A Primer to Molecular Phylogenetic Analysis in Plants

Reconstructing a tree of life by inferring evolutionary history is an important focus of evolutionary biology. Phylogenetic reconstructions also provide useful information for a range of scientific disciplines such as botany, zoology, phylogeography, archaeology and biological anthropology. Until the development of protein and DNA sequencing techniques in the 1960s and 1970s, phylogenetic reconstructions were based on fossil records and comparative morphological/physiological analyses. Since then, progress in molecular phylogenetics has compensated for some of the shortcomings of phenotype-based comparisons. Comparisons at the molecular level increase the accuracy of phylogenetic inference because there is no environmental influence on DNA/peptide sequences and evaluation of sequence similarity is not subjective. While the number of morphological/physiological characters that are sufficiently conserved for phylogenetic inference is limited, molecular data provide a large number of datapoints and enable comparisons from diverse taxa. Over the last 20 years, developments in molecular phylogenetics have greatly contributed to our understanding of plant evolutionary relationships. Regions in the plant nuclear and organellar genomes that are optimal for phylogenetic inference have been determined and recent advances in DNA sequencing techniques have enabled comparisons at the whole genome level. Sequences from the nuclear and organellar genomes of thousands of plant species are readily available in public databases, enabling researchers without access to molecular biology tools to investigate phylogenetic relationships by sequence comparisons using the appropriate nucleotide substitution models and tree building algorithms. In the present review, the statistical models and algorithms used to reconstruct phylogenetic trees are introduced and advances in the exploration and utilization of plant genomes for molecular phylogenetic analyses are discussed.     

Towards a phylogeny of chitons (Mollusca, Polyplacophora) based on combined analysis of five molecular loci

This study represents the first phylogenetic analysis of the molluscan class Polyplacophora using DNA sequence data. We employed DNA from a nuclear protein-coding gene (histone H3), two nuclear ribosomal genes (18S rRNA and the D3 expansion fragment of 28S rRNA), one mitochondrial protein-coding gene (cytochrome c oxidase subunit I), and one mitochondrial ribosomal gene (16S rRNA). A series of analyses were performed on independent and combined data sets. All these analyses were executed using direct optimization with parsimony as the optimality criterion, and analyses were repeated for nine combinations of parameters affecting indel and transversion/transition cost ratios. Maximum likelihood was also explored for the combined molecular data set, also using the direct optimization method, with a model equivalent to GTR + I + Γ that accommodates gaps. The results of all nine parameter sets for the combined parsimony analysis of all molecular data (as well as ribosomal data) and the maximum-likelihood analysis of all molecular data support monophyly of Polyplacophora. The resulting topologies mostly agree with a division of Polyplacophora into two major lineages: Lepidopleuridae and Chitonida (sensu Sirenko 1993). In our analyses the genus Callochiton is positioned as the sister group to Lepidopleuridae, and not as sister group to the remaining Chitonida (sensu Buckland-Nicks & Hodgson 2000), nor as the sister group to the remaining Chitonina (sensu Buckland-Nicks 1995). Chitonida (excluding Callochiton) is monophyletic, but conventional subgroupings of Chitonida are not supported. Acanthochitonina (sensu Sirenko 1993) is paraphyletic, or alternatively monophyletic, and is split into two clades, both with abanal gills only and cupules in the egg hull, but one has simple cupules whereas the other has more strongly hexagonal cupules. Sister to the Acanthochitonina clades is Chitonina, including taxa with adanal gills and a spiny egg hull. Schizochiton, the only genus with adanal gills that has an egg hull with cupules, is the sister-taxon to one of the Acanthochitonina clades plus Chitonina, or alternatively basal to Chitonina. Support values for either position are low, leaving this relationship unsettled. Our results refute several aspects of conventional classifications of chitons that are based primarily on shell characters, reinforcing the idea that chiton classification should be revised using additional characters.     

基于rDNA ITS序列对绒泡菌目黏菌系统发育的探讨

绒泡菌目Physarida是黏菌纲Myxogastria最大的一个目,对其系统发育关系的研究一直是根据形态特征。为了从分子水平探讨绒泡菌目乃至黏菌纲的系统发育关系,以黏菌r DNA ITS通用引物对绒泡菌目5属8种黏菌的r DNA ITS进行扩增和测序,结合Gen Bank中已有的黏菌r DNA ITS序列,利用贝叶斯推断法(Bayesian inference,BI)和最大似然法(Maximum likelihood,ML)构建系统发育树。结果表明:绒泡菌目不同物种的r DNA ITS区在碱基组成和长度上差异明显,长度为777–1 445bp,G+C mol%在53.4%–61.9%之间。绒泡菌目与发网菌目Stemonitida聚类为两个明显的分支,在绒泡菌目分支上,绒泡菌科Physaraceae和钙皮菌科Didymiaceae各聚为一支,支持了形态学上以孢丝是否具有石灰质为依据区分这两个科的观点。由多份不同地理来源的鳞钙皮菌Didymium squamulosum材料组成的钙皮菌科又形成3个分支,证实了这个形态种是由地域来源广泛、繁殖亲和性各异和遗传变异较大的不同生物种组成的复合体。     

The influence of phylogenetic uncertainty on the detection of positive Darwinian selection

The power of maximum likelihood tests of positive selection on protein-coding genes depends heavily on detecting and accounting for potential biases in the studied data set. Although the influence of transition:transversion and codon biases have been investigated in detail, little is known about how inaccuracy in the phylogeny used during the calculations affects the performance of these tests. In this study, 3 empirical data sets are analyzed using sets of simulated topologies corresponding to low, intermediate, and high levels of phylogenetic uncertainty. The detection of positive selection was largely unaffected by errors in the underlying phylogeny. However, the number of sites identified as being under positive selection tended to be overestimated.     

