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A Study on the RAPD and SCAR Molecular Markers of Piper Species

Keywords: Kava , Piper methysticum Forst. f. , Pepper , Piper nigrum L. , Random Amplified Polymorphic DNA (RAPD) , Sequence-characterized amplified region (SCAR)

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Abstract:

In order to compare the genetic relationships among Kava, Pepper and it’s wild relatives and to distinguish Kava from Pepper and it’s wild relatives, we conducted research on Kava by using RAPD and SCAR molecular markers. 20 random primers selected from 80 random primers were used for RAPD amplification to identify the genetic relationships among Kava, Pepper and it’s wild relatives. Total 170 bands were amplified by 20 random primers, in which 20 bands were polymorphic (12%). Cluster analysis grouped the 28 accessions into six groups at similarity coefficient of 0.36, where 6 materials of Kava formed a group, indicating that Kava was distantly relation to Pepper and its wild relatives. Kava had 562 bp and 355 bp specific fragments amplified by primers OPQ- 02 and OPQ-03, respectively, were recycled for cloning and sequencing analysis, and then converted to SCAR markers. Two pairs of specific SCAR primers for Kava, P4.1 and P4.2, P8.1 and P8.2 were designed. PCR amplification of 28 test materials were performed using the two pairs of the specific primers respectively, the specific bands of 562 bp and 355 bp with expected sizes were amplified in 6 Kava materials but not in other materials. The results showed that primers P4.1 and P4.2, P8.1 and P8.2 might be used as specific SCAR primers for Kava germplasm resources identification. This research provided the basis for selecting rootstocks, molecular identification and the fingerprint construction of Kava.

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