SCAR marker for identification and discrimination of specific medicinal Lycium chinense Miller from Lycium species from ramp-PCR RAPD fragments

Abstract

In the current study, ramp-PCR fragments from improved RAPD (random amplified polymorphic DNA) amplification of Lycium (Goji) species or cultivars were cut and cloned into the vector of pGEM-T. A positive clone 10-5 was screened by PCR amplification, enzymatic digestion, and Sanger sequencing. A SCAR (sequence-characterized amplified region) marker, named Goji 10-5, with 949 nucleotides in length, was identified. Goji 10-5 is specific to Goji species Lycium chinense Miller from Jiangxi in China and Texas in the USA. A BLAST search of this nucleotide sequence in the GenBank database indicated that it shows no identity with any other species, including no any other Lycium species. As a new sequence, we have deposited it in the GenBank database with accession No. MN862323. PCR assays were developed and converted the nucleotide sequence to become a novel molecular marker for Lycium chinense Miller, named Goji 10-5. This marker may be used for the genetic identification of other samples. This study has successfully developed Goji 10-5, a specific SCAR marker to identify L. chinense and distinguish it from other species, including other Lycium species from different locations.

Keywords: Authentication; Lycium species; Molecular marker; Random amplified polymorphic DNA; Sequence-characterized amplified region.