Abstract
A high-frequency and simple procedure for Agrobacterium tumefaciens-mediated genetic transformation of the medicinal plant Salvia miltiorrhiza was developed. Leaf discs were pre-cultured on MS medium supplemented with 6.6 μmol l−1 BAP and 0.5 μmol l−1 NAA for one day, then co-cultured with A. tumefaciens strain EHA105 harboring the plasmid pCAMBIA 2301 for three days on the same medium. Regenerated buds were obtained on selection medium (co-culture medium supplemented with 60 mg l−1 kanamycin and 200 mg l−1 cefotaxime) after two cycles’ culture of 10 days each and then transferred to fresh MS medium with 60 mg l−1 kanamycin for rooting. Fifteen days later, the rooted plantlets were obtained and then successfully transplanted to soil. The transgenic nature of the regenerated plants was confirmed by PCR, Southern hybridization analysis and GUS histochemical assay. Averagely, 1.1 independent verified transgenics per explant plated were obtained through this protocol. Adopting this procedure, positive transformed plants could be obtained within 2–3 months from mature seeds germination to transplant to soil, and more than 1,000 transgenic plants with several engineered constructs encoding different genes of interest were produced in our lab in the past two years.
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Acknowledgments
We thank Drs. Zhongmin Dong, Niu Dong, and all our colleagues in the laboratory for constructive discussions and technical support. This project was supported by the 10th “five-year-technique-project” of the Ministry of Technique and Science, P.R. China.
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Yan, Yp., Wang, Zz. Genetic transformation of the medicinal plant Salvia miltiorrhiza by Agrobacterium tumefaciens-mediated method. Plant Cell Tiss Organ Cult 88, 175–184 (2007). https://doi.org/10.1007/s11240-006-9187-y
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DOI: https://doi.org/10.1007/s11240-006-9187-y