cDNA yeast library construction of Dendrobium officinale Kimura et Migo seeds and screening and analysis of DELLA interacting proteins

Gibberellins (GAs) regulate diverse growth processes throughout the plant life cycle. DELLA proteins function as key repressors in the GA signaling pathway, inhibiting plant growth and development. The high expression of DELLA family genes in Dendrobium officinale Kimura et Migo seeds may be related to their poor natural germination. To identify interacting proteins of DoDELLA and elucidate its role in seed germination, a cDNA library was constructed from D. officinale seeds using Gateway recombination technology. A recombinant bait vector for DoDELLA4 was generated using homologous recombination and used to screen interacting proteins from the cDNA library. The constructed library exhibited a capacity of 1.20×10⁷ CFU, with an average insert size exceeding 1 000 bp and a recombination efficiency of 100%. The recombinant bait vector pGBKT7-DoDELLA4 effectively suppressed self-activation in SD/-Trp/-Leu/-His/-Ade medium supplemented with 20 mmol/L 3-AT. Screening identified 70 potential interacting proteins, and yeast two-hybrid assays confirmed interactions between DoDELLA4 and DoELIP1, 5807, DoTZF, DoUBQ, and DoGRPA3. These findings establish a foundation for further investigation into the molecular mechanisms by which DoDELLA regulates seed germination in D. officinale.