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

The entomological origin of honey was detected in the subtropical <i>Apis cerana cerana</i> using two sets of <i>Major Royal Jelly Protein 2</i> (<i>mrjp2</i>) primers. Due to the high genetic divergence of <i>A. cerana</i>, this study aimed to confirm that the primers can be used to detect honey origin from <i>A c. javana</i> and <i>Apis mellifera</i> in Indonesia. The <i>mrjp2</i> 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 <i>A. mellifera</i> and <i>A. c. javana</i> 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 <i>A. cerana</i> at 47 °C. This study confirmed that C-F and C-R primers are specifically applicable when amplifying <i>A. c. javana</i>.