Yang Qing, Sun Fanyue, Yang Zhi, Li Hongjun
Key Laboratory for Ecological Environment in Coastal Areas (SOA), National Marine Environmental Monitoring Center, Dalian 116023, China.
Department of Reconstructive Sciences, Center for Regenerative Medicine and Developmental Biology, University of Connecticut Health Center, Farmington, CT 06030, USA.
Biomed Res Int. 2014;2014:493825. doi: 10.1155/2014/493825. Epub 2014 May 20.
Calanus sinicus Brodsky (Copepoda, Crustacea) is a dominant zooplanktonic species widely distributed in the margin seas of the Northwest Pacific Ocean. In this study, we utilized an RNA-Seq-based approach to develop molecular resources for C. sinicus. Adult samples were sequenced using the Illumina HiSeq 2000 platform. The sequencing data generated 69,751 contigs from 58.9 million filtered reads. The assembled contigs had an average length of 928.8 bp. Gene annotation allowed the identification of 43,417 unigene hits against the NCBI database. Gene ontology (GO) and KEGG pathway mapping analysis revealed various functional genes related to diverse biological functions and processes. Transcripts potentially involved in stress response and lipid metabolism were identified among these genes. Furthermore, 4,871 microsatellites and 110,137 single nucleotide polymorphisms (SNPs) were identified in the C. sinicus transcriptome sequences. SNP validation by the melting temperature (T m )-shift method suggested that 16 primer pairs amplified target products and showed biallelic polymorphism among 30 individuals. The present work demonstrates the power of Illumina-based RNA-Seq for the rapid development of molecular resources in nonmodel species. The validated SNP set from our study is currently being utilized in an ongoing ecological analysis to support a future study of C. sinicus population genetics.
中华哲水蚤(Calanus sinicus Brodsky,桡足纲,甲壳亚门)是一种优势浮游动物物种,广泛分布于西北太平洋边缘海域。在本研究中,我们采用基于RNA测序的方法来开发中华哲水蚤的分子资源。使用Illumina HiSeq 2000平台对成年样本进行测序。测序数据从5890万个过滤后的 reads 中生成了69751个重叠群。组装后的重叠群平均长度为928.8 bp。基因注释使得能够在NCBI数据库中鉴定出43417个单基因匹配。基因本体论(GO)和KEGG通路映射分析揭示了与多种生物学功能和过程相关的各种功能基因。在这些基因中鉴定出了可能参与应激反应和脂质代谢的转录本。此外,在中华哲水蚤转录组序列中鉴定出4871个微卫星和110137个单核苷酸多态性(SNP)。通过熔解温度(Tm)偏移法进行的SNP验证表明,16对引物扩增出了目标产物,并在30个个体中表现出双等位基因多态性。本研究工作证明了基于Illumina的RNA测序在非模式物种分子资源快速开发方面的强大作用。我们研究中经过验证的SNP集目前正在用于正在进行的生态分析中,以支持未来对中华哲水蚤种群遗传学的研究。