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Selection and validation of appropriate reference genes for RT-qPCR analysis of Nitraria sibirica under various abiotic stresses

文献类型： 外文期刊

作者： Hu, Aishuang 1 ; Yang, Xiuyan 1 ; Zhu, Jianfeng 1 ; Wang, Xiuping 3 ; Liu, Jiaxin 1 ; Wang, Jiping 1 ; Wu, Haiwen 1 ; Zhang, Huilong 1 ; Zhang, Huaxin 1 ;

作者机构： 1.Chinese Acad Forestry, Inst Ecol Protect & Restorat, Beijing 10091, Peoples R China

2.Chinese Acad Forestry Yellow River Delta, Comprehens Expt Ctr, Dongying 257000, Peoples R China

3.Hebei Acad Agr & Forestry Sci, Inst Coastal Agr, Tangshan 063299, Peoples R China

4.Hebei Saline Alkali Land Greening Technol Innovat, Tangshan 063299, Peoples R China

关键词： Nitraria sibirica Pall; geNorm; NormFinder; BestKeeper; Comparative Delta Ct; RefFinder; Salt stress

期刊名称：BMC PLANT BIOLOGY （ 影响因子：5.26； 五年影响因子：5.761 ）

ISSN： 1471-2229

年卷期： 2022 年 22 卷 1 期

页码：

收录情况： SCI

摘要： Background: Nitraria sibirica Pall. is a halophytic shrub with strong environmental adaptability that can survive in extremely saline-alkali and drought-impacted environments. Gene expression analysis aids in the exploration of the molecular mechanisms of plant responses to abiotic stresses. RT-qPCR is the most common technique for studying gene expression. Stable reference genes are a prerequisite for obtaining accurate target gene expression results in RT-qPCR analysis. Results: In this study, a total of 10 candidate reference genes were selected from the transcriptome of N. sibirica, and their expression stability in leaves and roots under different treatment conditions (salt, alkali, drought, cold, heat and ABA) was evaluated with the geNorm, NormFinder, BestKeeper, comparative delta Ct and RefFinder programs. The results showed that the expression stability of the candidate reference genes was dependent on the tissue and experimental conditions tested. ACT7 combined with R3H, GAPDH, TUB or His were the most stable reference genes in the salt- or alkali-treated leaves, salt-treated roots and drought-treated roots, respectively; R3H and GAPDH were the most suitable combination for drought-treated leaves, heat-treated root samples and ABA-treated leaves; DIM1 and His maintained stable expression in roots under alkali stress; and TUB combined with R3H was stable in ABA-treated roots. TBCB and GAPDH exhibited stable expression in heat-treated leaves; TBCB, R3H, and ERF3A were stable in cold-treated leaves; and the three most stable reference genes for cold-treated roots were TBCB, ACT11 and DIM1. The reliability of the selected reference genes was further confirmed by evaluating the expression patterns of the NsP5CS gene under the six treatment conditions. Conclusion: This study provides a theoretical reference for N. sibirica gene expression standardization and quantification under various abiotic stress conditions and will help to reveal the molecular mechanisms that confer stress tolerance to N. sibirica.

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