Contributions of in situ hybridization of sRNA to the study on spatio-temporal gene expression in Hevea brasiliensis

Alemanno Laurence, Guilhaumon Claire, Marteaux Benjamin, Conejero Geneviève, Verdeil Jean-Luc, Dessailly Florence, Montoro Pascal. 2005. Contributions of in situ hybridization of sRNA to the study on spatio-temporal gene expression in Hevea brasiliensis. In : IRRDB Natural Rubber Conference, Cochin, Kerala, India, 7-8 November 2005. IRRDB. s.l. : s.n., 1-6. IRRDB Natural Rubber Conference, Kerala, Inde, 7 Novembre 2005/8 Novembre 2005.

Résumé : Since the dawn of the genomics era, much research has focused on functional studies of genes of interest. In situ hybridization is a method that can be used to precisely localize the expression of a gene in tissues and cells. This article describes how the method has been adapted to the analysis of Hevea tissues. Initially, the conventional method of digoxigenin detection with NBT/BCIP revealed the expression of strongly expressed genes in tissues of different differentiation intensity. A new digoxigenin detection method using Alexa488 fluorochrome-labelled antibodies has been used to detect the expression of more weakly expressed genes. This method, combined with observation under a confocal microscope, has enabled very precise localization of expression. Some examples of in situ hybridization use are described for Hevea gene expression in somatic plantlets and shoot bark: the uidA gene in callus and transgenic somatic plantlets, the HEV2.1 gene encoding hevein and the ACO-H5 gene involved in ethylene metabolism. Cell imaging methods therefore open up fundamental prospects for studying the different molecular mechanisms involved in some agronomic traits of Hevea.

Mots-clés Agrovoc : Hevea brasiliensis

Classification Agris : F30 - Génétique et amélioration des plantes

Auteurs et affiliations

• Alemanno Laurence, CIRAD-CP-UMR BEPC (FRA)
• Guilhaumon Claire, CIRAD-CP-UMR BEPC (FRA)
• Marteaux Benjamin
• Conejero Geneviève
• Verdeil Jean-Luc, CIRAD-CP-UMR BEPC (FRA)
• Dessailly Florence, CIRAD-CP-UMR BEPC (FRA) ORCID: 0000-0001-7873-3974
• Montoro Pascal, CIRAD-CP-UMR BEPC (FRA)

Source : Cirad - Agritrop (https://agritrop.cirad.fr/548938/)

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