Probing the Monophyly of the Sphaeropleales (Chlorophyceae) Using Data From Five Genes

Molecular phylogenetic analyses have had a major impact on the classification of the green algal class Chlorophyceae, corroborating some previous evolutionary hypotheses, but primarily promoting new interpretations of morphological evolution. One set of morphological traits that feature prominently in green algal systematics is the absolute orientation of the flagellar apparatus in motile cells, which correlates strongly with taxonomic classes and orders. The order Sphaeropleales includes diverse green algae sharing the directly opposite (DO) flagellar apparatus orientation of their biflagellate motile cells. However, algae across sphaeroplealean families differ in specific components of the DO flagellar apparatus, and molecular phylogenetic studies often have failed to provide strong support for the monophyly of the order. To test the monophyly of Sphaeropleales and of taxa with the DO flagellar apparatus, we conducted a molecular phylogenetic study of 16 accessions representing all known families and diverse affiliated lineages within the order, with data from four plastid genes (psaA, psaB, psbC, rbcL) and one nuclear ribosomal gene (18S). Although single‐gene analyses varied in topology and support values, analysis of combined data strongly supported a monophyletic Sphaeropleales. Our results also corroborated previous phylogenetic hypotheses that were based on chloroplast genome data from relatively few taxa. Specifically, our data resolved Volvocales, algae possessing predominantly biflagellate motile cells with clockwise (CW) flagellar orientation, as the monophyletic sister lineage to Sphaeropleales, and an alliance of Chaetopeltidales, Chaetophorales, and Oedogoniales, orders having multiflagellate motile cells with distinct flagellar orientations involving the DO and CW forms.     

DEEP DIVISION IN THE CHLOROPHYCEAE (CHLOROPHYTA) REVEALED BY CHLOROPLAST PHYLOGENOMIC ANALYSES1

The Chlorophyceae (sensu Mattox and Stewart) is a morphologically diverse class of the Chlorophyta displaying biflagellate and quadriflagellate motile cells with varying configurations of the flagellar apparatus. Phylogenetic analyses of 18S rDNA data and combined 18S and 26S rDNA data from a broad range of chlorophycean taxa uncovered five major monophyletic groups (Chlamydomonadales, Sphaeropleales, Oedogoniales, Chaetophorales, and Chaetopeltidales) but could not resolve their branching order. To gain insight into the interrelationships of these groups, we analyzed multiple genes encoded by the chloroplast genomes of Chlamydomonas reinhardtii P. A. Dang. and Chlamydomonas moewusii Gerloff (Chlamydomonadales), Scenedesmus obliquus (Turpin) Kütz. (Sphaeropleales), Oedogonium cardiacum Wittr. (Oedogoniales), Stigeoclonium helveticum Vischer (Chaetophorales), and Floydiella terrestris (Groover et Hofstetter) Friedl et O’Kelly (Chaetopeltidales). The C. moewusii, Oedogonium, and Floydiella chloroplast DNAs were partly sequenced using a random strategy. Trees were reconstructed from nucleotide and amino acid data sets derived from 44 protein‐coding genes of 11 chlorophytes and nine streptophytes as well as from 57 protein‐coding genes of the six chlorophycean taxa. All best trees identified two robustly supported major lineages within the Chlorophyceae: a clade uniting the Chlamydomonadales and Sphaeropleales, and a clade uniting the Oedogoniales, Chaetophorales, and Chaetopeltidales (OCC clade). This dichotomy is independently supported by molecular signatures in chloroplast genes, such as insertions/deletions and the distribution of trans‐spliced group II introns. Within the OCC clade, the sister relationship observed for the Chaetophorales and Chaetopeltidales is also strengthened by independent data. Character state reconstruction of basal body orientation allowed us to refine hypotheses regarding the evolution of the flagellar apparatus.     

