Analysis of expression level of floral-identity gene (SEPALLATA) in Epicalyx of Hibiscus rosa-sinensis L.

Abstract

Genetic identification of modified floral parts, such as epicalyx, may give new perspectives in flower development fundamentals and its applications. Research on floral-identity gene (SEPALLATA) expression level has been done in three parts of Hibiscus rosa-sinensis; they are leaves, epicalyx, and calyx. This research was conducted to observe the expression of the SEPALLATA gene in epicalyx. The expression level analysis was done qualitatively by the two-steps RT-PCR and visualized using agarose electrophoresis. Hibiscus rosa-sinensis RNA was isolated using the modified-CTAB method and continued by DNase-treatment to eliminate gDNA in the mixture. Furthermore, RNA was used to make cDNA using the Reverse Transcription method and amplified using the PCR method by specific primers. The result showed the presence of SEPALLATA amplification in epicalyx using GH7SEP1 primer, yet none on epicalyx using GHSEP1 primer. Confirmation using GH7SEP1 forward primer and GH1SEP1 reverse primer did not show any amplification. Sequencing and alignment results suggested that amplifications using GH1SEP1 or GH7SEP1 were allegedly, of which amplified from SEPALLATA gene.