A “Nonsterile” Method for Selecting and Growing Arabidopsis thaliana Transformants (T2 Transgenic Lines) Resistant to Kanamycin

Abstract

A breakthrough in transgenic Arabidopsis thaliana research was the development of the floral dip transformation protocol, a simple and reliable method of obtaining transformants, T1 transgenic lines, at high efficiency while avoiding the use of tissue culture. However, the traditional protocol (a “sterile” method) of obtaining T2 transgenic lines has not evolved along with improvements in transformation technology as it continues to be laborious and time-consuming. In this study, we report on the development of an improved protocol (a “nonsterile” method) for selecting and growing A. thaliana transformants (T2 transgenic lines) resistant to kanamycin under nonsterile conditions. This protocol involves the use of a simple yet specialized device that will aid in solium selection of T2 transgenic lines that can be rapidly grown in a hydroponic system. The “nonsterile” method reduces labor and time involved as compared to the “sterile” method; moreover, it is easy to set up and maintain. This method may also be applicable to other selecting agents, and perhaps to other plants.