基于线粒体Cyt b,16S rDNA和核28S rDNA的中国斑腿蝗科(直翅目,蝗总科)九亚科间的系统发育关系研究

基于线粒体Cytb、16S rDNA和核28S rDNA的部分序列的联合数据集,重建了中国斑腿蝗科9亚科26种蝗虫和癞蝗科笨蝗(外群)间的系统发育关系。结果显示:1)除黑蝗亚科、秃蝗亚科和切翅蝗亚科外,其余6亚科的单系性都得到支持;2)黑蝗亚科似乎应当与秃蝗亚科合并为一个亚科,且与其他几个亚科亲缘关系相对较远;3)凸额蝗属应该由切翅蝗亚科中独立出来;4)卵翅蝗亚科和稻蝗亚科是姐妹群;5)星翅蝗亚科、黑背蝗亚科、斑腿蝗亚科、刺胸蝗亚科及切翅蝗亚科中的凸额蝗属间的关系较近。     

棉铃虫18S核糖体RNA基因的序列分析及其分子系统学

克隆并分析了鳞翅目棉铃虫Helicoverpa armigera (Hübner)18S核糖体RNA基因(18S rDNA)的全序列,将该序列与鞘翅目、膜翅目、同翅目、双翅目、捻翅目和弹尾目各一种昆虫的同源保守区进行了比较。序列分析结果显示：鳞翅目和双翅目昆虫在18S rDNA结构上彼此较为相似,捻翅目昆虫的18S rDNA分子结构表现出与其它目昆虫有较大的差异,但相对与弹尾目昆虫的18S rDNA较为接近。该结果支持了有关捻翅目属于一个独立的目级分类阶元的论点。     

POLYPHYLETIC ORIGIN OF PARALLEL BASAL BODIES IN SWIMMING CELLS OF CHLOROPHYCEAN GREEN ALGAE (CHLOROPHYTA)1

The evolutionary affinities of Heterochlamydomonas Cox and Deason and Dictyochloris Vischer ex Starr were investigated using phylogenetic analyses of a combined data set of 18S and 28S rDNA sequences with those from 38 additional green algae. Previous ultrastructural studies have shown that motile cells of Heterochlamydomonas and Dictyochloris have an unusual flagellar apparatus organization in that the two flagella are of unequal length and the basal bodies are persistently parallel. Because of this similarity these taxa, along with Bracteacoccus Tereg, a third taxon with this same flagellar apparatus arrangement, are hypothesized to be closely related. We show, with maximum parsimony and Bayesian analyses, that the parallel basal bodies are not homologous in the three genera. Rather, Heterochlamydomonas is most closely related to Chlamydomonas baca in the clockwise flagellar apparatus clade, and Dictyochloris and Bracteacoccus are nested within the Sphaeropleales, which has the directly opposite flagellar absolute orientation. Surprisingly, Dictyochloris and Bracteacoccus are not supported as closest relatives. These relationships are supported by morphological features such as the presence or absence of a walled motile cell but not by the orientation of the basal bodies. In addition, our data are derived from multiple isolates of each study genera, and the analyses show that Heterochlamydomonas and Dictyochloris are each monophyletic.     

PHYLOGENETIC POSITION OF KOLIELLA (CHLOROPHYTA) AS INFERRED FROM NUCLEAR AND CHLOROPLAST SMALL SUBUNIT rDNA

The phylogenetic position of Koliella , a chlorophyte characterized by Klebsormidium type cell division, was inferred from analyses of partial 18S rDNA and partial 16S rDNA. Parsimony and distance analyses of separate and combined data sets indicated that the members of Koliella belonged to Trebouxiophyceae, and high decay indices and bootstrap values supported this affinity. However, the genus appeared to be polyphyletic. Koliella spiculiformis , the nomenclatural type of the genus, was allied with Nannochloris eucaryota and the "true" chlorellas ( Chlorella vulgaris , C. lobophora , C. sorokiniana , and C. kessleri ). The close relatives of Koliella longiseta (≡ Raphidonema longiseta ) and Koliella sempervirens appeared to be Stichococcus bacillaris and some species traditionally classified in Chlorella that were characterized by the production of secondary carotenoids under nitrogen-deficient conditions. This clade was also supported by the presence of a relatively phylogenetically stable group I intron (1506) in the 18S rRNA gene. Because of the presence of Klebsormidium type cell division, some authors regarded the members of Koliella as closely related to charophytes. Molecular analyses, however, did not confirm this affinity and suggested that a Klebsormidium type cell division is homoplastic in green plants.     

