Green Fluorescence Expression in Two Aquarium Plant Species by Agrobacterium-Mediated gfp Gene Transferred into the Target Tissues

Abstract

The development of plants containing novel characteristics can be done using gene transfer of the target gene into the plant genome. This study aimed to obtain information on the green fluorescence expression of the gfp gene in 2 aquarium plant species, Anubias nana and Hygrophila difformis. The Agrobacterium-mediated gene transfer was used to transfer the gfp gene and other necessary genes for the process of expression into the plant target tissues. Somatic callus was used as the target tissue for gene transfer in A. nana, while somatic callus and shoots were used in H. difformis. The results found that the transformation efficiency was 280 cells of the Agrobacterium with plasmid (pCAMBIA1304) inside per 1 µg of the plasmid used. The gfp gene with all necessary other genes was successfully transferred, confirmed by the blue spots by the GUS gene from histochemical analysis, the occurrence of the target 327 bp band of the gfp DNA by the PCR technique, and the gfp expression as green fluorescence of the GFP protein. However, in six-month-old T1 plants produced from tissue culture from the original gene-transferred plantlets (T0) of both A. nana and H. difformis, which the callus was used as the target tissue, did not show the gfp band, which was probably due to chimeras. But the T1 of H. difformis that the shoot was used as the target tissue showed the gfp band. It can be concluded that callus should not be used as the target tissue for gfp gene transfer in these two aquarium plant species, whereas shoots can be used as the target tissue in H. difformis.