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Screening of optimal reference genes for qRT-PCR and preliminary exploration of cold resistance mechanisms in Prunus mume and Prunus sibirica varieties

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Abstract

Prunus sibirica and Prunus mume are closely related plant species that differ in cold tolerance. Hybrids of P. sibirica and true mume, belonging to the apricot mei group, inherited strong cold resistance from P. sibirica. These materials are favourable for research on the molecular mechanisms of cold resistance. However, no suitable reference genes have been identified for analysing gene expression patterns between P. sibirica and P. mume. Ten candidate reference genes were assessed, namely, actins (ACT2-1, ACT2-2, ACT2-3, ACT2-4), protein phosphatase 2A-1 (PP2A-1), ubiquitins (UBQ2, UBQ3), ubiquitin extension protein (UBQ1) and tubulins (TUB1, TUB2), with four distinct algorithms (geNorm, NormFinder, BestKeeper and RefFinder). UBQ2 was recognized as the best reference gene in stems and buds across materials (P. sibirica; ‘Xiaohong Zhusha’, ‘Beijing Yudie’, and ‘Xiao Lve’ for true mume; and ‘Dan Fenghou’, ‘Fenghou’, and ‘Yanxing’ for apricot mei) under cold stress. In addition, the temporal and spatial expression patterns of PmCBF6 and PmLEA10 among seven varieties during winter periods were analysed using UBQ2 as a reference gene. The expression differed significantly among cultivars, which may contribute to their differences in cold tolerance. This paper confirmed the strong cold tolerance of apricot mei. And the best internal reference gene suitable for seven varieties was selected: UBQ2. Based on the above results, the expression of PmCBF6 and PmLEA10 genes during wintering in seven varieties was analysed. The molecular mechanisms of cold resistance were found to be possibly different in different varieties of P. sibirica and P. mume.

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Acknowledgements

This research was supported by the program for Science and Technology of Beijing (No. Z181100002418006) and Special Fund for Beijing Common Construction Project.

Funding

The program for Science and Technology of Beijing (No. Z181100002418006) and Special Fund for Beijing Common Construction Project.

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Author notes

  1. Anqi Ding and Fei Bao contributed equally to this work.

Authors and Affiliations

  1. Beijing Advanced Innovation Center for Tree Breeding By Molecular Design, Beijing Forestry University, No. 35 Tsinghua East Road, Haidian District, P.O. Box 155, Beijing, 100083, China

    Anqi Ding, Fei Bao, Tengxun Zhang & Qixiang Zhang

  2. Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding, Beijing Forestry University, Beijing, China

    Anqi Ding, Fei Bao, Tengxun Zhang, Weiru Yang, Jia Wang, Tangren Cheng & Qixiang Zhang

  3. National Engineering Research Center for Floriculture, Beijing Forestry University, Beijing, China

    Anqi Ding, Fei Bao, Tengxun Zhang, Weiru Yang, Jia Wang, Tangren Cheng & Qixiang Zhang

  4. Beijing Laboratory of Urban and Rural Ecological Environment, Beijing Forestry University, Beijing, China

    Anqi Ding, Fei Bao, Tengxun Zhang, Weiru Yang, Jia Wang, Tangren Cheng & Qixiang Zhang

  5. Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education, Beijing Forestry University, Beijing, China

    Anqi Ding, Fei Bao, Tengxun Zhang, Weiru Yang, Jia Wang, Tangren Cheng & Qixiang Zhang

  6. School of Landscape Architecture, Beijing Forestry University, Beijing, China

    Anqi Ding, Fei Bao, Tengxun Zhang, Weiru Yang & Qixiang Zhang

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  1. Anqi Ding
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Contributions

QZ supported resources. AD and FB conceived and designed research. AD conducted experiments, analyzed data and wrote the original draft. FB reviewed and edited the manuscript. TZ, WY, JW and TC supported resources. All authors have read and approved the final manuscript.

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Correspondence to Qixiang Zhang.

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Ding, A., Bao, F., Zhang, T. et al. Screening of optimal reference genes for qRT-PCR and preliminary exploration of cold resistance mechanisms in Prunus mume and Prunus sibirica varieties. Mol Biol Rep 47, 6635–6647 (2020). https://doi.org/10.1007/s11033-020-05714-x

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