Jin Yulin, Zhou Tao, Jiang Wansheng, Li Ning, Xu Xiaoyan, Tan Suxu, Shi Huitong, Yang Yujia, Yuan Zihao, Wang Wenwen, Qin Guyu, Liu Shikai, Gao Dongya, Dunham Rex, Liu Zhanjiang
The Fish Molecular Genetics and Biotechnology Laboratory, Aquatic Genomics Unit, School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL, 36849, USA.
Department of Marine Biology & Biotechnology, College of Ocean and Earth Sciences, Xiamen University, Xiamen, China.
Mar Biotechnol (NY). 2022 Mar;24(1):174-189. doi: 10.1007/s10126-022-10094-3. Epub 2022 Feb 15.
Identification of genetic markers associated with resistance against enteric septicemia of catfish (ESC) is of great interest for genetic enhancement programs of catfish. In the present study, bulk segregant RNA-Seq analysis was applied to determine differentially expressed genes and alleles after ESC infection. Here we report three genomic regions on LG1, LG12, and LG26, containing significant single-nucleotide polymorphisms (SNPs). These genomic regions aligned well with quantitative trait loci (QTL) previously identified. Within the QTL regions, eleven genes were found to be differentially regulated between phenotypic bulks. Importantly, the QTL on linkage group 1 (LG1) were found to be expressed in the liver, whereas the QTL on LG12 and LG26 were expressed in the intestine, suggesting multiple mechanisms of ESC resistance. It is apparent that apolipoproteins may be important for ESC resistance as the QTL on LG1 included the 14-kDa apolipoprotein genes that are both allelically expressed and differentially expressed between the resistant and susceptible bulks. Traf2 and NCK-interacting protein kinase (TNIK) were found in the QTL on LG12, and it was downregulated in resistant fish, suggesting the importance of NCK downregulation in ESC resistance, as previously reported. In addition, we observed divergent gene expression patterns between the liver and intestine after infection. Immune/inflammatory-related processes were overrepresented from liver DEGs, while those DEGs identified from intestine were enriched for proteolysis and wounding processes. Taken together, the BSR-Seq analysis presented here advanced the knowledge of ESC resistance, providing information of not only positions of QTL but also genes and their differential expression between resistant and susceptible fish, making it one step closer to the identification of the causal genes for ESC resistance.
鉴定与鲶鱼抗肠败血症(ESC)相关的遗传标记对于鲶鱼的遗传改良计划具有重要意义。在本研究中,采用混合分组RNA-Seq分析来确定ESC感染后差异表达的基因和等位基因。在此,我们报告了LG1、LG12和LG26上的三个基因组区域,其中包含显著的单核苷酸多态性(SNP)。这些基因组区域与先前鉴定的数量性状位点(QTL)高度吻合。在QTL区域内,发现11个基因在表型混合群体之间存在差异调控。重要的是,连锁群1(LG1)上的QTL在肝脏中表达,而LG12和LG26上的QTL在肠道中表达,这表明ESC抗性存在多种机制。显然,载脂蛋白可能对ESC抗性很重要,因为LG1上的QTL包含14 kDa载脂蛋白基因,这些基因在抗性和敏感混合群体之间既存在等位基因表达差异,也存在差异表达。在LG12上的QTL中发现了Traf2和NCK相互作用蛋白激酶(TNIK),并且它在抗性鱼类中下调,这表明如先前报道的那样,NCK下调在ESC抗性中很重要。此外,我们观察到感染后肝脏和肠道之间存在不同的基因表达模式。肝脏差异表达基因中免疫/炎症相关过程占比过高,而从肠道中鉴定出的差异表达基因则富集于蛋白水解和伤口愈合过程。综上所述,本文提出的BSR-Seq分析推进了对ESC抗性的认识,不仅提供了QTL的位置信息,还提供了抗性和敏感鱼类之间的基因及其差异表达信息,使其更接近鉴定ESC抗性的因果基因。