Molecular phylogeny of Phalaenopsis Blume (Orchidaceae) based on the internal transcribed spacer of the nuclear ribosomal DNA

The internal transcribed spacer (ITS1, 5.8S rDNA, and ITS2) region of nuclear ribosomal DNA (nrDNA) was sequenced from 53 species, which represent most of the living species diversity in the genus Phalaenopsis (Orchidaceae). A phylogeny was developed for the genus based on the neighbor-joining and maximum parsimony analyses of molecular data. Results of these analyses provided support for the monophyly of the genus Phalaenopsis and concurred in that the genera Doritis and Kingidium should be treated as being parts of the genus Phalaenopsis as suggested by Christenson (2001). Within the genus Phalaenopsis, neither subgenera Aphyllae nor Parishianae were monophyletic, and they were highly clustered with subgenus Proboscidioides plus sections Esmeralda and Deliciosae of subgenus Phalaenopsis based on ITS data. Those species also have the same characters of morphology of four pollinia and similar biogeographies. Furthermore, neither subgenus Phalaenopsis nor Polychilos was monophyletic. Within the subgenus Phalaenopsis, only section Phalaenopsis was highly supported as being monophyletic. As for the subgenus Polychilos, only section Polychilos was moderately supported as being monophyletic. In conclusion, the present molecular data obtained from the ITS sequence of nrDNA of the genus Phalaenopsis provide valuable information for elucidating the phylogeny of this genus.     

Molecular Phylogeny of Asian Meconopsis Based on Nuclear Ribosomal and Chloroplast DNA Sequence Data

The taxonomy and phylogeny of Asian Meconopsis (Himalayan blue poppy) remain largely unresolved. We used the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA) and the chloroplast DNA (cpDNA) trnL-F region for phylogenetic reconstruction of Meconopsis and its close relatives Papaver, Roemeria, and Stylomecon. We identified five main clades, which were well-supported in the gene trees reconstructed with the nrDNA ITS and cpDNA trnL-F sequences. We found that 41 species of Asian Meconopsis did not constitute a monophyletic clade, but formed two solid clades (I and V) separated in the phylogenetic tree by three clades (II, III and IV) of Papaver and its allies. Clade V includes only four Asian Meconopsis species, with the remaining 90 percent of Asian species included in clade I. In this core Asian Meconopsis clade, five subclades (Ia–Ie) were recognized in the nrDNA ITS tree. Three species (Meconopsis discigera, M. pinnatifolia, and M. torquata) of subgenus Discogyne were imbedded in subclade Ia, indicating that the present definition of subgenera in Meconopsis should be rejected. These subclades are inconsistent with any series or sections of the present classifications, suggesting that classifications of the genus should be completely revised. Finally, proposals for further revision of the genus Meconopsis were put forward based on molecular, morphological, and biogeographical evidences.     

Tribe Eritrichieae (Boraginaceae s.str.) in West Asia: a molecular phylogenetic perspective

A selection of Boraginaceae genera was used to obtain a framework for the phylogenetic position of some tribes belong to subfamily Boraginoideae and genera within tribe Eritrichieae (Heterocaryum, Rochelia, Eritrichium, Lappula, Lepechiniella, and Asperugo) and related species. Our results were produced on the basis of nrDNA ITS and cpDNAtrnL-F sequences. The combined nrDNA ITS trnL-F data confirm four main clades of Boraginoideae comprising Echiochileae, Boragineae, Lithospermeae, and Cynoglosseae s. l. (including Eritrichieae, Cynoglosseae s. str., and Myosotideae). The tribe Eritrichieae itself at the current status is paraphyletic; some members, for example Asperugo procumbens, Lepechiniella inconspicua, Myosotidium hortensia, and Cryptantha flavoculata are placed out of the core tribe Eritrichieae. The genus Heterocaryum is monophyletic and allied with a subclade of genera Lappula, Lepechiniella, Eritrichium, and Rochelia. Rochelia is monophyletic, but Eritrichium and Lappula are non-monophyletic. Lepechiniella is nested among a group of Lappula species.     

