Agrobacterium-mediated sunflower transformation (Helianthus annuus L.) in vitro and in Planta using strain of LBA4404 harboring binary vector pBi2E with dsRNA-suppressor proline dehydrogenase gene

To estimate the efficiency of proline dehydrogenase gene suppression towards increasing of sunflower (Helianthus annuus L.) tolerance level to water deficit and salinity, we employed strain LBA4404 harboring pBi2E with double-stranded RNA-suppressor, which were prepared on basis arabidopsis ProDH1 g...

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Published inTSitologiia i genetika Vol. 48; no. 4; p. 19
Main Authors Tishchenko, E N, Komisarenko, A G, Mikhal'skaia, S I, Sergeeva, L E, Adamenko, N I, Morgun, B V, Kochetov, A V
Format Journal Article
LanguageRussian
Published Ukraine 01.07.2014
Subjects
Agrobacterium - genetics
Gene Transfer Techniques
Genes, Plant
Genes, Suppressor
Genetic Vectors
Helianthus - enzymology
Helianthus - genetics
Plants, Genetically Modified - enzymology
Plants, Genetically Modified - genetics
Proline Oxidase - genetics
RNA, Double-Stranded - genetics
Seeds - enzymology
Seeds - genetics
Transformation, Genetic
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Summary:To estimate the efficiency of proline dehydrogenase gene suppression towards increasing of sunflower (Helianthus annuus L.) tolerance level to water deficit and salinity, we employed strain LBA4404 harboring pBi2E with double-stranded RNA-suppressor, which were prepared on basis arabidopsis ProDH1 gene. The techniques of Agrobacterium-mediated transformation in vitro and in planta during fertilization sunflower have been proposed. There was shown the genotype-depended integration of T-DNA in sunflower genome. PCR-analysis showed that ProDH1 presents in genome of inbred lines transformed in planta, as well as in T1- and T2-generations. In trans-genic regenerants the essential accumulation of free L-proline during early stages of in vitro cultivation under normal conditions was shown. There was established the essential accumulation of free proline in transgenic regenerants during cultivation under lethal stress pressure (0.4 M mannitol and 2.0% sea water salts) and its decline upon the recovery period. These data are declared about effectiveness of suppression of sunflower ProDH and gene participation in processes connected with osmotolerance.
ISSN:0564-3783