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Please use this identifier to cite or link to this item: http://www.erepo.iihr.ernet.in/handle/123456789/164

Title: Optimization of regeneration protocol and Agrobacterium mediated transformation in carnation (Dianthus caryophyllus L.)
Authors: H M, Kallesh Prasad
J B, Mythili
Lalitha, Anand
H J, Rashmi
C, Suneetha
Keywords: Carnation
Issue Date: 2009
Citation: H.M. Kallesh Prasad1, J.B. Mythili, Tejaswini , Lalitha Anand, H.J. Rashmi and C. Suneetha. 2009. Optimization of regeneration protocol and Agrobacterium mediated transformation in carnation (Dianthus caryophyllus L.). J.hort.sci. 4 (2):120-7.
Abstract: An efficient and reproducible regeneration protocol for carnation genotypes Arka Flame and IIHRS-1 has been developed from leaf and stem explants. Although IIHRS-1 showed slightly higher regeneration (55%) compared to Arka Flame (49.2%), there was no significant difference in their regeneration response. However, significant difference in regeneration potential was observed with leaf explant exhibiting higher regeneration potential (5.5 shoots/explant) as compared to (4.9) stem explant. Among the various plant growth regulator combinations tried for regeneration, the best regeneration response and maximum regeneration potential was obtained in MS medium supplemented with NAA (0.1 mg/l) and TDZ (1.0mg /l) for both the explants and genotypes used. The medium also proved suitable for inducing elongation of regenerated shoots. Rooting of in vitro formed shootlets could be induced at a greater frequency in MS medium supplemented with IAA (1.0 mg/l). Based on this protocol, transformation was carried out in genotype IIHRS-1 using leaf explants with the Agrobacterium tumefaciens LBA 4404 with binary vector pROK2 containing baculovirus chitinase gene under the control of 35S promoter and npt II serving as selectable marker. There was regeneration of putative transformants at a frequency 28.9 per cent. However, great difficulty was encountered in rooting of the shoots. Hence few shoots regenerated in selection medium at random were tested for transgene integration. Out of the three shoots tested for npt II amplification two shoots tested positive. The presence of transgene was confirmed through PCR amplification of npt II gene and dot blot analysis of chitinase gene.
URI: http://hdl.handle.net/123456789/164
Appears in Collections:Biotechnology

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