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Differential responses of microsomal proteins and metabolites in two contrasting cadmium (Cd)-accumulating soybean cultivars under Cd stress

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Abstract

While there are significant genotypic differences in cadmium (Cd) uptake and distribution in soybean cultivars, little attention has been paid to the underlying molecular mechanisms. We adopted a comparative proteomic approach coupled with metabolite analysis to examine Cd uptake and translocation in two contrasting Cd-accumulating soybean cultivars, Enrei and Harosoy, which accumulate higher amount of Cd in the roots and aerial parts, respectively. Proteins extracted from the root microsomal fraction were evaluated by immunoblot analysis using different subcellular marker proteins. Analysis of control and Cd-exposed samples by two-dimensional gel electrophoresis coupled with mass spectrometry revealed a total of 13 and 11 differentially expressed proteins in the Enrei and Harosoy cultivars, respectively. Metabolome profiling identified a total of 32 metabolites, the expression of 18 of which was significantly altered in at least in one cultivar in response to Cd stress. Analysis of the combined proteomic and metabolomic results revealed that proteins and amino acids associate with Cd-chelating pathways are highly active in the Enrei cultivar. In addition, proteins associated with lignin biosynthesis are significantly upregulated in the Enrei cultivar under Cd stress. Our results indicate that in the Enrei cultivar, Cd-chelating agents may bind excess free Cd ion and that translocation of Cd from the roots to the aerial parts might be prevented by increased xylem lignification.

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Acknowledgments

NA acknowledges the support of Japan Society for Promotion of Science. This work is also supported by the grant from National institute of Agriculture and Food Research Organization, Japan. Authors are thankful to Mr. MZ Nouri, National Institute of Crop Science for his technical help in MS analysis. We also thank to Dr. Y. Nanjo and Dr. K. Nishizawa, NICS for their valuable discussion.

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  1. Nagib Ahsan

    Present address: Department of Biochemistry and Interdisciplinary Plant Group, University of Missouri-Columbia, Columbia, MO, 65211, USA

Authors and Affiliations

  1. National Institute of Crop Science, 2-1-18 Kannondai, Tsukuba, 305-8518, Japan

    Nagib Ahsan, Takuji Nakamura & Setsuko Komatsu

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  1. Nagib Ahsan
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  2. Takuji Nakamura
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Correspondence to Setsuko Komatsu.

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726_2010_809_MOESM1_ESM.ppt

Representative 2-DE gel images of three biological replicates of microsomal proteins of control and Cd-treated soybean cultivars. A total of 100 μg protein was extracted, separated by 2-DE, and visualized with silver staining. (PPT 18584 kb)

726_2010_809_MOESM2_ESM.ppt

Representative 2-DE gel images of silver stained and CBB stained microsomal proteins of soybean roots. A total of 100 and 400 μg proteins were extracted, separated by 2-DE, and visualized with silver and CBB staining, respectively. Lower panel represents the magnified views of some spots that were common in both staining methods. (PPT 2351 kb)

726_2010_809_MOESM3_ESM.ppt

Graphical presentation of differentially expressed microsomal proteins in soybean cultivars in response to Cd stress. The height of each bar represents the average of three individual identical experiments (±SE). (PPT 150 kb)

Supplementary material 4 (DOC 33 kb)

Supplementary material 5 (DOC 51 kb)

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Ahsan, N., Nakamura, T. & Komatsu, S. Differential responses of microsomal proteins and metabolites in two contrasting cadmium (Cd)-accumulating soybean cultivars under Cd stress. Amino Acids 42, 317–327 (2012). https://doi.org/10.1007/s00726-010-0809-7

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