Identification of stingless bees (Hymenoptera: Apidae) in Kenya using morphometrics and DNA barcoding

Stingless bees are important pollinators of wild plants and crops. The identity of stingless bee species in Africa has not been fully documented. The present study explored the utility of morphometrics and DNA barcoding for identification of African stingless bee populations, and to further employ these tools to identify potential cryptic variation within species. Stingless bee samples were collected from three ecological zones, namely Kakamega Forest, Mwingi and Arabuko-Sokoke Forest, which are geographically distant and cover high, medium and low altitudes, respectively. Forewing and hind leg morphometric characters were measured to determine the extent of morphological variation between the populations. DNA barcodes were generated from the mitochondrial cytochrome <i>c</i>-oxidase I (<i>COI</i>) gene. Principal Component Analysis (PCA) on the morphometric measurements separated the bee samples into three clusters: (1) <i>Meliponula bocandei;</i> (2) <i>Meliponula lendliana</i> + <i>Plebeina hildebrandti;</i> (3) <i>Dactylurina schmidti</i> + <i>Meliponula ferruginea</i> black + <i>Meliponula ferruginea</i> reddish brown, but Canonical Variate Analysis (CVA) separated all the species except the two morphospecies (<i>M. ferruginea</i> reddish brown and black). The analysis of the <i>COI</i> sequences showed that DNA barcoding can be used to identify all the species studied and revealed remarkable genetic distance (7.3%) between the two <i>M. ferruginea</i> morphs. This is the first genetic evidence that <i>M. ferruginea</i> black and <i>M. ferruginea</i> reddish brown are separate species.