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Green Fluorescent Protein- and Discosoma sp. Red Fluorescent Protein-Tagged Organelle Marker Lines for Protein Subcellular Localization in Rice

文献类型： 外文期刊

作者： Chen, Ziqiang 1 ; Zheng, Wenhui 2 ; Chen, Longhai 1 ; Li, Chenlu 1 ; Liang, Tingmin 5 ; Chen, Zaijie 1 ; Xu, Huibing 5 ;

作者机构： 1.Fujian Acad Agr Sci, Biotechnol Res Inst, Fuzhou, Fujian, Peoples R China

2.Fujian Agr & Forestry Univ, State Key Lab Ecol Pest Control Fujian & Taiwan C, Coll Plant Protect, Fuzhou, Fujian, Peoples R China

3.Natl Univ Singapore, Temasek Life Sci Lab, Singapore, Singapore

4.Natl Univ Singapore, Dept Biol Sci, Singapore, Singapore

5.Minjiang Univ, Marine & Agr Biotechnol Lab, Inst Oceanog, Fuzhou, Fujian, Peoples R China

关键词： fluorescent protein; organelle marker lines; protein subcellular localization; rice; Magnaporthe oryzae

期刊名称：FRONTIERS IN PLANT SCIENCE （ 影响因子：5.753； 五年影响因子：6.612 ）

ISSN： 1664-462X

年卷期： 2019 年 10 卷

页码：

收录情况： SCI

摘要： The subcellular localization of proteins is a fundamental aspect of protein functions. Determining the subcellular localization is important for understanding the biological functions of proteins. Here, we developed a set of rice organelle marker lines, in which the expressing fluorescent organelle markers could be used as comparative standards in determining the subcellular localization of the protein of interest. We constructed green fluorescent protein (GFP)- and/or Discosoma sp. red fluorescent protein (DsRed)-tagged organelle markers targeted to the endoplasmic reticulum (ER), mitochondria, Golgi apparatus, peroxisome, actin cytoskeleton, plastid, tonoplast, plasma membrane, and nucleus, respectively. The utility of the rice marker lines for protein subcellular localization studies was demonstrated by detecting a nucleus-localized OsWRKY45 and a mitochondria-associated NbHxk1 in protoplasts of the GFP-OsH2B and the ScCOX4-DsRed lines, respectively. Using a sheath-inoculation method, followed by a live-cell imaging, we detected co-localization of a Magnaporthe oryzae PWL2:mCherry : NLS fusion with the nucleus marker in the GFP-OsH2B rice epidermal cells, confirming the translocation of the M. oryzae effector PWL2 into host cells, and further demonstrating the feasibility of using the organelle marker lines for studying dynamics of proteins in rice cells in the interactions between rice and pathogens. The set of organelle marker lines developed in the present study, provides a valuable resource for protein subcellular localization studies in rice.

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