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PEG-mediated transient gene expression and silencing system in maize mesophyll protoplasts: a valuable tool for signal transduction study in maize

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Abstract

Polyethylene glycol (PEG)-mediated transient gene expression and silencing in protoplasts is widely applied in model plants such as Arabidopsis thaliana and rice. Here, we developed an efficient transient gene expression system based on the PEG-mediated method both in etiolated and green maize mesophyll protoplasts. The results showed that both yellow fluorescent protein encoding gene and glucuronidase encoding gene were efficiently expressed in maize protoplasts. More importantly, double-stranded RNAs (dsRNAs) can also be transfected into maize protoplasts by the PEG-mediated method to specifically silence exogenous and endogenous genes. Our results showed that dsRNA can be used to knockdown both exogenous and endogenous gene expression. Furthermore, bimolecular fluorescence complementation system for the detection of protein–protein interactions in maize protoplasts was developed. We also overexpressed and knockdowned the mitogen-activated protein kinase encoding gene ZmMPK5 to investigate the role of ZmMPK5 in abscisic acid (ABA)-induced antioxidant defense in maize protoplasts. This method here we reported will be valuable for signal transduction study in maize.

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Acknowledgments

This work was supported by the National Basic Research Program of China (Grant No. 2012CB114300), the National Natural Science Foundation of China (Grant Nos. 30970238, 31070254, and 31271631), the Fundamental Research Funds for the Central Universities (Grant No. KYZ201157), the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, and the Research Fund for the Doctoral Program of Higher Education of China (Grant No. 20090097110017).

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Authors and Affiliations

  1. College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, China

    Jianmei Cao, Dongmei Yao, Fan Lin & Mingyi Jiang

  2. National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, 210095, China

    Jianmei Cao, Dongmei Yao & Mingyi Jiang

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  1. Jianmei Cao
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  2. Dongmei Yao
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  3. Fan Lin
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  4. Mingyi Jiang
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Correspondence to Mingyi Jiang.

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Communicated by P. Sowinski.

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Cao, J., Yao, D., Lin, F. et al. PEG-mediated transient gene expression and silencing system in maize mesophyll protoplasts: a valuable tool for signal transduction study in maize. Acta Physiol Plant 36, 1271–1281 (2014). https://doi.org/10.1007/s11738-014-1508-x

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  • DOI: https://doi.org/10.1007/s11738-014-1508-x

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