Establishment of Pcr Conditions for Determination of Silybum Marianum Genetic Diversity Using Issr Marker Publisher



Jamshidnia M¹ ; Asgary S² ; Rafieiankopaei M³ Show Affiliations
Source: Acta Horticulturae Published:2020

Abstract

Silybum marianum is used for treatment of toxic liver diseases. This activity of S. marianum is due to a mixture of flavonolignans, known as silymarin. S. marianum has economical and domestic worth, nevertheless there are low reports concerning the amount of its genetic diversity. ISSR marker is a PCR based marker that is a suitable technique for genetic variation studies. The purpose of this research is establishment of DNA extraction and PCR conditions that used in ISSR marker. The use of different reagents and the modification of their concentrations allowed obtaining white and clear DNA. Moreover, the modification of temperature and duration of hybridization resulted in clearer ISSR profiles. The optimized ISSR-PCR was used in the analysis of S. marianum genetic diversity. The use of genetic markers for the analysis of genetic material ISSR, carried out on 30 populations. Twenty-nine populations were collected from different regions of Iran following an introduced variety from Hungary. In this experiment 19 primers were used and clear banding pattern produced by 9 primers that totally a total of 85 repeatable polymorphic bands were detected. Among the primers, ISSR7 and ISSR8 identified the most number of PIC with 100%. In this study, protocol of DNA extraction and amplification by PCR for S. marianum were optimized. Large variability was found between the studied populations using ISSR marker. © 2020 International Society for Horticultural Science. All rights reserved.