ARS | Publication request: Biolistic transformation of cotton embryogenic cell suspension cultures

Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: August 14, 2012
Publication Date: January 7, 2013
Citation: Rajasekaran, K. 2013. Biolistic transformation of cotton embryogenic cell suspension cultures. In: Zhang, B-H. (editor). Transgenic Cotton: Methods and Protocols, Methods in Molecular Biology. Humana Press. 958:59-70.

Technical Abstract: Genetic transformation of cotton is highly dependent on the ability to regenerate fertile plants from transgenic cells through somatic embryogenesis. Induction of embryogenic cell cultures is genotype-dependant. However, once embryogenic cell cultures are available, they can be effectively used for transformation by Agrobacterium or biolistic bombardment methods. Here I describe a detailed procedure to transform cotton embryogenic cell suspension cultures by biolistic bombardment. Stable transformation at a high frequency (up to 4% of the transiently expressing cells) is possible. Regeneration of fertile transgenic plants from embryogenic cells takes only about two months. Another advantage of the embryogenic cell suspension cultures is that they are amenable for cryopreservation and long-term storage. It is highly preferable to transform commercial varieties of choice than obsolete varieties to avoid negative influence on agronomic and fiber quality traits due to backcrossing. This procedure will be useful to students, technicians, and scientists interested in cotton biotechnology.