Antifreeze activity of cold acclimated Japanese radish and purification of antifreeze peptide.
Japanese radish tuber and leaf produced antifreeze proteins (AFPs) having thermal hysteresis activity (TH) and ice recrystallization inhibiting activity (RI). Upon cold acclimation, the apoplastic fluid of the Japanese radish exhibited hexagonal crystal growth, indicating the presence of an antifreeze protein. The induction patterns of protein and the TH activity of apoplastic fraction from both samples were different. The TH activities of apoplastic fraction from tuber and leaves were 0.20 +/- 0.03 and 0.18 +/- 0.02 degree C, respectively. Also, the TH and RI activities of apoplastic fraction of leaves were activated by autoclave treatment at pH 10.0. An antifreeze peptide (molecular weight 1320), was purified using chromatography. Furthermore, the chitinase and beta-1, 3-glucanase activities in the apoplastic fraction of its tuber were induced by the cold acclimation. Some proteins in this apoplastic fraction were reacted with the anti-glucanase-like protein (GLP) antiserum and anti-chitinase-like protein (CLP) antiserum produced against isolated winter rye AFPs. This is the first report on the presence and characterization of AFPs from Japanese radish tuber.
Kawahara
Hidehisa
H
Department of Life Science and Biotechnology, Faculty of Chemistry, Materials and Bioengineering, Kansai University, Osaka, Japan. kawahara@ipcku.kansai-u.ac.jp
Fujii
Atsuko
A
Inoue
Michihiro
M
Kitao
Satoshi
S
Fukuoka
Jyoichi
J
Obata
Hitoshi
H
D000064:Acclimatization; D021301:Antifreeze Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D015153:Blotting, Western; D002688:Chitinases / Q000378:metabolism; D003080:Cold Temperature; D003460:Crystallization; D043326:Glucan 1,3-beta-Glucosidase / Q000378:metabolism; D007053:Ice; D018515:Plant Leaves / Q000737:chemistry / Q000502:physiology; D010940:Plant Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D035281:Plant Tubers / Q000737:chemistry / Q000502:physiology; D031224:Raphanus / Q000737:chemistry / Q000502:physiology*
D021301:Antifreeze Proteins; D007053:Ice; D010940:Plant Proteins; D002688:Chitinases; D043326:Glucan 1,3-beta-Glucosidase