Novel recombinant binary vectors harbouring Basta

Authors: Reham M Nada

Publish Date: 2016/06/08

Volume: 22, Issue: 2, Pages: 241-251

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Abstract

Genetic transformation is one of the most widely used technique in crop improvement However most of the binary vectors used in this technique especially cloning based contain antibiotic genes as selection marker that raise serious consumer and environmental concerns moreover they could be transferred to nontarget hosts with deleterious effects Therefore the goal of this study was reconstruction of the widely used pBI121 binary vector by substituting the harmful antibiotic selection marker gene with a lessharmful selection marker Basta herbicide resistance gene The generated vectors were designated as pBI121NB and pBI121CB in which Basta gene was expressed under the control of Nos or CaMV 35S promoter respectively The successful integration of the new inserts into both the vectors was confirmed by PCR restriction digestion and sequencing Both these vectors were used in transforming Arabidopsis Egyptian wheat and barley varieties using LBA4404 and GV3101 Agrobacterium strains The surfactant Tween20 resulted in an efficient transformation and the number of Arabidopsis transformants was about 6–9  Soaked seeds of wheat and barley were transformed with Agrobacterium to introduce the bacteria to the growing shoot apices The percentage of transgenic lines was around 16–17 and 14–15  for wheat and barley respectively The quantitative studies presented in this work showed that both LBA4404 and GV3101 strains were suitable for transforming Egyptian wheat and barleyI would like to thank Dr Gaber M Abogadallah Associate Professor of Plant Genetics Botany Department Faculty of Science Damietta University New Damietta 34517 Egypt for hosting this work in his laboratory Agrobacterium GV3101 strain was kindly provided from Dr Mohamed S Tawfik Agricultural Genetic Engineering Research Institute AGERI Agriculture Research Center ARC Giza Egypt 12619

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