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C E L L C U L T U R E FO R P R O D U C IN G P L A N T S R E S IS T A N T TO B IO T IC A N D A B IO T IC
FAC TO RS
INTRACELLULARMECHANISMSOF CADMIUMDETOXIFICATION IN
NICOTIANA PLUMBAGINIFOLIA CELL CULTURE
S.I.Fenik1'2, V.G.Solodushko1, T.B.Trofimyak1,2, Ya.B.BIume1
1Institute o fCellBiology and GeneticEngineering, acad. Zabolotny str., 148, KievGSP-
22,252650, Ukraine;
2Ternop il PedagogicalInstitute, Kryvonosa 2, Ternopil, 282009, Ukraine;
. м
Callus lines of N. plumbaginifolia tolerant to 500 mM o f CdCI2were
selected. One line (Cd-R) was selected on the MS medium containing 500
mM of CdCI2 (phytochelatin’s (PCs) synthesis wasn't inhibited). The second
line (Cd-Ri) was selected with elevating levels o f CdCI2from 400 to 500 mM
and with obligatory presence o f 100 mM buthionine sulfoximine (BSO) -
specific ihibitor o f PC’s synthesis. There were no differences in growth
characteristics of Cd-R and Cd-Ri lines growing in the presence o f 100-500
mM CdCI2. Both lines showed a slightly tolerance to Zn but not to Cu. In
contrast to wild line Cd-tolerant lines ac-cumulated lower levels o f several
organic acids. Citrate, malate and succinate were decreased to levels 26,
16,11-fold those o f the wild type in Cd-R cells and 1 0 ,3 ,2.7-fold those of
the wild type in Cd-Ri cells. At the same time all tested lines accumulated
almost equal levels o f fumarate. Treatment o f both lines with BSO (100 mM)
reduced Cd-tolerance o f Cd-R but not Cd-Ri line. In Cd-R line intracellular
PCs' concentration was about two times higher than in Cd-Ri line. In both
selected lines Cd induced accumulation o f Cd-binding proteins 42 ,40 ,37 ,
21 and 19 kDa. But there was a difference between tolerant lines in
accumulation o f low molecular weight Cd-binding proteins. In presents o f
500 mM CdCI2 Cd-R line accumulated of Cd-binding proteins 12.5,11.5,9
kDa. Instead these proteins Cd-Ri line accumulated two another Cd-binding
proteins with molecular weight 13 and 10 kDa. We suggest that intracellular
accumulation o f PCs may not correlate obligatory with Cd-tolerance of plants,
and the synthesis o f low molecular weight Cd-binding proteins is important
fo rdevelopment of the tolerance.
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