Agrobacterium-Mediated Stable Genetic Transformation of Populus angustifolia and Populus balsamifera

  • Maheshwari P
  • Kovalchuk I
N/ACitations
Citations of this article
16Readers
Mendeley users who have this article in their library.

Abstract

The present study demonstrates Agrobacterium tumefaciens-mediated stable genetic transformation of two species of Poplar - Populus angustifolia and Populus balsamifera. The binary vector pCAMBIA-Npro-long-Luc containing the luciferase reporter gene was used to transform stem internode and axillary bud explants. Putative transformants were regenerated on selection-free medium using our previously established in vitro regeneration method. Explant type, genotype, effect of pre-culture, Agrobacterium concentration, a time period of infection and varying periods of co-culture with bacteria were tested for the transformation frequency. The highest frequency of transformation was obtained with stem internode explants pre-cultured for 2 days, infected with Agrobacterium culture at the concentration of OD600=0.5 for 10 min and co-cultivated with Agrobacterium for 48 h. Out of the two genotypes tested, P. balsamifera exhibited a higher transformation rate in comparison to P. angustifolia. The primary transformants that exhibited luciferase activity in a bioluminescence assay under the CCD camera when subjected to PCR and southern blot analysis revealed a stable single-copy integration of luc in their genomes. The reported protocol is highly reproducible and can be applied to other species of poplar; it will also be useful for future genetic engineering of one of the most important families of woody plants for sustainable development.

Cite

CITATION STYLE

APA

Maheshwari, P., & Kovalchuk, I. (2016). Agrobacterium-Mediated Stable Genetic Transformation of Populus angustifolia and Populus balsamifera. Frontiers in Plant Science, 7. https://doi.org/10.3389/fpls.2016.00296

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free