Induction of double flowers in Pharbitis nil using a class-C MADS-box transcription factor with Chimeric REpressor gene-Silencing Technology

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Author(s)

    • Ozeki Yuko OZEKI Yuko
    • Gene Research Center, Graduate School of Life and Environmental Sciences, University of Tsukuba
    • HIGUCHI Yohei
    • National Institute of Floricultural Science, National Agriculture and Food Research Organization
    • KAMADA Hiroshi
    • Gene Research Center, Graduate School of Life and Environmental Sciences, University of Tsukuba
    • MITSUDA Nobutaka
    • Research Institute of Genome-Based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST)
    • OHME TAKAGI Masaru
    • Research Institute of Genome-Based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST)
    • ONO Michiyuki
    • Gene Research Center, Graduate School of Life and Environmental Sciences, University of Tsukuba

Abstract

The Chimeric REpressor gene-Silencing Technology (CRES-T) system is a novel reverse genetic method using a chimeric transcriptional repressor fusing an EAR transcriptional repression domain called SRDX. We sought to change the flower shape of <i>Pharbitis nil</i>, a model ornamental flower, using an <i>Arabidopsis</i> transcription factor fused with SRDX. For the first trial modulating flower shape we transformed with the class-C MADS-box transcription factor AGAMOUS (AG) fused with SRDX (<i>AGSRDX</i>). Defects in class-C genes cause double flowers in <i>Arabidopsis</i> and <i>Pharbitis</i>. However, when <i>AGSRDX</i> was expressed under the CaMV 35S promoter (<i>p35S</i>), the transgenic <i>Pharbitis</i> bore a malformed flower with a protruding pistil. We then used <i>DUPLICATED</i> (<i>DP</i>), one of the class-C genes in <i>Pharbitis</i>. The <i>p35S::DPSRDX</i>-introduced callus were difficult to regenerate during transgenic steps, but occasionally made a perfect double flower bud showing severe growth defects. The flower buds never developed to flower opening stage. These results indicate that CRES-T is functional in <i>Pharbitis</i> but even using a conserved transcription factor, some species-specific variation might exist. To avoid these unwanted effects, we recruited inducible promoters to control expression of the chimeric transcription repressors in combination with the DNA-binding domain of GAL4 in yeast fused with SRDX and the GAL4 upstream activator sequence (UAS). Normal regeneration was observed by inducible repression of <i>DPSRDX</i> during <i>in vitro</i> redifferentiation, and the double-flowered <i>Pharbitis</i> was generated. We successfully induced a non-transformant (NT)-like flower in <i>DPSRDX</i>-expressing double-flowered transformants. Our approach will enable us to breed transgenic horticultural plants with inducible fertility.

Journal

  • Plant Biotechnology

    Plant Biotechnology 28(2), 153-165, 2011-03-25

    Japanese Society for Plant Cell and Molecular Biology

References:  59

Cited by:  4

Codes

  • NII Article ID (NAID)
    10028258964
  • NII NACSIS-CAT ID (NCID)
    AA11250821
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    13424580
  • NDL Article ID
    11023496
  • NDL Source Classification
    ZR3(科学技術--生物学--植物)
  • NDL Call No.
    Z54-J126
  • Data Source
    CJP  CJPref  NDL  J-STAGE 
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