title: RoGFP1 is a quantitative biosensor in maize cells for cellular redox changes caused by environmental and endogenous stimuli.
abstract: Reduction-oxidation-sensitive green fluorescent proteins (roGFPs) have been demonstrated to be valuable tools in sensing cellular redox changes in mammalian cells and model plants, yet have not been applied in crops such as maize. Here we report the characteristics of roGFP1 in transiently transformed maize mesophyll protoplasts in response to environmental stimuli and knocked-down expression of ROS-scavenging genes. We demonstrated that roGFP1 in maize cells ratiometrically responds to cellular redox changes caused by H2O2 and DTT, as it does in mammalian cells and model plants. Moreover, we found that roGFP1 is sensitive enough to cellular redox changes caused by genetic perturbation of single ROS genes, as exemplified by knocked-down expression of individual ZmAPXs, in maize protoplasts under controlled culture conditions and under stress conditions imposed by H2O2 addition. These data provide evidence that roGFP1 functions in maize cells as a biosensor for cellular redox changes triggered by genetic lesion of single ROS genes even under stress conditions, and suggest a potential application of roGFP1 in large-scale screening for maize mutants of ROS signaling involved in development and stress resistance.
authors.affiliation: Faculty of Maize Functional Genomics, Department of Crop Genome Research and Genetic Improvement, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China; National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China.
authors.forename: Xiaoning
authors.initials: X
authors.lastname: Liu
authors.affiliation: Faculty of Maize Functional Genomics, Department of Crop Genome Research and Genetic Improvement, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China; National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China.
authors.forename: Jiamei
authors.initials: J
authors.lastname: Wu
authors.affiliation: Faculty of Maize Functional Genomics, Department of Crop Genome Research and Genetic Improvement, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China; National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China.
authors.forename: Hao
authors.initials: H
authors.lastname: Liu
authors.affiliation: Faculty of Maize Functional Genomics, Department of Crop Genome Research and Genetic Improvement, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China; National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China.
authors.forename: Na
authors.initials: N
authors.lastname: Zong
authors.affiliation: Faculty of Maize Functional Genomics, Department of Crop Genome Research and Genetic Improvement, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China; National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China. Electronic address: zhaojun01@caas.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhao
mesh_terms: D060387:Ascorbate Peroxidases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D015374:Biosensing Techniques; D004229:Dithiothreitol / Q000494:pharmacology; D018506:Gene Expression Regulation, Plant; D005978:Glutathione / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D010084:Oxidation-Reduction; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000378:metabolism*; D011523:Protoplasts / Q000187:drug effects / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000032:analysis* / Q000378:metabolism; D003313:Zea mays / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*
chemical_list: D010940:Plant Proteins; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D049452:Green Fluorescent Proteins; D006861:Hydrogen Peroxide; D060387:Ascorbate Peroxidases; D005978:Glutathione; D004229:Dithiothreitol
keywords: Maize; RNAi; ROS; Redox change; RoGFP1; ZmAPX