Peer-Reviewed Publications

    Impairing both HMA4 homeologs is required for cadmium reduction in tobacco

    Liedschulte, V.; Laparra, H.; Battey, J. N. D.; Schwaar, J. D.; Broye, H.; Mark, R.; Klein, M.; Goepfert, S.; Bovet, L.
    Published
    Nov 23, 2016
    DOI
    10.1111/pce.12870
    PMID
    27880006
    Link to publication
    Topic
    Summary

    In tobacco, the heavy metal P1B-ATPases HMA4.1 and HMA4.2 function in root-to-shoot zinc and cadmium transport. We present greenhouse and field data that dissect the possibilities to impact the two homeologous genes in order to define the best strategy for leaf cadmium reduction. In a first step, both genes were silenced using an RNAi approach leading to >90% reduction of leaf cadmium content. To modulate HMA4 function more precisely, mutant HMA4.1 and HMA4.2 alleles of a TILLING population were combined. As observed with RNAi plants, knockout of both homeologs decreased cadmium root-to-shoot transfer by >90%. Analysis of plants with segregating null and wild-type alleles of both homeologs showed that one functional HMA4 allele is sufficient to maintain wild type cadmium levels. Plant development was affected in HMA4 RNAi and double knockout plants which included retarded growth, necrotic lesions, altered leaf morphology and increased water content. The combination of complete functional loss (nonsense mutation) in one homeologous HMA4 gene and the functional reduction in the other HMA4 gene (missense mutation) is proposed as strategy to limit cadmium leaf accumulation without developmental effects.

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