<em>ZmPti1</em>基因正义和RNAi表达载体的构建及其转化玉米的研究

doi:10.7668/hbnxb.2008.05.001

华北农学报 ›› 2008, Vol. 23 ›› Issue (5): 1-5. doi: 10.7668/hbnxb.2008.05.001

所属专题： 玉米；生物技术；

• 论文 • 下一篇

ZmPti1基因正义和RNAi表达载体的构建及其转化玉米的研究

张森燕^1,2, 吴忠义², 张秀海^21,2, 刘占磊², 王永勤²

1. 首都师范大学生命科学学院;
2. 北京市农林科学院北京农业生物技术研究中心

收稿日期:2008-05-08 出版日期:2008-10-28
通讯作者: 王永勤(1968－)，男，山西兴县人，博士，副研究员，主要从事蔬菜遗传育种研究。黄丛林(1969－)，男，四川仁寿人，博士，副研究员，主要从事植物遗传育种研究。
作者简介:张森燕(1981－)，女，河南驻马店人，在读硕士，主要从事作物耐旱生物学分子机制研究。
基金资助:
国家“973”项目(2003CB114302)；北京市科玉米创新平台项目(YZPT02-06)

Construction ofZmPti1Sense and RNAi Expression Vectors and Their Transformation into Maize

ZHANG Sen-yan^1,2, WU Zhong-yi², ZHANG Xiu-hai^21,2, LIU Zhan-lei², WANG Yong-qin²

1. College of Life Science, Capital Normal University, Beijing 100037, China;
2. Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China

Received:2008-05-08 Published:2008-10-28

摘要/Abstract

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。

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

Abstract: ZmPti1(AY708048)cloned in our lab encodes a maize pti - like kinase.In order to efficiently identify the functions of Zm Pti1 gene,two plant - transformation vectors,pBPC - ZmPti1 - 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 Zm Pti1 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 Zm Pti1 overexpresssion lines,and 18 plants are Zm Pti1 RNAi lines.Furthermore,the transformation efficiency in our experiment is related to the introducing time.In T 0 generation,overexpression lines are more tolerant to disease stress than that of knockout lines.It indicates that Zm Pti1 is involved in disease resistant signal pathway.The transformants obtained can be used to study the mechanism of defense response for Zm Pti1 in maize.And the results demonstrate that the approach of pollen tube pathway is simple and effi - cient for maize transformation.

Key words: ZmPti1gene, Sense expression vector, RNAi vector, Pollen tube pathway, Transgenic plants

中图分类号:

S513.035.3

张森燕, 吴忠义, 张秀海, 刘占磊, 王永勤, 黄丛林. ZmPti1基因正义和RNAi表达载体的构建及其转化玉米的研究[J]. 华北农学报, 2008, 23(5): 1-5. doi: 10.7668/hbnxb.2008.05.001.

ZHANG Sen-yan, WU Zhong-yi, ZHANG Xiu-hai, LIU Zhan-lei, WANG Yong-qin, HUANG Cong-lin. Construction ofZmPti1Sense and RNAi Expression Vectors and Their Transformation into Maize[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(5): 1-5. doi: 10.7668/hbnxb.2008.05.001.

http://www.hbnxb.net/CN/Y2008/V23/I5/1

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