ЭлБиб

PLANT GENETIC TRANSFORMATION, CELL SELECTION AND SOMATIC HYBRIDIZATION

REGENERATIONAND GENETIC TRANSFORMATION OF SUGARB ЕЕТ

(BETA VULGARIS L.)

N.S.Zacharchenko. Ya.I.Buryanov

Branch ofSchemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Pushchino.

Moscow region, 142292 E-Mail:

phel@fibkh.serpukhov.su

Sugar beet plants were grown on phytohormon-free Murasige and Skoog

medium (MS). Leaves with petioles formed regenerants with frequency 30-

40% (depending on cultivar) on a MS medium containing hormones 6-

benzylaminopurine (BAP) and gibberelicacid (MS-1). Calli formed from leaves

after 35 days of incubation on a MS medium, containing hormones BAP and

triiodobenzoic acid (TIBA). A fter transfer these calli on MS medium with

BAP (1,0 mg/i) they formed differentiated shoots. The normal plants were

obtained from all 5 cultivars of sugar beet. Sugar beet genetic transformation

was carried out by method of cocultivation of petioles with different straines

Agrobacterium tumefaciens.

Sugar beet explants transformation with strain

C58 containing" undisarmed'' C5S plasmid, or C58/21 plasmid harbouring

methyltransferase M.EcoRIl bacterial gene in T-DNA region, induced tumor

formation. Analysis of tumors growing on hormone-free medium revealed

presence of nopaline synthase activity. DNA from C58 tumor was subjected

to hydrolysis by R.EcoRl endonuclease much more deaper, than DNA from

C58/21 tissue, but both DNA were subjected in equal degree to hydrolysis

by R.BstNI endonuclease, which recognizes the same site, but cut DNA

independently of internal cytosine methylation. These data indicate on

additional DNA modification of transformants by DNA methyltransferase

EcoRII. Sugar beet explants transformation with Agrobacterium tumefaciens

GV3101 with pMP90RK helper plasmid and rolC gene on a binary vector

pPCV002 led to root development on MS-1 medium, contained 50 mg/l

kanamycin. The presence of neomycin phosphotransferase activity II in a

root tissues of sugar beet transformants was shown.

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