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PLA NT GENETIC TRANSFO RMATION, CELL SELECTION AND SOMATIC h y b rid iz a tio n
RIBONUCLEASE ANDGLYCOSIDASEACTIVITIESINGINSENG
CULTURES TRANSGENIC FOR THE AGROBACTERIUMRHIZOGENES
rolCONCOGENE
V.P.Bulgakov, L.I.Fedoreeva, E.G.YasnetskayaandYu.N.Zhuravlev
The Institute o fBiology and Soil Science, Far EastBranch o fRussianAcademy o f
Sciences, Vladivostok, 690022Russia Pacific Institute ofBioorganic Chemistry, Far East
Branch o fRussian Academy ofSciences,
Ribonuclease activity was measured in the cell culture of Рапах ginseng
(strain 1c) and transgen ic fo r the rolC gene 1c-derived tissues by
spectrophotometry. It has been found that RNAse activity of all analysed 1c -
rolC root cultures was 1.5-3.5 times higher than that of the respective control,
being in the range of 1600-3500 Un/g FW. Since the rolC gene encodes for
an enzyme able to release active cytokinins from their glucoside conjugates
(Estruch et al., 1991), glycosidase activity of 1c-rolC roots was studied too.
This activity was foud to vary from 28 to 39 Un/g FW fo r the 1c culture and
from 55 Un/g FW to 98 Un/g FW for the 1c-rolC roots. High correlation
dependence (r=0.9147) was found for the increase in glycosidase and
ribonuclease activities for the 1c-rolC ginseng root lines. This result may be
explained by presumption that rolC protein likely to be possess glycosidase
and ribonuclease activities simultaneously. Indeed, the protein with an
apparent molecular mass of 23 kDa, which correspond to the mass of the
rolC gene product (Estruch et al., 1991), was visualized on the SDS PAG
electrophoregramms of the 1c-rolC root extracts. This protein hydrolysed p-
nitrophenyl-glucopyranosid as well as the yeast RNA. Efforts are being made
to determining the specific activities of the pure rolC encoded protein.
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