ЭлБиб

PLANT GENETIC TRANSFORMATION, CELL SELECTION AND SOMATIC HYBRIDIZATION

THEINVESTIGATIONOFSUGARBEETEXPLANTSTOLERANCE

TOWARDA.

TUMEFAIENCE

ST.A281

A .G.Bogdaev,A.V.Kornienko

All-Russia research instityte o fsugarbeetandsugar. Ramon. Russia.

Agrobacterium

transformation o f sugarbeet culture tissue

in vitro

was

performed with using o f strain A281 containing binary vector system with

the predetermined gene of entomotoxine under constituent promoter. We

studied some factors were influencing transgenesis o f shoots in three

genotypes of sugarbeet obtainment: Ramonskaya 09 (sort); inbreed lines -

95155,95196. The composition of nutrient medium for cocultivation based

on MS, was modified: the quantity of saccharose -1 0 g/l, ascorbic acid - 50

mg/l. Fragments of leafstalk were used as explants being more suitable for

agrobacterium transformation.

The transformation was made more effective by the longer duration of

cocultivation up to 3 days and nights. In this connection the direct testing of

best samples was made to check their tolerance to high titres of agrobacteria.

It was found out that under long (to 20 days) contamination of leafstalks

tissues with strain A281 a weak negative reactions was witnessed - in a

form o f some layers of cells dying off a long perimeter o f thin cutting. But the

appearance of necrotic breeding grounds in greater part of explants was not

witnessed, and this fact allowed to stimulate the induction of adventive

offspring formation. As a result of testing o f the obtained regenerants in

selective media with Km it was proved that the samples possessing

resistance to Km100 demonstrated stability of the given feature. The

differences in resistance levels among samples also was stable under all

variants of testing.

Thus the tolerance to the presence of transforming strain agrobacteria

promoted viability of explants during morphogenesis of adventiv shoots. Owing

to this the production o f shoots resistant to Km100 was increased from 6%

to 15%.

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