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Summary: On the species Arabidopsis thaliana, Cucurbita pepo and
some conifers (black pine, redwood) plant cell transformation with plasmids
(bearing chimeric genes or wild ones) is investigated. Also, it is studied
regeneration of transgenic plants, and activity of chimeric genes depending
on plant developmental stage.
Keywords: Transformation, transgenic plant regeneration, chimeric genes, beta-glucuronidase gene as reporter gene, regulation of gene expression during plant development, Agrobacterium sp.
Research goals: The aim is to comprehend the gene activity
pattern(s) crucial in higher plant development, primarly that ones which
are active during embryogenesis and regeneration in general, also to
investigate mechanisms of their activation. Results - Agrobacterium
tumefaciens - mediated transformation of root and stem explants was
performed using the strain LBA4404 containing the binary plasmid pBI121 or
its constructs with the fragments of the DC-8 promotor. - It was examined
a number of parameters (ecotypes, number of inoculations, different
transformation markers) as well as physiological conditions (photoperiod,
temperature) in relation to "in planta" transformation efficiency for A.
thaliana. In order to determine the segregation of phenotypic markers (GUS
activity, kanamycin resistance, chlor sulfuron resistance) more than a
hundred T2 lines were analyzed. The segregation pattern of transformation
markers as well as Southern hybridization analysis showed that more than
50% of the lines had one copy of inserted T-DNA while the others had more
the one per genome. - Regeneration of hairy roots in black pine (Pinus
nigra Arn.) and red wood (Sequoia sempervirens) explants (two-month-old
epicotyl and stem segment has achieved after infection with the wild
strain of A. rhizogenes 8196 or trans conjugant strain A.tumefaciens
bearing Ri plasmid C58CI (pArA4abc). The presence of Ri T-DNA (pRi8196) in
regerated black pine roots was confirmed by mannopine content.
COOPERATION - PROJECTS
Name of project
: 1-08-268 Regeneracija i transformacija biljnih
stanica Name of institution: Prirodoslovno-matematički fakultet,
Prirodoslovni odjeli, Sveučilište u Zagrebu City: 10000 - Zagreb, Croatia Other information about the project.