Tu Pin-Yang, Huang Shin-Jie, Rajanbabu Venugopal, Wu Jen-Leih, Chen Jyh-Yih
Marine Research Station, Institute of Cellular and Organismic Biology, Academia Sinica, 23-10 Dahuen Rd., Jiaushi, Ilan, 262, Taiwan.
Institute of Cellular and Organismic Biology, Academia Sinica, 128 Academia Road, Section 2, Nankang, Taipei, 115, Taiwan.
BMC Genomics. 2021 Mar 22;22(1):200. doi: 10.1186/s12864-021-07521-5.
Tilapia (Oreochromis niloticus) cultures are frequently infected by Vibrio vulnificus, causing major economic losses to production units. Previously, tilapia expressing recombinant delta-5 desaturase and delta-6 desaturase (D56) were found to be resistant to V. vulnificus infection. In this report, we profile the D56-mediated molecular changes underlying this resistance in tilapia. A comparative transcriptome analysis was performed on V. vulnificus-infected wild-type and D56-transgenic tilapia using Illumina's sequencing-by-synthesis approach. Gene enrichment analysis on differentially expressed unigenes was performed, and the expression patterns were validated by real-time PCR.
Comparative transcriptome analysis was performed on RNA-sequence profiles obtained from wild-type and D56-transgenic tilapia at 0, 6 and 24 h post-infection with V. vulnificaus. GO and KEGG gene enrichment analyses showed that D56 regulates several pathways and genes, including fatty acid (FA) metabolism associated, and inflammatory and immune response. Expression of selected FA metabolism-associated, inflammatory and immune responsive genes was validated by qPCR. The inflammatory and immune responsive genes that are modulated by FA-associated D56 likely contribute to the enhanced resistance against V. vulnificus infection in Tilapia.
Transcriptome profiling and filtering for two-fold change variation showed that 3795 genes were upregulated and 1839 genes were downregulated in D56-transgenic tilapia. These genes were grouped into pathways, such as FA metabolism, FA elongation, FA biosynthesis, biosynthesis of unsaturated FA, FA degradation, inflammation, immune response, and chemokines. FA-associated genes and immune-related genes were modulated by D56 at 6 h and 24 h post infection with V. vulnificus. The expression patterns of FA-related genes, inflammatory genes, antimicrobial peptide genes and immune responsive genes at 0, 3, 6, 12, 24 and 48 h post-infection suggests these genes are involved in the enhanced resistance of D56 transgenic tilapia to V. vulnificus.
罗非鱼(尼罗罗非鱼)养殖常受创伤弧菌感染,给生产单位造成重大经济损失。此前发现,表达重组Δ-5去饱和酶和Δ-6去饱和酶(D56)的罗非鱼对创伤弧菌感染具有抗性。在本报告中,我们剖析了罗非鱼中这种抗性背后由D56介导的分子变化。使用Illumina的合成测序方法,对感染创伤弧菌的野生型和D56转基因罗非鱼进行了比较转录组分析。对差异表达的单基因进行了基因富集分析,并通过实时PCR验证了表达模式。
对野生型和D56转基因罗非鱼在感染创伤弧菌后0、6和24小时获得的RNA序列图谱进行了比较转录组分析。GO和KEGG基因富集分析表明,D56调节多种途径和基因,包括脂肪酸(FA)代谢相关途径以及炎症和免疫反应。通过qPCR验证了所选的与FA代谢相关、炎症和免疫反应基因的表达。由与FA相关的D56调节的炎症和免疫反应基因可能有助于增强罗非鱼对创伤弧菌感染的抗性。
转录组分析和筛选两倍变化差异显示,D56转基因罗非鱼中有3795个基因上调,1839个基因下调。这些基因被归类为脂肪酸代谢、脂肪酸延长、脂肪酸生物合成、不饱和脂肪酸生物合成、脂肪酸降解、炎症、免疫反应和趋化因子等途径。在感染创伤弧菌后6小时和24小时,与FA相关的基因和免疫相关基因受到D56的调节。感染后0、3、6、12、24和48小时,FA相关基因、炎症基因、抗菌肽基因和免疫反应基因的表达模式表明这些基因参与了D56转基因罗非鱼对创伤弧菌抗性的增强。