Application of the RAPD and miRNA markers in the genotyping of Silybum marianum (L.) Gaertn.

Abstract

DOI: 10.15414/afz.2015.18.04.83–89

Received 7. July2015 ǀ Accepted 30. September 2015 ǀ Available online 7. December 2015

Two types of molecular markers based on randomly amplified DNA by RAPD-based assay and amplified microRNA by
miRNA-based assay were applied for the genotyping of five accessions of Silybum marianum (L.) Gaertn. It was also verified
the effectiveness of the isolation of genomic DNA by three commercially available isolation kits and two similar complex
methods and their applicability for this medicinal herb. None of the commercial isolation kits provided the genomic DNA
of sufficient quality and quantity. Applied originally designed CTAB method and modified CTAB method for purpose of
DNA isolation from medicinal plants allowed to isolate DNA of required quality and of sufficient yield. RAPD-based DNA
fingerprints, allowed to distinguish the individual genotypes of Silybum marianum. MicroRNA-based markers showed the
cross-genera transferability potential and displayed sufficient level of polymorphism. Both types of molecular markers
could be used as suitable tool for genotyping of milk thistle. However, because of the size of miRNA amplicons is the
efficiency of miRNA-based markers PCR amplifications preferred and consequently less DNA quality-dependent which may
be advantageous in view of the content of undesirable secondary metabolites in medicinal plants.

Keywords: molecular markers, RAPD, miRNA, Silybum marianum

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