Title Genetic studies, in vitro propagation and conservation of some pomegranate cultivars grown in Taif province, Kingdom of Saudi Arabia Paper ID UQh1w Keywords Pomegranate, genetic variations, RAPD, ISSR, In vitromicro-propagation,In vitro conservation. Abstract
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Pomegranate (Punicagranatum L.) was recorded as one of the most important medicinal plant. It has several pharmaceutical and ornamental use due to bioactive compounds from its parts. Therefore, this study focused on determination of the genetic variations between two cultivars (Taify and Yemeni) of pomegranate common agriculture in the Taif area. This was followed by establishment of in vitro micro-propagation system using shoot tips as explants. In vitro conservation of the two cultivars on tissue culture media using the same explants was also conducted. A number of 20 pomegranate samples cultivated in Taif area and belonging to two cultivars were collected (10 sample for each cultivar). DNA extractions were prepared from the pomegranate plant samples and both of random amplified polymorphic of DNA-polymerase chain reaction (RAPD-PCR) and Inter specific sequence repeat (ISSR) tools were used for determination of the genetic variations between the two cultivars. In vitro micro-propagation system using shoot tips as explants was established for the two pomegranate cultivars under investigation. Results showed that 10 out of 20 RAPD-PCR and ISSR primers showed DNA polymorphisms between the 20 pomegranate samples collected from Taif area. In the present study, good success on MS medium supplemented with 2.0 mg/L BAP, and 0.1 mg/L Kin was obtained in the three stages of pomegranate micro-propagation. This was obvious by the highest % of explants initiated shoots; number of initiated shoots per explant and length of produced shoots using shoot tip explants. Results also showed that root producing was obtained best on MS medium containing 1.0 mg/L NAA plus 2.00 mg/L IBA among the highest % of rooted shoots (83.3 and 79.6) for the two pomegranate cultivars (Taify and Yemeni), respectively. These auxin combinations were the most effective in number of proliferated shoots per explant and root length. Results showed that the use of full strength MS in combination of 2% of each of sucrose and mannitol as carbon sources was the most effective treatment. As a conclusion, in vitro conservation (slow growth technique) using full strength MS media supplemented with low sucrose and sorbitol concentrations was a suitable method for germplasm conservation. The applied RAPD-PCR and/or ISSR tools could be used to analyze and identify the genetic relationships among pomegranate genotypes.
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