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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 9, No. 4, 2006, pp. 349-357
Bioline Code: ej06053
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 9, No. 4, 2006, pp. 349-357

 en Agrobacterium rhizogenes check for this species in other resources mediated genetic transformation resulting in hairy root formation is enhanced by ultrasonication and acetosyringone treatment
Kumar, Vinod; Sharma, Ashwani; Prasad, Bellur Chayapathy Narasimha; Gururaj, Harishchandra Bhaskar & Ravishankar, Gokare Aswathanarayana

Abstract

The phytopathogenic bacteria Agrobacterium rhizogenes check for this species in other resources genetically transforms plants by transferring a portion of the resident Ri plasmid, the T-DNA to the plant. Plant species differ in their temporal competence for transformation. But various physical and chemical methods are found to enhance transformation frequency. Agrobacterium rhizogenes mediated transformation efficiency was assessed under the influence of sonication, calcium treatment, acetosyringone and macerating enzymes in suitable combinations in Nicotiana tabacum check for this species in other resources as a model system. Manual wounding resulted in 21% transformation frequency. Where as sonication resulted in 2.2 fold increase, followed by sonication with CaCl2 treatment resulted in 2.5 fold increase and sonication with acetosyringone treatment resulted in 4.1 fold increase in transformation frequency. However, sonication with macerating enzyme treatment resulted in 1.5 to 5.25-fold decrease in transformation frequency. Micro wounding through sonication followed by acetosyringone treatment enhanced transformation frequency substantially. The results of this study may be very useful in genetic manipulation of plants by Agrobacterium rhizogenes mediated gene delivery to higher plants, which are recalcitrant to A. tumefaciens mediated genetic manipulation.

Keywords
Agrobacterium rhizogenes, hairy roots, Nicotiana tabacum, transformation frequency, ultra-sonication.

 
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