摘要
通过遗传转化技术研究了拟南芥脂转移蛋白AtDHyPRP1在细胞中的定位及其对真菌病原体的抗性。采用PCR方法从拟南芥Ws生态型克隆了AtDHyPRP1基因,构建产生pRI101-AN-AtDHyPRP1植物双元表达载体和pCAMBIA1302-AtDHyPRP1-GFP融合表达载体,经农杆菌介导的叶盘和浸花法得到烟草和拟南芥转基因植株。AtDHyPRP1基因能够明显增加烟草对灰霉菌的抗性,转AtDHyPRP1烟草叶片的被侵染部位有大量H2O2积累,激光共聚焦显微观察发现AtDHyPRP1蛋白定位于细胞表面。说明AtDHyPRP1蛋白在合成后被分泌到细胞外执行特殊的功能,与植物抗病防御机制有关。
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.
出处
《生物工程学报》
CAS
CSCD
北大核心
2012年第5期602-612,共11页
Chinese Journal of Biotechnology
基金
国家自然科学基金(No.30870194)
陕西省重点实验室科研计划(Nos.08JZ70
2010JS090)
陕西省教育厅科研计划(No.11JK0612)
陕西省科学技术研究发展计划(社发攻关
No.2010K16-04-01)
西北大学研究生创新计划(Nos.10YSY12
10YSY13)资助~~
关键词
AtDHyPRP1
秦烟95
拟南芥
蒜薹灰霉菌
DAB染色
台酚兰染色
亚细胞定位
AtDHyPRP1
Nicotiana tabacum qinyan 95
Arabidopsis thaliana
Botrytis cinerea Pers.ex of Garlic Sprout
DAB staining
trypan blue staining
subcellular localization
