title: Characterization of vegetative storage protein (VSP) and low molecular proteins induced by water deficit in stolon of white clover.
abstract: In stolon of white clover (Trifolium repens L.), the 17.3 kDa protein has been newly identified as a vegetative storage protein (VSP) which has preponderant roles in N accumulation and mobilization to sustain growth when capacity of N uptake is strongly reduced. To characterize the water deficit effect on this protein, the kinetic pattern of soluble protein, SDS-PAGE, Western blotting, and proteomic analysis was studied in the stolon of white clover during 28 days of water-deficit. Water deficit led to decrease protein concentration. SDS-PAGE revealed that two major proteins of 17.3 and 16 kDa were accumulated to high level in response to water stress. These proteins cross-reacted positively with antibodies raised against the 17.3 kDa VSP, a protein which shared biochemical features with stress proteins implied in dehydration tolerance. Using two-dimensional electrophoresis (2-DE) gel and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF-MS) analysis, it was demonstrated that 19.5 and 17.3 kDa protein spots were up-regulated by water stress, and both spots were identical to nucleoside diphosphate kinase (NDPK) and lipid transfer proteins (LTPs), respectively. These results suggest that low molecular proteins induced by water-deficit in the stolon of white clover act as an alternative N reserves or play significant roles in plant protection against water-deficit stress.
authors.affiliation: Department of Animal Science, Institute of Agricultural Science and Technology, College of Agriculture and Life Science, Chonnam National University, P.O. Box 205, Buk-Gwangju, South Korea; Bioenergy Research Center, Chonnam National University, Gwangju 500-757, South Korea.
authors.forename: Bok-Rye
authors.initials: BR
authors.lastname: Lee
authors.affiliation: Division of Life Science, Korea Basic Science Institute, Daejeon 305-806, South Korea.
authors.forename: Dong-Gi
authors.initials: DG
authors.lastname: Lee
authors.affiliation: INRA, UMR INRA/UCBN 950 Ecophysiologie Végétale, Agronomie et Nutritions NCS, IFR 146 ICORE, Institut de Biologie Fondamentale et Appliquée, Université de Caen Basse-Normandie, F-14032 Caen, France.
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Avice
authors.affiliation: Department of Animal Science, Institute of Agricultural Science and Technology, College of Agriculture and Life Science, Chonnam National University, P.O. Box 205, Buk-Gwangju, South Korea. Electronic address: grassl@chonnam.ac.kr.
authors.forename: Tae-Hwan
authors.initials: TH
authors.lastname: Kim
mesh_terms: D055864:Droughts; D004591:Electrophoresis, Polyacrylamide Gel; D010940:Plant Proteins / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013312:Stress, Physiological; D029921:Trifolium / Q000378:metabolism / Q000502:physiology*; D014867:Water / Q000502:physiology*
chemical_list: D010940:Plant Proteins; D014867:Water
keywords: 2-DE; LTPs; MALDI-TOF-MS; NDPK; Proteomics; ROS; SDS–PAGE; VSP; Vegetative storage protein; Vegetative storage protein (VSP); Water-deficit; White clover; lipid transfer proteins; matrix-assisted laser desorption/ionization time-of-flight mass spectrometer; nucleoside diphosphate kinase; reactive oxygen species; sodium dodecylsulfate polyacrylamide gel electrophoresis; two-dimensional electrophoresis