Development and functional annotation of an 11303

Authors: Guylaine Keller Thibault Marchal Hélène SanClemente Marie Navarro Nathalie Ladouce Patrick Wincker Arnaud Couloux Chantal Teulières Christiane Marque

Publish Date: 2008/10/31

Volume: 5, Issue: 2, Pages: 317-327

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Abstract

A cDNA library was constructed from leaves of a coldacclimated Eucalyptus gunnii elite clone and 13056 ESTs were sequenced When the 5457 unique sequences were compared with Arabidopsis rice and poplar datasets more than 38 of nohits were found and the best annotation was provided by the woody perennial poplar database However among the 62 of sequences that could be annotated the vast majority 875 was found to be common to the four compared plant species which shows how highly conserved this gene pool is across the plant kingdom When the distribution of the annotated sequences was studied according to the Gene Ontology classification some typical features of the dehydrative stress transcriptome were observed In particular genes from sugar metabolism and above all those induced by stress or external stimuli were well represented 6 in total In addition the library was found to contain a substantial number of ESTs encoding putative transcription factors including CBF CRTBinding Factor The enrichment of this library with stressrelated genes was strongly suggested by the high redundancy up to 300 ESTs of several genes known to be involved in cell protection such as the PCP Pollen Coat Protein Lti6b Low temperature induced or metallothionein It was further confirmed by demonstrating the cold induction of a set of representative genes through Real TimePCR All these characteristics make this Eucalyptus EST collection very suitable for investigating the molecular basis of cold tolerance in a woody perennial and for isolating important or new candidate frost tolerance genesWe thank AFOCEL for supplying the Eucalyptus cuttings Research supports salaries and grants were provided by French Ministry of Research and Technology MNRT the “Midi Pyrénées French Council” and TEMBEC SA RD KRAFT St GAUDENS France Thanks to Victoria McBride for accurate English editing of the manuscript

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