《植物生理学报》 2010, 46(7): 731-736

农杆菌介导的大麦Mlo 反义基因转化小麦获得抗白粉病后代

赵同金^1,*, 刘恒^1,2,*, 赵双宜¹, 陈惠民¹, 夏光敏^1,**

1山东大学生命科学学院, 植物细胞工程与种质创新教育部重点实验室, 济南250100; 2山东中医药大学科研中心, 济南250355

通信作者：赵同金；E-mail: xiagm@sdu.edu.cn；Tel: 0531-88564 525

摘　要：

从大麦 ‘ 斯特林 ’ 幼叶总 RNA 中分离 Mlo 基因 cDNA 完整编码区, 反向连接到植物双元载体(pBI-121.2) 35S 启动子 下游, 通过农杆菌介导的苗端转化法获得两种小麦基因型(‘ 烟优2801’ 和‘ 烟优361’)的转基因小麦。T0 代405 株中有 55 株 PCR 检测阳性, 平均转化率达到 13.58%, T0 和 T1 基因组 DNA Southern 杂交可以证明大麦 Mlo 基因片段已整合到小麦基因 组中并可传递到后代。两种基因型的转基因小麦T0 和T1 植株在温室及大田中均表现出对白粉病抗性的提高。农杆菌介导 的苗端转化法可以简单、快速、高效地获得转基因株系; 排除体细胞变异对转基因植株的影响; 克服基因型对农杆菌转化 的限制, 是小麦遗传转化的一种实用方法。

关键词：转基因小麦; 农杆菌转化; 白粉病; 大麦Mlo反义基因

收稿：2010-05-03   修定：2010-05-11

资助：国家 “863” 项目(2001AA241032)。

Transgenic Wheat Progenies Resistant to Powdery Mildew Generated by Agrobacterium Transformation of Barley Mlo Anti-Sense Gene

ZHAO Tong-Jin^1,*, LIU Heng^1,2,*, ZHAO Shuang-Yi¹, CHEN Hui-Min¹, XIA Guang-Min^1,**

¹The Key Laboratory of Plant Cell Engineering and Germplasm Innovation, Ministry of Education, School of Life Sciences, Shandong University, Jinan 250100, China; ²Centre of Scientific and Research, Shandong University of Traditional Chinese Medicine, Jinan 250355, China

Corresponding author: ZHAO Tong-Jin; E-mail: xiagm@sdu.edu.cn; Tel: 0531-88564 525

Abstract:

Complete coding region sequence of Mlo cDNA from barley cultivar ‘Stirling’ was cloned and inserted into the BamHI site of binary vector pBI121.2, driven by 35S promotor in reverse direction. Two wheat (Triticum aestivum L.) genotypes ‘Yanyou2801’ and ‘Yanyou361’ were transformed by Agrobacterium tumefaciens with shoot apical meristem transformation method. A total of 55 T₀ transgenic plants were obtained, and the average transformation frequency was as high as 13.58%, Southern blot proved Mlo gene was integrated into T₀ wheat genome and inherited to T₁ progenies. The T₀ and T₁ transgenic plants of both genotypes showed higher resistance to powdery mildew than the controls in both green house and field. The shoot apical meristem transformation with A. tumefaciens is a simple, dependable and efficient method to generate transgenic wheat. It can overcome the difficult of the restriction to transformation of different wheat genotypes, and can get rid of the somoclonal variations due to tissue culture of immature embryos and calli.

Key words: transgenic wheat; Agrobacterium transformation; powdery mildew; barley Mlo anti-sense gene