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Genetic transformation of a hepatoprotective plant, Phyllanthus amarus

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Abstract

Phyllanthus amarus Schum & Thonn. is a source of various pharmacologically active compounds such as phyllanthin, hypophyllanthin, gallic acid, catechin, and nirurin, a flavone glycoside. A genetic transformation method using Agrobacterium tumefaciens was developed for this plant species for the first time. Shoot tips of full grown plants were used as explants for Agrobacterium-mediated transformation. Transgenic plants were obtained by co-cultivation of shoot tips explants and A. tumefaciens strain LBA4404 containing the pCAMBIA 2301 plasmid harboring neomycin phosphotransferase II (NPT II) and β-glucuronidase encoding (GUS) genes in the T-DNA region in the presence of 200 μM acetosyringone. Integration of the NPT II gene into the genome of transgenic plants was verified by PCR and Southern blot analyses. Expression of the NPT II gene was confirmed by RT-PCR analysis. An average of 25 explants was used, out of which an average of 19 explants produced kanamycin-resistant shoots, which rooted to produce 13 complete transgenic plants.

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Acknowledgments

This work received financial support from Council of Scientific and Industrial Research and Department of Biotechnology, Government of India. We express our gratitude to the Director of IICB for his constant support and encouragement. A.B. is thankful to CSIR, New Delhi for providing Junior Research Fellowship. We are grateful to Prof. Asis Datta, Director, NCPGR, New Delhi for providing Agrobacterium tumefaciens strain LBA 4404 and Dr. D. Sengupta, Bose Institute, Kolkata for the vector pCAMBIA 2301.

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  1. Drug Development, Diagnostics and Biotechnology Division, Indian Institute of Chemical Biology, 4 Raja S. C. Mullick Road, Kolkata, 700 032, India

    Anindita Banerjee & Sharmila Chattopadhyay

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  1. Anindita Banerjee
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  2. Sharmila Chattopadhyay
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Correspondence to Sharmila Chattopadhyay.

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Editor: Gregory C. Phillips

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Banerjee, A., Chattopadhyay, S. Genetic transformation of a hepatoprotective plant, Phyllanthus amarus . In Vitro Cell.Dev.Biol.-Plant 45, 57–64 (2009). https://doi.org/10.1007/s11627-008-9160-z

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