Conversion of RAPD markers to co-dominant based Sequence Characterized Amplified Region (SCAR) Markers in three paphiopedilum species

Thun Evonne, (2007) Conversion of RAPD markers to co-dominant based Sequence Characterized Amplified Region (SCAR) Markers in three paphiopedilum species. University Malaysia Sabah. (Unpublished)

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Abstract

The genus Paphiopedilum is endangered species protected under Convention on International Trade in Endangered Species (CITES). However, the discrimination between the species is difficult due to their similar morphological characters. Therefore, RAPD assay were applied to detect polymorphisms between the three species: P. rothschildianum, P. dayanum and P. lowii. Diagnostic fragments identified for discrimination for each species were converted into SCAR markers by cloning and sequencing the diagnostic fragments and designing SCAR primer pairs to amplify the diagnostic fragments. The reproducibility of these markers was analysed by performing diagnostic PCRs. In this study, PCR amplification with RAPD primers was successful in detecting polymorphisms between species. Three diagnostic fragments were converted into SCAR markers and all primer pairs amplified single distinct bands whose sizes were the same as expected fragments. One of the SCAR primers amplified the expected product size in P. dayanum and in one of the P. rothschildianum samples. When digested with restriction enzyme DpnII found polymorph isms in fragment length between the two samples. In the other two cases, the products of the expected fragments were amplified in P. lowii and all samples of P. rothschildianum and subsequent RE digestion failed to detect polymorph isms between the species. The overall findings had shown discovery of two SCAR markers to identify P. rothschildianum and P. lowii apart from P. dayanum and one SCAR marker to identify P. dayanum and its possible hybrids. Hence, more studies and screenings need to be done in order to develop species-specific markers for the Paphiopedilum species.

Item Type: Academic Exercise
Uncontrolled Keywords: Paphiopedilum, International Trade in Endangered Species (CITES), discrimination, morphological character, RAPD assay, SCAR, PCR amplification
Subjects: S Agriculture > SD Forestry
Depositing User: MDM SITI AZIZAH IDRIS
Date Deposited: 23 May 2014 02:59
Last Modified: 17 Oct 2017 04:26
URI: http://eprints.ums.edu.my/id/eprint/9072

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