ЭлБиб

PLANT GENETIC TRANSFORMATION, CELL SELECTION AND SOMATIC HYBRIDIZATION

TRANSFORMATION O FBRASSICA NAPUS

USINGAGROBACTERIUM

VECTORS

E.Calle Cossio1& A.A Shadenkov2, S.I.Ivashuta1.

1AllRussianReseach Institute o ffodder crops, Lugovaya, Russia.

2All RussianReseach InstituteforAgriculturalBiotechnology,Moscow, Russia.

Modem varietes o f

B.napus

possess impoverished genetic potential

because they were bred on the base o f two parental genotypes cv. Oro

(Canada) and cv. Bronowski (Poland). Oilseed population can be saturated

with novel genes by genetic transformation techniques. S ix background

genotypes o f

Brassica napus

(ssp.oleifera, var.annua) were used as

recipients. Transformation procedure undergo in accordance with protocol

developed by Moloney (Moloney et al. ,1989). Shoot regeneration frequencies

fo r cotyledonary petioles and h y p o c o ty ls were II -61% and 0-56%

respectively. Multiple shoots often formed from a single explant. The binary

p la sm id pH22Kneo w ith p lan t de fens ins gene (ac) m ob ilised to

A.tumefaciens strain C58CI (pGV3850) employed in this study. The first

transformed shoots developed after about 14 days and after 21 days vary

few new shoots were produced. Transformation frequencies for petioles was

0,9-7%. Hypocotyles produced Km-resistant shoots in low frequencies (<3%)

and only when cultured on regeneration medium for 24h before transformation.

The fact of plant transformation was confirmed by marker NPT activity test.

The NPT actively was present in leaves from two out of 25 putative

transformants. Analogous to this result large number o f escapes was

observed with B. napus by Fry (Fry et al., 1987). This fact can be explaind

by endogenous resistanse o f B. napus to a range of aminoglycosides,

including kanamicin.

273

Научная электронная библиотека ЦНСХБ