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Development of an efficient in-planta Agrobacterium-mediated transformation method for Iranian purslane (Portulaca oleracea L.) using sonication and vacuum infiltration

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Abstract

Purslane (Portulaca oleracea L.), a valuable medicinal herb, is used as a source of pharmaceutical components such as flavonoids, alkaloids, fatty acids, terpenoids, and sterols. Regeneration of transgenic purslane plantlets from transformed cells is a time-consuming procedure and needs hard work. In this study, in-planta transformation of purslane, using sonication and vacuum infiltration, is reported. The purslane seeds were infected through Agrobacterium tumefaciens strain LBA4404 having the uidA gene on pBI121 vector. Effective selection of transformants was performed by supplementing MS media with 250 mg l−1 kanamycin. Several factors affecting the in-planta procedure including pre-culture duration, acetosyringone dose, sonication, and vacuum infiltration duration were investigated. The results demonstrated that the highest number of GUS-positive purslane plantlets was obtained when the pre-cultured seeds were sonicated and vacuum-infiltered for 2 min in agro-bacterial cell suspension, and then co-cultivated in MS media having 100 µM acetosyringone. The integration and expression of uidA gene in transgenic purslane was successfully corroborated by southern blot and GUS histochemical analyses.

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Acknowledgements

This work was supported by the Iranian National Science Foundation [grant number: 90003004].

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Authors and Affiliations

  1. Department of Biotechnology, Faculty of Agriculture and Natural Resources, Imam Khomeini International University, Qazvin, Iran

    Behnam Sedaghati & Raheem Haddad

  2. Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Behnam Sedaghati & Mojgan Bandehpour

  3. Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Mojgan Bandehpour

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  1. Behnam Sedaghati
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  2. Raheem Haddad
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  3. Mojgan Bandehpour
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Correspondence to Behnam Sedaghati or Raheem Haddad.

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Sedaghati, B., Haddad, R. & Bandehpour, M. Development of an efficient in-planta Agrobacterium-mediated transformation method for Iranian purslane (Portulaca oleracea L.) using sonication and vacuum infiltration. Acta Physiol Plant 43, 17 (2021). https://doi.org/10.1007/s11738-020-03185-y

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