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Aquaculture Environment Interactions

    AEI prepress abstract   -  DOI:

    Characterization of a novel ADP ribosylation factor gene from Macrobrachium nipponense and its response to ammonia nitrogen stress

    Leifeng Guo, Bing Sun, Duanduan Chen, Cao Yi, Jian Teng, Jielun Yu, Shouquan Wang, Yuanyuan Ru, Hui Wang*

    *Corresponding author:

    ABSTRACT: ADP-ribosylation factors (Arfs) are known as GTP-binding proteins that play essential roles in membrane trafficking, and they have been recently reported to be involved in innate immunity in crustaceans. However, there is little information on Arfs in Macrobrachium nipponense and their response to ammonia nitrogen stress to date. In this study, a novel M. nipponense Arfn gene (MnArfn) was identified. The full-length cDNA of MnArfn was 1076 bp. It contained a 537 bp open reading frame (ORF) and encoded a 178-amino acid protein with a predicted molecular weight of 19.85 kDa. Sequence and phylogenetic analyses showed that MnArfn was an unidentified Arf, sharing 55–61% identity with other known Arfs. Quantitative real-time PCR (qRT-PCR) indicated that all the examined tissues, namely, the hepatopancreas, stomach, gill, heart, muscle, and eyestalk, expressed MnArfn. Hepatopancreas and gills, 2 organs involved in environmental stress management, had the highest expression. Under conditions of ammonia nitrogen stress, MnArfn expression in hepatopancreas and gills was significantly up-regulated at 6, 12, and 24 h. Western blotting experiments revealed that MnArfn was distributed in all the examined tissues, with the highest expression in hepatopancreas and gills, consistent with qRT-PCR results. The findings from this study indicate that MnArfn may have important roles in the response to ammonia nitrogen stress in M. nipponense, which provides a new idea to study the resistance mechanism of crustaceans to ammonia nitrogen and to screen individuals with resistance to unfavorable environment.