非生物胁迫下苗期白羊草荧光定量PCR内参基因的筛选及验证

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中图分类号：S543.9 文献标识码：A 文章编号：1007-0435（2025）11-3559-12

引用格式：，等.非生物胁迫下苗期白羊草荧光定量PCR内参基因的筛选及验证[J].草地学报，2025， 33（11） ：3559—3570 GAO Shou-yu， XIANG Qing-yuan，MA Jing-ru，et al.Selection and Validation of Reference Genes for qRT-PCR in Bothriochloa ischaemum seedlingsunderAbiotic Stresses[J].Acta Agrestia Sinica，2O25，33（11）：3559-3570

Selection and Validationof Reference Genes for qRT -PCRinBothriochloa ischaemum seedlings under Abiotic Stresses

GAO Shou-yu，XIANG Qing-yuan，MA Jing-ru，LI Yu-ying*，XIA Fang-shan （College of Grassand Science，Shanxi Agricultural University，Taigu，ShanxiProvince O3o8Ol，China）

Abstract：Due to its high sensitivity，strong specificity，and simple operation，real-time fluorescence quantita tive PCR（qRT-PCR） is an important tool for gene expression analysis.In order to identify suitable reference genes for qRT-PCR in Bothriochloa ischaemum under abiotic stresses，this study selected seven candidate reference genes（18S，GAPDH，PAL， EF -la，ACT，UBQ and TUB） based on previous transcriptome data. The expresson levels （Ct values）of these seven candidates in leaves and roots were determined under salt， drought，and cold stresses using qRT-PCR. Their stability was then evaluated using the comparative ΔCt （2号 method and four software programs （geNorm，NormFinder，BestKeeper and RefFinder）.The results showed that the optimal reference genes varied with diffrent stresses and tissues.Underdrought stress，the best reference genes were ACT and PAL in leaves，and 18S and GAPDH in roots. Under salt stress， PAL and EF -la weremost stableinleaves，whereas TUB and UBQ were most stable in roots.Under cold stress， ACT and GAPDH were the top candidates in leaves，while PAL and ACT were the best in roots. Finally，the expression of the salt tolerance-related gene BiADH28 was validated using the selected reference genes，confirming the reliability of the screening results. This study provides a solid foundation for gene expression analysis of B ischaemumunderabiotic stresses. KeyWords：Bothriochloa ischaemum;Reference genes;Quantitative real-time PCR（qRT-PCR）;Abiotic stress

荧光定量PCR技术因其高灵敏度、强特异性、操作简洁等优点，已成为基因表达分析的重要工具[1-2]。（剩余17912字）