Genetic diversity of stingless bees (Apidae: Meliponinae) in Thailand detected by PCR-RFLP of mitochondrial dna

Abstract

Stingless bees (Trigona spp.) are the highly eusocial insects belonging to subfamily Meliponinae. These bees have no sting which the honey bees use for defensive purposes. Genetic variation of five species of Trigona in a total of 407 colonies (T. collina (224 colonies), T. fuscobalteata (18 colonies), T. laevicep (131 colonies), T. terminate (18 colonies), and T. thoracica (6 colonies)) from various parts of Thailand (North, North-East, East, Central and South) were determined. PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) analysis was used to detect variation at 16S rRNA gene of mitochondrial DNA. The size of PCR product at 16S rRNA gene is about 550 bp. Variation in 16S rRNA gene of T. collina and T. laevicep were found when digested with Dra ∣, Hpy188 ∣∣∣, and Ssp ∣. Variation in 16S rRNA gene of T. fuscobalteata, T. terminate, and T. thoracica were found when digested with Dra ∣ but there were no variation with Hpy188 ∣∣∣ and Ssp ∣ digestion. Digestion of these PCR products of five species (T. collina, T. fuscobalteata, T. laeviceps, T. terminate, and T. thoracica) with Dra ∣, Hpy188 ∣∣∣, and Ssp ∣ revealed 3, 4, and 4 patterns, 2, 1, and 1 patterns, 4, 2, and 3 patterns, 2, 1, and 1 patterns, 2, 1, and 1 patterns, respectively. Cluster analysis of T. collina, T. fuscobalteata, T. laeviceps, T. terminate, and T. thoracica populations (3 enzymes) by UPGMA (unweighted pair group method with arithmetic mean) could be separated into 3, 2, 2, 2, and 2 groups, respectively. These results indicated that genetic variation of five species of Trigona in Thailand could be detected by using PCR-RFLP analysis at 16S rRNA gene of mitochondrial DNA.