Abstract
Genetic manipulation of gymnosperms using Agrobacterium—mediated system, a powerful field in forest biotechnology, has a potential to accelerate forest tree improvements with desired attributes. However, most conifers such as pine have been considered recalcitrant to genetic transformation. The present study reports the successful production of transgenic Pinus massoniana from zygotic embryos using Agrobacterium tumefaciens harboring the pBI121:CslA2 binary vector. Various independent parameters were tested for their effects on transformation efficiency in P. massoniana. According to our results, combination of Agrobacterium density at OD600 of 0.5, cold treatment of Agrobacterium suspension at 4 °C for approximately 5 h, inoculation period of 5 h, and the addition of 100 µM acetosyringone in co-cultivation medium significantly enhanced the transformation efficiency. The stable integration of CslA2 gene into the genome of putative transgenic plants was confirmed by polymerase chain reaction and Southern blot hybridization, and the expression levels were determined using quantitative reverse transcription-PCR. Our optimized transformation procedure could provide an opportunity for transferring economically important genes into P. massoniana and other conifer species as well.
Published Version
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