Entomological origin detection of honey from Apis mellifera and Apis cerana javana in Indonesia based on the Major Royal Jelly Protein 2 (mrjp2) gene

Major Royal Jelly Protein 2 (<i>mrjp2</i>) Gene Detection in <i>Apis dorsata</i> Fabricius, 1793, <i>Apis dorsata binghami</i> Cockerell, 1906, <i>Apis florea</i> Fabricius, 1787, and <i>Apis nigrocincta</i> Smith, 1860

“…In this study, following the previous study conducted by (Raffiudin et al 2023;Zhang et al 2019) this research will provide new information about the mrjp2 gene present in honey bee species found in Indonesia such as A. dorsata, A. dorsata binghami, A. florea, and A. nigrocincta. New information provided by this research can be used to determine the entomological origin of honey sold in Indonesia as one way to avoid honey adulteration.…”

“…This research also succeeded in yielding DNA from two honey bee species, which are A. nigrocincta and A. cerana using species-specific primers of mrjp2 gene for A. cerana (CF-CR) at the annealing temperature 47℃ (Figure 2). Raffiudin et al (2023) was successfully amplified the DNA of A. mellifera using the MF-MR primer at the annealing temperature 50, 53, 55, 57, and 59℃, while the DNA of A. cerana was successfully amplified at the annealing temperature 47, 50, 53, and 55℃. The amplicon of the mrjp2 gene in MF-MR primers was obtained at a size about 600 bp, while in CF-CR primers it was obtained at a size about 200 bp.…”

“…The research method is designed based on Zhang et al (2019) and Raffiudin et al (2023) with some modification. This research was conducted from August 2022 to February 2023 in Laboratory of Genetic Engineering, Inter-University Centre, Universitas Gadjah Mada which included DNA extraction and isolation, DNA amplification and visualisation, and data analysis.…”

“…PCR is carried out by making a 25 μL PCR cocktail consisting of 12.5 μL PCR Promega GoTaq Green Master Mix, 9.5 μL Nuclease Free Water, 1.5 μL primer, and 1.5 μL DNA template. The mrjp2 gene was amplified with an initial pre-denaturation at 94ºC for 2 min, 35 cycles of denaturation at 94ºC for 30 s, annealing at 53ºC (for samples with A. mellifera primer) and 47°C (for samples with A. cerana primer) for 30 s, extension at 72ºC for 30 seconds, and final extension at 72ºC for 5 minutes (Zhang et al 2019;Raffiudin et al 2023). After going through the PCR process, 1.2% agarose gel electrophoresis was used to separate the amplicon, which was then visualised with a UV transilluminator.…”

“…Zhang et al (2019) could determine the entomological origin of honey using speciesspecific primers of A. mellifera (MF-MR) and A. cerana (CF-CR) based on differences in mrjp2 gene sequences in A. mellifera and A. cerana. Raffiudin et al (2023) have conducted research on the mrjp2 gene in A. mellifera and A. c. javana from Indonesia and showed that CF and CR primers previously used in Zhang et al (2019) research can amplify honey bee species A. c. javana.…”