Optimization of Regeneration Protocol and Agrobacterium Mediated Transformation in Carnation (Dianthus caryophyllus L.)

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 resp...

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Autores principales:	H M Kallesh Prasad, J B Mythili, Tejaswini ., Lalitha Anand, H J Rashmi, C Suneetha
Formato:	article
Lenguaje:	EN
Publicado:	Society for Promotion of Horticulture - Indian Institute of Horticultural Research 2009
Materias:	carnation genotype morphogenesis agrobacterium mediated transformation Plant culture SB1-1110
Acceso en línea:	https://doaj.org/article/28faf25edff241d2b2d7c1cb1e2472bf
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spelling	oai:doaj.org-article:28faf25edff241d2b2d7c1cb1e2472bf2021-12-02T12:10:31ZOptimization of Regeneration Protocol and Agrobacterium Mediated Transformation in Carnation (Dianthus caryophyllus L.)0973-354X2582-4899https://doaj.org/article/28faf25edff241d2b2d7c1cb1e2472bf2009-12-01T00:00:00Zhttps://jhs.iihr.res.in/index.php/jhs/article/view/528https://doaj.org/toc/0973-354Xhttps://doaj.org/toc/2582-4899An 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 various plant growth regulator combinations tested 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 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 Agrobacterium tumefaciens LBA 4404 with binary vector pROK2 containing baculovirus chitinase gene under the control of 35S promoter with npt II serving as selectable marker. There was regeneration of putative transformants at a frequency of 28.9%. However, great difficulty was encountered in rooting of shoots. Hence a few shoots regenerated on 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.H M Kallesh PrasadJ B MythiliTejaswini .Lalitha AnandH J RashmiC SuneethaSociety for Promotion of Horticulture - Indian Institute of Horticultural Researcharticlecarnationgenotypemorphogenesisagrobacterium mediated transformationPlant cultureSB1-1110ENJournal of Horticultural Sciences, Vol 4, Iss 2, Pp 120-127 (2009)
institution	DOAJ
collection	DOAJ
language	EN
topic	carnation genotype morphogenesis agrobacterium mediated transformation Plant culture SB1-1110
spellingShingle	carnation genotype morphogenesis agrobacterium mediated transformation Plant culture SB1-1110 H M Kallesh Prasad J B Mythili Tejaswini . Lalitha Anand H J Rashmi C Suneetha Optimization of Regeneration Protocol and Agrobacterium Mediated Transformation in Carnation (Dianthus caryophyllus L.)
description	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 various plant growth regulator combinations tested 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 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 Agrobacterium tumefaciens LBA 4404 with binary vector pROK2 containing baculovirus chitinase gene under the control of 35S promoter with npt II serving as selectable marker. There was regeneration of putative transformants at a frequency of 28.9%. However, great difficulty was encountered in rooting of shoots. Hence a few shoots regenerated on 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.
format	article
author	H M Kallesh Prasad J B Mythili Tejaswini . Lalitha Anand H J Rashmi C Suneetha
author_facet	H M Kallesh Prasad J B Mythili Tejaswini . Lalitha Anand H J Rashmi C Suneetha
author_sort	H M Kallesh Prasad
title	Optimization of Regeneration Protocol and Agrobacterium Mediated Transformation in Carnation (Dianthus caryophyllus L.)
title_short	Optimization of Regeneration Protocol and Agrobacterium Mediated Transformation in Carnation (Dianthus caryophyllus L.)
title_full	Optimization of Regeneration Protocol and Agrobacterium Mediated Transformation in Carnation (Dianthus caryophyllus L.)
title_fullStr	Optimization of Regeneration Protocol and Agrobacterium Mediated Transformation in Carnation (Dianthus caryophyllus L.)
title_full_unstemmed	Optimization of Regeneration Protocol and Agrobacterium Mediated Transformation in Carnation (Dianthus caryophyllus L.)
title_sort	optimization of regeneration protocol and agrobacterium mediated transformation in carnation (dianthus caryophyllus l.)
publisher	Society for Promotion of Horticulture - Indian Institute of Horticultural Research
publishDate	2009
url	https://doaj.org/article/28faf25edff241d2b2d7c1cb1e2472bf
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Optimization of Regeneration Protocol and Agrobacterium Mediated Transformation in Carnation (Dianthus caryophyllus L.)

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