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Published on: March 21, 2014
Comprehensive analysis of the transcriptome implicated in the immune response of Procambarus clarkii to Aeromonas
Xi-Rong Zhu1, Wen-Hui Tang2, Qin Peng2
1Jiangsu Key Laboratory for Bioresources of Saline Soils, Jiangsu Synthetic Innovation Center for Coastal Bio-agriculture, Jiangsu Provincial Key Laboratory of Coastal Wetland Bioresources and Environmental Protection, School of Wetlands, Yancheng Teachers University, Yancheng 224007, People's Republic of China; College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, People's Republic of China.
Abstract:
In order to better understand the immune response of Procambarus clarkii to Aeromonas hydrophila injection, the transcriptome data of the gill tissue of P. clarkii were compared and analyzed. 1008 significant DEGs were identified in A. hydrophila infected and PBS control groups, including 411 up-regulated genes and 597 down-regulated genes. Endocytosis, phagocyte and lysosome were the most clustered pathways of DEGs in KEGG database. RNA-Seq results were validated through the verification of immune-related differentially expressed genes (DEGs), whose expression levels were assessed using quantitative real-time PCR (qRT-PCR). With the increase of treatment time of A. hydrophila, the total protein in gill of the treatment group showed a trend of increasing first, then decreasing and then increasing and decreasing. ACP and AKP both show a trend of rising first, then falling and then rising. The results of comprehensive research showed that crayfish infected with A. hydrophila caused damage to gill tissue, and the related immune genes were up-regulated and the immune mechanism was operated to protect the body from A. hydrophila. The differential gene MRC1 was screened through the transcriptome, and to further understand its impact, it was disrupted by RNAi technology, which showed a significant down-regulation of immune genes (TAB2, TLR3, ALF6, Lyso3, clotting factor G beta subunit-like and coagulation factor X-like) as well as genes downstream of the pathway (AP4E1, ARSB and TUBA1A). This study provides a theoretical basis for further exploring the immune adaptability of aquatic animals under bacterial infection.
