MIR396-GRF/GIF enhances in planta shoot regeneration of Dendrobium catenatum
Zhenyu Yang,
Meili Zhao,
Xiaojie Zhang,
Lili Gu,
Jian Li,
Feng Ming,
Meina Wang,
Zhicai Wang
Affiliations
Zhenyu Yang
Shenzhen Key Laboratory for Orchid Conservation and Utilization, the National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen
Meili Zhao
Shenzhen Key Laboratory for Orchid Conservation and Utilization, the National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen
Xiaojie Zhang
Shenzhen Key Laboratory for Orchid Conservation and Utilization, the National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen
Lili Gu
Xinjiang Key Laboratory of Grassland Resources and Ecology, College of Grassland Sciences, Xinjiang Agricultural University
Jian Li
Shenzhen Key Laboratory for Orchid Conservation and Utilization, the National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen
Feng Ming
Shanghai Key Laboratory of Plant Molecular Sciences, College of Life Sciences, Shanghai Normal University
Meina Wang
Shenzhen Key Laboratory for Orchid Conservation and Utilization, the National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen
Zhicai Wang
Shenzhen Key Laboratory for Orchid Conservation and Utilization, the National Orchid Conservation Center of China and the Orchid Conservation & Research Center of Shenzhen
Summary Recent studies on co-transformation of the growth regulator, TaGRF4-GIF1 chimera (Growth Regulating Factor 4-GRF Interacting Factor 1), in cultivated wheat varieties (Triticum aestivum), showed improved regeneration efficiency, marking a significant breakthrough. Here, a simple and reproducible protocol using the GRF4-GIF1 chimera was established and tested in the medicinal orchid Dendrobium catenatum, a monocot orchid species. TaGRF4-GIF1 from T. aestivum and DcGRF4-GIF1 from D. catenatum were reconstructed, with the chimeras significantly enhancing the regeneration efficiency of D. catenatum through in planta transformation. Further, mutating the microRNA396 (miR396) target sites in TaGRF4 and DcGRF4 improved regeneration efficiency. The target mimicry version of miR396 (MIM396) not only boosted shoot regeneration but also enhanced plant growth. Our methods revealed a powerful tool for the enhanced regeneration and genetic transformation of D. catenatum.
