Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
Biochem Genet. 2010 Apr;48(3-4):181-92. doi: 10.1007/s10528-009-9300-0. Epub 2009 Dec 18.
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.
成功开发了一种用于鉴定无刺蜜蜂（Trigona pagdeni Schwarz）的种诊断性 SCAR 标记。最初，使用 64 对引物组合对 12 种无刺蜜蜂的代表进行了扩增片段长度多态性分析。在 T. pagdeni 中发现了一个 284 bp 的限制性带，对其进行了克隆和测序。设计并测试了一对引物（CUTP1-F/R）在 15 种无刺蜜蜂中的种特异性。在所有检查的 T. pagdeni 个体中（129/129）都成功扩增了预期的 163 bp 片段。然而，在 T. fimbriata（1/3）、T. collina（11/112）、T. laeviceps（1/12）和 T. fuscobalteata（15/15）中也观察到了交叉种扩增，但在其他种中没有。CUTP1 的 SSCP 分析进一步将 T. fuscobalteata 和 T. collina 与 T. pagdeni 区分开来。尽管 T. laeviceps、T. fimbriata 和 T. pagdeni 具有相同的 SSCP 基因型，但它们在分类上并不是有问题的物种。