Comparisons of phylogenetic hypotheses among different data sets in dwarf dandelions (Krigia,Asteraceae): Additional information from internal transcribed spacer sequences of nuclear ribosomal DNA

The internal transcribed spacer (ITS) region of the 18 S–25 S nuclear ribosomal DNA repeat was sequenced from 19 populations of the tribeLactuceae, including all species of dwarf dandelion (Krigia) and five outgroup genera. The incidence of length changes and base substitutions was at least two times higher for ITS 1 than ITS 2. Interspecific sequence divergence withinKrigia averaged 9.62% (1.61%–15.19%) and 4.26% (0%–6.64%) in ITS 1 and ITS 2, respectively. Intergeneric sequence divergence ranged from 15.6% to 44.5% in ITS 1 and from 8.0% to 28.6% in ITS 2. High sequence divergence and homoplasy among genera of tribeLactuceae suggest that the phylogenetic utility of ITS sequence data is limited to interspecific studies or comparisons among closely related genera. Trees generated from ITS sequences are essentially identical to those from restriction site comparisons of the entire nuclear ribosomal (nr) DNA region. The degree of tree resolution differed depending on how gaps were treated in phylogenetic analyses. The ITS trees were congruent with the chloroplast DNA and morphological phylogenies in three major ways: 1) the sister group relationship betweenKrigia andPyrrhopappus; 2) the recognition of two monophyletic sections,Krigia andCymbia, in genusKrigia; and 3) the monophyly of theK. occidentalis-K. cespitosa clade in sect.Cymbia. However, the two nrDNA-based trees are not congruent with morphology/chloroplast DNA-based trees for the interspecific relationships in sect.Krigia. An average of 22.5% incongruence was observed among fourKrigia data sets. The relatively high degree of incongruence among data sets is due primarily to conflict between trees based on nrDNA and morphological/cpDNA data. The incongruence is probably due to the concerted evolution of nrDNA repeating units. The results fromKrigia and theLactuceae suggest that nrDNA data may have limited utility in phylogenetic studies of plants, especially in groups which exhibit high levels of sequence divergence. Our combined phylogenetic analysis as a total evidence shows the least conflict to each of the individual data sets.     

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 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 systematics of the genus Astragalus L. (Fabaceae): Phylogenetic analyses of nuclear ribosomal DNA internal transcribed spacers and chloroplast gene ndhF sequences

Comparative sequencing of internal transcribed spacers (ITS) and 5.8S gene of nuclear ribosomal DNA was carried out to examine phylogenetic relationships among subgenera and sections of Old World Astragalus as well as the recent segregate genera Barnebyella and Ophiocarpus. For a subset of these taxa (43 accessions), the nrDNA ITS data were supplemented by sequences from the chloroplast ndhF gene. Phylogenetic trees resulting from separate analyses of the nrDNA ITS and ndhF sequences were in conflict mainly on the position and relationships of Ophiocarpus aitchisonii, Astragalus hemsleyi, A. grammocalyx, A. coelicolor, A. capito, A. epiglottis and A. annularis. Excluding these taxa, phylogenetic analysis of a combined nrDNA ITS-ndhF data matrix was also conducted, so that in the resulting tree, most clades were more resolved and better statistically supported than those were in the separate analyses. Our results indicate that the monotypic segregate genera Barnebyella (= A. migpo), Ophiocarpus (= A. ophiocarpus) and morphologically isolated annual species A. dipelta (= Didymopelta turkestanica), A. schmalhausenii (= Sewerzowia turkestanica) and A. vicarius (= S. vicaria) are clearly nested within Astragalus. Our results confirm earlier studies that shows A. vogelii is allied with the genera Colutea and Oxytropis rather than with any Astragalus species. It is therefore excluded from Astragalus and elevated to the new generic rank and named as Podlechiella Maassoumi and Kazempour Osaloo. None of the eight traditionally recognized Astragalus subgenera Epiglottis, Trimeniaeus, Phaca, Hypoglottis, Calycophysa, Tragacantha, Cercidothrix and Calycocystis are monophyletic. Similarly, the monophyly of Podlechs new subgenera Trimeniaeus, Astragalus and Cercidothrix is not supported. Among the many species-rich sections analyzed here, only Cenanthrum, Chronopus, Laxiflori, Lotidium, Incani and Amodendron are monophyletic.     

