Functional analysis of transgenic rice plants expressing a novel mutated ALS gene of rice

Abstract

We performed functional analysis of transgenic plants expressing the W548L/S627I mutated ALS gene from rice as well as studies on inheritance of the mutated gene, phenotype and fertility of transgenic plants to ensure availability of the mutated gene as a selectable marker for plant genetic transformation. Expression levels of the ALS (endogenousϩmutated ALS) gene of transgenic rice plants were correlated with the resistance of transgenic plants to bispyribac-sodium (BS). The BS-resistant trait of a transgenic plant was stably inherited by the progeny in a Mendelian manner. A homozygote of transgenic plants harboring the mutated gene was normal in its growth and fertility compared with the wild type. These results ensured that the W548L/S627I mutated gene from rice can be efficiently used as a selectable marker for genetic transformation of rice in combination with BS. © Pesticide Science Society of Japan

Application of Mutated Acetolactate Synthase Genes for Herbicide Resistance to Crop Improvement

“…We have isolated a double-mutated ALS gene from rice cells (OsALS-W548L/ S627I) (Figure 2), which confers a high level of resistance to the PC-type ALS-inhibiting herbicide bispyribac-sodium (BS) (Figure 1), and demonstrated that it could be used as a selectable marker for generating transgenic rice plants (Kawai et al, 2007a;Kawai et al, 2007b). We also found that the single amino acid substitution S627I in the ALS gene (OsALS-S627I) imparts high levels of resistance to the PCs pyrithiobac-sodium (PS) and pyriminobac (PM).…”

Herbicide sensitivities of mutated enzymes expressed from artificially generated genes of acetolactate synthase

“…We have also shown that a transgenic rice plant into which this double-mutated ALS gene was introduced expressed resistance to BS. 2,3) Herbicide-resistant ALS genes have been shown to be useful not only for the generation of transgenic plants that express resistance to the corresponding herbicides but also for introducing foreign traits into plants as a selectable marker. 4) Indeed, the W548L/S627I double-mutated ALS gene was shown to be useful as a selectable marker for plant genetic transformation when used together with BS.…”

Transgenic Tall Fescue and Maize with Resistance to ALS-Inhibiting Herbicides

“…A two-point mutated rice ALS gene, OsALS (dm), was created when two-point mutations, W548L and S627I, in the OsALS gene were introduced by site-directed mutagenesis. The OsALS ( d m ) g e n e w a s i n t r o d u c e d i n t o r i c e b y Agrobacterium-mediated transformation (Kawai et al, 2007a). After spraying over the leaves and stems of transgenic rice carrying the OsALS (dm) gene, they grew normally, indicating that the OsALS (dm) gene acted functionally in rice plants.…”

“…We introduced the OsALS (dm) gene into turf-type tall fescue to confer herbicide resistance using Agrobacterium-mediated transformation (Sato et al, 2009). Agrobacterium tumefaciens strain EHA105 carries the binary vector pMLH7133-OsALS (dm) consisting of the OsALS (dm) gene and hygromycin phosphotransferase gene (hpt) under the control of the enhanced cauliflower mosaic virus (CaMV) 35S promoter (Kawai et al, 2007a). Embryogenic calli were induced from shoot tips of the turf-type tall fescue cultivar Tomahawk germinated in vitro.…”