浙江大学学报（农业与生命科学版）2025，Vol.51Issue(3)：402-413,12.DOI:10.3785/j.issn.1008-9209.2023.12.211
高温及抗生素处理下四纹豆象四龄幼虫内参基因的筛选
Screening of reference genes in Callosobruchus maculatus(Coleoptera:Bruchidae)fourth-instar larvae under high-temperature and antibiotic treatments
摘要
Abstract
To identify stable reference genes for Callosobruchus maculatus fourth-instar larvae under high-temperature and antibiotic treatments,four candidate reference genes(α-tubulin_1,α-tubulin_2,RpL39_1,and RpL39_2)were selected on the basis of transcriptomic data.The expression levels of these genes were measured by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR)in whole-body tissues of larvae exposed to high-temperature treatment(30℃control vs.37℃treatment)and in midgut tissues of larvae subjected to antibiotic treatments(10%acetic acid control vs.0.01%ciprofloxacin+0.01%tetracycline hydrochloride treatment)at 30℃and 37℃.Gene stability was evaluated by ΔCT approach,NormFinder,and GeNorm,with HSP70 as a target gene for validation.The results demonstrated that RpL39_1 exhibited the highest stability across all the treatment groups,while the combination of RpL39_1 and RpL39_2 also met the normalization requirements.HSP70 expression levels showed significant differences(P＜0.05)between the high-temperature-treated group and the control group after normalization to RpL39_1 or RpL39_2.Therefore,RpL39_1 is recommended as a single reference gene for gene expression analysis in C.maculatus larvae under high-temperature and antibiotic treatments.This study provides methodological insights into the gut microbiota-mediated regulation of heat tolerance-related genes in the host.关键词
四纹豆象/四龄幼虫/高温/抗生素/内参基因Key words
Callosobruchus maculatus/fourth-instar larvae/high temperature/antibiotic/reference gene分类
农业科技引用本文复制引用
耿加美,何云川,陈秋霖,高扬,周瀛,祝增荣..高温及抗生素处理下四纹豆象四龄幼虫内参基因的筛选[J].浙江大学学报（农业与生命科学版）,2025,51(3):402-413,12.基金项目
浙江省"三农六方"科技协作计划项目(CTZB-F190625LWZ-SNY1) （CTZB-F190625LWZ-SNY1）
海南省重点研发计划项目(ZDYF2021XDNY194) （ZDYF2021XDNY194）
海南省自然科学基金项目(323RC521) （323RC521）
海南省三亚市科技创新专项(2022KJCX19) （2022KJCX19）
海南省三亚崖州湾科技城海南专项博士研究生科学研究基金项目(HSPHDSRF-2022-04-006). （HSPHDSRF-2022-04-006）
