Expression of the vacuolar Ca2+/H+ exchanger, OsCAX1a, in rice: cell and age specificity of expression, and enhancement by Ca2+.
Calcium is an essential macronutrient for plants and functions in signal transduction. Regulation of the cytosolic calcium concentration is required for normal cell growth. In calcium homeostasis in plant cells, Ca(2+)/H(+) exchangers are involved in Ca(2+) compartmentalization into intracellular compartments. Here, we examine the intracellular localization of a rice Ca(2+)/H(+) exchanger, OsCAX1a, fused to a green fluorescent protein and transiently expressed in onion epidermis and rice protoplasts. Green fluorescence was observed in the vacuolar membrane. After sucrose gradient centrifugation of the homogenate of rice plants, OsCAX1a was detected in the same fraction as the vacuolar membrane aquaporin gamma-TIP. We then quantified the mRNA and protein of OsCAX1a in plants grown with metal ions. OsCAX1a mRNA was induced in roots by high concentrations of Ca(2+). The protein level in shoots was also increased in the presence of high concentrations of Ca(2+). Furthermore, transgenic rice plants transformed with the OsCAX1a promoter fused to beta-glucuronidase showed reporter expression in vascular bundles, stomata, trichomes, steles, flowers, embryos and aleurone layers. In the case of stomata and trichomes, transcription of OsCAX1a was particularly high in aged organs. These results suggest that OsCAX1a transports Ca(2+) into vacuoles and is involved in Ca(2+) homeostasis in cells that suffer from high concentrations of Ca(2+).
Kamiya
Takehiro
T
Laboratory of Cell Dynamics, Graduate School of Bioagricultural Sciences, Nagoya University, Japan.
Akahori
Taro
T
Ashikari
Motoyuki
M
Maeshima
Masayoshi
M
D017920:Antiporters / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D027682:Cation Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D018744:DNA, Plant / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D017343:Genes, Plant; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D012275:Oryza / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D009997:Osmotic Pressure; D030821:Plants, Genetically Modified; D011092:Polyethylene Glycols / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution; D014617:Vacuoles / Q000378:metabolism
D017920:Antiporters; D002135:Calcium-Binding Proteins; D027682:Cation Transport Proteins; D018744:DNA, Plant; D008670:Metals; D012333:RNA, Messenger; D018749:RNA, Plant; D011993:Recombinant Fusion Proteins; C027614:calcium-hydrogen antiporters; D049452:Green Fluorescent Proteins; D011092:Polyethylene Glycols; D002118:Calcium