An approach to identify cold-induced low-abundant proteins in rice leaf.
A proteomic approach has been adopted to investigate the low-abundant proteins in rice leaf in response to cold stress. Rice seedlings were exposed to different temperatures, such as 5 or 10 degrees C, and samples were collected after different time course. To eliminate the high-abundant proteins in leaf tissues such as ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), proteins were fractionated by polyethylene glycol (PEG). The elimination of Rubisco from the protein samples was confirmed by Western blot analysis. The PEG fractionated protein samples were separated by 2-DE and visualized by silver or CBB staining. A total 12 up-regulated protein spots were identified using the analysis of MALDI-TOF mass spectrometry or ESI MS/MS. We identified some novel proteins such as cysteine proteinase, thioredoxin peroxidase, a RING zinc finger protein-like, drought-inducible late embryogenesis abundant, and a fibrillin-like protein that had not yet been reported in the earlier reports on cold proteomic analysis. The identification of some novel low-abundant proteins in response to cold stress may provide a new homeostasis to develop enhanced cold tolerance transgenic plants. Thus, we propose that a PEG fractionation system can be used as an influential protein extraction method from the leaf samples, which can lead to knowledge of the expression pattern of low-abundant proteins in response to various biotic or abiotic stresses.
Lee
Dong-Gi
DG
Division of Applied Life Science (BK21 program), PMBBRC, Gyeongsang National University, Jinju 660-701, Republic of Korea.
Ahsan
Nagib
N
Lee
Sang-Hoon
SH
Kang
Kyu Young
KY
Lee
Jeung Joo
JJ
Lee
Byung-Hyun
BH
D000704:Analysis of Variance; D003080:Cold Temperature; D004190:Disasters; D015180:Electrophoresis, Gel, Two-Dimensional; D004798:Enzymes / Q000096:biosynthesis / Q000302:isolation & purification; D007723:Korea; D013058:Mass Spectrometry; D012275:Oryza / Q000502:physiology*; D018515:Plant Leaves / Q000502:physiology*; D010940:Plant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D020543:Proteome; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D004798:Enzymes; D010940:Plant Proteins; D020543:Proteome