalescent framework) was reconstructed and dates the divergence time between the two Hymenonema species to the Pleistocene (around 1.3 Ma ago). Maximum likelihood-based biogeographical reconstructions suggest a Miocene (pre-Messinian) differentiation of the subtribe on the northern Tethyan platform, followed by Miocene/Pliocene dispersal events to the western Mediterranean and North-African platforms and final, small-scale vicariance events within the genera in the Pleistocene.