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Functional characterization of tomato Sl-IAA3 and Sl-hls genes. Role ...

Functional characterization of tomato Sl-IAA3 and Sl-hls genes. Role ...

CHAPITRE IV Isolation

CHAPITRE IV Isolation and Functional Characterization of Tomato Hookless Genes by Complementation of Arabidopsis hookless 1 mutant (Manuscript in preparation)

Chapitre IV: Functional Characterization of Tomato Hookless Genes Title: Isolation and Functional Characterization of Tomato Hookless Genes by Complementation of Arabidopsis hookless 1 mutant Authors : Salma Chaabouni, Alain Latche, Jean-Claude Pech, and Mondher Bouzayen ABSTARCT The apical hook of dark-grown dicotyledonous plants results from asymmetric growth of its inner and outer side of the upper part of the hypocotyl. This protective structure prevents damage of the shoot apical meristem and the young leaves as the seedlings pushes through the soil. HOOKLESS (HLS1) of Arabidopsis was recognised as an ethylene response gene whose product is required for hook formation. Tow hormones, ethylene and auxin are through to be involved in regulating apical hook formation. We cloned two cDNA from tomato, Sl-HLS1 and Sl-HLS2, and showed that the encoded proteins are functional homologs to At-HLS1. Both Sl-HLS1 and Sl-HLS2 complement the hls1 mutation. Ectopic ewpression of both tomato genes were able to restaure wild type phenotypes such as de-etiolation, enhanced tolerance to glucose and to elevated ABA concentrations and agravitropic growth. The genomic clones of Sl-HLS1 and Sl-HLS2 showed similar structure with two introns and three exons. While these data indicate complete functional redundancy between the two tomato HLs genes, only the expression of Sl-HLS2 is enhanced by ethylene and auxin. The expression of Sl-HLS2 gene, assessed by Sl-HLS2 promoter-driven GUS revealed its up-regulation by ethylene restricted to the outer side of the apical hook. These data is in agreement with the important role of HLS gene in the integration of multiple signalling pathways. 101

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