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<p><i>Corydalis saxicola</i> Bunting is a rare perennial species in the <i>Corydalis</i> genus of the Papaveraceae family. It has been traditionally used for treating liver diseases for over a millennium. Research on molecular biology of <i>C. saxicola</i> is just beginning, and there is an urgent need to establish a mature genetic transformation system for conducting functional genomics and germplasm innovation. In this study, we established an <i>Rhizobium rhizogenes</i>-mediated transformation protocol for <i>C. saxicola</i>, exhibiting high efficiency. After screening <i>R. rhizogenes</i> strains, optimizing the root induction medium, evaluating various explants, and standardizing infection time and co-cultivation duration, we found that infecting adventitious buds with the AR1193 strain at an OD<sub>600</sub> of 0.6 for 15&#xa0;min, followed by three days of co-cultivation and sub-culturing on root induction medium supplemented with 1-Naphthaleneacetic acid (NAA), resulted in highly efficient induction of hairy roots containing the reporter gene <i>DsRed</i>. The transformation efficiency reached up to 52.63% in our experiments. This protocol serves as a valuable tool for molecular biology investigations, particularly in the research of secondary metabolic pathways, and will for sure accelerate the development of CRISPR/Cas9-based genome editing in <i>C. saxicola</i>.</p>

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Developing an efficient protocol for hairy root transformation of Corydalis saxicola

  • 期刊论文

  • 发表日期：

    2025年1月23日

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  • Yanlin Pan,
  • Yibin Zhang,
  • Hongbo Qin,
  • Xingui Wang,
  • Lunfa Guo

摘要

Corydalis saxicola Bunting is a rare perennial species in the Corydalis genus of the Papaveraceae family. It has been traditionally used for treating liver diseases for over a millennium. Research on molecular biology of C. saxicola is just beginning, and there is an urgent need to establish a mature genetic transformation system for conducting functional genomics and germplasm innovation. In this study, we established an Rhizobium rhizogenes-mediated transformation protocol for C. saxicola, exhibiting high efficiency. After screening R. rhizogenes strains, optimizing the root induction medium, evaluating various explants, and standardizing infection time and co-cultivation duration, we found that infecting adventitious buds with the AR1193 strain at an OD600 of 0.6 for 15 min, followed by three days of co-cultivation and sub-culturing on root induction medium supplemented with 1-Naphthaleneacetic acid (NAA), resulted in highly efficient induction of hairy roots containing the reporter gene DsRed. The transformation efficiency reached up to 52.63% in our experiments. This protocol serves as a valuable tool for molecular biology investigations, particularly in the research of secondary metabolic pathways, and will for sure accelerate the development of CRISPR/Cas9-based genome editing in C. saxicola.