利用农杆菌介导法将反义Wx基因导入水稻的研究

利用根癌农杆菌将含多基因(hpt选择标记基因、反义Wx基因、GFP和GUS报告基因)的pCAMBIA1304载体导入水稻品种(501R、中花9号和日本晴)的幼胚愈伤组织,分别在含35mg/L、45mg/L和65mg/L潮霉素浓度的筛选培养基上筛选获得抗性愈伤。后经PCR检测,从415株T0代再生植株中选出92株(其中501R、中花9号和日本晴分别为4株、43株和45株)。对这些转基因后代植株进行Southern blotting分析表明：hpt、反义Wx基因已经整合进植物基因组中。绝大多数转基因植株后代的表型正常。成熟种子直链淀粉含量分析表明,部分转基因植株的T1代种子中的直链淀粉含量有不同程度的下降,最低的已下降至6．3％,与对照相比下降了9．8％。

A Study on Transformation of the Antisense Waxy Gene into Rice Mediated by A grobacterium

The pCAMBIA1304 vector that carried multi-gene(hpt gene?antisense waxy gene?GFP and GUS genes)was transferred into calli which were derived from immature embryos of three rice varieties(indica rice 501R?japonica rice Zhonghua 9 and Niponbare)mediated by Agrobacterium tumefaciens. These calli were respectively cultived on selective media which containing 65mg/L?45mg/L and 35mg/L hygromycin and obtained resistant calli. Ninety-two positive transgenic plants(forty-three Zhonghua 9?forty-five Niponbare and four 501R)were identified by PCR from 415 T0 regeneration plants. Southern blotting assay for these transgenic progenies indicated that the antisense waxy gene and hpt gene have been successfully introduced and integrated into the genome of the receptors. The phynotype of great majority transgenic progenies was normal. The amylase content in the endosperm of mature seeds from some T1 transgenic plants was measured and the results showed that the amylase content in some grains reduced. The lowest was reduced to 6.3%, which was 9.8% lower than that of the untransferred grains.