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Plant and Cell Physiology Advance Access originally published online on June 11, 2009
Plant and Cell Physiology 2009 50(7):1305-1318; doi:10.1093/pcp/pcp074
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

This article appears in the following Plant and Cell Physiology issue: Special Issue Articles: Omics and Bioinformatics [View the issue table of contents]

Mitochondrial Damage in the Soybean Seed Axis During Imbibition at Chilling Temperatures

Guangkun Yin1,2, Hongmei Sun1,2, Xia Xin1,2, Guozheng Qin3, Zheng Liang1 and Xinming Jing1,4,*

1Laboratory of Seed Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, PR China
2Graduate School of the Chinese Academy of Sciences, Beijing 100039, PR China
3Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, PR China

*Corresponding author: E-mail, xmjing{at}ibcas.ac.cn; Fax, +86-10-62836009.


  Abstract

The development of mitochondria during seed germination is essential for plant growth. However, the developmental process is still poorly understood. Temperature plays a key role in soybean germination, and in this study we characterized the mitochondrial ultrastructure and proteome after imbibition at 22, 10 and 4°C for 24 h. The mitochondria from the soybean seed axis can be divided into light and heavy mitochondria by Percoll density gradient centrifugation. The axes imbibed at 4°C mainly contained light mitochondria, which had lower levels of specific mitochondrial enzymes and oxidative phosphorylation activity. In contrast, the axes imbibed at 22°C mainly contained heavy mitochondria, which exhibited higher metabolism. Electron microscopy revealed that mitochondria in the axes imbibed at 4°C had a poorly developed internal membrane system with few cristae, while the mitochondria in the axes imbibed at 22°C developed more normally. Furthermore, we compared the axis mitochondrial proteomes during imbibition at different temperatures. The differentially expressed proteins were identified using ESI-Q-TOF-MS/MS (electrospray ionization quadrupole time-of-flight tandem mass spectrometry). Proteins involved in mitochondrial metabolites including malate dehydro-genase (tricarboxylic acid cycle enzyme), putative ATP synthase subunit (oxidative phosphorylation complex subunits), mitochondrial chaperonin-60 (heat shock protein), arginase (urea cycle enzyme) and mitochondrial elongation factor Tu (mitochondrial genome transcript enzyme) were identified. The reduced expression of these proteins might not support normal mitochondrial metabolism. We conclude that chilling during imbibition causes mitochondrial damage at both ultrastructural and metabolic levels.

Keywords: Glycine max (L.) Merr. - Imbibition - Mitochondria - Proteome - Ultrastructure

Abbreviations: APX, ascorbate peroxidase; BSA, bovine serum albumin; CAT, catalase; CBB, colloidal Coomassie blue; CHAPS, 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate; COX, cytochrome c oxidase; 2D-GE, two-dimensional gel electrophoresis; DTT, dithiothreitol; ESI-Q-TOF-MS/MS, electrospray ionization quadrupole time-of-flight tandem mass spectrometry; G, ; i, germination index; IEF, isoelectric focusing; KCN, potassium cyanide; MDA, malondialdehyde; pI, isoelectric point; PVP, polyvinylpyrrolidone; ROS, reactive oxygen species; SHAM, salicylhydroxamic acid; SOD, superoxide dismutase; TCA, tricarboxylic acid; TEM, transmission electron microscopy; TFA, trifluoroacetic acid.

4Present address: Institute of Botany, Chinese Academy of Sciences, Xiangshan, Beijing100093, PR China.

(Received April 16, 2009; Accepted May 25, 2009)
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