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Isolation of novel types of Arabidopsis mutants with altered reactions to cadmium: cadmium-gradient agar plates are an effective screen for the heavy metal-related mutants

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Abstract

We are interested in elucidating the molecular mechanisms underlying plant reactions to the toxic heavy metal cadmium (Cd). To this end, we devised a new screening strategy using agar plates with a gradient of Cd concentrations, termed Cd-gradient agar plates (CGAPs), to isolate Arabidopsis mutants that displayed altered reactions to the metal. Arabidopsis M2 seeds, derived from ethyl methanesulfonate (EMS) treated seeds, were germinated on the CGAPs such that the primary root of each seedling elongated against increasing concentrations of Cd on the surface of the plate. Under these conditions, the lengths of the primary roots reliably demonstrated the degree of Cd tolerance of individual seedlings. The use of CGAPs also allowed close observation of the root reaction of each seedling to Cd without causing lethal damage. The screen identified three mutant lines, MRC-32, MRC-22 and MRC-26, which showed distinctly different characteristics. MRC-32 plants exhibited enhanced tolerance to Cd and contained Cd at higher concentrations than wild-type (WT) plants treated with the heavy metal. The whole root system of MRC-22 plants showed a Cd-phobic response. MRC-26 plants accumulated less Cd in their aboveground tissues than WT plants, suggesting that they were defective in transporting the heavy metal from roots to aboveground tissues. We also determined the likely chromosomal location of each mutation.

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Abbreviations

Cd:

Cadmium

CGAP:

Cadmium gradient agar plate

EMS:

Ethyl methanesulfonate

Mbp:

Mega base pair

Mes:

2-Morpholinoethanesulfonic acid

MRC:

Mutant altered in the reaction to Cd

MS:

Murashige and Skoog

SDS:

Sodium dodecyl sulfate

TAIR:

The Arabidopsis information resource

WT:

Wild type

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Authors and Affiliations

  1. Laboratory of Plant Genetics and Breeding, Department of Biological Production, Faculty of Bioresource Sciences, Akita Prefectural University, Kaidobata-Nishi 241-438, Shimoshinjo-Nakano, Akita, 010-0195, Japan

    Akio Watanabe, Hirono Shimizu, Namiko Satoh-Nagasawa, Hidekazu Takahashi, Kenji Sakurai & Hiromori Akagi

  2. Laboratory of Plant Nutrition and Fertilizer Science, Department of Biological Production, Faculty of Bioresource Sciences, Akita Prefectural University, Kaidobata-Nishi 241-438, Shimoshinjo-Nakano, Akita, 010-0195, Japan

    Hitomi Ito, Megumi Chiba, Azumi Ito, Shin-ichi Nakamura, Hiroyuki Hattori & Mitsuo Chino

  3. Laboratory of Plant Protection, Department of Biological Production, Faculty of Bioresource Sciences, Akita Prefectural University, Kaidobata-Nishi 241-438, Shimoshinjo-Nakano, Akita, 010-0195, Japan

    Shin-ichi Fuji

Authors
  1. Akio Watanabe
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  2. Hitomi Ito
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  3. Megumi Chiba
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  4. Azumi Ito
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  8. Hiroyuki Hattori
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  10. Namiko Satoh-Nagasawa
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  11. Hidekazu Takahashi
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  12. Kenji Sakurai
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  13. Hiromori Akagi
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Correspondence to Akio Watanabe.

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Watanabe, A., Ito, H., Chiba, M. et al. Isolation of novel types of Arabidopsis mutants with altered reactions to cadmium: cadmium-gradient agar plates are an effective screen for the heavy metal-related mutants. Planta 232, 825–836 (2010). https://doi.org/10.1007/s00425-010-1217-7

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  • DOI: https://doi.org/10.1007/s00425-010-1217-7

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