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 degrees C, 53 degrees C, 55 degrees C, 57 degrees C, and 59 degrees C. However, caution should be placed on using these primers when used to amplify A. cerana at 47 degrees C. This study confirmed that C-F and C-R primers are specifically applicable when amplifying A. c. javana.