水滴伪康纤虫交替氧化酶基因(<em>AOX3</em>)的克隆及其基因家族鉴定与表达分析

doi:10.3969/j.issn.1674-7968.2026.05.015

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Abstract Pseudocohnilembus persalinus is a pathogenic scuticociliate parasite in mariculture fish species. In this study, the alternative oxidase (AOX) gene from Pseudocohnilembus persalinus was cloned. The AOX gene family was systematically identified at the genomic level, its biological functions and expression characteristics were predicted and analyzed. The full-length cDNA of the alternative oxidase 3 (AOX3) gene from P. persalinus was cloned using the rapid amplification of cDNA ends (RACE) technique. The AOXs gene family members in P. persalinus was systematically identified by screening the genome using the conserved Pfam domain PF01786. Bioinformatics approaches were applied to analyze the physicochemical properties, subcellular localization, gene structure, Scaffold localization, promoter cis-regulatory elements, and phylogenetic relationships of these genes. Expression analysis of P. persalinus AOXs gene family members were carried out by qRT-PCR during the trophic and cyst stages and under temperature stress. The result showed that the full-length cDNA of the AOX3 gene from P. persalinus was cloned, with a total length of 1 091 bp (GenBank No. PV953026), including a 47 bp 5'-UTR, a 150 bp 3'-UTR, and an ORF of 894 bp. Four AOXs gene family members (designated as PpAOX1~PpAOX4) were identified in the whole genome; All PpAOXs were predicted to be hydrophilic proteins without signal peptides and were primarily localized in the cytoplasm and mitochondria. Cis-acting regulatory element analysis of the promoter regions suggested that PpAOXs should be involved in responses to hypoxia, temperature stress, and defense-related stimuli. Phylogenetic analysis of 33 AOXs genes from 13 protist species revealed that the 4 AOXs members of P. persalinus clustered within a clade of Oligohymenophorean ciliates. qRT-PCR results showed that PpAOX1 was highly expressed during the trophic stage, whereas PpAOX2~PpAOX4 exhibited significant upregulation during the cyst stage. The PpAOXs gene family members displayed distinct expression patterns in response to temperature stress. This study provides critical insights for further investigating the biological functions of PpAOXs and deciphering the adaptive strategies of P. persalinus in response to environmental stress.

Key words： Pseudocohnilembus persalinus Alternative oxidase (AOX) Gene cloning Gene family Expression analysis

Received: 13 August 2025

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S943

Corresponding Authors: ^*zhbio@fjnu.edu.cn; lnfeng@hotmail.com

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	LIN Tong
	ZENG Hong
	PAN Ying
	LYU Yan-Hong
	LIN Neng-Feng

Cite this article:

LIN Tong,ZENG Hong,PAN Ying, et al. Cloning of the Alternative Oxidase Gene (AOX3) from Pseudocohnilembus persalinus and Identification and Expression Analysis of the AOX Gene Family[J]. 农业生物技术学报, 2026, 34(5): 1104-1117.

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https://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2026.05.015 OR https://journal05.magtech.org.cn/Jwk_ny/EN/Y2026/V34/I5/1104

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