水稻蛋白二硫键异构酶基因沉默载体构建及其转基因后代的-作物学报.PDF

作物学报 ACTA AGRONOMICA SINICA 2013, 39(5): 816−826 /zwxb/ ISSN 0496-3490; CODEN TSHPA9 E-mail: xbzw@ DOI: 10.3724/SP.J.1006.2013.00816 水稻蛋白二硫键异构酶基因沉默载体构建及其转基因后代的高温结实 特性分析 刘光快 曹珍珍 韦克苏 潘 刚 苏 达 张春娇 程方民* , 310058 : RT-PCR PDI 450 bp , hpRNA (ihpRNA)pTCK303-RiOsPDI, , ; T0 (Hyg) PCR , T-DNA , T1 3 ∶1 PCR PCR , PDI PDI , , 80% T2 , PDI , , , PDI , , : ; ; ; RNAi ; ; RNAi Vector Construction for Protein Disulfide Isomerase Gene and Seed Set- ting Characteristics in Offspring of Transgenic Rice under High Temperature Treatment LIU Guang-Kuai, CAO Zhen-Zhen, WEI Ke-Su, PAN Gang, SU Da, ZHANG Chun-Jiao, and CHENG * Fang-Min College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China Abstract: To clarify the effect of PDI gene silence on rice yield traits and grain quality, we sub-cloned a 450 bp RNAi fragment from Nipponbare by RT-PCR, and successfully constructed a binary-vector pTCK303-RiOsPDI containing intron hpRNA (ih- pRNA), then transformed it into the callus of wild type Nipponbare mediated by EHA105. The T-DNA region for PDI RNA inter- ference in regenerating rice plants was integrated with rice genome via single copy in T0 generation transgenic plants, and showed a 3:1 genetic mode in T1 transgenic population, which could be conformed by PCR amplifying analysis and Southern blotting identification. The PCR an