Isolation and characterization of maize ZmPP2C26 gene promoter in drought-response
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RESEARCH ARTICLE
Isolation and characterization of maize ZmPP2C26 gene promoter in drought-response Fengzhong Lu1 • Kexin Wang1 • Lamei Yan1 • Yalin Peng1 • Jingtao Qu1 Jing Wu1 • Yang Cao1 • Qingqing Yang1 • Fengling Fu1 • Haoqiang Yu1
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Received: 27 July 2020 / Revised: 20 October 2020 / Accepted: 4 November 2020 / Published online: 18 November 2020 Ó Prof. H.S. Srivastava Foundation for Science and Society 2020
Abstract The clade A members of serine/threonine protein phosphatase 2Cs (PP2Cs) play crucial roles in plant growth, development, and stress response via the ABA signaling pathway. But little is known about other PP2C clades in plants. Our previous study showed that maize the ZmPP2C26, a clade B member of ZmPP2Cs, negatively regulated drought tolerance in transgenic Arabidopsis. However, the upstream regulatory mechanism of ZmPP2C26 remains unclear. In the present study, the expression of ZmPP2C26 gene in maize was analyzed by quantitative real time PCR (qRT-PCR). The results showed that the expression of ZmPP2C26 in shoot and root was both significantly inhibited by drought stress. Subsequently, a 2175 bp promoter of ZmPP2C26 was isolated from maize genome (P2175). To validate whether the promoter possess some key cis-element and negatively drive ZmPP2C26 expression in drought stress, three 5-deletion fragments of 1505, 1084 and 215 bp was amplified from P2175 and were fused to b-glucuronidase (GUS) and luciferase gene (LUC) to produce promoter::GUS and promoter::LUC constructs, and transformed into tobacco, respectively. Transient expression assays indicated that all promoters could drive GUS and LUC expression. The GUS and LUC activity were both significantly inhibited by PEGFengling Fu and Haoqiang Yu have contributed equally to this work. & Fengling Fu [email protected] & Haoqiang Yu [email protected] 1
Key Laboratory of Biology and Genetic Improvement of Maize in Southwest Region, Ministry of Agriculture, Maize Research Institute, Sichuan Agricultural University, Chengdu 611130, People’s Republic of China
6000 treatment. Notably, the - 1084 to - 215 bp promoter possess one MBS element and inhibits the expression of GUS and LUC under drought stress. Meanwhile, we found that the 215 bp length is enough to drive ZmPP2C26 expression. These findings will provide insights into understanding the transcription-regulatory mechanism of ZmPP2C26 negatively regulating drought tolerance. Keywords Maize Serine/threonine protein phosphatase 2C Promoter Drought stress
Introduction Protein phosphorylation and dephosphorylation is a dynamic balance process and widespread in prokaryotes and eukaryotes. Plants can select phosphorylation or dephosphorylation of substrate molecules to realize the cascade of signals and enable plants to respond to any stimuli (Luan et al. 2003; Ma et al. 2009; Schweighofer et al. 2004). The protein phosphatases (PPs) catalyze the dephosphorylation and play crucial roles in plant growth and development. According to the difference of homology and substrate specificity, PPs ar
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