Abstract
Poplar is a model organism for high in vitro regeneration in woody plants. We have chosen a hybrid poplar Populus davidiana Dode × Populus bollena Lauche. By optimizing the Murashige and Skoog medium with (0.3 mg/L) 6-benzylaminopurine and (0.08 mg/L) naphthaleneacetic acid, we have achieved the highest frequency (90%) for shoot regeneration from poplar leaves. It was also important to improve the transformation efficiency of poplar for genetic breeding and other applications. In this study, we found a significant improvement of the transformation frequency by controlling the leaf age. Transformation efficiency was enhanced by optimizing the Agrobacterium concentration (OD600 = 0.8–1.0) and an infection time (20–30 min). According to transmission electron microscopy observations, there were more Agrobacterium invasions in the 30-day-old leaf explants than in 60-day-old and 90-day-old explants. Using the green fluorescent protein (GFP) marker, the expression of MD–GFP fusion proteins in the leaf, shoot, and root of hybrid poplar P. davidiana Dode × P. bollena Lauche was visualized for confirmation of transgene integration. Southern and Northern blot analysis also showed the integration of T-DNA into the genome and gene expression of transgenic plants. Our results suggest that younger leaves had higher transformation efficiency (~30%) than older leaves (10%).
Highlights
Poplar has several exceptional qualities, such as a high capacity for vegetative propagation and a fast growth rate
Several factors were systematically analyzed to improve transformation efficiency, including poplar genotype, Agrobacterium tumefaciens strain for transformation, bacterial concentration, acetosyringone (AS) [20,29,30,31,32], and different explants including leaf discs and stems [23,33]
Leaf explants of a hybrid poplar Populus davidiana Dode × P. bollena Lauche were cultured in media with 0.1~0.5 mg/L BA and 0.01~0.1 mg/L naphthaleneacetic acid (NAA)
Summary
Poplar has several exceptional qualities, such as a high capacity for vegetative propagation and a fast growth rate. Parsons et al [19] first reported the genetic transformation of poplar This technology has been applied to various Populus species to improve their transformation efficiency [20,21,22,23,24,25,26,27,28]. Several factors were systematically analyzed to improve transformation efficiency, including poplar genotype, Agrobacterium tumefaciens strain for transformation, bacterial concentration, acetosyringone (AS) [20,29,30,31,32], and different explants including leaf discs and stems [23,33]. In order to improve the transformation efficiency in this work, we optimized the conditions of the leaf explants age, as well as Agrobacterium concentration
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
