Arabidopsis metallothioneins 2a and 3 enhance resistance to cadmium when expressed in Vicia faba guard cells.
The Arabidopsis metallothionein genes AtMT1 and AtMT2 confer Cd(II) resistance to Cd(II)-sensitive yeast, but it has not been directly shown whether they or other metallothioneins provide the same protection to plants. We tested whether AtMT2a and AtMT3 can confer Cd(II) resistance to plant cells by introducing GFP- or RFP-fused forms into guard cells of Vicia faba by biolistic bombardment. AtMT2a and AtMT3 protected guard cell chloroplasts from degradation upon exposure to Cd(II), an effect that was confirmed using an FDA assay to test the viability of the exposed guard cells. AtMT2a- and AtMT3-GFP were localized in the cytoplasm both before and after treatment of V. faba guard cells or Arabidopsis protoplasts with Cd(II), and the levels of reactive oxygen species were lower in transformed guard cells than in non-transformed cells after Cd(II)-treatment. These results suggest that the Cd(II)-detoxification mechanism of AtMT2a and AtMT3 may not include sequestration into vacuoles or other organelles, but does involve reduction of the level of reactive oxygen species in Cd(II)-treated cells. Increased expression of AtMT2a and AtMT3 was observed in Arabidopsis seedlings exposed to Cd(II). Together, these data support a role for the metallothioneins AtMT2a and AtMT3 in Cd(II) resistance in intact plant cells.
Lee
Joohyun
J
National Research Laboratory of Phytoremediation, Division of Molecular Life Sciences, Pohang University of Science and Technology, Korea.
Shim
Donghwan
D
Song
Won-Yong
WY
Hwang
Inhwan
I
Lee
Youngsook
Y
D029681:Arabidopsis Proteins / Q000235:genetics / Q000502:physiology*; D002104:Cadmium / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D002736:Chloroplasts / Q000187:drug effects / Q000502:physiology; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D004351:Drug Resistance; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008668:Metallothionein / Q000235:genetics / Q000502:physiology*; D008856:Microscopy, Fluorescence; D019441:Plant Epidermis / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D018515:Plant Leaves / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D021381:Protein Transport / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D014170:Transformation, Genetic; D031307:Vicia faba / Q000187:drug effects / Q000235:genetics / Q000502:physiology*
D029681:Arabidopsis Proteins; C100789:MT2A protein, Arabidopsis; D017382:Reactive Oxygen Species; D011993:Recombinant Fusion Proteins; D002104:Cadmium; D049452:Green Fluorescent Proteins; D008668:Metallothionein