title: A rapid and efficient method to study the function of crop plant transporters in Arabidopsis.
abstract: Iron (Fe) is an essential micronutrient for humans. Fe deficiency disease is widespread and has led to extensive studies on the mechanisms of Fe uptake and storage, especially in staple food crops such as rice. However, studies of functionally related genes in rice and other crops are often time and space demanding. Here, we demonstrate that transgenic Arabidopsis suspension culture cells and Arabidopsis plants can be used as an efficient expression system for gain-of-function study of selected transporters, using Fe transporters as a proof-of-principle. The vacuolar membrane transporters OsVIT1 and OsVIT2 have been described to be important for iron sequestration, and disruption of these two genes leads to Fe accumulation in rice seeds. In this study, we have taken advantage of the fluorescent-tagged protein GFP-OsVIT1, which functionally complements the Fe hypersensitivity of ccc1 yeast mutant, to generate transgenic Arabidopsis suspension cell lines and plants. GFP-OsVIT1 was shown to localize on the vacuolar membrane using confocal microscopy and immunogold EM. More importantly, the Fe concentration, as well as the concentration of Zn, in the transgenic cell lines and plants were significantly increased compared to that in the WT. Taken together, our study shows that the heterologous expression of rice vacuolar membrane transporter OsVIT1 in Arabidopsis system is functional and effectively enhances iron accumulation, indicating an useful approach for studying other putative transporters of crop plants in this system.
authors.affiliation: School of Life Sciences, Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
authors.forename: Xiangfeng
authors.initials: X
authors.lastname: Wang
authors.affiliation: School of Life Sciences, Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
authors.forename: Fudi
authors.initials: F
authors.lastname: Zhong
authors.affiliation: School of Life Sciences, Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
authors.forename: Cheuk Hang
authors.initials: CH
authors.lastname: Woo
authors.affiliation: School of Life Sciences, Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
authors.forename: Yansong
authors.initials: Y
authors.lastname: Miao
authors.affiliation: Department of Pediatrics, United States Department of Agriculture/Agricultural Research Service Children's Nutrition Research Center, Baylor College of Medicine, 1100 Bates Street, Houston, TX, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Grusak
authors.affiliation: Institute of Crop Science, Zhejiang University, No. 866 Yuhangtang Road, Hangzhou, 310058, China.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Institute of Crop Science, Zhejiang University, No. 866 Yuhangtang Road, Hangzhou, 310058, China.
authors.forename: Jumin
authors.initials: J
authors.lastname: Tu
authors.affiliation: School of Life Sciences, Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
authors.forename: Yum Shing
authors.initials: YS
authors.lastname: Wong
authors.affiliation: School of Life Sciences, Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China. ljiang@cuhk.edu.hk.|CUHK Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, 518057, China. ljiang@cuhk.edu.hk.
authors.forename: Liwen
authors.initials: L
authors.lastname: Jiang
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D001692:Biological Transport; D001901:Botany / Q000379:methods*; D027682:Cation Transport Proteins / Q000378:metabolism*; D018556:Crops, Agricultural / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D007501:Iron / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011523:Protoplasts / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism
chemical_list: D027682:Cation Transport Proteins; D010940:Plant Proteins; D049452:Green Fluorescent Proteins; D007501:Iron
keywords: Arabidopsis suspension culture cells; Iron accumulation; OsVIT1