Identification of an anti-lipopolysaccharide factor AV-R isoform (LvALF AV-R) related to Vp_PirAB-like toxin resistance in Litopenaeus vannamei.
Identification of an anti-lipopolysaccharide factor AV-R isoform (LvALF AV-R) related to Vp_PirAB-like toxin resistance in Litopenaeus vannamei.
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DOI：
10.1016/j.fsi.2018.10.005
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发表时间：
2019
影响因子：
4.7
通讯作者：
S. Tinwongger;J. Thawonsuwan;H. Kondo;I. Hirono
中科院分区：
文献类型：
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作者：
S. Tinwongger;J. Thawonsuwan;H. Kondo;I. Hirono
急性肝胰腺坏死病（AHPND）是一种虾养殖疾病，由致病性副溶血性弧菌携带编码vp_pirab样毒素的质粒引起。福尔马林杀死v细胞。用致ahpnd菌株D6 （FKC-VpD6）口服筛选vp_pirab样耐毒凡纳滨对虾。对存活一周的虾的胃和肝胰脏组织进行RNA测序。鉴定了存活对虾、感染ahpnd对虾和正常对虾之间的差异表达基因（DEGs）。通过qPCR验证10个deg的表达。只有一个基因（同源toL基因）。凡纳米抗脂多糖因子AV-R异构体（LvALF AV-R）在活虾肝胰腺中的表达明显强于其他各组。在另外两个FKC-VpD6选择试验中幸存的对虾中，flvalf AV-R的表达也显著增加。重组ALF AV-R在elisa中结合LPS、PGN、革兰氏阴性菌和部分革兰氏阳性菌。在ELISA和Far-Western blot中，ALF AV-R重组蛋白不与天然vp_pirab样毒素相互作用。ForL。饲喂ALF AV-R蛋白3 d后，vpd6浸渍攻毒后的成活率与饲喂2种对照饲料的成活率无显著差异。这些结果表明，lvalf AV-R在虾肝胰脏中表达是对vp_pirab样毒素的响应，尽管其他因素也可能参与了抗性机制。
Acute hepatopancreatic necrosis disease (AHPND) is a shrimp farming disease, caused by the pathogenic Vibrio parahaemolyticus carrying a plasmid encoding Vp_PirAB-like toxins. Formalin-killed cells ofV. parahaemolyticusAHPND-causing strain D6 (FKC-VpD6) were used to select Vp_PirAB-like toxin-resistant Litopenaeus vannamei by oral administration. Stomach and hepatopancreas tissues of shrimps that survived for one week were subjected to RNA sequencing. Differentially expressed genes (DEGs) between surviving shrimp, AHPND-infected shrimp, and normal shrimp were identified. The expressions of 10 DEGs were validated by qPCR. Only one gene (a gene homologous toL. vannameianti-lipopolysaccharide factor AV-R isoform (LvALF AV-R)) was expressed significantly more strongly in the hepatopancreas of surviving shrimp than in the other groups. Significantly higher expression ofLvALF AV-R was also observed in shrimp that survived two other trials of FKC-VpD6 selection. Recombinant ALF AV-R bound to LPS, PGN, Gram-negative bacteria, and some Gram-positive bacteria in ELISAs. ALF AV-R recombinant protein did not interact with native Vp_PirAB-like toxin in an ELISA or a Far-Western blot. ForL. vannameiorally fed ALF AV-R protein for 3 days, the survival rate following challenge with VpD6-immersion was not significantly different from that of shrimp fed two control diets. These results suggest thatLvALF AV-R expression was induced in the hepatopancreas of shrimp in response to the presence of Vp_PirAB-like toxin, although other factors might also be involved in the resistance mechanism.