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Int. J. Mol. Sci. 2014, 15(6), 10780-10793; doi:10.3390/ijms150610780

An Efficient Agrobacterium-Mediated Transformation System for Poplar

1
The Cooperative Innovation Center of Southern Modern Forestry, Nanjing Forestry University, Nanjing 210037, China
2
Department of Genomics, Agricultural Biotechnology Research Institute, Central Region of Iran (ABRICI), Najafabad Road, Isfahan 85135-487, Iran
*
Author to whom correspondence should be addressed.
Received: 1 March 2014 / Revised: 3 June 2014 / Accepted: 4 June 2014 / Published: 13 June 2014
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Abstract

Poplar is a model system for the regeneration and genetic transformation of woody plants. To shorten the time required for studies of transgenic poplar, efforts have been made to optimize transformation methods that use Agrobacterium tumefaciens. In this study, an Agrobacterium infective suspension was treated at 4 °C for at least 10 h before infecting explants. By transforming the Populus hybrid clone “Nanlin895” (Populus deltoides × P. euramericana) with Agrobacterium harboring the PBI121:CarNAC6 binary vector, we showed that the transformation efficiency was improved significantly by multiple independent factors, including an Agrobacterium infective suspension with an OD600 of 0.7, an Agrobacterium infection for 120 min, an Agrobacterium infective suspension at a pH of 5.0, an acetosyringone concentration of 200 µM, a cocultivation at 28 °C, a cocultivation for 72 h and a sucrose concentration of 30 g/L in the cocultivation medium. We also showed that preculture of wounded leaf explants for two days increased the regeneration rate. The integration of the desired gene into transgenic poplars was detected using selective medium containing kanamycin, followed by southern blot analysis. The expression of the transgene in the transgenic lines was confirmed by northern blot analysis.
Keywords: transformation system; Populus; Agrobacterium-mediated; efficient transformation system; Populus; Agrobacterium-mediated; efficient
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This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Movahedi, A.; Zhang, J.; Amirian, R.; Zhuge, Q. An Efficient Agrobacterium-Mediated Transformation System for Poplar. Int. J. Mol. Sci. 2014, 15, 10780-10793.

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