Phylogeny of coral-inhabiting barnacles (Cirripedia; Thoracica; Pyrgomatidae) based on 12S, 16S and 18S rDNA analysis

The traditional phylogeny of the coral-inhabiting barnacles, the Pyrgomatidae, is based on morphological characteristics, mainly of the hard parts. It has been difficult to establish the phylogenetic relationships among Pyrgomatidae because of the apparent convergence of morphological characteristics, and due to the use of non-cladistic systematics, which emphasize ancestor-descendant relationships rather than sister-clade relationships. We used partial sequences of two mithochondrial genes, 12S rDNA and 16S rDNA, and a nuclear gene, 18S rDNA, to infer the molecular phylogeny of the pyrgomatids. Our phylogenetic results allowed us to reject previous classifications of Pyrgomatidae based on morphological characteristics. Our results also suggested the possibility of paraphyly of the Pyrgomatidae. The hydrocoral barnacle Wanella is not found on the same clade as the other pyrgomatids, but rather, with the free-living balanids. The basal position of Megatrema and Ceratoconcha is supported. The archeaobalanid Armatobalanus is grouped with Cantellius at the base of the Indo-Pacific pyrgomatines. Fusion of the shell plate and modification of the opercular valves are homoplasious features that occurred more than three times on different clades. The monophyly of the "Savignium" group, comprising four nominal genera, is also not supported, and the different taxa are placed on different clades.     

从核rDNA序列探讨隙蛛亚科Coelotinae的分类地位

隙蛛亚科Coelotinae主要分布于东亚地区,其中我国的已有种类占到全世界种数的一半以上,因此对于我国隙蛛类蜘蛛的研究已经成为世界暗蛛科研究的重点之一。隙蛛亚科属于无筛器类群,于1893年,由Cambridge以隙蛛属为模式属而建立,归属于无筛器的漏斗蛛科。之后,虽然经历了数次修订     

15 Phylogeny of the chlorophyta: inferences from 18S and 26S rDNA

Recent studies of the Chlorophyceae using 18S and 26S rDNA data in meta‐analysis have demonstrated the power of combining these two sets of rDNA data. Furthermore, the 26S rDNA data complement the more conserved 18S gene for many chlorophycean lineages. Consequently, this data approach was pursued in an expanded taxon‐sampling scheme for the Chlorophyta, with special reference to the classes Chlorophyceae and Trebouxiophyceae. Results of these new phylogenetic analyses identify Microspora sp. (UTEX LB 472) and Radiofilum transversale (UTEX LB 1252) as sister taxa which, in turn, form a basal clade in the Cylindrocapsa alliance (Treubaria, Trochiscia, Elakatothrix). The relative position of the “Cylindrocapsa” clade within the Chlorophyceae remains uncertain. The enhanced taxon‐sampling has not resolved the relative positions of the Oedogoniales, Chaetophorales or Chaetopeltidales. Furthermore, the Sphaeropleaceae are supported as members of the Sphaeropleales in only some analyses, raising concerns about the status of the order. Although based on a limited set of taxa (currently <10), a combined data approach reveals support for a monophyletic Trebouxiophyceae that includes the distinctive organisms, Geminella and Eremosphaera. The goal of a well‐resolved phylogeny for the Chlorophyta remains just that, a goal. Achieving that goal obviously will require additional taxon sampling in the Prasinophyceae and Ulvophyceae, as well as, the Trebouxiophyceae. Moreover, it is clear that other genes (e.g., cp‐atpB, cp‐rbcL, cp‐16S, mt‐nad5) will be needed to help address problems of resolution based on the rDNA data alone. Supported by NSF DEB 9726588 and DEB 0129030.     

