Efficient transient gene expression system using buckwheat hypocotyl protoplasts for large-scale experiments

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  • Sakamoto Shingo
    Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
  • Matsui Katsuhiro
    Institute of Crop Science, National Agriculture and Food Research Organization (NARO)
  • Oshima Yoshimi
    Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
  • Mitsuda Nobutaka
    Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)

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<p>Buckwheat (Fagopyrum esculentum) is cultivated worldwide and its flour is used in a variety of food products. Although functional analyses of genes in buckwheat are highly desired, reliable methods to do it have yet to be developed. In this study we established a simple and efficient transient gene expression system using buckwheat protoplasts isolated from young hypocotyls using 96-well plates as a high-throughput platform. The transformation efficiency was comparable with that of similar systems, such as Arabidopsis mesophyll protoplasts. Stable results were obtained in a typical example of the experiment to examine transcription factor activity. This system shows potential for the large-scale analysis of gene function using protoplast isolated from fewer and younger plants than the conventional system and may provide novel information for efficient buckwheat breeding.</p>

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