ЭлБиб

PLANT GENETIC TRANSFORMATION, CELL SELECTION AMD SOMATICHYBRIDIZATION

CONSTRUCTION OF TRANSGENIC RAPESEED PLANTS

(BRASSICA

NAPUSL.)

CONTAININGCRUCIFERINFECTINGTOBAMOVIRUS

MOVEMENT PROTEIN GEN

S.D.Zvereva, G.N.Raldugina, *Yu.L.Dorokhov, *O.V.Borisova

Institute o f Plant PhysiologyRAS, Botanicheskayastr. 35, Moskow 127276,

*AN.BelozerskyInstitute Physico-ChemicalBiology, MSU, Moscow 119899,

It have been constructed transgenic rapeseeds plants containing crucifer

infecting tobamovirus movement protein gene. The crTMV 29K movement

protein gene was cloned into the plant expression vector pRT101, carrying

the 35S promoter and the polyA signal of CaMV strain Cabb B-D. Next step

was cloning the construction flanked by 35S promoter and polyA signal into

binary

Agrobacterium

vector for plant transformation. Resulting plasmid BL208

and binary vector alone were transferred to A. tumefaciense BPA1459 by

direct transformation. Restriction analysis of A.

tumefaciense-

purified BL208

plasmid confirmed the positive clones. 5-day age

B.napus L.

cotyledons

were transformed by BPA1459 containing BL208 or binary vector alone

(control plants). The regenerated, tranformed seedlings (Ro) ( frequency o f

regeneration is 10% ) were identified by the NPTII activity and by western

blot with 29K movement protein antibody. The transgenic plants in the

phytotron conditions have normal phenotype and fertility.The next step o f

this work w ill be the determination o f genetic status (copy number o f

insertions, the obtaining R1 plants) and the ir ability o f system ically

reproduction o f TMV Ul that can not be able to infect system ically

nontransgenic rapeseeds plants.

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