Hybridization drives speciation in Gagea (Liliaceae)

Hybridization seems to play an important role in speciation of Gagea Salisb., a genus which is characterised by polyploid taxa lines and in which diploids (2n = 24) appear only to be common in basal sections. Hybrid detection was applied utilising direct and cloning nrDNA ITS data (ITS1, 5.8S rDNA, ITS2) combined with neighbour and ribotype networks and discussed in connection with previously published cpDNA, morphological and karyological data of the authors. We have evidence of the hybrid origin of taxa within the section Gagea (G. pomeranica, G. megapolitana) and the monophyletic clade of sections Didymobulbos and Fistulosae (G. microfistulosa, G. polidorii, G. cf. bohemica). Morphologically and karyologically differentiated Gagea megapolitana and G. pomeranica, adapted to synanthropic habitats, represent both hybrids of G. pratensis × G. lutea. Gagea microfistulosa represents a hybrid of G. villosa × G. fragifera; Gagea polidorii could represent the reverse hybrid. G. glacialis is also closely related to the latter complex.     

Lindgomyces angustiascus, (Lindgomycetaceae,Pleosporales, Dothideomycetes), a new lignicolous species from freshwater habitats in the USA

In this paper, we describe and illustrate a new species, Lindgomyces angustiascus, from submerged wood in lotic and lentic habitats from Florida, North Carolina and Wisconsin, USA. The new species is characterized by black, partially immersed, flattened, globose ascomata; fissitunicate, long, slender, obclavate asci; and one-septate, hyaline, fusiform ascospores with bipolar appendages, each covered with a gelatinous cap. Maximum Likelihood and Bayesian analyses of partial 18S nrDNA and 28S nrDNA, as well as the entire nuclear ribosomal internal transcribed spacer (ITS1, 5.8S and ITS2) region support the placement and establishment of this new species in the Lindgomycetaceae, Pleosporales, Dothideomycetes. Chemical analysis of the organic extract of L. angustiascus revealed the presence of 6E,9E-octadecadienoic acid and ergosterol peroxide as major secondary metabolites.     

基于核糖体DNA内转录间隔区ITS序列的广义蓼属及近缘属分子系统学研究(英文)

通过对广义蓼属及近缘属共32个代表种内转录间隔区ITS序列的分子系统学分析,尝试研究备受争议的广义蓼属及近缘属的物种族、属、组级的划分问题,结果显示,广义蓼属在系统发育树上并不能形成一个单系类群,这些物种共聚为3大支,分别对应春蓼族、蓼族及荞麦族,其中荞麦属与翅果蓼属形成了一支独立于春蓼族及蓼族之外的类群。在春蓼族中,冰岛蓼属与分叉蓼组形成一个单系类群。     

基于核糖体DNA内转录间隔区ITS序列的广义蓼属及近缘属分子系统学研究

通过对广义蓼属及近缘属共32个代表种内转录间隔区ITS序列的分子系统学分析,尝试研究备受争议的广义蓼属及近缘属的物种族、属、组级的划分问题,结果显示,广义蓼属在系统发育树上并不能形成一个单系类群,这些物种共聚为3大支,分别对应春蓼族、蓼族及荞麦族,其中荞麦属与翅果蓼属形成了一支独立于春蓼族及蓼族之外的类群。在春蓼族中,冰岛蓼属与分叉蓼组形成一个单系类群。     

