Zhu L, Li C H, Su X R, Guo C Y, Wang Z, Jin C H, Li Y, Li T W
School of Marine Sciences, Ningbo University, Ningbo, Zhejiang Province, China.
Genet Mol Res. 2013 Aug 20;12(3):3028-37. doi: 10.4238/2013.August.20.4.
Rapid and efficient growth is a major consideration and challenge for global mariculture. The differential growth rate of the sea cucumber, Apostichopus japonicus, has significantly hampered the total production of the industry. In the present study, forward and reverse suppression subtractive hybridization libraries were constructed and sequenced from a fast-growth group and a slow-growth group of the sea cucumber. A total of 142 differentially expressed sequence tags (ESTs) with insertions longer than 150 bp were identified and further analyzed. Fifty-seven of these ESTs (approximately 40%) were functionally annotated for cell structure, energy metabolism, immunity response, and growth factor categories. Six candidate genes, arginine kinase, cytochrome c oxidase subunit I, HSP70, β-actin, ferritin, and the ADP-ribosylation factor, were further validated by quantitative PCR. Significant differences were found between the fast- and slow-growth groups (P < 0.05) for the expression levels of arginine kinase, cytochrome c oxidase, HSP70, the ADP-ribosylation factor, and β-actin. However, no significant difference was observed for ferritin. Our results provide promising candidate gene markers for practical size screening, and also further promote marker-assisted selective breeding of this species.
快速高效的生长是全球海水养殖的主要考量因素和挑战。刺参（Apostichopus japonicus）生长速度的差异显著阻碍了该行业的总产量。在本研究中，从刺参的快速生长组和缓慢生长组构建并测序了正向和反向抑制消减杂交文库。共鉴定出142个插入片段长度超过150 bp的差异表达序列标签（EST），并进行了进一步分析。其中57个EST（约40%）在细胞结构、能量代谢、免疫反应和生长因子类别方面进行了功能注释。通过定量PCR进一步验证了6个候选基因，即精氨酸激酶、细胞色素c氧化酶亚基I、热休克蛋白70（HSP70）、β-肌动蛋白、铁蛋白和ADP-核糖基化因子。在快速生长组和缓慢生长组之间，精氨酸激酶、细胞色素c氧化酶、HSP70、ADP-核糖基化因子和β-肌动蛋白的表达水平存在显著差异（P < 0.05）。然而，铁蛋白未观察到显著差异。我们的结果为实际的大小筛选提供了有前景的候选基因标记，也进一步推动了该物种的分子标记辅助选择育种。
