Gel-based proteomics reveals potential novel protein markers of ozone stress in leaves of cultivated bean and maize species of Panama.
We examined responses of cultivated bean (Phaseolus vulgaris L. cv. IDIAP R-3) and maize (Zea mays L. cv. Guarare 8128) plants exposed to ozone (O(3)) using a leaf injury assessment and proteomics approach. Plants grown for 16 days in greenhouse were transferred to an O(3) chamber and exposed continuously to 0.2 ppm O(3) or filtered pollutant-free air for up to 72 h. CBB-stained gels revealed changes in ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) protein. By Western analysis changes in marker proteins for O(3) damage in leaves by 1-DE were checked. In bean leaves, two superoxide dismutase (SOD) protein (19 and 20 kDa) were dramatically decreased, while ascorbate peroxidase (APX, 25 kDa), small heat shock protein (HSP, 33 kDa), and a naringenin-7-O-methyltransferase (NOMT, 42 kDa) were increased by O(3). In maize leaves, expression levels of catalase (increased), SOD (decreased), and APX (increased) were drastically changed by O(3) depending on the leaf stage, whereas crossreacting HSPs (24 and 30 kDa) and NOMT (41 kDa) proteins were strongly increased in O(3)-stressed younger leaves. These results indicated a clear modulation of oxidative stress-, heat shock-, and secondary metabolism-related proteins by O(3). Finally, 2-DE at 72 h after O(3) exposure revealed changes (induction/suppression) in expression levels of 25 and 12 protein spots in bean and maize leaves, respectively. Out of these, ten and nine nonredundant proteins in bean and maize, respectively, were identified by MS. A novel pathogenesis-related protein 2 may serve as a potential marker for O(3) stress in bean.
Torres
Nilka Lineth
NL
University Regional Center of Azuero, Chitre, Herrera, Republic of Panama.
Cho
Kyoungwon
K
Shibato
Junko
J
Hirano
Misato
M
Kubo
Akihiro
A
Masuo
Yoshinori
Y
Iwahashi
Hitoshi
H
Jwa
Nam-Soo
NS
Agrawal
Ganesh Kumar
GK
Rakwal
Randeep
R
D060387:Ascorbate Peroxidases; D002374:Catalase / Q000032:analysis / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004784:Environmental Monitoring / Q000379:methods*; D004789:Enzyme Activation; D004790:Enzyme Induction; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D005819:Genetic Markers; D006360:Heat-Shock Proteins / Q000032:analysis / Q000378:metabolism; D008780:Methyltransferases / Q000032:analysis / Q000378:metabolism; D010126:Ozone / Q000633:toxicity*; D010176:Panama; D010544:Peroxidases / Q000032:analysis / Q000378:metabolism; D027805:Phaseolus / Q000378:metabolism*; D018515:Plant Leaves / Q000187:drug effects / Q000201:enzymology*; D010940:Plant Proteins / Q000032:analysis* / Q000378:metabolism; D020543:Proteome / Q000187:drug effects; D040901:Proteomics / Q000379:methods; D012273:Ribulose-Bisphosphate Carboxylase / Q000032:analysis / Q000378:metabolism; D013482:Superoxide Dismutase / Q000032:analysis / Q000378:metabolism; D003313:Zea mays / Q000378:metabolism*
D005819:Genetic Markers; D006360:Heat-Shock Proteins; D010940:Plant Proteins; D020543:Proteome; D010126:Ozone; D010544:Peroxidases; D060387:Ascorbate Peroxidases; D002374:Catalase; D013482:Superoxide Dismutase; D008780:Methyltransferases; C099725:naringenin 7-O-methyltransferase; D012273:Ribulose-Bisphosphate Carboxylase