Phylogenetic analyses of Cornales based on 26S rRNA and combined 26S rDNA-MATK-RBCL sequence data

Nuclear 26S rDNA sequences were used to corroborate and test previously published matK-rbcL-based hypotheses of phylogenetic relationships in Cornales. Sequences were generated for 53 taxa including Alangium, Camptotheca, Cornus, Curtisia, Davidia, Diplopanax, Mastixia, Nyssa, and four families: Grubbiaceae, Hydrangeaceae, Hydrostachyaceae, and Loasaceae. Fifteen taxa from asterids were used as outgroups. The 26S rDNA sequences were initially analyzed separately and then combined with matK-rbcL sequences, using both parsimony and maximum likelihood methods. Eight strongly supported major clades were identified within Cornales by all analyses: Cornus, Alangium, nyssoids (Nyssa, Davidia, and Camptotheca), mastixioids (Mastixia and Diplopanax), Hydrangeaceae, Loasaceae, Grubbia-Curtisia, and Hydrostachys. However, relationships among the major lineages are not strongly supported in either 26S rDNA or combined 26S rDNA-matK-rbcL topologies, except for the sister relationships between Cornus and Alangium and between nyssoids and mastixioids in the tree from combined data. Discrepancies in relationships among major lineages, especially the placement of the long-branched Hydrostachys, were found between parsimony and maximum likelihood trees in all analyses. Incongruence between the 26S rDNA and matK-rbcL data sets was suggested, where Hydrangeaceae was found to be largely responsible for the incongruence. The long branch of Hydrostachys revealed in previous analyses was reduced significantly with more sampling. Maximum likelihood analysis of combined 26S rDNA-matK-rbcL sequences suggested that Hydrostachys might be sister to the remainder of Cornales, that Cornus-Alangium are sisters, that nyssoids-mastixioids are sisters, and that Hydrangeaceae-Loasaceae are sisters, consistent with previous analyses of matK-rbcL sequence data.     

MOLECULAR PHYLOGENETIC RELATIONSHIPS IN THE FRESHWATER FAMILY HYDRODICTYACEAE (SPHAEROPLEALES,CHLOROPHYCEAE), WITH AN EMPHASIS ON PEDIASTRUM DUPLEX1

The freshwater green algal family Hydrodictyaceae (Sphaeropleales, Chlorophyta) has traditionally consisted of four coenobial genera, Pediastrum Meyen 1829, Hydrodictyon Roth 1797, Sorastrum Kützing 1845, and Euastropsis Lagerheim1894. Two recent molecular phylogenetic studies demonstrated the need for reevaluation of the generic and species boundaries in this morphology‐rich family. This study expands the previous work to include phylogenetic analyses of 103 ingroup isolates representing North America, Europe, and Australia, with an emphasis on the common and geographically widespread species Pediastrum duplex. Nucleotide sequence data were collected from the nuclear LSU (26S rDNA) and the chloroplast RUBISCO LSU (rbcL) genes, totaling >3,000 aligned characters. The 26S and rbcL data sets were analyzed using maximum‐likelihood (ML) and Bayesian phylogenetic methods. In addition, SEM was used to examine the wall morphology of a majority of the isolates. The results supported previous indications that the P. duplex Meyen 1829 morphotype is nonmonophyletic and resolved some previously ambiguous relationships recovered in earlier phylogenetic estimations using fewer isolates. These new data allowed testing of the recent taxonomic revisions of the family that split Pediastrum into five genera. Some of the previous revisions by Buchheim et al. (2005) were well supported (erection of Stauridium and Monactinus), while others were not (Pediastrum, Pseudopediastrum, Parapediastrum).     

