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Plant Physiol, August 2001, Vol. 126, pp. 1519-1526
Enhanced Copper Tolerance in Silene vulgaris (Moench)
Garcke Populations from Copper Mines Is Associated with
Increased Transcript Levels of a 2b-Type Metallothionein
Gene1
Nathalie A.L.M.
van Hoof,
Viivi H.
Hassinen,
Henk W.J.
Hakvoort,
Koos F.
Ballintijn,
Henk
Schat,*
Jos A.C.
Verkleij,
Wilfried H.O.
Ernst,
Sirpa O.
Karenlampi, and
Arja I.
Tervahauta
Department of Ecology and Ecotoxicology of Plants, Faculty of
Biology, Vrije Universiteit, De Boelelaan 1087, 1081 HV Amsterdam, The
Netherlands (N.A.L.M.v.H., H.W.J.H., K.F.B., H.S., J.A.C.V., W.H.O.E.);
and Department of Biochemistry, University of Kuopio, P.O. Box 1627, 70211 Kuopio, Finland (V.H.H., S.O.K., A.I.T.)
Silene vulgaris (Moench) Garcke has evolved
populations with extremely high levels of copper tolerance. To evaluate
the role of metallothioneins (MTs) in copper tolerance in S.
vulgaris, we screened a cDNA library derived from a highly
copper-tolerant population using Arabidopsis-based MT probes and
identified an MT2b-like gene. When expressed in yeast, this gene,
SvMT2b, restored cadmium and copper tolerance in
different hypersensitive strains. Northern-blot analysis and
quantitative reverse transcriptase-PCR showed that plants from the
copper-tolerant S. vulgaris populations had
significantly higher transcript levels of SvMT2b than
plants from the copper-sensitive populations, both in roots and shoots and with and without copper exposure. Southern-blot analysis suggested that the higher expression of the latter allele was caused by gene
amplification. Segregating families of crosses between copper-sensitive and copper-tolerant plants exhibited a 1 to 3 segregation for SvMT2b expression. Allele-specific PCR showed that
low-expression F3 plants were homozygous for the allele
inherited from the copper-sensitive parent, whereas high-expression
plants possessed at least one allele from the tolerant parent.
SvMT2b expression did not cosegregate with copper
tolerance in crosses between sensitive and tolerant plants. However, a
significant cosegregation with copper tolerance did occur in families
derived from crosses between moderately tolerant F3 plants
with different SvMT2b genotypes. Thus, overexpression of
SvMT2b conferred copper tolerance although only within
the genetic background of a copper tolerant plant.
1
This work was supported by the European
Community (Environment Research Program: Environment and Climate,
contract no. ENV 4-CT95-0083 [Phytorehab]).
*
Corresponding author; e-mail hschat{at}bio.vu.nl; fax
31-20-4447123.
© 2001 American Society of Plant Physiologists
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