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The effect of an elevated cytokinin level using the ipt gene and N 6-benzyladenine on single node and intact potato plant tuberization in vitro
Authors:Ivan Gális  Ji?í Macas  Josef Vlasák  Milo? Ond?ej  Henri A Van Onckelen
Institution:(1) Institute of Plant Molecular Biology, Academy of Sciences of the Czech Republic, Braniscaronovská 31, 370 05 Ccaroneské Budecaronjovice, Czech Republic;(2) Department of Biology, Universitaire Instelling Antwerpen, Universiteitsplein 1, B-2610 Wilrijk, Belgium
Abstract:Two models of potato (Solanum tuberosum L.) tuberization in vitro (intact plants and single nodes) were used to study the role of cytokinins in this process. We applied hormone in two different ways. The exogenous addition of 10 mg · L-1 N 6-benzyladenine (BA) into the tuberization medium resulted in advanced tuber formation in intact plants, and microtubers appeared 10–20 days earlier than in the experiments in which no cytokinin was supplied. Transformation with the Agrobacterium tumefaciens ipt gene provided potato clones with endogenously elevated cytokinin levels (3–20 times higher zeatin riboside content in different clones). The onset of tuberization in intact ipt-transformed plants with low transgene expression was advanced in comparison with control material, and exogenously applied BA further promoted the tuberization process. On the contrary, tuberization was strongly inhibited in ipt-transformed nodes, and an external increase of the cytokinin level caused complete inhibition of expiant growth. In untransformed (control) nodes cytokinin application resulted in primary and secondary tuber formation, which depended on the BA concentration in cultivation media.Abbreviations BA N 6-benzyladenine - PCR polymerase chain reaction - HPLC high performance liquid chromatography - ELISA enzyme-linked immunosorbent assay - NAA agr-naphthylacetic acid
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