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Molecular cloning and development of RAPD-SCAR markers for Dimocarpus longan variety authentication

Luquan Yang1, Shelly Fu2, Md Asaduzzaman Khan1, Weimin Zeng3* and Junjiang Fu1*

Author Affiliations

1 Research Center for Preclinical Medicine, Luzhou Medical College, Luzhou, Sichuan 646000, China

2 Michael E. DeBakey High School for Health Professions, 3100 Shenandoah Street, Houston, TX 77021, USA

3 Department of Biochemistry, School of Life Sciences, Central South University, Changsha, Hunan 410013, China

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SpringerPlus 2013, 2:501  doi:10.1186/2193-1801-2-501

Published: 3 October 2013

Abstract

As an edible fruit and source of traditional medicine, D. longan is grown in most areas of Southern China. Identification of D. longan cultivars by using molecular markers is important genetically. In this study, we cloned fragments from improved randomly amplified polymorphic DNA (RAPD), and developed stably diagnostic sequence-characterized amplified region (SCAR) markers. The specific RAPD bands of D. longan cultivars from Guangxi, with size ranging from 500 bp to 900 bp were gel-purified, cloned and sequenced. Four clones named LY2-1, LY4-7, LY4-8 and LY5-2 were identified. In order to investigate whether the fragments were specific for the species, four pairs of SCAR primers were then designed. PCR amplifications were conducted to analyze 18 samples including different D. longan cultivars and other species. The specific bands with expected sizes were amplified in five D. longan samples but not in others. To identify and characterize the difference between D. longan and D. confinis, PCR amplifications were performed again. The specific bands with expected sizes were found in D. longan but not in D. confinis by SCAR markers LY2-1, LY4-7 and LY5-2, respectively. These results showed that our developed SCAR markers could be very useful as a specific D. longan variety authentication. Therefore, our study provides an effective and precise PCR-based diagnostic method and markers to identify D. longan species.

Keywords:
Dimocarpus longan Lour; Dimocarpus confinis; Improved RAPD; Cloning; SCAR marker; Authentication