Functional characterization of ampd1 and its SNPs in the muscle IMP synthesis of hybrid bream (Megalobrama amblycephala ♀ × Culter alburnus ♂)

IF 3.9 1区农林科学 Q1 FISHERIES

Aquaculture Pub Date : 2025-05-28 DOI:10.1016/j.aquaculture.2025.742772

Zhehua Xu , Qinqin Xiao , Mingyu Zhang , Yimiao He , Qianting Zheng , Junchi Zhao , Anli Zuo , Jianzhou Tang , Yonghua Zhou , Shenping Cao , Fufa Qu , Zhen Liu

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引用次数: 0

Abstract

Adenosine monophosphate deaminase 1 (AMPD1) plays a key role in the formation of inosine monophosphate (IMP), which is an important umami substance in fish muscle. The BBTB is a new type of hybrid bream derived from two rounds of backcrossing within an established hybrid lineage resulting from the crossing of blunt snout bream (Megalobrama amblycephala, BSB, ♀) × topmouth culter (Culter alburnus, TC, ♂). In this study, the role of ampd1 in IMP synthesis was first investigated in muscle cells of the BBTB. The content of IMP and the expression of genes involved in the IMP salvage and metabolism pathway (aprt, hprt, impdh, and gmps) were inhibited after the knockdown of ampd1. The cDNA sequence of ampd1 was subsequently cloned from BBTB, and it contained a full open reading frame (ORF) of 2160 bp encoding 719 amino acids. The analysis of sequence identity and the phylogenetic tree revealed that BBTB is closely related to BSB. Tissue expression analysis revealed that the mRNA expression of ampd1 was highest in the muscle of BSB and BBTB. Sanger sequencing was employed for the identification of single nucleotide polymorphisms (SNPs) of ampd1 from BBTB and BSB, revealing four SNPs located in the 8th exon (g.9801 T > G), 16th exon (16,920 T > C), and 3’UTR (g.16947 T > C, g.16984 G > T). These SNPs are significantly associated with IMP content, with individuals of the homozygous genotype showing higher IMP content than those of the heterozygous genotype. Moreover, the proportion of homozygous genotypes in BBTB was significantly greater than that in BSB. Finally, three SNP loci (g.16920 T > C, g.16947 T > C and g.16984 G > T) exhibited strong pairwise linkage disequilibrium. These findings suggest a strong correlation between ampd1 polymorphisms and IMP content in BBTB and BSB, and the ampd1 gene could be used as a candidate marker for the genetic selection of BBTB with high IMP content.

查看原文本刊更多论文

杂交鳙鱼（Megalobrama amblycephala♀× Culter alburnus♂）肌肉IMP合成中ampd1及其snp的功能表征

单磷酸腺苷脱氨酶1 （AMPD1）在鱼肌肉中重要鲜味物质单磷酸肌苷（IMP）的形成中起关键作用。BBTB是由钝口鲷（Megalobrama amblycepphala， BSB，♀）与顶口鲷（culter alburnus， TC，♂）杂交而成的一种新型杂交鲷。本研究首次在BBTB肌肉细胞中研究了ampd1在IMP合成中的作用。敲低ampd1后，IMP的含量以及IMP挽救和代谢途径相关基因（aprt、hprt、impdh、gmp）的表达均受到抑制。ampd1 cDNA序列克隆自BBTB，全长2160 bp，编码719个氨基酸。序列同源性分析和系统进化树分析表明，BBTB与BSB亲缘关系密切。组织表达分析显示，ampd1 mRNA在BSB和BBTB肌肉中表达量最高。采用Sanger测序对BBTB和BSB的ampd1基因进行单核苷酸多态性（snp）鉴定，发现4个snp位于第8外显子（g.9801）T比;G)，第16外显子(16,920 T >；C)和3'UTR （g.16947）T比;C, g.16984G比;T)。这些snp与IMP含量显著相关，纯合子基因型个体的IMP含量高于杂合子基因型个体。此外，纯合基因型在BBTB中的比例显著高于BSB。最后，三个SNP位点(g.16920T比;C, g.16947T比;C、g.16984G比;T)表现出强烈的两两联动不平衡。上述结果提示，在BBTB和BSB中ampd1基因多态性与IMP含量之间存在较强的相关性，ampd1基因可作为BBTB高IMP含量遗传选择的候选标记。

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来源期刊

Aquaculture 农林科学-海洋与淡水生物学

CiteScore

8.60

自引率

17.80%

发文量

1246

审稿时长

56 days

期刊介绍： Aquaculture is an international journal for the exploration, improvement and management of all freshwater and marine food resources. It publishes novel and innovative research of world-wide interest on farming of aquatic organisms, which includes finfish, mollusks, crustaceans and aquatic plants for human consumption. Research on ornamentals is not a focus of the Journal. Aquaculture only publishes papers with a clear relevance to improving aquaculture practices or a potential application.