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Title of article :
Agrobacterium-Mediated Transformation of Pomegranate (Punica granatum L.) ‘Yousef Khani’ Using the gus Reporter Gene
Author/Authors :
وليزاده كاجي، بابك نويسنده Bu-Ali Sina University Valizadeh Kaji, Babak , ارشادي، احمد نويسنده Bu-Ali Sina University Ershadi , Ahmad , توحيدفر، مسعود نويسنده ,
Issue Information :
فصلنامه با شماره پیاپی 0 سال 2014
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Abstract :
In this study, an efficient Agrobacterium-mediated transformation method was developed for pomegranate (Punica granatum L.), a difficult-to-transform plant. In vitro shoot segments were inoculated with Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pBI121 carrying the neomycin phosphotransferase (nptII) gene as a selectable marker and B-glucuronidase (gus) gene as a reporter. After 28 d in WPM selection medium containing 50 mg L-1 kanamycin, 59 new shoots proliferated. gus analysis was performed on these putative transgenic shoots, of which 32 stained positive. Positive staining shoots were cut and cultured in selection medium for 2 subsequent subcultures until final gus analysis. After three months of the selection period, 6 putative transgenic shoots were obtained. Presence of the gus and nptII genes was confirmed by polymerase chain reaction. Southern blot analysis confirmed that T-DNA was stably integrated into the genome of three out of six PCR-positive plants. The transgenic plants were rooted and successfully acclimatized.
Journal title :
International Journal of Horticultural Science and Technology
Journal title :
International Journal of Horticultural Science and Technology
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