International Journal of Farming and Allied Sciences Available online at www.ijfas.com ©2014 IJFAS Journal-2014-3-5/538-542/ 31 May, 2014 ISSN 2322-4134 ©2014 IJFAS Reduction of negative effects of cefotaxime in tomato transformation by using FeEDDHA Elham Mohajel Kazemi 1* , Parisa Jonoubi 1 , Ahmad Majd 1 and Maghsoud Pazhouhandeh 2 1. Department of Plant Biology, Faculty of Life Science, Kharazmi University, Tehran, Iran 2. Department of Biotechnology, Faculty of Agriculture, Azarbaijan Shahid Madani University, Km35 Tabriz- Azarshahr Road, Iran Corresponding author: Elham Mohajel Kazemi ABSTRACT: Tomato (Lycopersicon esculentum) is one of the most important vegetable crops and a genetic model for improving other dicotyledonous crop plants. Agrobacterium mediated gene transformation method is considered as an effective method for transferring foreign genes into plants. cefotaxime, commonly used for elimination of Agrobacterium tumefaciens during plant transformation. In high concentrations of cefotaxime, a decrease in regeneration, reducing the number of shoots and chlorosis were observed. We examined the effect of FeEDDHA on shoot regeneration from cotyledon explants in Falat CH cultivar. When applied during multiplication, FeEDDHA reduced chlorosis, increased the percentage of regeneration and the number of adventitious shoots. Therefore replacement of FeEDTA by FeEDHHA in media reduced the negative effects of cefotaxime. Keywords: Agrobacterium, cefotaxime, chelate Fe, transformation INTRODUCTION Agrobacterium-mediated transformation is an effective and widely used method to introduce DNA into dicotyledonous plants. The successful elimination of Agrobacterium from regeneration media in transformation protocols is important for the successful recovery of transgenic cells and tissues. Carbenicillin and cefotaxime have been commonly used as effective antibiotics for elimination or suppression of Agrobacterium cells (Mathias and Boyd, 1986; Tang, 2000; Alsheikh, 2002). They have a broad spectrum of activity against both Gram-positive and Gram-negative organisms, and function by blocking the cell wall mucopeptide biosynthesis, by inhibiting the cross- linking of peptidoglycan by binding and inactivating of transpeptidases; cefotaxime is a cephalosporin antibiotic with a broad spectrum of activity, low eukaryote toxicity, and effective at low doses (Mathias and Boyd, 1986), making it particularly attractive as a selective agent in plant tissue cultures. But both carbenicillin and cefotaxime reduced callus growth, shoot regeneration and transformation efficiency when they were used in high concentration either alone or in combination, e. g., cefotaxime up to 500 mg P 1 and carbenicillin at 250-1000 mg P 1 (Lin, 1995; Vergauwe al., 1996a;Nauerby, 1997). Silva and Fukai (2001) reported that the success in transgenic plant regeneration relies on the fine balance of four points, such as (1) the plant, (2) the Agrobacterium, (3) selection agent and (4) the agent for eliminating Agrobacterium. Low concentration of cefotaxime enhanced shoot regeneration in wheat (Mathias and Boyd, 1986; Borrelli et al., 1992) maize (Danilova. and Dolgikh, 2004) apple (James et al., 1989) and strawberry (James, 1990). However regeneration efficiency was reduced by increasing the concentration of cefotaxime. Reduction of regeneration in high concentration of cefotaxime was reported in various plants including tomato (Ling, 1998), chrysanthemum (Silva and Fukai 2001), maize, apple, rice and wheat