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PLANT GENETIC TRANSFORMATION. CELL SELECTION AND SOMATIC HYBRIDIZATION
USE OF ANTISENSERNAFOR INHIBITION OFTUMOUR FORMATION IN
PLANTS
V.V.Alekseeva, E.B.Rukavtsova, Ia.I.Buryanov
Branch ofInstitute o fBioorganic Chemistry, PushinoE-Mail: r
Infection of plants by Agrobacterium causes tumour formation in plants.
It was shown expression of agrobacterium genes in tumour cells such as
phytohormone genes.
The aim o f this work was production of tobacco plants with cytokinin
tm r gene in sense and antisense orientation.
It is known, antisense RNA strategy have been used in genetic
engeneering o f plants in orderto inhibit definite genes .
The tm r gene was cloned to produce plants resistent to Agrobacterium
tumours. Plasm id pTiBo542 was used as donor of gene. Gene was
synthesized by PCR and inserted into plant vector pRT103 containing 35S
promotor and polyadenylation site. This cassette carrying the tm r gene under
control of 35S promoter was inserted into binary vector pGA482. This
construction was introduced into Agrobacterium tumefaciensstrain LBA4404
(pAL.4404).
DNA hybridization of obtaining clones was confirmed the presence of
tm r gene.
The resulting Agrobacterium was used for the leaf disk transformation
o f tobacco plants and shoots were obtained on medium with 50 mg/ml
kamamycin. In the case o f himeric tm r gene in sense orientation, shoot
formation was produced on hormonfree medium. Transgenic plants had
another phenotype and were enable to root formation. It was detected onto
gene expression. It was nesessary addition of hormones into medium for
obtaining shoots with tm r gene in antisense orientation.
It was demonstrated presence of tm r gene in transgenic plants using
PCR.
We plan infection of transformed plants by Agrobacterium strains
containing tm r gene in opposite orientasion. We suppose that expression of
agrobacterium phytohormone gene in transgenic plants will be suppressed
by antisense RNA.
Thise strategy will be useful for the defence of plants from Agrobacterium
tumours.
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