Proteome analysis of differentially displayed proteins as a tool for investigating ozone stress in rice (Oryza sativa L.) seedlings.
Employing classical two-dimensional electrophoresis (2-DE), amino acid sequencing and immunoblot analysis, we examine for the first time the effect of ozone, a highly notorious environmental pollutant, on rice seedling proteins. Drastic visible necrotic damage to leaf by ozone and consequent increase in ascorbate peroxidase protein(s) was accompanied by rapid changes in the 2-DE protein profiles, over controls. Out of a total of 56 proteins investigated, which were reproducible in repeated experiments, 52 protein spots were visually identified as differentially expressed over controls. Six proteins were N-terminally blocked, and the sequence of 14 proteins could not be determined, whereas 36 proteins were N-terminally and one was internally sequenced. Ozone caused drastic reductions in the major leaf photosynthetic proteins, including the abundantly present ribulose-1, 5-bisphosphate carboxylase/oxygenase, and induction of various defense/stress related proteins. Most prominent change in leaves, within 24 h post-treatment with ozone, was the induced accumulation of a pathogenesis related (PR) class 5 protein, three PR 10 class proteins, ascorbate peroxidase(s), superoxide dismutase, calcium-binding protein, calreticulin, a novel ATP-dependent CLP protease, and an unknown protein. Present results demonstrate the highly damaging effect of ozone on rice seedlings at the level of the proteome.
Research Laboratory for Agricultural Biotechnology and Biochemistry (RLABB), Kathmandu, Nepal.
Ganesh Kumar
GK
Agrawal
Randeep
R
Rakwal
Masami
M
Yonekura
Akihiro
A
Kubo
Hikaru
H
Saji
D000393:Air Pollutants / Q000633:toxicity; D000595:Amino Acid Sequence; D060387:Ascorbate Peroxidases; D015180:Electrophoresis, Gel, Two-Dimensional; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D012275:Oryza / Q000187:drug effects / Q000502:physiology*; D010083:Oxidants, Photochemical / Q000494:pharmacology* / Q000633:toxicity; D018384:Oxidative Stress; D010126:Ozone / Q000633:toxicity*; D010544:Peroxidases / Q000378:metabolism; D010940:Plant Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020543:Proteome; D012273:Ribulose-Bisphosphate Carboxylase / Q000378:metabolism; D036226:Seedlings / Q000187:drug effects / Q000502:physiology
D000393:Air Pollutants; D010083:Oxidants, Photochemical; D010940:Plant Proteins; D020543:Proteome; C053376:pathogenesis-related proteins, plant; D010126:Ozone; D010544:Peroxidases; D060387:Ascorbate Peroxidases; D012273:Ribulose-Bisphosphate Carboxylase