Stepwise Optimization of the RT-qPCR Protocol and the Evaluation of Housekeeping Genes in Pears (Pyrus bretschneideri) under Various Hormone Treatments and Stresses

文献类型： 外文期刊

第一作者： Zhou, Peng

作者： Zhou, Peng;Huang, Linlin;Feng, Xinxin;Li, Liulin;Wang, Yingtao;Li, Xiao

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关键词： housekeeping gene; RT-qPCR; hormone; biotic and abiotic stress; pear

期刊名称：HORTICULTURAE （ 影响因子：3.1； 五年影响因子：3.4 ）

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年卷期： 2023 年 9 卷 2 期

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收录情况： SCI

摘要： Real-time quantitative PCR (RT-qPCR) has become a widely used method for exploring plant gene expression level. The method requires using some stably expressed genes as a reference to accurately normalize the RT-qPCR data. However, under various stresses and hormone treatments, the levels of most reference genes vary. Environmental variations also influence their expression levels. The lack of validated, stably expressed reference genes can mislead the study of gene function in pears. "Huangguan" pears have recently become the focus of research on stress resistance mechanisms, such as high resistance. Therefore, the aim of this study was to select the optimal reference genes in Huangguan pears, and we analyzed the expression of the genes EF1 alpha, ACT, SKD1, YLS8, UBQ, GAPDH, TUB, and WDP in a series of pear leaf sets under various stresses and hormone treatments. Using different statistical algorithms, we found that under various treatments, the WDP gene had more stable expression, ACT was the most stable under MeJA treatment, YLS8 was the most valuable reference gene under ABA hormone and heat stress conditions, and GAPDH showed worst results compared to other housekeeping genes, except under heat stress. These results will supply valuable and updated information for the selection of housekeeping genes in pears under biotic and abiotic stresses in the future.

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