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 degrees 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 degrees C mainly contained light mitochondria, which had lower levels of specific mitochondrial enzymes and oxidative phosphorylation activity. In contrast, the axes imbibed at 22 degrees C mainly contained heavy mitochondria, which exhibited higher metabolism. Electron microscopy revealed that mitochondria in the axes imbibed at 4 degrees C had a poorly developed internal membrane system with few cristae, while the mitochondria in the axes imbibed at 22 degrees 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 dehydrogenase (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.
F Fi ig gu ur re e 1 1 The effects of ultradry storage on Eucommia seeds. No difference in viability or vigour was evident in seeds stored for two years either at ambient temperature after ultradrying or at low temperature. a a, , Ultradried seeds stored at ambient temperature: moisture content (m.c.) was 3.3%, germination was 97%, vigour index was 26.7. b b, , Seeds stored under conventional conditions at -20 °C: m.c. was 8.4%, germination was 96%, vigour index was 24. 1. c c, , Control stored at ambient temperature: m.c. was 7.5%, viability was lost.
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