Abstract
Among the plant species cultivated for human consumption, tobacco is one that accumulates cadmium to a significant degree. In order to reduce the Cd levels in tobacco leaves, we have introduced into the tobacco genome a gene encoding a mammalian metallothionein, since these low-molecular-weight cysteine-rich proteins fix Cd and other heavy metals. Here we describe the Cd accumulation characteristics observed during two years of greenhouse tests and one year of field trial of tobacco plants expressing a metallothionein gene. In all three tests, leaf Cd levels were markedly decreased. For instance, in the field trial, Cd levels in the leaf lamina tissue of the transformed line were decreased by 73% compared to controls. The decrease in leaf Cd was correlated with an increase in Cd in the roots and stems. The plants had normal growth characteristics, and the distribution of other ions was not affected by the expression of the metallothionein gene. Even though the transformation/expression vector used here can cause frequent post-transcriptional gene silencing, comparison of hemizygous and homozygous individuals of the plant line expressing the metallothionein gene gave little evidence for silencing.
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References
Brandle JE, Labbe H, Hattori J, Miki BL: Field performance and heavy metal concentrations of transgenic fluecured tobacco expressing a mammallian metallothionein βglucuronidase gene fusion. Genome 36: 255–260 (1993).
Burkitt A, Lester P, Nickless G: Distribution of heavy metals in the vicinity of an industrial complex. Nature 238: 327–328 (1972).
Cunningham SD, Ow DW: Promises and prospects of phytoremediation. Plant Physiol 110: 715–719 (1996).
Dellaporta SL, Wood J, Hicks, J.B: A plant DNA minipreparation: version II. Plant Mol Biol Rep 1: 19–21 (1983).
Dorlhac de Borne F, de Roton C, Delon R, Chupeau Y: Etude du comportement agronomique de tabacs industriels transgéniques présentant une activité nitrate réductase élevée. Ann Tabac 26: 19–38 (1984).
Elmayan T, Tepfer M: Synthesis of a bifunctional metallothionein/β-glucuronidase fusion protein in transgenic tobacco plants as a means of reducing leaf cadmium levels. Plant J 6: 433–440 (1994).
Elmayan T, Tepfer M: Evaluation in tobacco of the organ specificity and strength of the rolD promoter, domain A of the 35S promoter and the 35S2 promoter. Transgen Res 4: 388–396 (1995).
Elmayan T, Vaucheret H: Expression of single copies of a strongly expressed 35S transgene can be silenced posttranscriptionally. Plant J 9: 787–797 (1996).
Feinberg AP, Vogelstein B: A technique for radiolabelling restriction endonuclease fragments to a high specific activity. Anal Biochem 132: 6–13 (1983).
Gramsick C, Huel G, Moreau T, Sarmini H: The combined effect of tobacco and alcohol comsumption on the level of lead and cadmium in blood. Sci Total Environ 41: 207–217 (1985).
Hamer DH: Metallothioneins. Annu Rev Biochem 55: 913–951 (1986).
Jarvis SC, Jones LHP, Hopper MJ: Cadmium uptake from solution by plants and its transport from roots to shoots. Plant Soil 44: 179–191 1976.
Karin M, Richards RI: Human metallothionein genes: molecular cloning and sequence analysis of the mRNA. Nucl Acids Res 10: 3165–3173 (1982).
Kay R, Chan A, Daly M, McPherson J: Duplication of CaMV 35S promoter sequences creates a strong enhancer for plant genes. Science 236: 1299–1302 (1987).
Kuboi T, Noguchi A, Yazaki J: Family-dependent cadmium accumulation characteristics in higher plants. Plant Soil 92: 405–415 (1986).
Maiti IB, Wagner GJ, Yeargan R, Hunt AG: Inheritance and expression of the mouse metallothionein gene in tobacco. Plant Physiol 91: 1020–1024 (1989).
Maiti IB, Murphy JF, Shaw JG, Hunt AG: Plants that express a potyvirus proteinase gene are resistant to virus infection. Proc Natl Acad Sci USA 90: 6110–6114 (1993).
Mench M, Tancogne J, Gomez A, Juste C: Cadmium bioavailability to Nicotiana tabacum L., Nicotiana rustica L., and Zea mays L. grown in soil amended or not amended with cadmium nitrate. Biol Fert Soils 8: 48–53 (1989).
Misra S, Gedamu L: Heavy metal tolerant transgenic Brassica napus L. and Nicotiana tabacum L. plants. Theor Appl Genet 78: 161–168 (1989).
Nielsen KB, Atkins CL, Winge DR: Distinct metalbinding configurations in metallothionein. J Biol Chem 260: 5324–5350 (1985).
Palauqui J-C, Vaucheret H: Field trial anaysis of nitrate reductase co-suppression: a comparative study of 38 combinations of transgene loci. Plant Mol Biol 29: 149–159 (1995).
Pan A, Yang M, Tie F, Li L, Chen Z, Ru B: Expression of mouse metallothioneinI gene confers cadmium resistance in transgenic tobacco plants. Plant Mol Biol 24: 341–351 (1994).
Phu-Lich N, Truhaut R, Claude JR, Tancogne J, Schiltz P: Le cadmium et le tabac. Bull Ass Rech Nicotian 1990: 50–64 (1990).
Salt DE, Blaylock M, Kumar N, Dushenkov V, Ensley BD, Chet I, Raskin I: Phytoremediation: a novel strategy for the removal of toxic metals from the environment using plants. Bio/technology 13: 468–474 (1995).
Sambrook J. Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Sweeney RA, Rexroad PR: Comparison of LECO FP228 ‘nitrogen determinator’ with AOAC copper catalyst Kjeldahl method for crude protein. J Ass Off Anal Chem 70: 1028–1030 (1987).
Wagner GJ: Accumulation of cadmium in crop plants and its consequences to human health. Adv Agron 51: 173–212 (1993).
Wagner GJ, Yeargan R: Variation in cadmium accumulation potential and tissue distribution of cadmium in tobacco. Plant Physiol 82: 274–279 (1986).
Walinga I, van Vark W, Houba VJG, van der Lee JJ: Soil and plant analysis, a series of syllabi, part 7: Plant analysis procedures.Wageningen Agricultural University, Wageningen, Netherlands (1989).
Yeargan R, Maiti IB, Nielsen MT, Hunt AG, Wagner GJ: Tissue partitioning of cadmium in transgenic tobacco seedlings and field grownplants expressing themousemetallothionein I gene. Transgen Res 1: 261–267 (199).
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de Borne, F.D., Elmayan, T., de Roton, C. et al. Cadmium partitioning in transgenic tobacco plants expressing a mammalian metallothionein gene. Molecular Breeding 4, 83–90 (1998). https://doi.org/10.1023/A:1009669412489
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DOI: https://doi.org/10.1023/A:1009669412489