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Misexpression Approaches for the Manipulation of Flower Development

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Flower Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2686))

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Abstract

The generation of dominant gain-of-function mutants through activation tagging is a forward genetic approach that can be applied to study the mechanisms of flower development, complementing the screening of loss-of-function mutants. In addition, the functions of genes of interest can be further analyzed through reverse genetics. A commonly used method is gene overexpression, where ectopic expression can result in an opposite phenotype to that caused by a loss-of-function mutation. When overexpression is detrimental, the misexpression of a gene using tissue-specific promoters can be useful to study spatial-specific function. As flower development is a multistep process, it can be advantageous to control gene expression, or its protein product activity, in a temporal and/or spatial manner. This has been made possible through several inducible promoter systems as well as inducible proteins by constructing chimeric fusions between the ligand-binding domain of the glucocorticoid receptor (GR) and the protein of interest. The recently introduced CRISPR-Cas9-based platform provides a new way of bioengineering transcriptional regulators in plants. By fusing a catalytically inactive dCas9 with functional activation or repression domains, the CRISPR-Cas9 module can achieve transcriptional activation or repression of endogenous genes. All these methods allow us to genetically manipulate gene expression during flower development. In this chapter, we describe methods to produce the expression constructs, method of screening, and more general applications of the techniques.

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Acknowledgments

This work was supported by Grant-in-Aid for Scientific Research (JP20H00470, JP21K19266, and JP22H05176) to T.I., and Startup Fund (680-804006) and the Jiangsu Specially-Appointed Professor Program to Y.X.

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Authors and Affiliations

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

    Yifeng Xu

  2. Republic Polytechnic, School of Applied Science (SAS), Singapore, Singapore

    Eng-Seng Gan

  3. Nara Institute of Science and Technology, Biological Sciences, Plant Stem Cell Regulation and Floral Patterning Laboratory, Ikoma, Nara, Japan

    Toshiro Ito

Authors
  1. Yifeng Xu
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  2. Eng-Seng Gan
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  3. Toshiro Ito
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Correspondence to Yifeng Xu or Toshiro Ito .

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Editors and Affiliations

  1. Centre for Research in Agricultural Genomics CSIC-IRTA-UAB-UB, Barcelona, Spain

    José Luis Riechmann

  2. Instituto de Biología Molecular y Celular de Plantas CSIC-UPV, Valencia, Spain

    Cristina Ferrándiz

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Xu, Y., Gan, ES., Ito, T. (2023). Misexpression Approaches for the Manipulation of Flower Development. In: Riechmann, J.L., Ferrándiz, C. (eds) Flower Development . Methods in Molecular Biology, vol 2686. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3299-4_21

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  • DOI: https://doi.org/10.1007/978-1-0716-3299-4_21

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  • Publisher Name: Humana, New York, NY

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