Wound-inducible expression of the Bacillus thuringiensis CrylB gene in transgenic rice

• Main Authors: Breitler, J.C, Marfa, V, Meynard, D, Vila, L, Murillo, I, Dominguez Rodriguez, M.J, Royer, M, San Segundo, B, Martinez-Izquierdo, J.A, Messeguer, J, Vassal, J.M, Guiderdoni, E
• Format: Publication
• Language: English
• Published: Los Baños (Philippines) International Rice Research Institute 2003
• Subjects: ARROZ; BIOLISTICA; BIOLISTICS; BIOLISTIQUE; EXPRESION GENICA; EXPRESSION DES GENES; GENE; GENE EXPRESSION; GENE TRANSPOSITION; GENES; GLICOPROTEINAS; GLYCOPROTEINE; GLYCOPROTEINS; INHIBIDORES DE PROTEINASAS; INHIBITEUR DE PROTEINASES; PEST RESISTANCE; PROTEINASE INHIBITORS; RESISTANCE AUX ORGANISMES NUISIBLES; RESISTENCIA A LAS PLAGAS; RICE; RIZ; TRANSPOSICION DE GENES; TRANSPOSITION DE GENES

To investigate whether wound-induced production of Cry endotoxin protects transgenic rice against striped stem borer (SSB), we prepared constructs bearing a synthetic cry1B coding sequence controlled by the promoter region of either the constitutive maize ubiquitin gene or wound-inducible maize genes - the maise proteinase inhibitor gene (mpi) and the hydroxyproline-rich glycoprotein (hrgp) gene. The mpi-cry1B-nos and Ubi-cry1B-nos gene cassettes on the one hand and the hrgp-cry1B-nos and Ubi-cry1B-nos gene cassettes on the other hand were cloned in pBKS+ vectors and pCAMBIA1300 binary vectors, respectively, for microprojectile bombardment- and Agrobacterium-mediated transformation of cv. Ariete. Accumulation of Cry1B from an undetectable level to 0.2% of total soluble protein, 8-12 h after mechanical wounding, was demonstrated in leaf and pith tissues of lines harboring the pmpi-cry1B construct. In these plants, Cry1B was not detected in pollen, endosperm, and embryo, unlike in pUbi-cry1B plants. Two out of 43 pmpi-cry1B plants were found to exhibit full protection against SSB damage. Seventeen and 66 plants harboring the Hrgp-cry1B and Ubi-cry1B T-DNA, respectively, were obtained through co-culture of embryogenic calli with strain EHA105. Some 58.8% and 42.2% of Hrgp-cry1B and Ubi-cry1B plants, respectively, accumulated Cry1B above 0.1% total soluble protein in leaf tissue. Wound-stimulated expression of cry1B was demonstrated in young leaf tissue of most Hrgp-cry1B plants.