Screening of optimal reference genes for qRT-PCR and preliminary exploration of cold resistance mechanisms in Prunus mum
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ORIGINAL ARTICLE
Screening of optimal reference genes for qRT‑PCR and preliminary exploration of cold resistance mechanisms in Prunus mume and Prunus sibirica varieties Anqi Ding1,2,3,4,5,6 · Fei Bao1,2,3,4,5,6 · Tengxun Zhang1,2,3,4,5,6 · Weiru Yang2,3,4,5,6 · Jia Wang2,3,4,5 · Tangren Cheng2,3,4,5 · Qixiang Zhang1,2,3,4,5,6 Received: 25 April 2020 / Accepted: 2 August 2020 © Springer Nature B.V. 2020
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. Keywords P. sibirica · P. mume · True mume group · Apricot mei group · Reference gene · Cold resistance Anqi Ding and Fei Bao contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11033-020-05714-x) contains supplementary material, which is available to authorized users.
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
2
Fei Bao [email protected]
Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding, Beijing Forestry University, Beijing, China
3
Tengxun Zhang [email protected]
National Engineering Research Center for Floriculture, Beijing Forestry University, Beijing, China
4
Weiru Yang [email protected]
Beijing Laboratory of Urban and Rural Ecological Environment, Beijing Forestry University, Beijin
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