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

The entomological origin of honey was detected in the subtropical Apis cerana cerana using two sets of Major Royal Jelly Protein 2 (mrjp2) primers. Due to the high genetic divergence of A. cerana, this study aimed to confirm that the primers can be used to detect honey origin from A c. javana and Apis mellifera in Indonesia. The mrjp2 gene was amplified using M-F M-R and C-F C-R primers at various annealing temperatures to find the optimal temperature based on the presence-absence of the amplicon. Honey was successfully differentiated from 11 colonies of A. mellifera and A. c. javana using M-F and M-R primers at annealing temperatures of 50 °C, 53 °C, 55 °C, 57 °C, and 59 °C. However, caution should be placed on using these primers when used to amplify A. cerana at 47 °C. This study confirmed that C-F and C-R primers are specifically applicable when amplifying A. c. javana.