Regeneration and genetic transformation of acacia mangium

C - Chemistry – Metallurgy – 12 – N

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C12N 15/29 (2006.01) A01H 4/00 (2006.01) A01H 5/00 (2006.01) C12N 15/82 (2006.01)

Patent

CA 2362767

The present invention is directed to a method of Acacia mangium regeneration through organogenesis and a method of genetic transformation of Acacia mangium. The method of regeneration comprises inducing callus from different parts of seedlings and vegetatively micropropagated plantlets as explants; adventitious bud induction followed by pinnate leaf and bud elongation and eventually, elongated shoots were induced to root. Based on the regeneration system, a marker gene GUS under cauliflower mosaic virus promoter was introduced to Acacia mangium via Agrobacterium infection. GUS staining in the regenerated plants and Southern blot hybridization prove the incorporation of the foreign gene into the host genome and expression of the foreign gene.

L'invention concerne un procédé de régénération d' Acacia mangium par le biais d'une organogenèse, et elle concerne également un procédé de transformation génétique d' Acacia mangium . Le procédé de régénération consiste à induire un cal à partir de différentes parties de semis, et de plantules micropropagées de manière végétative en tant qu'explants, à induire des bourgeons adventifs, puis une feuille pennée et un allongement des bourgeons, et éventuellement à induire la prise en racine de rejets de forme allongée. En fonction de ce système de régénération, on a introduit, dans Acacia mangium , un gène marqueur GUS sous la commande d'un promoteur du virus de la mosaïque du chou-fleur, par le biais d'une infection à une agrobactérie. Le marquage de GUS dans les plantes régénérées, ainsi que l'hybridation à l'aide d'une technique de Southern, prouvent l'incorporation du gène étranger dans le génome hôte et l'expression dudit gène.

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