Journal of Experimental Botany 52: 1753-1759 (2001)

Antisense-transformation reveals novel roles for class I ß-1,3-glucanase in tobacco seed after-ripening and photodormancy

Gerhard Leubner-Metzger and Frederick Meins, Jr.

glucanase transformation constructs

Figure 1.
Schematic representation of sense and antisense class I ß-1,3-glucanase (ßGLU I) expression vectors used for tobacco transformation. Transcriptional fusions of the castor bean Cat1 promoter (PCat1), the tobacco ßGLU I B gene (Glb) full length promoter (1.7 kb PGlb), and the truncated proximal Glb promoter (0.5 kb PGlb) with a 1.3 kb ßGLU I cDNA in reverse orientation and the CaMV 35S terminator (T35S) constitute the antisense ßGLU I constructs of pKAG4, pGAG2, and pGAG3, respectively. The chimeric sense transgene construct pTKSG7 contained 2.7 kb of a genomic DNA fragment of Glb with the entire coding sequence; and for empty-vector pCIB200 was used to generate control transformants. The chimeric neomycin phosphotransferase gene (NPT II) confers kanamycin resistance and is under the control of the nopaline synthase promoter (PNOS) and terminator (TNOS). The right (RB) and left (LB) T-DNA borders are indicated. Transformant tobacco plants were named TKSG7 (sense ßGLU I), TCIB1 (empty-vector control), and TKAG4, TGAG2, TGAG3 for antisense ßGLU I. For publications on seed germination of these transgenic plants see:

Sense constructs transformed: Leubner-Metzger and Meins, Plant J 23: 215-221 (2000)
Antisense constructs transformed: Leubner-Metzger and Meins, J Experimental Botany (2001)
Sense constructs transformed: Manz et al., Plant Physiol 138: 1538-1551 (2005)
Sense constructs transformed: Leubner-Metzger, Planta 215: 959-968 (2002)

Article in PDF format (160 KB) Abstract          Fig. 1          Fig. 2          Table 1          Table 2          Glucanase constructs
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