A Simple Method of Plant Sectioning Using Agarose Embedding Technique for Screening Intracellular Green Fluorescent Protein

Abstract

The transformed plant tissue sections through the agroinfiltration method are difficult to be observed under a fluorescent microscope. It is due to the softness of the post-transformation explant. This research was conducted to optimize the sectioning of the transformed plant using the agarose embedding technique. The optimization method was carried out in various agarose concentrations: 2%, 4%, and 6%, followed by five minutes of incubation in various temperatures: -80 oC, 4 oC, and 25 oC. The results showed that the embedding method using 6% agarose produced a transformed plant section was better than 2% and 4% agarose. Meanwhile, the incubation method at 25 oC was more suitable for the transformed plant tissue than 4 oC and -80 oC. Green Fluorescent Protein (GFP) could be determined using these methods under a fluorescent microscope. Thus, the optimum method for making sections of transformed plants by embedding was using 6% agarose then incubating at 25 oC for 5 minutes.