超级稻DAD1基因正反义植物表达载体的构建及转化菊花的研究.pdfVIP

第33卷第6期 湖南农业大学学报 (自然科学版) V01．33 No．6 2007年l2月 Journal of Hunan Agricultural University(Natural Sciences) Dec．2007 文章编号：1007．1032(2007)06-0656-06 超级稻DAD 1基因正反义植物表达 载体的构建及转化菊花的研究 周 涌 ，洪亚辉 ．b ，王若仲 ，欧阳琳 ，周树良 (湖南农业大学a．生物科学技术学院；b．湖南省植物激素与生长发育重点实验室，湖南长沙 410128) 摘 要：将水稻DADl基因插入Ti质粒，构建成该基因的正反义植物表达载体pWM．DADI，pWM．antisense DADI，转入根癌农杆菌LBA4404后，利用农杆菌介导的叶盘法转化菊花，经Hyg抗性筛选，并诱导愈伤组织 和分化成苗，得到转基因菊花植株，通过PCR检测，从部分转基因植株中检测出预期的目的基因的存在． 关 键 词：DADl基因；菊花；转化；根癌农杆菌 中图分类号：Q786 文献标识码：A Research on construetion and transformation of sense and antisense expression vector of rice DAD1 gene into chrysanthemmn ZHOU Y0n ，HONE,Ya-lmP一，WANG如 zh叩 ，OUYANG Lin，ZHOU li柚 (a．College of Bioscience and Technology；b．Hunan Provincial Key Laboratory of Phytohormones and Growth Development，HNAU，Changsha 410128，China) Ab《 mct：We first inserted the DADI gene from rice into Ti plasmid．then constructed sense and antisense DADI gene expression vector which were named as pWM·DADI and pWM‘antisense DADI respectively．Afetr that two recombinated plasmids were transformed into Agrobacterium tumefaciens strain LBA4404 and infected chrysanthemum leaves via leaf disc co-cultivation．By the process of Hyg sreceening，the infected leaf discs were induced to form callus and differentiated to shoots further more．At last 28 transgenic chrysanthemum plants were obtained．The PCR results indicated DADI gene exists in part of transgenic plants，which confirmed the fact that the sense and antisense fragment of DAD I gene have been integrated into chrysanthemum genome successfully． Key words：DADI gene；chrysanthemum；transformation；Agrobacterium tumefaciens 常规的育种方法在菊花良种选育中已经取得了 导入菊花中，使其在花色、花期