Bromotyrosine Alkaloids from the Australian Marine Sponge <i>Pseudoceratina verrucosa</i>

Two new bromotyrosine alkaloids, pseudoceralidinone A (<b>1</b>) and aplysamine 7 (<b>2</b>), along with three known compounds were isolated from the Australian sponge <i>Pseudoceratina verrucosa</i>. Their structures were characterized by NMR and MS data and the synthetic route. Their cytotoxicity was evaluated against cancer cell lines (HeLa and PC3) and a noncancer cell line (NFF)

Rhodocomatulin-Type Anthraquinones from the Australian Marine Invertebrates <i>Clathria hirsuta</i> and <i>Comatula rotalaria</i>

Chemical investigations of an Australian sponge, <i>Clathria hirsuta</i>, from the Great Barrier Reef, have resulted in the isolation of two known anthraquinones, rhodocomatulin 5,7-dimethyl ether (<b>1</b>) and rhodocomatulin 7-methyl ether (<b>2</b>). Additionally, four new anthraquinone metabolites, 6-methoxy­rhodocomatulin 7-methyl ether, 3-bromo-6-methoxy-12-desethyl­rhodocomatulin 7-methyl ether, 3-bromo-6-methoxy­rhodocomatulin 7-methyl ether, and 3-bromo­rhodocomatulin 7-methyl ether (<b>3</b>–<b>6</b>), were also isolated and characterized. This is the first report of the rhodocomatulin-type anthraquinones from a marine sponge, as <b>1</b> and <b>2</b> were previously isolated from the marine crinoid genus <i>Comatula</i>. An additional chemical investigation of the marine crinoid <i>Comatula rotalaria</i> enabled the isolation of further quantities of <b>1</b> and <b>2</b>, as well as two additional new crinoid metabolites, 12-desethyl­rhodocomatulin 5,7-dimethyl ether and 12-desethyl­rhodocomatulin 7-methyl ether (<b>7</b> and <b>8</b>). An NMR spectroscopic analysis of compounds <b>7</b> and <b>8</b> provided further insight into the rhodocomatulin planar structure and, together with the successful implementation of DFT-NMR calculations, confirmed that the rhodocomatulin metabolites existed as <i>para</i> rather than <i>ortho</i> quinones

Supplementary Material for Dohrmann et al. (2017) Front. Zool.

Supplementary Material for:<br><br>Dohrmann, M., Kelley, C., Kelly, M., Pisera, A., Hooper, J.N.A., Reiswig, H.M. 2017. An integrative systematic framework helps to reconstruct skeletal evolution of glass sponges (Porifera, Hexactinellida). Frontiers in Zoology. <br

Trikentramides A–D, Indole Alkaloids from the Australian Sponge <i>Trikentrion flabelliforme</i>

Chemical investigations of two specimens of <i>Trikentrion flabelliforme</i> collected from Australian waters have resulted in the identification of four new indole alkaloids, trikentramides A–D (<b>9</b>–<b>12</b>). The planar chemical structures for <b>9</b>–<b>12</b> were established following analysis of 1D/2D NMR and MS data. The relative configurations for <b>9</b>–<b>12</b> were determined following the comparison of ¹H NMR data with data previously reported for related natural products. The application of a quantum mechanical modeling method, density functional theory, confirmed the relative configurations and also validated the downfield carbon chemical shift observed for one of the quaternary carbons (C-5a) in the cyclopenta­[<i>g</i>]­indole series. The indole-2,3-dione motif present in trikentramides A–C is rare in nature, and this is the first report of these oxidized indole derivatives from a marine sponge

Barcoding Sponges: An Overview Based on Comprehensive Sampling

<div><h3>Background</h3><p>Phylum Porifera includes ∼8,500 valid species distributed world-wide in aquatic ecosystems ranging from ephemeral fresh-water bodies to coastal environments and the deep-sea. The taxonomy and systematics of sponges is complicated, and morphological identification can be both time consuming and erroneous due to phenotypic convergence and secondary losses, etc. DNA barcoding can provide sponge biologists with a simple and rapid method for the identification of samples of unknown taxonomic membership. The Sponge Barcoding Project (<a href="http://www.spongebarcoding.org">www.spongebarcoding.org</a>), the first initiative to barcode a non-bilaterian metazoan phylum, aims to provide a comprehensive DNA barcode database for Phylum Porifera.</p> <h3>Methodology/Principal Findings</h3><p>∼7,400 sponge specimens have been extracted, and amplification of the standard COI barcoding fragment has been attempted for approximately 3,300 museum samples with ∼25% mean amplification success. Based on this comprehensive sampling, we present the first report on the workflow and progress of the sponge barcoding project, and discuss some common pitfalls inherent to the barcoding of sponges.</p> <h3>Conclusion</h3><p>A DNA-barcoding workflow capable of processing potentially large sponge collections has been developed and is routinely used for the Sponge Barcoding Project with success. Sponge specific problems such as the frequent co-amplification of non-target organisms have been detected and potential solutions are currently under development. The initial success of this innovative project have already demonstrated considerable refinement of sponge systematics, evaluating morphometric character importance, geographic phenotypic variability, and the utility of the standard barcoding fragment for Porifera (despite its conserved evolution within this basal metazoan phylum).</p> </div

