Detection of the reporter and selection genes in transformed hop (Humulus lupulus L.)

Agrobacterium-mediated transformation of hop nodal explants with meristems was used for the introduction of a gus reporter gene and nptII plant selection gene into Slovenian hop cv. Aurora. Emerging hop regenerants were previously tested for the gus gene expression by histochemical analysis of β-glu...

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Main Authors: Suzana ŠKOF, Zlata LUTHAR
Format: Article
Language:English
Published: University of Ljubljana Press (Založba Univerze v Ljubljani) 2005-10-01
Series:Acta Agriculturae Slovenica
Subjects:
Online Access:https://journals.uni-lj.si/aas/article/view/15236
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author Suzana ŠKOF
Zlata LUTHAR
author_facet Suzana ŠKOF
Zlata LUTHAR
author_sort Suzana ŠKOF
collection DOAJ
description Agrobacterium-mediated transformation of hop nodal explants with meristems was used for the introduction of a gus reporter gene and nptII plant selection gene into Slovenian hop cv. Aurora. Emerging hop regenerants were previously tested for the gus gene expression by histochemical analysis of β-glucoronidase (GUS) activity. Approximately six months after the transformation procedure, PCR molecular analysis of shoots originating from previously GUS positive regenerants was performed to check integration of the reporter and selection genes into the hop genome. We also compared whether there were any differences in transgene integration in relation to the intensity of gus gene expression (intensive blue coloration on a larger proportion of the leaf surface or just a few blue spots) revealed by GUS-assay. In both cases, the majority of shoots had both transgenes integrated (47.7 or 55.3%) and in smaller number of shoots both transgenes were missing (38.6 or 18.8%). The fewest shoots analyzed showed just gus (2.3 or 8.9%) and slightly more nptII (11.4 or 17.0%) gene presence.
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spelling doaj.art-51ab6854ecca431f9b92f0231087029f2023-12-14T22:15:42ZengUniversity of Ljubljana Press (Založba Univerze v Ljubljani)Acta Agriculturae Slovenica1854-19412005-10-01852Detection of the reporter and selection genes in transformed hop (Humulus lupulus L.)Suzana ŠKOF0Zlata LUTHAR1SI-1111 Ljubljana, Jamnikarjeva 101SI-1111 Ljubljana, Jamnikarjeva 101Agrobacterium-mediated transformation of hop nodal explants with meristems was used for the introduction of a gus reporter gene and nptII plant selection gene into Slovenian hop cv. Aurora. Emerging hop regenerants were previously tested for the gus gene expression by histochemical analysis of β-glucoronidase (GUS) activity. Approximately six months after the transformation procedure, PCR molecular analysis of shoots originating from previously GUS positive regenerants was performed to check integration of the reporter and selection genes into the hop genome. We also compared whether there were any differences in transgene integration in relation to the intensity of gus gene expression (intensive blue coloration on a larger proportion of the leaf surface or just a few blue spots) revealed by GUS-assay. In both cases, the majority of shoots had both transgenes integrated (47.7 or 55.3%) and in smaller number of shoots both transgenes were missing (38.6 or 18.8%). The fewest shoots analyzed showed just gus (2.3 or 8.9%) and slightly more nptII (11.4 or 17.0%) gene presence. https://journals.uni-lj.si/aas/article/view/15236hoptransformationAgrobacterium tumefaciensgus genenptII genePCR
spellingShingle Suzana ŠKOF
Zlata LUTHAR
Detection of the reporter and selection genes in transformed hop (Humulus lupulus L.)
Acta Agriculturae Slovenica
hop
transformation
Agrobacterium tumefaciens
gus gene
nptII gene
PCR
title Detection of the reporter and selection genes in transformed hop (Humulus lupulus L.)
title_full Detection of the reporter and selection genes in transformed hop (Humulus lupulus L.)
title_fullStr Detection of the reporter and selection genes in transformed hop (Humulus lupulus L.)
title_full_unstemmed Detection of the reporter and selection genes in transformed hop (Humulus lupulus L.)
title_short Detection of the reporter and selection genes in transformed hop (Humulus lupulus L.)
title_sort detection of the reporter and selection genes in transformed hop humulus lupulus l
topic hop
transformation
Agrobacterium tumefaciens
gus gene
nptII gene
PCR
url https://journals.uni-lj.si/aas/article/view/15236
work_keys_str_mv AT suzanaskof detectionofthereporterandselectiongenesintransformedhophumuluslupulusl
AT zlataluthar detectionofthereporterandselectiongenesintransformedhophumuluslupulusl