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Nepenthes species in Thailand, namely N. mirabilis Druce, N. gracilis Korth., N. smilesii Hemsl., N. ampullaria Jack and N. kampotiana Lecomte, were collected for development of Sequence Characterized Amplified Region (SCAR) marker, a genotype identification tool. Forty Random Amplified Polymorphic DNA (RAPD) primers were screened and three successful primers produced different banding patterns including five candidate species-specific markers. The candidate markers were cloned and sequenced. The marker sequences are 602, 379, 420, 473 and 1017 bp for N. mirabilis, N. gracilis, N. smilesii, N. ampullaria and N. kampotiana, respectively. Then the sequences were used to design primers for development of a species-specific band being a SCAR marker, including Mir 1, Mir 2 and Mir 3 for N. mirabilis; Gra 1 and Gra 2 for N. gracilis; Smi 1, Smi 2 and Smi 3 for N. smilesii; Amp 1 and Amp 2 for N. ampullaria and Kam 1 and Kam 2 of N. kampotiana. The primers were evaluated with each other Nepenthes species. Finally, species-specific SCAR markers were successfully developed for N. gracilis, N. ampullaria and N. kampotiana. Application of these markers is feasible for identification of Nepenthes species in Thailand. |
