Genetic Diversity of Promised Russian Grapevine Genotype and Varieties by Using Inter Single Sequence Repeat (ISSR) DNA Marker

Background and Aim: Genetic diversity determination and its preservation in a plant species are important in breeding programs and plant protection. The key to the success of any plant-breeding program is the existence of genetic diversity. Knowledge of diversity at the molecular level of DNA is important for the study of evolutionary and phylogenetic relationships, the development of a genetic map and the evaluation of ecological and environmental traits and adaptations.Methods: This study carried out to investigate the genetic diversity of Russian grapevine varieties by using inter simple sequence repeats (ISSR) molecular marker. The DNA extraction was done by the Cetyltrimethyl Ammonium Bromide (CTAB) method. Fifteen ISSR primers were used which included UBC-۸۱۵, UBC-۸۱۷, UBC-۸۲۳, UBC-۸۲۵, UBC-۸۲۶, UBC-۸۳۴, UBC-۸۴۱, UBC-۸۴۶, UBC-۸۴۹, UBC-۸۵۰, UBC-۸۵۵, UBC-۸۵۶, UBC-۸۷۳, UBC-۸۷۷ and UBC-۸۸۹. Polymerase chain reaction (PCR) samples were electrophoresed in ۳.۱% agarose gel. Molecular indexes were included: polymorphic percentage index, polymorphic information content (PIC), Marker Index (MI), Effective Multiplex Ratio (EMR) and Resolving Power (RP). Cluster analysis with the UPGMA method was used based on the Jaccard coefficient because of the proper dendrograms presenting (without chains), higher Cophenetic correlation, and especially proper grouping and in accordance with morphological traits. The linear discriminant function was used to determine the accuracy of the grouping of cultivars and genotypes. The calculations were performed using GenAlex, NTSYS, and SPSS ver.۲۱ software.Results: The UBC-۸۱۵ and UBC-۸۳۴ primer had no clear and concise bands. Overall ۷۳ bands were obtained from the ۱۳ primers of the ISSR molecular marker which ۶۴ bands were polymorphic and the total polymorphism was ۸۸.۱%. The size of the bands varied between ۱۰۰ and ۱۵۰۰ base pairs. UBC-۸۲۳, UBC-۸۲۵, UBC-۸۴۱, UBC-۸۴۶, UBC-۸۵۵, and UBC-۸۷۳ primers had ۱۰۰%polymorphism. The lowest polymorphism (۶۶.۷%) also belonged to the UBC-۸۱۷ primer. The highest and lowest polymorphic content indexes were ۰.۴۸۷ and ۰.۲۱۶ for UBC-۸۲۵ and UBC-۸۵۶ primers, respectively. The cluster analysis by unweighted pair group method with arithmetic means (UPGMA) method based on Jaccard's coefficient of similarity led to the classification of ۷۲ grape varieties with a similarity value of ۰.۱۲ to ۰.۸۹ in seven different groups.Conclusion: In the present study, a high variation was observed among grapevine cultivars. Since the ISSR molecular marker could well differentiate and classify different cultivars in separate groups, it can be said that the ISSR markers are an efficient and reliable marker system for analyzing the genetic diversity of grape varieties.

کلیدواژه ها:

grapevine ، genetic polymorphism ، genetic affinity.