Zhang Chen, Li Lan, Xu Ziqin
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Provincial Key Laboratory of Biotechnology of Shaanxi, College of Life Sciences, Northwest University, Xi'an 710069, Shaanxi, China.
Sheng Wu Gong Cheng Xue Bao. 2012 May;28(5):602-12.
Genetic transformation was adopted to analyze the subcellular localization and the resistance to fungal pathogens of Arabidopsis lipid transfer protein AtDHyPRP1. The coding sequence of AtDHyPRP1 amplified by PCR from Ws ecotype was used to construct the plant binary expression vector pRI101-AN-AtDHyPRP1 and the fusion expression vector pCAMBIA1302-AtDHyPRP1-GFP. Transgenic tobacco and Arabidopsis plants were produced by leaf disc and floral dip protocols, respectively. AtDHyPRP1 could improve the resistance of tobacco to Botrytis cinerea remarkably and the infection sites on transgenic tobacco leaves accumulated large amounts of H2O2. Observation under laser scanning confocal microscope showed that AtDHyPRP1 was localized to cell surface. It suggested that AtDHyPRP1 might play special function after secretion to outside of the cell and was involved in plant defense system against pathogens.
采用遗传转化方法分析拟南芥脂质转移蛋白AtDHyPRP1的亚细胞定位及对真菌病原体的抗性。通过PCR从Ws生态型中扩增出AtDHyPRP1的编码序列,用于构建植物双元表达载体pRI101-AN-AtDHyPRP1和融合表达载体pCAMBIA1302-AtDHyPRP1-GFP。分别采用叶盘法和花浸染法培育出转基因烟草和拟南芥植株。AtDHyPRP1能显著提高烟草对灰葡萄孢的抗性,转基因烟草叶片上的感染部位积累了大量H2O2。激光扫描共聚焦显微镜观察表明,AtDHyPRP1定位于细胞表面。这表明AtDHyPRP1分泌到细胞外后可能发挥特殊功能,并参与植物对病原体的防御系统。