Application of Microsatellite Markers for Identification and Registration of Grape Cultivars
2011
Mohammadi, Seyed Abolghasem | Nejatian, Mohammad Ali | Jamali, Seyed Hossein | Oni Kanamouei, Amir | Kavand, Abdolreza | Razavi, Alireza | Sadeghi, Leila | Tahernezhad, Zahra | Dolati Baneh, Hamed | Alavikia, Siamak | Asdaghi, Gholamreza
To develop molecular descriptor and identification key for 12 commercial grape cultivars, leaf samples were taken from 79 trees consisted of 5 to 10 clones for each cultivar growing at Oroomieh and Takestan research stations. Genomic DNA was extracted using CTAB method and DNA quality and quantity were assessed based on spectrophotometer and 0.8% agaros gel electrophoresis. Preliminary evaluation of accessions was done using 47 SSR primer pairss and 20 polymorphic primers with reproducible banding pattern were used to develop identification keys for the cultivars. The number of allels amplified in each locus varied from 2 (VVIB19 and ISV3) to 12 (ISV4). Grouping of the cultivars was performed based on each marker data using cluster analysis to determine marker differentiation power. Among the marker, ISV4 showed maximum differentiation power and could provide specific banding patterns for eight cultivars including Khoshnov, Paykani, Kashmar, Fakhri, Yaghoti, Shast-Aros, Mish-pestan, Shahrodi and Rish-baba-ghermez. To identify differentiating marker combinations, grouping of cultivars was performed using various marker combinations and grouping based on ISV4 and ISV2 could separate all the cultivars except Bidaneh Ghermez and Sefied. Based on pairwise comparison of cultivars at microsatellite loci, Mish-pestan and Shahrodi as well as Mish-pestan and Fakhri with differences at 13 out 20 loci showed maximum dissimilarity. Asghari-Shahrodi and Mish-pestan, Rish-baba-ghermez cultivar pairs were differed at 12 microsatellite loci. Bidaneh Ghermez and Sefied cultivars showed similar genotypes at the all studied loci and Rish-baba-ghermez was differed from Bidaneh Ghermez and Sefied at two loci.
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