166 Phylogeny of the Chlorophyceae: Inferences from the Chloroplast atpB Gene

Phylogenetic reconstruction using the 18S and 26S rDNA genes has proven successful for many chlorophycean lineages, but some nodes remain poorly-resolved. The chloroplast-encoded atpB gene has been used to examine phylogenetic problems in several green algal lineages, including the basal streptophytes. However, data for other Chlorophyta are largely unavailable. The class Chlorophyceae, in particular, has not been examined (save for dense sampling among colonial and unicellular flagellates) using the atpB gene. An investigation of variability in the atpB gene for various chlorophycean exemplars was undertaken. Intergeneric distances (p) range from 0.08 ( Chlamydomonas reinhardtii vs. Volvox carteri ) to 0.24 ( Haematococcus lacustris vs. Chloromonas radiata ). Distance analysis also reveals high levels of divergence (as compared to the 18S and 26S rDNA data) for the family Sphaeropleaceae. Preliminary phylogenetic reconstructions fail to support a monophyletic Sphaeropleales, but this observation may be due to inadequate taxon sampling. The relative positions of other examplars (e.g., Chaetophora, Elakatothrix, Ascochloris) are largely consistent with interpretations of 18S and 26S rDNA data, although the position of Elakothrix is not robust. These preliminary observations suggest that the atpB gene will provide a powerful complement to current phylogenetic assessments of the Chlorophyceae using 18S and 26S rDNA data. Supported by NSF DEB 9726588 and DEB 0129030.     

PHYLOGENY OF LOBOCHARACIUM (CHLOROPHYCEAE) AND ALLIES: A STUDY OF 18S AND 26S rDNA DATA1

The phylogenetic affinities of Lobocharacium coloradoense were investigated by analysis of combined 18S and 26S rDNA data. Results from both parsimony and likelihood methods supported a close alliance among Lobocharacium, Characiosiphon, and Characiochloris. These three taxa formed a clade near the base of the “Dunaliella” group within the chlamydomonad lineage. Protosiphon, which exhibits a siphonous habit similar to Characiosiphon and Lobocharacium, was not resolved as a close ally of the latter two taxa. The Lobocharacium alliance was characterized by the presence of an attachment pad associated with the nonmotile vegetative stage and pyrenoids that possess cytoplasmic invaginations. The pyrenoid feature is an ultrastructural trait that has now been observed in five different chlorophycean lineages. The Lobocharacium–Characiosiphon–Characiochloris clade is not predicted by any classifications of green algae. Additional taxon and data sampling need to be completed to resolve inconsistencies between the molecular phylogenetic evidence and at least some of the current family‐level taxa.     

基于16S rDNA和28S rDNA D2基因序列与形态特征联合分析的中国角顶叶蝉亚科系统发育研究（半翅目： 叶蝉科）

首次在国内利用28S rDNA D2区段和16S rDNA基因序列,结合50个形态特征对角顶叶蝉亚科(Deltocephalinae)［半翅目（Hemiptera）: 叶蝉科（Cicadellidae）］19个属进行系统发育分析研究。从无水乙醇浸泡保存的标本中提取基因组DNA并扩增了19个内群和1种外群Typhlocybinae［半翅目(Hemiptera): 叶蝉科(Cicadellidae)］种类的28S rDNA D2基因片段并测序,同时扩增了16S rDNA基因片段并测序11条,采用了GenBank中1个种类的16S rDNA同源序列。采用PAUP*4.0和MrBayes3.0两个分析软件和3种建树方法,利用同源28S D2 rDNA和16S rDNA两个基因序列与形态特征结合进行系统发育分析研究。分析结果表明,二叉叶蝉族Macrostelini是一个单系,并在角顶叶蝉亚科的系统发育中处于基部的位置,是内群中最原始的族;角顶叶蝉族Deltocephalini中除了纹翅叶蝉属Nakaharanus,其余各属构成单系;殃叶蝉族Euscelini内属的归属比较混乱,可能是一个并系群,属间差异有待进一步研究。隆额叶蝉族Paralimnini与顶带叶蝉族Athysanini是姐妹群。带叶蝉属Scaphoideus与纹翅叶蝉属Nakaharanus是姐妹群,二者与木叶蝉属Phlogotettix的关系最近,三者构成一个单系,建议将三者归为带叶蝉族Scaphoideini。研究结果还表明,小眼叶蝉族Xestocephalini和Balcluthini的系统发育位置不明,有待进一步研究。     

