Abstract
Carotenoids are plastidic isoprenoid pigments of great biological and biotechnological interest. The precursors for carotenoid production are synthesized through the recently elucidated methylerythritol phosphate (MEP) pathway. Here we have identified a tomato (Lycopersicon esculentum Mill.) cDNA sequence encoding a full-length protein with homology to the MEP pathway enzyme hydroxymethylbutenyl 4-diphosphate synthase (HDS, also called GCPE). Comparison with other plant and bacterial HDS sequences showed that the plant enzymes contain a plastid-targeting N-terminal sequence and two highly conserved plant-specific domains in the mature protein with no homology to any other sequence in the databases. The ubiquitous distribution of HDS-encoding expressed sequence tags (ESTs) in the tomato collections suggests that the corresponding gene is likely expressed throughout the plant. The role of HDS in controlling the supply of precursors for carotenoid biosynthesis was estimated from the bioinformatic and molecular analysis of transcript abundance in different stages of fruit development. No significant changes in HDS gene expression were deduced from the statistical analysis of EST distribution during fruit ripening, when an active MEP pathway is required to support a massive accumulation of carotenoids. RNA blot experiments confirmed that similar transcript levels were present in both the wild-type and carotenoid-depleted yellow ripe (r) mutant fruit independent of the stage of development and the carotenoid composition of the fruit. Together, our results are consistent with a non-limiting role for HDS in carotenoid biosynthesis during tomato fruit ripening.
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Abbreviations
- EST:
-
expressed sequence tag
- DXR:
-
deoxyxylulose 5-phosphate reductoisomerase
- DXS:
-
deoxyxylulose 5-phosphate synthase
- GCPE:
-
hydroxymethylbutenyl 4-diphosphate synthase
- HDS:
-
hydroxymethylbutenyl 4-diphosphate synthase
- MEP:
-
methylerythritol phosphate
- PSI:
-
plant-specific internal (domain)
- PSN:
-
plant-specific N-terminal (domain)
- UTR:
-
untranslated region
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Acknowledgements
We thank D. Frisch (Clemson University Genomics Institute) for the tomato cLED4F3 clone. The help and advice of the staff of the Serveis de Camps Experimentals (greenhouse facilities) of the Universitat de Barcelona are greatly appreciated. We also thank A. Orozco for valuable technical assistance, and the laboratory members for their collaboration. This work was supported in part by grant BIO2002-1653 (to M.R.-C.) from the Spanish Ministerio de Ciencia y Tecnología and grant 2001SGR00109 from the Generalitat de Catalunya (to A.B.). L.M.L. received a doctoral fellowship from the Spanish Ministerio de Educación y Cultura.
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Rodríguez-Concepción, M., Querol, J., Lois, L.M. et al. Bioinformatic and molecular analysis of hydroxymethylbutenyl diphosphate synthase (GCPE) gene expression during carotenoid accumulation in ripening tomato fruit. Planta 217, 476–482 (2003). https://doi.org/10.1007/s00425-003-1008-5
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DOI: https://doi.org/10.1007/s00425-003-1008-5