تراريختي كلزا (Brassica napus) با ژن 5- انول پيروويل شيكيمات -3- فسفات سنتاز (EPSPS) با منشاء آرابيدوپسيس در راستاي ايجاد مقاومت به علف كش گليفوسيت

Transaformation of Oilseed rape (B. napuss L.) with Arabidopsis 5- enol- pyruvylshikimic acid 3-phosphate synthase (EPSPS) Gene towards Conferring Glyphosate Resistance

Glyphosate IS a broad spectrum herbicide that inhibits the 5- enolpyruvylshikimic acid 3-phosphate synthsae (EPSPS) enzyme which is the key enzyme of shikimate pathway. One of the major approaches taken to obtain glyphosate tolerant plants is to increase the target enzyme EPSPS. A cDNA clone encoding A. thaliana EPSPS was cloned into the pBI121 binary vector under the control ofCaMV 35S promoter to abtain high level expression of the enzyme. The At EPSPS and NPT II transgenes were transformed into cotyledonary explants of rapeseed via A. tumefacience. The green transformed plantlets were obtained on selective medium suplemented with kanamicin. These plants were evaluated by PCR, DNA dot blotting and RT-PCR for continning transgene integration and expression. Tl plants were obtained from seeds of TO transgenic plants. Phenotypic assay for glyphosate resistance and quantitative analysis of shikimate were performed after spraying the TO, T1·and wild type plants with 0, 2.5 and 5 mM glyphosate twice a week. The results indicated increased glyphosate tolerance in TO and Tl transgenic plants