运用农杆菌介导法将PTA和反义Wx双基因导入水稻的研究

利用根癌农杆菌将含多基因(hyg选择标记基因、反义Wx 基因、PTA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 和 PTA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部分程度的下降，最低的已下降至6.3%，与对照相比下降了9.8%。

Study on Transformation of the Pinellia Tenata Agglutinin(pta)Gene and Waxy Antisense Gene into Rice Mediated by Agrobacterium

The pCAMBIA1304 vector that carried multi-gene(hygromycin gene、waxy antisense gene、pta gene、GFP and GUS genes)was transforred into calli which were derived from immature embryos of three rice varieties(japonica rice NO.9 of zhonghua and ribenqing and indica rice 501)by agrobacterium tumefaciens. These calli were respectively cultived on selective media which containing 65 mg/L, 45 mg/L and 35 mg/L hygromycin and obtained resistant calli. Ninety-two positive transgenic plants(forty-three NO.9 of zhonghua、forty-five ribenqing and four 501)were identified by PCR from 415 T0 regeneration plants. Southern blotting assay for these transgenic progenies indicated that the pta gene and waxy antisense 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 untransfored grains.