最大似然法和贝叶斯推论与似然比检验探讨泽泻目系统发育关系

应用最大似然法(ML)、贝叶斯推论(BI)、邻接法(NJ)和似然比检验(hLRTs)进行泽泻目分子系统学研究。所用的rbcL基因序列代表了泽泻目14科46属以及作为外类群的6相关属。研究结果表明,*等级制似然比检验表明泽泻目rbcL序列最适合的DNA进化模型为GTR+I+G,最大似然法、贝叶斯法和邻接法构建的系统发育树拓扑结构相似,没有显著的差异,但贝叶斯树支持率较高;泽泻目为一单系类群,由两个主要谱系分支构成,深层分布格局由5个主要分支构成。基于分子系统发育树,文中对泽泻目科间、水鳖科+茨藻科、泽泻科+花蔺科+黄花蔺科、和\"Cymodoeaceae complex\"的系统发育关系进行了讨论。研究结果还表明,泽泻目系统发育关系可能还需要更多的证据进一步的澄清。     

Consideration of RNA secondary structure significantly improves likelihood-based estimates of phylogeny: examples from the bilateria

Sequences from ribosomal RNA (rRNA) genes have made a huge contribution to our current understanding of metazoan phylogeny and indeed the phylogeny of all of life. That said, some parts of this rRNA-based phylogeny remain unresolved. One approach to increase the resolution of these trees would be to use more appropriate models of sequence evolution in phylogenetic analysis. RNAs transcribed from rRNA genes have a complex secondary structure mediated by base pairing between sometimes distant regions of the rRNA molecule. The pairing between the stem nucleotides has important consequences for their evolution which differs from that of unpaired loop nucleotides. These differences in evolution should ideally be accounted for when using rRNA sequences for phylogeny estimation. We use a novel permutation approach to demonstrate the significant superiority of models of sequence evolution that allow stem and loop regions to evolve according to separate models and, in common with previous studies, we show that 16-state models that take base pairing of stems into account are significantly better than simpler, 4-state, single-nucleotide models. One of these 16-state models has been applied to the phylogeny of the Bilateria using small subunit rRNA (SSU) sequences. Our optimal tree largely echoes previous results based on SSU in particular supporting the tripartite Bilaterian tree of deuterostomes, lophotrochozoans, and ecdysozoans. There are also a number of differences, however, perhaps most important of which is the observation of a clade consisting of the gastrotrichs plus platyheminthes that is basal to all other lophotrochozoan taxa. Use of 16-state models also appears to reduce the Bayesian support given to certain biologically improbable groups found using standard 4-state models.     

150 years beyond Darwin's Origin of species: finding new approaches to reconstruct early animal phylogeny

The conference ‘Celebrating Darwin: From the Origin of Species to Deep Metazoan Phylogeny’ was held at the Humboldt University in Berlin, from 3 to 6 March 2009. Specialists from the fields of bioinformatics, molecular biology, developmental biology, comparative morphology and paleontology joined forces to present and discuss novel approaches in reconstructing the still unresolved early branching patterns of the metazoan tree of life.     

Illumination or confusion? Dinoflagellate molecular phylogenetic data viewed from a primarily morphological standpoint

Recent molecular sequencing results involving multiple genes require evaluation in the light of preexisting morphological data, particularly as different methodologies and genes produce trees that are incongruent in some respects or have major areas with poorly supported branch resolution. The present paper summarizes the current situation, primarily from a morphologist's perspective. Most of the tabulation‐based groups are coherent in small subunit (SSU) and large subunit (LSD) trees; but some, notably the prorocentroids and peridinioids, are not. In prorocen‐troids this is primarily because of intrinsic inadequacies of the molecules to resolve their phylogeny. In peridinioids it seems to be because of paraphyly of the group. Other artefacts are noted, such as the drastically different positions of Oxyrrhis in phylogenetic trees based on SSU and protein genes, and of Noctiluca in SSU trees that include analyses with different numbers of nucleotides. Polyphyly in non‐tabulate or poorly known groups has been confirmed, as has been the presence of cryptic thecae in members of those groups (group misattribution). Whether or not some extant groups of athecate, wholly dinokaryotic forms originated prior to polytabulate groups, like the suessioids, peridinioids and gonyaula‐coids, remains unclear. Gymnodinioids with a spiral acrobase seem to have given rise to the more complex athecate forms, whereas morphological features of the genus Gymnodinium are consistent with it being a sister group to polytabulate taxa such as Woloszynskia and the suessioids. Peridinioids and gonyaulacoids appear to have originated after that split. Dinophysoid and prorocentroid dinoflagellates appear to be derived from peridinioid forms. Trees based on protein genes, such as actin or α‐ and β‐tubulin, may help resolve some of the positions of key groups, but they do not include enough taxa to be widely useful as yet.