Identification, Functional Analysis of the Rh Gene Family in Sinonovacula constricta and Screening of It's SNPs Associated with Ammonia Nitrogen Tolerance
LAI Cong-Ying1,2, LIU Zi-Dai1,2, DONG Ying-Hui1,3, LYU Li-Yuan1,2,*, LIN Zhi-Hua1,*
1 Zhejiang Key Laboratory of Aquatic Germplasm Resources/College of Biological & Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China; 2 Ninghai Institute of Mariculture Breeding and Seed Industry, Zhejiang Wanli University, Ninghai 315604, China; 3 College of Advanced Agricultural Sciences, Zhejiang Wanli University, Ningbo 315101, China
Abstract：Rhesus proteins (Rh) belong to the ammonia transporter gene family and play an important role in the ammonia excretion of animals. Sinonovacula constricta, as a main economic shellfish in mixed culture in confined ponds, often faces severe high ammonia nitrogen environment due to its buried lifestyle. In this study, the gene family of Rh transporters in S. constricta (Sco-Rhs) was identified at the genome-wide level by bioinformatics methods. Subsequently, their expression patterns under ammonia stress, gene functions, and correlation with ammonia-nitrogen tolerance were investigated. The results showed that 2 Sco-Rhs genes (Sco-Rhbg-1和Sco-Rhbg-2) were identifed from the S. constricta genome. Moreover, the homology between Sco-Rhs was 57.96%. The tissue expression results showed that Sco-Rhs genes expressed in all tissues, with the highly specific expression in gills (P<0.01). And Sco-Rhs had different expression patterns under ammonia stress. With the increase of stress time, the expression of Sco-Rhbg-1 gene in gills showed a downward trend and had a stronger response to ammonia stress. The expression of Sco-Rhbg-2 gene firstly down-regulated and then increased, and finally approached to the initial level. In addition, heterologous yeast complementation was used to study the function of Sco-Rhs. The results showed that the Sco-Rhbg-1 protein is a functional ammonia transporter, and its ability to transport NH4+ was not affected by external pH. And 15 SNPs in the CDS region of Sco-Rhbg-1 were significantly associated with the ammonia tolerance of S. constricta. These results indicated that the Sco-Rhbg-1 gene played an important role in the regulation of ammonia stress in S. constricta. This study provides an important theoretical basis for further exploration of the molecular regulation of Sco-Rhs in high ammonia nitrogen adaptation and molecular marker-assisted breeding.
赖聪颖, 刘耔岱, 董迎辉, 吕丽媛, 林志华. 缢蛏Rh基因家族的鉴定、功能分析及其与氨氮耐受性相关SNPs位点筛选[J]. 农业生物技术学报, 2025, 33(10): 2209-2223.
LAI Cong-Ying, LIU Zi-Dai, DONG Ying-Hui, LYU Li-Yuan, LIN Zhi-Hua. Identification, Functional Analysis of the Rh Gene Family in Sinonovacula constricta and Screening of It's SNPs Associated with Ammonia Nitrogen Tolerance. 农业生物技术学报, 2025, 33(10): 2209-2223.
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