Development of a species-diagnostic marker for identification of the stingless bee Trigona pagdeni in Thailand.
A species-diagnostic SCAR marker for identification of the stingless bee (Trigona pagdeni Schwarz) was successfully developed. Initially, amplified fragment length polymorphism analysis was carried out across representatives of 12 stingless bee species using 64 primer combinations. A 284 bp band restrictively found in T. pagdeni was cloned and sequenced. A primer pair (CUTP1-F/R) was designed and tested for species-specificity in 15 stingless bees. The expected 163 bp fragment was successfully amplified in all examined individuals of T. pagdeni (129/129). Nevertheless, cross-species amplification was also observed in T. fimbriata (1/3), T. collina (11/112), T. laeviceps (1/12), and T. fuscobalteata (15/15), but not in other species. SSCP analysis of CUTP1 further differentiated T. fuscobalteata and T. collina from T. pagdeni. Although T. laeviceps, T. fimbriata, and T. pagdeni shared an identical SSCP genotype, they are not taxonomically problematic species.
Thummajitsakul
Sirikul
S
Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand. sirikul.thum@gmail.com
Klinbunga
Sirawut
S
Sittipraneed
Siriporn
S
D054458:Amplified Fragment Length Polymorphism Analysis / Q000379:methods; D000818:Animals; D001483:Base Sequence; D001516:Bees / Q000235:genetics*; D005819:Genetic Markers; D025202:Molecular Diagnostic Techniques / Q000379:methods; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000379:methods; D013045:Species Specificity; D013785:Thailand
D005819:Genetic Markers