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Advances in the Transformation of Cyclamen persicum Mill. Through Direct Regeneration Based on an Optimized Kanamycin Selection Scheme

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Abstract

Gene transfer technology has great value in ornamental plants toward the generation of varieties with new ornate characteristics. In the previous studies through the transformation of cyclamen, hygromycin was mainly used as a selective marker. However, there have been some drawbacks associated with hygromycin usage as a selecting agent. Therefore, in the current study, the optimization of kanamycin concentration in the regeneration media has been considered. Subsequently, the plant transformation using three different in vitro explants from three Cyclamen persicum cultivars using three Agrobacterium tumefaciens strains has been examined. Accordingly, the optimal kanamycin concentrations for regeneration from root and leaf explants were determined as 10 mg/L and for microtuber explants as 30 mg/L. The successful gene transformation in the antibiotic-resistant shoots were examined by PCR and UV-equipped microscopes. The gfp reporter gene transfer resulted in the highest efficiency of transformation (60%) to date, from the leaf explants of cv. Pure White inoculated with Agrobacterium tumefaciens strain LBA4404. In contrast, the lowest gene transfer efficiency (25%) was observed in root explants of cv. Dark Violet and cv. Neon Pink inoculated with strains GV3101 and AGL-1, respectively. The results of the current project are expandable to the subsequent investigations of Cyclamen persicum transformation.

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Data Availability

The authors declare that the major data supporting the results of the current study are available within the article. The other data are available on request from the corresponding author.

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Acknowledgements

The authors acknowledge Ms. Monireh Shahabi for technical support during in vitro propagation of the cyclamen explants. Additionally, the authors thank Ms. Maryam Ebrahimi for support in some data analysis and initial drafting.

Funding

The current study was financially supported as a Master’s thesis by the faculty of agriculture at Tarbiat Modares University (TMU) (Approved Proposal No.: 78186).

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Authors and Affiliations

  1. Department of Biotechnology, Faculty of Agriculture, Tarbiat Modares University, 1497713111, Tehran, Iran

    Mohammad Hossein Mirzaee & Masoumeh Emadpour

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  1. Mohammad Hossein Mirzaee
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  2. Masoumeh Emadpour
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Contributions

MHM contributed to formal analysis, resources, data curation, writing—original draft, and visualization; ME contributed to conceptualization, methodology, validation, writing—original draft, writing—review & editing, and supervision.

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Correspondence to Masoumeh Emadpour.

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Mirzaee, M.H., Emadpour, M. Advances in the Transformation of Cyclamen persicum Mill. Through Direct Regeneration Based on an Optimized Kanamycin Selection Scheme. Mol Biotechnol 66, 311–320 (2024). https://doi.org/10.1007/s12033-023-00758-3

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