Molecular characterization of Oxyspirura mansoni and Philophthalmus gralli collected from the eyes of domestic chickens in Bangladesh

A variety of helminths have been found in domestic chickens in Bangladesh, but little is known about their gene sequences. Here, parasitic nematodes and trematodes were collected from the eyes of domestic chickens and analyzed for their morphological and morphometric characteristics, and characterized molecularly. The helminths were identified as Oxyspirura mansoni and Philophthalmus gralli. The ITS1 and ITS2 sequences of O. mansoni were 532 bp and 306 bp in length, respectively, and showed low identity (50.7–62.7%) with those of O. petrowi and O. conjunctivalis. Furthermore, the O. mansoni CO1 sequences (393 bp) showed five haplotypes (97.5–99.5% similarity) that formed a monophyletic clade. With respect to P. gralli, the ITS1 (452 bp) and ITS2 (736 bp) sequences showed 100% similarity with the reference sequences in GenBank. Both the ND1 and CO1 phylograms showed that P. gralli from Bangladesh, Costa Rica and Peru form a monophyletic clade, distinct from the clades of P. lucipetus and P. lacrymosus. Our data show that, Philophthalmus gralli isolates from Bangladesh, Costa Rica and Peru are genetically close to each other.     

栝楼属基于核糖体ITS序列的系统发育分析

栝楼属（Trichosanthes）是葫芦科(Cucurbitaceae)中一个种类较多、药用价值较大的属。本文基于ITS序列分析了栝楼属16个种的系统发育关系。聚类分析表明：在组的划分上,具有分类争议的截叶组(sect. Truncata)与大苞组(sect. Involucraria)形成一大支,处于一亚分枝位置,不支持独立成组。在属内分类地位上,在核酸水平不支持贵州栝楼(T.guizhouensis)为一独立种,结合形态特征建议并入双边栝楼(T.rosthornii)。大方油栝楼(T.dafangensis)与小苞组(sect. Trichosanthes)成为姊妹群,处于一个向叶苞组(sect. Foliobracteola)过渡位置,结合其形态特征,建议作为小苞组(sect. Trichosanthes)成员。     

Analysis of nrDNA polymorphism in closely related diploid sexual, tetraploid sexual and polyploid agamospermous species

Nuclear sequences of ITS1-5.8S-ITS2 region of rDNA may be an important source of phylogenetically informative data provided that nrDNA is cloned and the character of sequence variation of clones is properly analyzed. nrDNA of selected Taraxacum sections was studied to show sequence variation differences among diploid sexual, tetraploid sexual and polyploid agamospermous species. We examined nucleotide characteristics, substitution pattern, secondary structure, and the phylogenetic utility of ITS1-5.8S-ITS2 from 301 clones of 32 species representing 11 sections. The most divergent sequences of ITS1&2 differed by 17.1% and in 5.8S only by 3.7%. The ITS1-5.8S-ITS2 characteristics, integrity and also stability of secondary structures confirmed that pseudogenes are not responsible for the above variation. The within-individual polymorphism of clones implies that the concerted evolution of ITS cistron of agamospermous polyploid Taraxacum is remarkably suppressed. Sequences of ITS clones proved to be a useful tool for mapping pathways of complex reticulation (polyploid hybridity) in agamospermous Taraxacum.     