黄颡单尾虫(粘体门,双壳目)的重描述及基于28S rDNA和ITS-5.8S序列的系统地位分析

采用形态分类学方法与以28S rDNA和ITS-5.8S序列为基础的分子系统学研究方法,对采自嘉陵江重庆市磁器口江段的黄颡单尾虫Unicauda pelteobagrusMa,1998进行了形态学和分子生物学的研究。基于28S rDNA数据探讨了黄颡单尾虫以及单尾虫属与相邻种属粘孢子虫间的系统地位;基于5.8S rDNA数据比较分析了粘孢子虫的系统地位。补充了黄颡单尾虫重庆种群形态学信息和28S rDNA、ITS-5.8S rDNA序列的分子信息。     

速足目主要类群系统发育的分子证据

文章基于速足目现生主要类群18S rDNA、28S rDNA和COI基因序列,采用贝叶斯法、邻接法和最大简约法,尝试构建速足目的分子系统树;结合形态特征和化石记录,主要对速足目各超科级分类阶元的系统发育关系进行探讨。结果表明,速足目现生超科Bairdiacea、Darwinulacea、Cypridacea和Cytheracea均为单系群,支持形态学上关于上述4个超科的的界定;3种基因均支持形态学上Darwinulacea和Cypridacea具有较近的亲缘关系的观点。18S rDNA序列分析在较显著水平上支持Darwinulacea和Bairdiacea为姐妹群,Darwinulacea可能从Bairdia-cea中的一支演化而来;Bairdiacea和Darwinulacea组成的分支是Cypridacea的姐妹群,支持将三者合并为Bairdio-copina亚目的观点;Cytheracea是Cypridacea(Darwinulacea Bairdiacea)的姐妹群,可提升为Cytheracopina亚目。     

16 Comparisons of macrophyte cover and community primary productivity on two southern california shores

Recent studies of the Chlorophyceae using 18S and 26S rDNA data in meta-analysis have demonstrated the power of combining these two sets of rDNA data. Furthermore, the 26S rDNA data complement the more conserved 18S gene for many chlorophycean lineages. Consequently, this data approach was pursued in an expanded taxon-sampling scheme for the Chlorophyta, with special reference to the classes Chlorophyceae and Trebouxiophyceae. Results of these new phylogenetic analyses identify Microspora sp. (UTEX LB 472) and Radiofilum transversale (UTEX LB 1252) as sister taxa which, in turn, form a basal clade in the Cylindrocapsa alliance (Treubaria, Trochiscia, Elakatothrix). The relative position of the "Cylindrocapsa" clade within the Chlorophyceae remains uncertain. The enhanced taxon-sampling has not resolved the relative positions of the Oedogoniales, Chaetophorales or Chaetopeltidales. Furthermore, the Sphaeropleaceae are supported as members of the Sphaeropleales in only some analyses, raising concerns about the status of the order. Although based on a limited set of taxa (currently <10), a combined data approach reveals support for a monophyletic Trebouxiophyceae that includes the distinctive organisms, Geminella and Eremosphaera. The goal of a well-resolved phylogeny for the Chlorophyta remains just that, a goal. Achieving that goal obviously will require additional taxon sampling in the Prasinophyceae and Ulvophyceae, as well as, the Trebouxiophyceae. Moreover, it is clear that other genes (e.g., cp-atpB, cp-rbcL, cp-16S, mt-nad5) will be needed to help address problems of resolution based on the rDNA data alone. Supported by NSF DEB 9726588 and DEB 0129030.     

