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Specific promoters for genetic engineering of rubber tree

Pujade-Renaud V., Montoro P., Sanier C., Phuangkosol N., Kongsawadworakul P., Chrestin H.. 2001. In : Sainte-Beuve Jérôme (ed.). Biotechnology and rubber tree : Proceedings of IRRDB symposium, 25-28 September 2001, Montpellier, France. Montpellier : CIRAD. IRRDB Symposium, 2001-09-25/2001-09-28, Montpellier (France).

Cloning of ethylene-inducible and/or laticifer-specific promotors from genes expressed in rubber tree latex was untertaken with the objective to optimize transgene expression in genetically engineered rubber tree. Glutamine synthetase (GS) and hevein (hev) gene promotors were sought for, based on the fact that (1) overexpression of the gs genes was observed after ethylene treatment in latex, and (2) the hevein protein has been found in the laticifers only. Several genomic clones were obtained. Partial sequencing revealed that both hevein and gs are encoded by multigene families. Hevein genes could be classed in two groups, based on sequence homology. Three different gs genes (gs1, gs2 and gs3) were identified. Gene expression analysis revealed that: (1) both gs1 and gs2/gs3 (without possible distinstion) were responsive to ethylene in latex, with gs1 apparently strictly induced and gs2/gs3 overexpressed; (2) the very high expression level of the hev gene in latex masked any potential effect of ethylene; (3) gs1 and gs2/gs3 were differentially expressed in tissues from in vitro culture at various stages of development; (4) both gs and hev genes were expressed in callus; (5) hv gene expression increased concomitantly with the development of differentiated laticifers. Two different hevein promoters (hev1 and hev4) and two gs promoters (gs2 and gs3) were isolated and cloned in fusion with the gus reporter gene in a transformation vector. The different strategies envisaged to analyse the functionality of the isolated promoters are presented. The choice of a promoter, for a genetic engineering programme, needs to take into account both the physiology of the plant and the application being sought, in order to optimize the expression of the transgene. In the case of rubber tree (Hevea brasiliensis), genetic engineering is being considered in order to improve existing varieties, but also as a physiological study tool intended to analyse or confirm the function of certain gen

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