Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Published on January 13, 2006; 10.1104/pp.105.072066

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Received September 27, 2005
Returned for revision November 4, 2005
Accepted December 20, 2005

An Arabidopsis mutant impaired in CoA biosynthesis is sugar-dependent for seedling establishment

Silvia Rubio , Tony R. Larson , Miguel Gonzalez-Guzman , Santiago Alejandro , Ian A. Graham , Ramón Serrano , and Pedro L. Rodriguez *

Instituto de Biología Molecular y Celular de Plantas, Universidad Politécnica de Valencia-Consejo Superior de Investigaciones Científicas, Camino de Vera, E-46022 Valencia, Spain
CNAP, Department of Biology, University of York, PO Box 373, York YO10 5YW, UK

* Corresponding author; email: prodriguez{at}ibmcp.upv.es.

Once the plant coenzyme A (CoA) biosynthetic pathway has been elucidated by comparative genomics, it is feasible to analyze the physiological relevance of CoA biosynthesis in plant life. To this end, we have identified and characterized Arabidopsis T-DNA knockout mutants of two CoA biosynthetic genes, HAL3A and HAL3B. The HAL3A gene encodes a 4'-phosphopantothenoylcysteine decarboxilase that generates 4'-phosphopantetheine. A second gene, HAL3B, whose gene product is 86% identical to that of HAL3A, is present in the Arabidopsis genome. HAL3A appears to have a predominant role over HAL3B according to their respective mRNA expression levels. The hal3a-1, hal3a-2 and hal3b mutants were viable and showed a similar growth rate than wt plants; in contrast, a hal3a-1 hal3b double mutant was embryo lethal. Unexpectedly, seedlings that were null for HAL3A and heterozygous for HAL3B (aaBb genotype) displayed a sucrose-dependent phenotype for seedling establishment, which is in common with mutants defective in {beta}-oxidation. This phenotype was genetically complemented in aaBB siblings of the progeny and chemically complemented by pantethine. In contrast, seedling establishment of Aabb plants was not sucrose-dependent, proving a predominant role of HAL3A over HAL3B at this stage. Total fatty acid and acyl CoA measurements of 5-day-old aaBb seedlings in medium lacking sucrose revealed stalled storage lipid catabolism and impaired CoA biosynthesis; in particular, acetyl-CoA levels were reduced by ~80 %. Taken together, these results provide in vivo evidence for the function of HAL3A and HAL3B, and they point out the critical role of CoA biosynthesis during early post-germinative growth.




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