Transformation of Agrobacterium Tumefactions by Conjugation Technique

Abstract

Two genera of Rhizobiaceae ,wildstrain Agrobacterium tumefaciens (AtMcol) and standerd strain Agrobacterium tumefaciens (C58C1), in addition toSinrhizobium melilti are used in this study. The results showed, these twostrains ofA. tumifaciens bacteria were resistant to Trimetheprim and sensitiveto chloromphinicol while the S.melilti bacteria gave the opposite of theseresults,and used as genetic markers for conjugation as one of the methods usedin the technique of Horizontal gene transfer, with the use of acardine orangeto cure the bacteria from plasmid.Two conjugation procces were performed,between Agrobacterium tumefaciens (AtMcol) and S. melilti, and betweenAgrobacterium tumefaciens (C58C1)and S.melilti, result showed succecful ofconjugation procces with conjugation frequency 1.8x10-2 and 2.1x10-2respectively, and the transconjugant bacteria result from procces succeded ininfecting the root tip of alfalfa seedlings to form root nodules.Characterization of plasmid DNA content for bacterial  isolates belonging to conjugated cells, werecarried out by using agarose gel electrophorsis and the results showed isolatethe presence of discrete clear and isolated band of similar size present inagaros gel.