Centre for Marine Bioproducts Development, Flinders University, Adelaide, South Australia, SA 5042, Australia.
Department of Medical Biotechnology, School of Medicine, Faculty of Medicine, Nursing and Health Sciences, Flinders University, Adelaide, South Australia, SA 5042, Australia.
Sci Rep. 2017 Feb 2;7:41422. doi: 10.1038/srep41422.
For sponges (phylum Porifera), there is no reliable molecular protocol available for species identification. To address this gap, we developed a multilocus-based Sponge Identification Protocol (SIP) validated by a sample of 37 sponge species belonging to 10 orders from South Australia. The universal barcode COI mtDNA, 28S rRNA gene (D3-D5), and the nuclear ITS1-5.8S-ITS2 region were evaluated for their suitability and capacity for sponge identification. The highest Bit Score was applied to infer the identity. The reliability of SIP was validated by phylogenetic analysis. The 28S rRNA gene and COI mtDNA performed better than the ITS region in classifying sponges at various taxonomic levels. A major limitation is that the databases are not well populated and possess low diversity, making it difficult to conduct the molecular identification protocol. The identification is also impacted by the accuracy of the morphological classification of the sponges whose sequences have been submitted to the database. Re-examination of the morphological identification further demonstrated and improved the reliability of sponge identification by SIP. Integrated with morphological identification, the multilocus-based SIP offers an improved protocol for more reliable and effective sponge identification, by coupling the accuracy of different DNA markers.
对于海绵动物门（Porifera），目前尚无可靠的分子鉴定协议可用于物种鉴定。为了解决这一差距，我们开发了一种基于多基因座的海绵鉴定协议（SIP），该协议通过来自南澳大利亚的 10 个目 37 种海绵物种的样本进行了验证。我们评估了通用条码 COI mtDNA、28S rRNA 基因（D3-D5）和核 ITS1-5.8S-ITS2 区，以确定它们是否适合和能够用于海绵鉴定。应用最高比特评分来推断身份。通过系统发育分析验证了 SIP 的可靠性。28S rRNA 基因和 COI mtDNA 在对各种分类水平的海绵进行分类方面优于 ITS 区。主要限制是数据库没有很好地填充，多样性低，使得难以进行分子鉴定协议。分子鉴定还受到已提交到数据库的海绵序列的形态分类准确性的影响。对形态分类的重新检查进一步证明并提高了 SIP 对海绵鉴定的可靠性。结合形态学鉴定，基于多基因座的 SIP 通过结合不同 DNA 标记的准确性，为更可靠和有效的海绵鉴定提供了一种改进的协议。