Sequencing success rates per sponge family.

<p>Grey and black colours represent sequences corresponding to non-target organisms and poriferans, respectively. The included families correspond to the families used for the analysis of PCR success, and were analised in the generalised linear model. Asterisks on the right side correspond to the different family groups analysed: *: Acarnidae, Ancorinidae, Axinellidae, Dictyonellidae, Dysideidae, Halichondridae, Iotrochotidae, Microcionidae, Mycalidae, Plakinidae, Raspailidae, Tedaniidae, Tetillidae and Thorectidae; **: Chalinidae, Clionaidae and Suberitidae; ***: Chondropsidae, Coelosphaeridae, Crellidae, Desmacellidae, Isodictyidae and Podospongiidae.</p

Generalised linear model (binomial errors, logit link) of the effect of sample taxonomic affiliation and sample age over sequencing success.

<p>The reported values, are for the core family group (Acarnidae, Ancorinidae, Axinellidae, Dyctionellidae, Dysideidae, Halichondridae, Iotrochotidae, Microcionidae, Mycalidae, Plakinidae, Raspailidae, Tedaniidae, Tetillidae and Thorectidae; upper value) and for this group together with the families Chalinidae, Clionaidae and Suberitidae (middle value), and with the families Chondropsidae, Coelosphaeridae, Crellidae, Desmacellidae, Isodictyidae and Podospongiidae (lower value). N.S. = not significant.</p

Generalised linear model (binomial errors, logit link) of the effect of sample taxonomic affiliation and sample age over PCR success.

<p>The reported values, are for the core family group (Acarnidae, Ancorinidae, Axinellidae, Dyctionellidae, Dysideidae, Halichondridae, Iotrochotidae, Microcionidae, Mycalidae, Plakinidae, Raspailidae, Tedaniidae, Tetillidae and Thorectidae; upper value) and for this group together with the families Chalinidae, Clionaidae and Suberitidae (middle value), and with the families Chondropsidae, Coelosphaeridae, Crellidae, Desmacellidae, Isodictyidae and Podospongiidae (lower value).</p

Amplification success of the standard barcoding <i>COI</i> partition per sponge family.

<p>Grey and black colours represent failed and positive reactions, respectively. Only families with more than 30 documented PCRs are included in the figure, these taxa correspond to the families analised in the generalised linear model. Asterisks on the right side correspond to the different family groups analysed: *: Acarnidae, Ancorinidae, Axinellidae, Dictyonellidae, Dysideidae, Halichondridae, Iotrochotidae, Microcionidae, Mycalidae, Plakinidae, Raspailidae, Tedaniidae, Tetillidae and Thorectidae; **: Chalinidae, Clionaidae and Suberitidae; ***: Chondropsidae, Coelosphaeridae, Crellidae, Desmacellidae, Isodictyidae and Podospongiidae.</p

Modifications to the genomic DNA protocol of Ivanova et al. 2006 used for sponge barcoding.

1<p>Lysis mix: 100 mM NaCl, 50 mM Tris-HCl pH 8.0, 10 mM EDTA pH 8.0, 0.5% SDS, Proteinase K 10% v/v.</p>2<p>Binding buffer: 6 M GuSCN, 20 mM EDTA pH 8.0, 10 mM Tris-HCl pH 6.4, Triton X-100 4% v/v. The Binding mix is a 50% v/v solution of Binding Buffer in ethanol 96%.</p>3<p>Protein wash buffer is a 30% v/v solution of Binding Buffer in ethanol 96%.</p>4<p>Wash buffer: 50 mM NaCl, 10 mM Tris-HCl pH 7.4, 0.5 mM EDTA pH 8.0, ethanol 60%.</p