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Changes in phytochelatins and their biosynthetic intermediates in red spruce (Picea rubens Sarg.) cell suspension cultures under cadmium and zinc stress

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Abstract

Cell suspension cultures of red spruce (Picea rubens Sarg.) were selected to study the effects of cadmium (Cd) and zinc (Zn) on phytochelatins (PCs) and related metabolites after 24 h exposure. The PC2 and its precursor, γ-glutamylcysteine (γ-EC) increased two to fourfold with Cd concentrations ranging from 12.5 to 200 μM as compared to the control. However, Zn-treated cells showed a less than twofold increase in γ-EC and PC2 levels as compared to the control even at the highest concentration of 800 μM. In addition, unidentified higher chain PCs were also found in both the Cd and Zn treated cells and they increased significantly with increasing concentrations of Cd and Zn. The cellular ratio of PC2 : Cd or Zn content clearly indicated that Cd (with ratios ranging from 0.131 to 0.546) is a more effective inducer of PC2 synthesis/accumulation than Zn (with ratios ranging from 0.032 to 0.102) in red spruce cells. A marginal decrease in glutathione (GSH) was observed in both Cd and Zn treated cells. However, the GSH precursor, cysteine, declined twofold with all Cd concentrations while the decrease with Zn was 1.5–2-fold only at the higher treatment concentrations of Zn as compared to control. In addition, changes in other free amino acids, polyamines, and inorganic ions were also studied. These results suggest that PCs and their biosynthetic intermediates play a significant role in red spruce cells protecting against Cd and Zn toxicity.

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Abbreviations

ACN:

Acetonitrile

γ-EC:

γ-glutamylcysteine

GSH:

Glutathione

HPLC:

High-performance liquid chromatography

mBBr:

Monobromobimane

NAC:

N-acetyl-l-cysteine

PCs:

Phytochelatins

TFA:

Trifluroacetic acid

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Acknowledgments

The authors are grateful to Dr. Beth A. Ahner and Dr. James E. Gawel for their valuable suggestions. The authors are thankful to Prof. Curtis Givan, Dr. Kevin T. Smith, Prof. Leland Jahnke, and Prof. Subhash C. Minocha for critical reviews of this manuscript. Thanks are also extended to Benjamin Mayer and Kenneth R. Dudzik for technical assistance.

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  1. Northern Research Station, USDA Forest Service, 271 Mast Road, Durham, NH, 03824, USA

    P. Thangavel, Stephanie Long & Rakesh Minocha

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  1. P. Thangavel
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  2. Stephanie Long
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Correspondence to P. Thangavel.

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Thangavel, P., Long, S. & Minocha, R. Changes in phytochelatins and their biosynthetic intermediates in red spruce (Picea rubens Sarg.) cell suspension cultures under cadmium and zinc stress. Plant Cell Tiss Organ Cult 88, 201–216 (2007). https://doi.org/10.1007/s11240-006-9192-1

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