Application of Microsatellite Markers for Identification and Registration of Olive Cultivars

Oleaceae family contains approximately 20 species of small trees and ancient world of the Mediterranean, North Africa, Southeast Asia, north to southern China, Scotland and East Australia have a wide dispersion. They are evergreen and have small leaves that are integrated have been met and the fruit is drup. Olive trees resistant to drought and adapt to poor soils and produce high-value, low-cost Kmbazdh and economically very important that the product is well-known in poor soils. In the past, methods for cultivar identification was based on morphological characteristics of leaves, fruits and kernels, but except in specific cases identification using morphological traits alone are not enough. Due to complex identification of young olive cultivars (Olea europaea L.) by morphological traits, advance molecular tools have provided a new prospect for DNA fingerprinting.
For this study, sampling was done after receiving information from Horticulture Science Research Institute (HSRI) and Seed and Plant Certification and Registration Institute (SPCRI). Sampling was done from young and fresh shoot of trees. Approximately, 30 mg of young leaf were ground in liquid nitrogen and total DNA was isolated using the procedure described Core Bio DNA extraction kit and PCR-amplified using 22 pairs of primers flanking SSR sequences.
Specific molecular keys were identified for 11 Iranian olive cultivars (Olea europaea L.) using 10 out of 22 SSR markers. The results showed that 10 SSR markers produced polymorphism or polymorphic bands for studied Iranian olive cultivars. SSR markers MOL 1 in Tokhmekabki, MOL 7 in Dezfooli, Mavi and Roghani, MOL 8 in Dezfooli and Shangeh, MOL 9 in Dezfooli, Golooleh, Mari and Roghani, MOL 10 in Tokhmekabki, MOL 11 in Dezfooli and Fishemi, MOL 14 in Dehghan, MOL 18 in Golooleh, MOL 20 in Mari produced specific molecular keys. Out of 12 Iranian olive cultivars, four cultivars (Dezfooli, Golooleh, Mavi and Roghani) were identified by SSR marker MOL 9.
Off-type trees were identified using cluster analysis based on SSR data. The specific molecular keys were verified on 64­ olive mother's trees and the results were confirmed at two independent laboratories (SPCRI and Agriculture Biotechnology Research Institute of Iran). Most mother trees of different varieties were classified in separate groups. Dendrogram of Splits Tree 4 software separated mother trees of evaluated olive cultivar with 11 SSR markers. The results demonstrated that cluster analysis and Structure software, was very appropriate for study of genetic relationships among olive cultivars. The reported specific molecular keys can be efficiently used for identification of olive cultivars for Genetic authentication