Molecular authentication of Anthemis deserti Boiss. (Asteraceae) based on ITS2 region of nrDNA gene sequence

The dried plant material of medicinally important Anthemis deserti Boiss. (family: Asteraceae) especially when it remains in the powdered form often look similar to Anthemis melampodina Del.; and therefore, difficult to distinguish, finally lead to chances of adulteration. The adulteration in medicinal plants effects on the efficacy of the drugs. The molecular authentication of herbal plant materials such as based on the internal transcribed spacer 2 (ITS2) sequences of nuclear ribosomal DNA (nrDNA) is considered as more reliable method compared to other the biochemical or histological methods. The present study aims to molecular authentication ofA. deserti based on molecular phylogenetic analyses of ITS2 gene sequence of nrDNA region. The ITS2 region of nrDNA of A. deserti were sequenced, and the molecular phylogenetic analyses were performed together with the GenBank sequences. The Maximum Parsimony tree revealed the close relationships of A. deserti with A. melampodina; however, the Neighbor-Joining and Maximum Likelihood tree clearly revealed that A. deserti is distinct from A. melampodina, which is also supported by the differences in nucleotides at five diffident positions (i.e. 22, 28, 87, 175 and 198) in the DNA sequence alignment.     

PHYLOGENY OF THEPARMELIACEAE—DNA DATA VERSUS MORPHOLOGICAL DATA

In order to identify monophyletic groups within the familyParmeliaceae, eleven taxa (Bryoria capillaris,Cetraria islandica,Evernia pruniastri,Hypogymnia physodes,Parmelia saxatilis,Platismatia glauca,Pleurosticta acetabulum,Usnea florida,Vulpicida juniperina,V. pinastri, andXanthoparmelia conspersa) were studied using newly produced nuclear rDNA sequence data from the ITS and 5·8S regions. The resulting evolutionary hypothesis was compared with results from previous phylogenetic analyses based on anatomy, morphology, and chemistry. The outcome of this comparison does not support the earlier proposed phylogenies but is not stable enough for identifying monophyletic groups, with one exception. The results indicate a close relationship betweenCetrariaandVulpicida, which is contradictory to previous published analyses. The variation in ascus structures in theParmeliaceaeis discussed and it is questioned whether the earlier distinguished ‘ forms ’ of ascus types represent synapomorphies, if they are based on poorly supported analyses, or if they are exaggerations of relatively slight variation in shape. Further interpretations of the results are discussed and areas of future studies based on DNA-data are suggested.     

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.     

Comparison of methods for rooting phylogenetic trees: A case study using Orcuttieae (Poaceae: Chloridoideae)

DNA sequence data (cpDNA trnL intron and nrDNA ITS1 and ITS2) were analyzed to identify relationships within Orcuttieae, a small tribe of endangered grasses endemic to vernal pools in California and Baja California. The tribe includes three genera: Orcuttia, Tuctoria, and Neostapfia. All three genera carry out C₄ photosynthesis but aquatic taxa of Orcuttia lack Kranz anatomy. The unusual habitat preference of the tribe is coupled with the atypical development of C₄ photosynthesis without Kranz anatomy. Furthermore, the tribe has no known close relatives and has been noted to be phylogenetically isolated within the subfamily Chloridoideae. In this study we examine the problem of inferring the root of the tribe in the absence of an identified outgroup, analyze the phylogenetic relationships of the constituent taxa, and evaluate the evolutionary development of C₄ photosynthesis. We compare four methods for inferring the root of the tree: (1) the outgroup method, (2) midpoint rooting, the imposition of a molecular clock for both (3) maximum likelihood (ML) and (4) Bayesian analysis. We examine the consequences of each method for the inferred phylogenetic relationships. Three of the methods (outgroup rooting and the ML and Bayesian molecular clock analyses) suggest that the root of Orcuttieae is between Neostapfia and the Tuctoria/Orcuttia lineage, while midpoint rooting gives a different root. The Bayesian method additionally provides information about probabilities associated with other possible root locations. Assuming that the true root of Orcuttieae is between Neostapfia and the Tuctoria/Orcuttia lineage, our data indicate Neostapfia and Orcuttia are both monophyletic, while Tuctoria is paraphyletic (with no synapomorphies in either dataset) and forming a grade between the other two genera and needs taxonomic revision. Our data support the hypothesis that Orcuttieae was derived from a terrestrial ancestor and evolved specializations to an aquatic environment, including C₄ photosynthesis without Kranz anatomy.