Journal of Stress Physiology & Biochemistry, Vol. 9 No. 3 2013, pp. 98-112 ISSN 1997-0838
Original Text Copyright (cc) 2013 by   Magdum

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QueryDate : 2016-12-24
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Effect of Agrobacterium Induced Necrosis, Antibiotic Induced Phytotoxicity and Other Factors in Successful Plant Transformation

Sandip S. Magdum

Amity Institute of Biotechnology, Amity University, Noida 201303, India


Received  February 16, 2013

Agrobacterium tumefaciens infection and antibiotic wash are the critical steps of Agrobacterium mediated plant transformation procedure, most time responsible for lower transformation efficiency due to necrosis and phytotoxicity caused by biotic stress of Agrobacterium and abiotic stress by antibiotics respectively.
Ammi majus Egyptian origin medicinal plant and Pearl millet cereal grain crop were studied for their stress responses to Agrobacterium mediated transformation (AMT). Agrobacterium strains LBA4404 (O.D.=0.6-0.8) and EHA105 (O.D.=0.2-0.4) were used for transformation experiments to infect calli of Ammi majus and embryogenic calli of Pearl millet respectively. Incase of antibiotic wash, Cefotaxime 500 mg L-1 was used for LBA4404 infected Ammi majus calli and Timentin 300 mg L-1 was used for EHA105 infected embryogenic calli of Pearl millet.
Effects of Agrobacterium infection, antibiotic and NaOCl washes on Agrobacterium removal and both explants physiological changes during transformation experimental procedures were studied. At the end of the experiments explants survival efficiency of Ammi majus and pearl millet were 8% and 5% respectively. Biotic and abiotic stress factors responsible for lower efficiency were investigated with various other factors and strategies were discussed which are need to be considered for higher transformation events and target tissue survival.

Key words:    Abiotic, Agrobacterium tumefaciens, Ammi majus, Biotic, Pearl millet, Phytotoxicity

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