PHYLOGENY OF THE HYDRODICTYACEAE (CHLOROPHYCEAE): INFERENCES FROM rDNA DATA1

The hydrodictyacean green algal lineage has been the focus of much research due to the fossil record of at least some members, their ornamented cell walls, and their distinctive reproductive strategies. The phylogeny of the family was, until recently, exclusively morphology based. This investigation examines hydrodictyacean isolates from several culture collections, focusing on sequences from ribosomal data: 18S rDNA, 26S rDNA (partial), and internal transcribed spacer (ITS)‐2 data. Results from phylogenetic analyses of independent and combined data matrices support the Hydrodictyaceae as a monophyletic lineage that includes isolates of Chlorotetraedron, Hydrodictyon, Pediastrum, Sorastrum, and Tetraedron. Phylogenetic analyses of rDNA data indicate that the three‐dimensional coenobium of Hydrodictyon is evolutionarily distinct from the three‐dimensional coenobium of Sorastrum. The more robust aspects of the ITS‐2 data corroborate the 18S+26S rDNA topology and provide a structural autapomorphy for the Hydrodictyaceae and Neochloridaceae, that is, an abridgment of helix IV in the secondary structure. The rDNA data do not support monophyly of Pediastrum but rather suggest the existence of four additional hydrodictyacean genera: Monactinus, Parapediastrum, Pseudopediastrum, and Stauridium.     

THE EVOLUTION OF ELONGATE SHAPE IN DIATOMS1

Diatoms have been classified historically as either centric or pennate based on a number of features, cell outline foremost among them. The consensus among nearly every estimate of the diatom phylogeny is that the traditional pennate diatoms (Pennales) constitute a well‐supported clade, whereas centric diatoms do not. The problem with the centric–pennate classification was highlighted by some recent analyses concerning the phylogenetic position of Toxarium, whereby it was concluded that this “centric” diatom independently evolved several pennate‐like characters including an elongate, pennate‐like cell outline. We performed several phylogenetic analyses to test the hypothesis that Toxarium evolved its elongate shape independently from Pennales. First, we reanalyzed the original data set used to infer the phylogenetic position of Toxarium and found that a more thorough heuristic search was necessary to find the optimal tree. Second, we aligned 181 diatom and eight outgroup SSU rDNA sequences to maximize the juxtapositioning of similar primary and secondary structure of the 18S rRNA molecule over a much broader sampling of diatoms. We then performed a number of phylogenetic analyses purposely based on disparate sets of assumptions and found that none of these analyses supported the conclusion that Toxarium acquired its pennate‐like outline independently from Pennales. Our results suggest that elongate outline is congruent with SSU rDNA data and may be synapomorphic for a larger, more inclusive clade than the traditional Pennales.     

A REASSESSMENT OF PHYLOGENETIC RELATIONSHIPS WITHIN THE PHAEOPHYCEAE BASED ON RUBISCO LARGE SUBUNIT AND RIBOSOMAL DNA SEQUENCES

To better assess the current state of phaeophycean phylogeny, we compiled all currently available rbc L, 18S, and 26S rDNA sequences from the EMBL/GenBank database and added 21 new rbc L sequences of our own. We then developed three new alignments designed to maximize taxon sampling while minimizing information loss due to partial sequences. Phylogenetic analyses were performed on separate and combined data sets (with and without taxa from the sister classes Tribophyceae and Phaeothamniophyceae as outgroups) using a variety of assumption sets, tree-drawing algorithms (parsimony, neighbor joining, and likelihood), and resampling methods (bootstrap, decay, jackknife). Partition homogeneity testing (PHT) by codon position within rbc L showed that all positions could be used despite mild third position saturation. PHT by gene and domain within rDNA showed that the 26S D1 and D2 regions do not enhance phylogenetic signal even when combined with the 18S. The rbc L and rDNA (excluding the 26S D1 and D2) could be combined under PHT. The topology of the combined tree was the same as that of the rbc L tree alone, but bootstrap support was consistently higher in the combined analysis, applied to more branches, and enabled the establishment of sister group relationships among six orders. Although the taxon sampling for the combination tree was lower ( n = 22) than for individual gene analyses ( n = 58 for rbc L and n = 59 for rDNA), results show that the Laminariales (previously reported) and Sphacelariales (new) are both paraphyletic. Choristocarpus tenellus (Kützing) Zanardini is the most basal phaeophyte and the Dictyotales the most basal order. In contrast, the Laminariales sensu stricto ( s.s. ) and Ectocarpales sensu lato ( s.l. ) are the most derived. For phylogenetic studies in the Phaeophyceae, rbc L has more resolving power than rDNA, though the reason for this is unclear based on the fact that both genes are highly conserved.