Development of a species-diagnostic marker and its application for population genetics studies of the stingless bee Trigona collina in Thailand

Abstract

ABSTRACT. A molecular maker for authenticating species origin of the stingless bee (Trigona collina) was developed. Initially, amplified fragment length polymorphism analysis was made of 11 stingless bee species using 64 primer combinations. A 316-bp band found only in T. collina was cloned and sequenced. A primer pair (CUTc1-F/R) was designed and tested for species-specificity in 15 stingless bee species (239 nests). The expected 259-bp fragment was consistently amplified in all T. collina individuals (134/134 nests, 100%). Cross-species amplification was observed in T. pagdeni (43/51 nests; 84.3%), but not in other species. SSCP analysis of CUTc1 unambiguously differentiated T. collina from T. pagdeni. CUTc1 generated three genotypes in Thai T. collina (134 nests). An AA (259/259 bp) genotype was found in all stingless bees from the north (21 nests) and northeast (32 nests), and 23/28 nests from the Central region, whereas a BB (253/253 bp) genotype was observed in most samples from peninsular Thailand (42/53 nests). Heterozygotes exhibiting the AB (253/259 bp) genotype were observed in 5 of 28 nests from Prachuap Khiri Khan located slightly above the Kra ecotone and 11 of 53 nests originated further south of the Kra ecotone. Genotype distribution patterns of CUTc1 clearly indicated intraspecific population differentiation of Thai T. collina.

Genetic Differentiation of the Stingless Bee Tetragonula pagdeni in Thailand Using SSCP Analysis of a Large Subunit of Mitochondrial Ribosomal DNA

“…In this study, relatively high genetic diversity and strong degrees of geographic differentiation of T. pagdeni in Thailand were identified by polymorphism of the 16S RNA gene. This demonstrates the potential of SSCP analysis for assessing intraand inter-specific genetic variations among Tetragonula species in Thailand (Theeraapisakkhun et al 2010). AMOVA revealed significant genetic differentiation among all pairwise comparisons of T. pagdeni.…”

“…Recently, Theeraapisakkhun et al (2010) developed a molecular marker in T. collina based on SSCP analysis of an AFLP-derived marker, CUTc1. Genotypic distribution patterns of CUTc1 were different in stingless bee samples from the North-to-Central region (alleles of 259/259 bp corresponding to the AA genotype found in 76/81 nests) and most individuals of T. collina from Peninsular Thailand (alleles of 253/253 bp corresponding to the BB genotype found in 42/53 nests).…”

Multilocus species delimitation in Mesoamerican Scaptotrigona stingless bees (Apidae: Meliponini) supports the existence of cryptic species

“…It is suggested that extant diversity corresponds to a third of actual native bee species richness in the Neotropics (Freitas et al, 2009), including many cryptic species yet to be discovered (Michener, 2007). In fact, previous morphological and molecular studies have revealed cryptic species (Melo, 2003;Rasmussen & Cameron, 2007;Tavares et al, 2007;Koch, 2010) or distinct evolutionary lineages within stingless bees species (Quezada-Euán et al, 2007;Tavares et al, 2007;May-Itzá et al, 2010;Theeraapisakkun et al, 2010;Bonatti et al, 2014;Duarte et al, 2014).…”

Southeast Asian clearwing moths buzz like their model bees

“…H. hyaloptera occurs in Thailand and further North up to China but is not known from Malaysia. T. collina , which is widespread in both Malaysia and Thailand [ 19 ] could potentially serve as a model for both studied sesiid species. T. collina lack functional stings and thus cannot inject venom, however they can still bite and chase away intruders [ 20 ].…”