Mitogen-activated protein kinase （MAPK） cascade has been shown to be important components in stress signal transduction pathway. In the present study, protection of maize seedlings （Zea mays L.） against paraquat-generated oxidative toxicity by abscisic acid （ABA）, its association with MAPK and ZmMPK5, a candidate for MAPK were investigated. Treatment of maize leaves with exogenous ABA led to significant decreases in the content of malondialdehyde, the percentage of ion leakage and the level of protein oxidation （in terms of carbonyl groups） under paraquat （PQ） stress. However, such decreases were blocked by the pretreatment with two MAPK kinase inhibitors PD98059 and U0126. The damage caused by PQ was further aggravated by inhibitors. Two inhibitors also suppressed the total activities of the antioxidant enzymes superoxide dismutase （SOD, EC 1.15.1.1）, catalase （CAT, EC 1.11.1.6）, ascorbate peroxidase （APX, EC 1.11.1.tl）, and glutathione reductase （GR, EC 1.6.4.2）. Besides, treatment with PQ stimulated the activation of a 46 kDa MAPK, which was identified as ZmMPK5 by in-gel kinase assay with immunoprecipitation. These results reveal that ABA-induced protection against PQ-generated oxidative damage is mediated through MAPK cascade in maize leaves, in which ZmMPK5, a candidate for MAPK, is demonstrated to be involved.
Hai-Dong DingXiao-Hua ZhangShu-Cheng XULi-Li SunMing-Yi JiangA-Ying ZhangYin-Gen Jin
With the exception of rice, little is known about the existence of respiratory burst oxidase homolog （rboh） gene in cereals. The present study reports the cloning and analysis of a novel rboh gene, termed ZmrbohB, from maize （Zea mays L.）. The full-length cDNA of ZmrbohB encodes a 942 amino acid protein containing all of the respiratory burst oxidase homolog catalytically critical motifs. Alternative splicing of ZmrbohB has generated two transcript isoforms, ZmrbohB-α and -β. Spliced transcript ZmrbohB-β retains an unspliced intron 11 that carries a premature termination codon and probably leads to nonsense-mediated mRNA decay. Expression analysis showed that two splice isoforms were differentially expressed in various tissues and at different developmental stages, and the major product was ZmrbohB-e. The transcripts of ZmrbohB-α accumulated markedly when the maize seedlings were subjected to various abiotic stimuli, such as wounding, cold （4℃）, heat （40℃）, UV and salinity stress. In addition, several abiotic stimuli also affected the alternative splicing pattern of ZmrbohB except wounding. These results provide new insight into roles in the expression regulation of plant rboh genes and suggest that ZrnrbohB gene may play a role in response to environmental stresses.
