水稻转座子Pong特异性RNAi载体TPAS的构建和农杆菌介导的遗传转化条件的优化  

Specific RNAi vector construction of transposon Pong in rice and Agrobacterium tumefaciens-mediated genetic transformation optimization

作　　者：高立宏[1,2] 张春玉[3] 柴娜[2] 路云侠[2] 刘宝[2] 刘立侠[2] 

机构地区：[1]长春师范学院生命科学学院,吉林长春130032 [2]东北师范大学生命科学学院,吉林长春130024 [3]长春职业技术学院,吉林长春130033

出　　处：《东北师大学报（自然科学版）》2011年第4期122-127,共6页Journal of Northeast Normal University(Natural Science Edition)

基　　金：国家重点基础研究发展计划(973)项目(2005CB120805)

摘　　要：为研究水稻转座子Pong的功能,构建了Pong的特异RNAi表达载体TPAS,并利用农杆菌介导法将这个载体转化到水稻的成熟胚愈伤组织,获得了PCR阳性的再生植株.同时通过对农杆菌侵染途径、侵染浓度和侵染时间等的研究,建立并优化了水稻的遗传转化程序.结果表明:在农杆菌侵染成熟胚愈伤组织的时间为3min,共培养2～4d,预培养7d,G418质量浓度为150～200mg/L条件下筛选7d,转化率可达到30%,为最佳的遗传转化参数.研究结果为今后水稻转座子Pong功能的进一步研究提供了一定的基础和参考.To study the function of rice transposon Pong,the research constructed the specific RNAi expressed vector TPAS of transposon Pong in rice,and transferred the vectors into the mature embryo callus of the rice cultivar Matsumae,and then obtained the regeneration of the positive PCR results by Agrobacterium tumefaciens-mediated method.To establish and optimize genetic transformation process,the different infected ways,concentrations and time of Agrobacterium were chosen.The result showed that：the infection time of Agrobacterium to mature embryo callus is 3 min,the time of co-culture is 2~4 d,the pre-culture period is 7 d,all the dates were the best genetic transformation parameters,making the basis for the function of rice Tn Pong.

关 键 词：水稻 转座子 RNAI 农杆菌介导 遗传转化 

分 类 号：Q943.2[生物学—植物学]