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Genetic Modification of Lignin Biosynthetic Pathway in Populus ciliata Wall. via Agrobacterium-Mediated Antisense CAD Gene Transfer for Quality Paper Production

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Abstract

The present studies describe the development of transgenic Himalayan poplar (Populus ciliata Wall.) plants via Agrobacterium-mediated antisense cinnamyl alcohol dehydrogenase (CAD) gene transfer for genetic modification of lignin biosynthetic pathway for lowering down the lignin content by post-transcriptional silencing of CAD gene. Agrobacterium tumefaciens strain C58C1rifR(pMP90) harboring CAD gene in antisense orientation and npt-II gene in a binary vector p35SASCAD was used for genetic transformation experiments. By combining the best treatments of 48 h pre-culturing and 48 h co-cultivation time periods, a transformation frequency of 14 % was obtained from petiole explant of P. ciliata. Out of total 9 randomly selected putative transgenic plants of P. ciliata, five were found to show the presence and integration of npt-II gene in poplar genome as confirmed by PCR using npt-II gene specific primers. npt-II gene which is transferred along with antisense CAD gene (in the same gene construct), got amplified with gene specific primers at 0.7 Kb. The lignin content of transgenic plants of P. ciliata was found to be reduced by a fraction of 3–4 % as compared to the control plants after 4 months of transfer to the greenhouse conditions.

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Acknowledgments

The authors are thankful to Dr. W. Boerjan, Plant Biology System, Gent University, Belgium for kindly providing the Agrobacterium tumefaciens strain containing antisense CAD gene construct for carrying out genetic transformation experiments.

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  1. A. K. Thakur

    Present address: Directorate of Rapeseed-Mustard Research (ICAR), Bharatpur, 321 303, Rajasthan, India

Authors and Affiliations

  1. Department of Biotechnology, Dr. Yashwant Singh Parmar University of Horticulture and Forestry, Solan, 173 230, Himachal Pradesh, India

    A. K. Thakur, G. Aggarwal & D. K. Srivastava

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  1. A. K. Thakur
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  2. G. Aggarwal
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  3. D. K. Srivastava
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Correspondence to A. K. Thakur.

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Thakur, A.K., Aggarwal, G. & Srivastava, D.K. Genetic Modification of Lignin Biosynthetic Pathway in Populus ciliata Wall. via Agrobacterium-Mediated Antisense CAD Gene Transfer for Quality Paper Production. Natl. Acad. Sci. Lett. 35, 79–84 (2012). https://doi.org/10.1007/s40009-012-0018-x

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