Browsing by Author "Wickett, N. J."
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- Deep sequencing of Ptilidium (Ptilidiaceae) suggests evolutionary stasis in liverwort plastid genome structurePublication . Forrest, L. L.; Wickett, N. J.; Cox, C. J.; Goffinet, B.Background and aims – Organellar genome sampling is patchy for non-vascular groups, with the earliest land plants poorly represented; currently only two liverworts, two mosses and one hornwort have sequenced, annotated plastid genomes. This is in part due to methodological difficulties that have hampered attempts to generate plastid genome data from liverworts. In this paper we present a method that overcomes some of the inherent difficulties by circumventing the need for plastid enrichment, but that also provides other valuable information from nuclear and mitochondrial regions including sequences from loci that may be phylogenetically useful, and potential population-level markers such as single nucleotide polymorphisms and microsatellites. Methods – A shotgun library developed from total genomic liverwort DNA was subjected to high-throughput pyrosequencing using the Roche 454 platform. Plastid reads were bioinformatically identified, assembled and annotated. To maximize usage of the vast number of reads generated using 454 sequencing technology, combined nuclear, mitochondrial and plastid contigs were also screened for microsatellite markers, and presumed nuclear contigs were scanned for protein domains. Key Results – This is the first plastid genome to be assembled for a leafy liverwort (i.e. Ptilidium) and also the first such genome to be sequenced using next generation technology for any bryophyte. The 119,007 base long plastid genome of Ptilidium pulcherrimum contains 88 protein-coding genes, four rRNAs and thirty tRNAs. The Inverted Repeat occurs between trn V-GAC and trn N-GUU. Functional copies of the two plastid-encoded sulphate import protein-coding genes (cysA and cysT) are absent, although pseudogenes are present in the same position that the functional genes occupy in Marchantia. Microsatellites: 197 novel potential primer pairs for P. pulcherrimum were found. Presumed nuclear Ptilidium contigs gave multiple hits to Class I transposable elements. Conclusions – The arrangement of genes is identical to the plastid of the complex thalloid liverwort Marchantia, suggesting that structural rearrangements are rare in hepatics. This dataset represents a valuable resource for novel phylogenetic and population level marker design in hepatics.
- Distribution and phylogenetic significance of the 71-kb inversion in the plastid genome in Funariidae (Bryophyta)Publication . Goffinet, B.; Wickett, N. J.; Werner, O.; Ros, R. M.; Shaw, A. J.; Cox, C. J.Background and Aims The recent assembly of the complete sequence of the plastid genome of the model taxon Physcomitrella patens (Funariaceae, Bryophyta) revealed that a 71-kb fragment, encompassing much of the large single copy region, is inverted. This inversion of 57% of the genome is the largest rearrangement detected in the plastid genomes of plants to date. Although initially considered diagnostic of Physcomitrella patens, the inversion was recently shown to characterize the plastid genome of two species from related genera within Funariaceae, but was lacking in another member of Funariidae. The phylogenetic significance of the inversion has remained ambiguous. Methods Exemplars of all families included in Funariidae were surveyed. DNA sequences spanning the inversion break ends were amplified, using primers that anneal to genes on either side of the putative end points of the inversion. Primer combinations were designed to yield a product for either the inverted or the non-inverted architecture. Key Results The survey reveals that exemplars of eight genera of Funariaceae, the sole species of Disceliaceae and three generic representatives of Encalyptales all share the 71-kb inversion in the large single copy of the plastid genome. By contrast, the plastid genome of Gigaspermaceae (Funariales) is characterized by a gene order congruent with that described for other mosses, liverworts and hornworts, and hence it does not possess this inversion. Conclusions The phylogenetic distribution of the inversion in the gene order supports a hypothesis only weakly supported by inferences from sequence data whereby Funariales are paraphyletic, with Funariaceae and Disceliaceae sharing a common ancestor with Encalyptales, and Gigaspermaceae sister to this combined clade. To reflect these relationships, Gigaspermaceae are excluded from Funariales and accommodated in their own order, Gigaspermales order nov., within Funariideae.
- Moss diversity: a molecular phylogenetic analysis of generaPublication . Cox, C. J.; Goffinet, B.; Wickett, N. J.; Boles, S. B.; Shaw, A. J.In this study we present phylogenetic and molecular phylogenetic diversity analyses of moss taxa from a total of 655 genera of mosses. Three loci were sampled: chloroplast ribosomal small protein 4, the intronic region of the mitochondrial NADH dehydogenase subunit 5, and partial sequences of the nuclear 26S ribosomal RNA. Maximum likelihood and Bayesian phylogenetic analyses were performed on individual loci and on multilocus data sets. A measure of phylogenetic diversity was calculated and constrasted among major lineages of mosses. We reveal many instances of incongruence among genomic partitions, but, overall, our analyses describe relationships largely congruent with previous studies of the major groups of mosses. Moreover, our greater sampling highlights the possible non-monophyly of many taxonomic families, particularly in the haplolepideous and pleurocarpous mosses. Comparisons of taxic and phylogenetic diversity among genera indicate that the Dicranidae (haplolepideous taxa) include about 15% of moss genera, but nearly 30% of the phylogenetic diversity. By contrast, the Hypnanae (hypnalian pleurocarps) contain about 45% of moss genera, but a lower percentage of phylogenetic diversity. Agreement between numbers of genera and phylogenetic diversity within other moss clades are remarkably consistent.
- Phylogenetic significance of the rpoA loss in the chloroplast genome of mossesPublication . Goffinet, B.; Wickett, N. J.; Shaw, A. J.; Cox, C. J.A recent survey of arthrodontous mosses revealed that their chloroplast genome lacks the gene encoding the alpha subunit of the RNA polymerase (i.e., rpoA), and that at least in Physcomitrella patens the gene has been transferred to the nuclear genome. Subsequently the gene was recorded from the cytoplasmic genome in Takakia and Sphagnum. Here we extend the survey to representatives of all major lineages of mosses to determine when in the evolutionary history of the Bryophyta the loss took place. Amplifications using primers annealing to the flanking regions of the rpoA gene yield a product that contains the gene in Takakia, Sphagnum, Andreaea, Oedipodium, Polytrichaceae, and Buxbaumia. The gene is lacking in all arthrodontous mosses, including Diphyscium but also in both species of Tetraphis. Reconstruction of the transfer on the phylogeny of mosses suggests (a) that the rpoA gene was lost twice and (b) that the gene was lost after the divergence of Buxbaumiidae and prior to the divergence of Diphyscium from the remaining Bryopsida.