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Rol genes alter hormonal requirements for protoplast growth and modify the expression of an auxin responsive promoter
Authors:Richard Walden  Inge Czaja  Thomas Schmülling  Jeff Schell
Affiliation:(1) Max-Planck-Institut für Züchtungsforschung, Carl von Linne Weg 10, D-50837 Köln, Germany;(2) Present address: Universität Tübingen, Lehrstuhl für Allgemeine Genetik, Auf der Morgenstelle 28, D-72076 Tübingen, Germany
Abstract:Summary Growth characteristics of tobacco protoplasts containing rolA linked to its own promoter, or the rolB, or rolC genes of Agrobacterium rhizogenes linked to the Cauliflower Mosaic Virus 35S RNA promoter were compared with those from untransformed plants. RolA protoplasts require auxin and cytokinin for callus formation. Protoplasts overexpressing rolB and C form callus in the absence of exogenously applied auxin and cytokinin, respectively. Long term callus growth requires auxin, but the requirement for cytokinin is not critical. Optimal transient expression of an auxin responsive promoter element occurred at lower external levels of auxin in rolB and rolC protoplasts compared with untransformed protoplasts. Addition of putrescine was required for auxin responsive transient gene expression in rolA protoplasts suggesting that polyamines, or their products affect gene expression in rolA plants.Abbreviations T-DNA transferred DNA - TL-DNA left transferred DNA - NAA naphthalene acetic acid - PEG polyethylene glycol - GUS glucuronidase - CaMV cauliflower mosaic virus
Keywords:Auxin  Cytokinin  Rol genes  Protoplasts  CaMV 35S RNA promoter
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