ZmPti1基因正义和RNAi表达载体的构建及其转化玉米的研究  

Construction of ZmPti1 Sense and RNAi Expression Vectors and Their Transformation into Maize

作　　者：张森燕[1,2] 吴忠义[2] 张秀海[2] 刘占磊[1,2] 王永勤[2] 黄丛林[2] 

机构地区：[1]首都师范大学生命科学学院,北京100037 [2]北京市农林科学院北京农业生物技术研究中心,北京100097

出　　处：《华北农学报》2008年第5期1-5,共5页Acta Agriculturae Boreali-Sinica

基　　金：国家“973”项目(2003CB114302);北京市科玉米创新平台项目(YZPT02-06)

摘　　要：构建了ZmPti1基因的正义表达载体和RNAi载体,以bar基因为抗性筛选标记,通过花粉管通道法将构建的两个表达载体分别转化到玉米自交系178。收获的种子播种于营养钵中,出苗后,经0.1%的草丁膦筛选得到40株草丁膦抗性植株,进一步用PCR鉴定与PCR-Southern检测得到33株转基因植株,其中15株是转化正义表达载体的转基因植株,18株是转化RNAi表达载体的转基因植株。并对转化方法和ZmPti1基因的功能进行了讨论。ZmPtil （AY708048）cloned in our lab encodes a maize pti-like kinase. In order to efficiently identify the functions of ZmPtil gene, two plant-transformation vectors, pBPC-ZmPtil-bar（sense expression vector）and pBPC-PtS-GFP-PtAs-bar（RNAi vector）were prepared. And promoters used in these two constructs were maize ubiquitin promoter. These two vectors contain herbicide resistance gene （bar gene）that can facilitate large scale selection of transformant in farm land. Sense expression and RNAi vectors of ZmPtil were delivered into maize inbred lines178 by pollen tube path- way method respectively, and 33 transformed plants were obtained through glufosinate screening, PCR analysis and PCR- southern identification. Among these transformants, fifteen plants are ZmPtil overexpresssion lines, and 18 plants are ZmPtil RNAi lines. Furthermore, the transformation efficiency in our experiment is related to the introducing time. In TO generation, overexpression lines are more tolerant to disease stress than that of knockout lines. It indicates that ZmPtil is involved in disease resistant signal pathway. The transformants obtained can be used to study the mechanism of defense response for ZmPtil in maize. And the results demonstrate that the approach of pollen tube pathway is simple and efficient for maize transformation.

关 键 词：ZmPti1基因 正义表达载体 RNAI载体 花粉管通道法 转基因植株 

分 类 号：S513.035.3[农业科学—作物学]