Abstract Procambarus clarkii is an important aquatic organism in China but its aquaculture has suffered great economic losses due to pathogen infections. To better understand the P. clarkii immune response, we used RNA sequencing (RNA-seq) to examine the expression responses of its hemocyte transcriptome to lipopolysaccharide (LPS). Through assembly and annotation, a total of 53,910 unigenes were identified, with an average length of 1246 bp. 589 differentially in total expressed genes (DEGs) were obtained by the injection of LPS with 310 upregulated genes and 279 downregulated genes. Kyoto Encyclopedia of Genes and Genomes enrichment analysis identified several immune response pathways. Additionally, a lot of DEGs to do with the Ras signaling pathway, lysosome, Rap1 signaling pathway, and mitogen-activated protein kinase signaling pathway were upregulated after LPS challenge. Results of Real-time quantitative reverse transcription PCR revealed that 11 randomly selected immune response genes were upregulated after LPS stimulation compared to phosphate-buffered saline stimulation, and this result also validated the RNA-seq data. Our data further enrich the transcriptome databases of P. clarkii and provide a basis for further analysis of the immune system and defense mechanisms of P. clarkii against LPS challenge.

中文翻译：

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脂多糖攻击后红沼泽小龙虾原螯虾血细胞中差异表达的基因

摘要 克拉克原螯虾是我国重要的水生生物，但其水产养殖业因病原菌感染而遭受了巨大的经济损失。为了更好地理解 P. clarkii 免疫反应，我们使用 RNA 测序 (RNA-seq) 来检查其血细胞转录组对脂多糖 (LPS) 的表达反应。通过组装和注释，共鉴定出53,910个unigenes，平均长度为1246 bp。通过注射具有 310 个上调基因和 279 个下调基因的 LPS，获得了 589 个差异表达基因 (DEG)。京都基因和基因组富集分析百科全书确定了几种免疫反应途径。此外，很多 DEG 与 Ras 信号通路、溶酶体、Rap1 信号通路、LPS 攻击后，丝裂原活化蛋白激酶信号通路上调。实时定量逆转录 PCR 的结果显示，与磷酸盐缓冲盐水刺激相比，LPS 刺激后 11 个随机选择的免疫反应基因上调，该结果也验证了 RNA-seq 数据。我们的数据进一步丰富了 P. clarkii 的转录组数据库，并为进一步分析 P. clarkii 对抗 LPS 攻击的免疫系统和防御机制提供了基础。