The effect of different antibiotics on the elimination of Agrobacterium and high frequency Agrobacterium-mediated transformation of indica rice (Oryza sativa L.)

A. Mohana Priya, S. Karutha Pandian and Ramesh Manikandan
Additional contact information
A. Mohana Priya: Department of Biotechnology, Alagappa University, Karaikudi, Tamil Nadu, India
S. Karutha Pandian: Department of Biotechnology, Alagappa University, Karaikudi, Tamil Nadu, India
Ramesh Manikandan: Department of Biotechnology, Alagappa University, Karaikudi, Tamil Nadu, India

Czech Journal of Genetics and Plant Breeding, 2012, vol. 48, issue 3, 120-130

Abstract: We report here the suitable explant with high efficiency of transformation and the positive effects of timentin over other antibiotics like carbenicillin and cefotaxime on the elimination of Agrobacterium tumefaciens during the genetic transformation of popular indica rice (Oryza sativa L.). The tissues assayed were embryogenic calli, embryos with endosperm contamination, intact seeds, leaf blades, leaf bases and coleoptiles. The frequency of transient β-glucuronidase (GUS) expression as revealed by histochemical assay was 90% for embryogenic calli, which was the highest among the explants used. On the basis of disc-diffusion assay, the maximum zone of inhibition (29 mm) at 250 mg/l was obtained for timentin. In tissue culture conditions the frequency of Agrobacterium recurrence after 20 days of infection was minimum (2.3%) at 200 mg/l of timentin. At 250 mg/l of timentin there was no Agrobacterium growth, besides, there were no negative effects on the callus growth unlike other antibiotics, hence it was selected as the optimum concentration for high frequency callus proliferation and regeneration.The effect of the parameters evaluated was determined by the callus proliferation during selection, reduction in browning, transient GUS expression and stable transformation efficiency (23.3%). The resultant plants were stable transformants as confirmed by a molecular analysis of the gus and hpt genes. The developed transformation protocol will be very helpful for the information on indica rice cultivars in general and on IR 64 in particular.

Keywords: Agrobacterium-mediated transformation; Agrobacterium elimination; gus reporter gene; hygromycin phosphotransferase selectable marker gene (hpt); rice (search for similar items in EconPapers)
Date: 2012
References: View complete reference list from CitEc
Citations: View citations in EconPapers (1)

Downloads: (external link)
http://cjgpb.agriculturejournals.cz/doi/10.17221/77/2011-CJGPB.html (text/html)
http://cjgpb.agriculturejournals.cz/doi/10.17221/77/2011-CJGPB.pdf (application/pdf)
free of charge

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:caa:jnlcjg:v:48:y:2012:i:3:id:77-2011-cjgpb

DOI: 10.17221/77/2011-CJGPB

Access Statistics for this article

Czech Journal of Genetics and Plant Breeding is currently edited by Ing. Markéta Knížková, (Executive Editor)

More articles in Czech Journal of Genetics and Plant Breeding from Czech Academy of Agricultural Sciences
Bibliographic data for series maintained by Ivo Andrle ().