Olasard Praewrung, Suksri Phassorn, Taneerat Chanikan, Rungrassamee Wanilada, Sathapondecha Ponsit
Center for Genomics and Bioinformatics Research, Division of Biological Science, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand.
Biosensing and Bioprospectiing Technology Research Group, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 111 Thailand Science Park, Phahonyothin Road, Khlong Luang, Pathum Thani 12120, Thailand.
Fish Shellfish Immunol. 2024 Sep;152:109768. doi: 10.1016/j.fsi.2024.109768. Epub 2024 Jul 14.
Acute hepatopancreatic necrosis disease (AHPND) caused by toxin-producing Vibrio parahaemolyticus (Vp) has severely affected shrimp production. Long non-coding RNA (lncRNA), a regulatory non-coding RNA, which can play important function in shrimp disease responses. This study aimed to identify and investigate the role of lncRNA involved in Vp infection in Pacific white shrimp, Litopenaeus vannamei. From a total of 368,736 de novo assembled transcripts, 67,559 were identified as putative lncRNAs, and only 72 putative lncRNAs showed differential expression between Vp-infected and normal shrimp. The six candidate lncRNAs were validated for their expression profiles during Vp infection and tissue distribution using RT-qPCR. The role of lnc2088 in response to Vp infection was investigated through RNA interference. The result indicated that the suppression of lnc2088 expression led to an increase in shrimp mortality after Vp infection. To explore the set of genes involved in lnc2088 knockdown, RNA sequencing was performed. A total of 275 differentially expressed transcripts were identified in the hepatopancreas of lnc2088 knockdown shrimp. The expression profiles of five candidate metabolic and immune-related genes were validated in lnc2088 knockdown and Vp-infected shrimp. The result showed that the expression of ChiNAG was significantly increased, while that of NCBP1, WIPF2, and NFKB1 was significantly downregulated in ds2088-injected shrimp. Additionally, the expression of NFKB1, NCBP1 and WIPF2 was significantly increased, whereas that of ChiNAG and CUL5 were significantly decreased after infection with Vp. Our work identified putative lncRNA profiles in L. vannamei in response to Vp infection and investigated the role of lncRNA in shrimp immunity.
由产毒素副溶血性弧菌（Vp）引起的急性肝胰腺坏死病（AHPND）严重影响了对虾养殖。长链非编码RNA（lncRNA）是一种调控性非编码RNA，在对虾疾病反应中发挥重要作用。本研究旨在鉴定并研究参与太平洋白对虾凡纳滨对虾Vp感染过程的lncRNA的作用。从总共368,736条从头组装的转录本中，鉴定出67,559条为假定lncRNA，其中只有72条假定lncRNA在Vp感染的对虾和正常对虾之间表现出差异表达。使用RT-qPCR验证了6条候选lncRNA在Vp感染期间的表达谱及其组织分布。通过RNA干扰研究了lnc2088在应对Vp感染中的作用。结果表明，抑制lnc2088表达会导致Vp感染后对虾死亡率增加。为了探索参与lnc2088敲低的基因集，进行了RNA测序。在lnc2088敲低的对虾肝胰腺中总共鉴定出275条差异表达的转录本。在lnc2088敲低和Vp感染的对虾中验证了5条候选代谢和免疫相关基因的表达谱。结果显示，注射ds2088的对虾中，几丁质酶N-乙酰葡糖胺酶（ChiNAG）的表达显著增加，而核仁结合蛋白1（NCBP1）、Wiskott-Aldrich综合征蛋白家族成员2（WIPF2）和核因子κB1（NFKB1）的表达显著下调。此外，Vp感染后，NFKB1、NCBP1和WIPF2的表达显著增加，而ChiNAG和Cullin 5（CUL5）的表达显著降低。我们的工作鉴定了凡纳滨对虾对Vp感染的假定lncRNA谱，并研究了lncRNA在对虾免疫中的作用。
