AOBPreview originally published online on July 18, 2005
Annals of Botany 2005 96(4):703-715; doi:10.1093/aob/mci222
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Protein Synthesis by Rice Coleoptiles During Prolonged Anoxia: Implications for Glycolysis, Growth and Energy Utilization
1 School of Plant Biology, Faculty of Natural and Agricultural Sciences and 2 Plant Molecular Biology Group, School of Biomedical and Chemical Sciences, The University of Western Australia, 35 Stirling Highway, Crawley, 6009 WA, Australia
* For correspondence. E-mail hmillar{at}cyllene.uwa.edu.au
Received: 29 October 2004 Returned for revision: 2 December 2004 Accepted: 21 January 2005 Published electronically: 18 July 2005
• Background and Aims Anoxia-tolerant plant tissues synthesize a number of proteins during anoxia, in addition to the ‘classical anaerobic proteins’ involved in glycolysis and fermentation. The present study used a model system of rice coleoptile tips to elucidate patterns of protein synthesis in this anoxia-tolerant plant tissue.
• Methods Coleoptile tips 7–11 mm long were excised from intact seedlings exposed to anoxia, or excised from hypoxically pre-treated seedlings and then exposed to anoxia for 72 h. Total proteins or 35S-labelled proteins were extracted, separated using two-dimensional isoelectric focusing/SDS–polyacrylamide gel electrophoresis and analysed using mass spectrometry.
• Key Results The coleoptile tips excised after intact seedlings had been exposed to anoxia for 72 h had a similar proteome to tips that were first excised and then exposed to anoxia. After 72 h anoxia, Bowman–Birk trypsin inhibitors and a glycine-rich RNA-binding protein decreased in abundance, whereas a nucleoside diphosphate kinase and several proteins with unknown functions were strongly enhanced. Using [35S]methionine as label, proteins synthesized at high levels in anoxia, and also in aeration, included a nucleoside diphosphate kinase, a glycine-rich RNA-binding protein, a putative elicitor-inducible protein and a putative actin-depolymerizing factor. Proteins synthesized predominately in anoxia included a pyruvate orthophosphate dikinase (PPDK), alcohol dehydrogenase 1 and 2, fructose 1,6-bisphosphate aldolase and a protein of unknown function.
• Conclusion The induction of PPDK in anoxic rice coleoptiles might, in combination with pyruvate kinase (PK), enable operation of a ‘substrate cycle’ producing PPi from ATP. Production of PPi would (a) direct energy to crucial transport processes across the tonoplast (i.e. the H+-PPiase); (b) be required for sucrose hydrolysis via sucrose synthase; and (c) enable acceleration of glycolysis, via pyrophosphate:fructose 6-phosphate 1-phosphotransferase (PFP) acting in parallel with phosphofructokinase (PFK), thus enhancing ATP production in anoxic rice coleoptiles; ATP production would need to be increased if there was a substantial requirement for PPi.
Key words: Anoxia, coleoptile, ethanol production, glycolysis, protein synthesis, proteomics, pyruvate orthophosphate dikinase, pyrophosphate, Oryza sativa
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