| As an important grass, rice is a model plant in monocots. Since 1994, research of agro-bacterium-mediated transgenic rice has been continuously improved and advanced. This study was designed to establish agrobacterium-mediated genetic transformation system of rice and to study whether the over-expression of TMM and FK in rice affect the phenotype of rice, and to further understand the functions of these two genes.(1) Rice mature seeds without hull could be effectively sterilized by soaking in 70% ethanol for 0.5-1 min and 0.1% mercuric chloride for 15 min.(2) Callus could be successfully induced by N6D medium supplemented with 2 mg/L 2,4-D; embryog-enic callus were cultured on MS medium supplemented with 0.5 mg/L NAA,2 mg/L KT, sorbitol and casein acid hydrolysis can be successfully regenerated.(3) Acetosyingone could successfully promote the transformation of rice embryogenic callus by EHA105.(4) The resistant callus were obtained through selecting with 50 mg/L hygromycin B, and the transgenic plants were effectively acquired by selecting with 25 mg/L hygromycin B.(5) The hygromycin resistance gene, encoding hygromycin B phosphotransferase, was successfully cloned in transgenic plants.Compared with the control, the epidermal structure of overexpressed TMM plants was normal. However, abnormal blades were found in two overexpressed TMM transgenic plants. Their blades were involute towards axial planes, straight. And the angles between leaves and stems were smaller. In the cross section, there were error distributions of vesicular cells and vascular bundles. The error distribution of vascular bundles and vesicular cells may lead to involution of blades. Involute leaves would result in straight blades. Why the abnormal phenotypes was not found in the other TMM transgenic plants, whether it could be stably inherited, and the reasons were yet further studied. However, the overexpressed FK transgenic plants were normal compared with the control. |
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