Ozone is now considered to be the second most important gaseous pollutant in our environment. The phytotoxic potential of O₃ was first observed on grape foliage by B.L. Richards and coworkers in 1958 (Richards et al. 1958). To date, unsustainable resource utilization has turned this secondary pollutant into a major component of global climate change and a prime threat to agricultural production. The projected levels to which O₃ will increase are critically alarming and have become a major issue of concern for agriculturalists, biologists, environmentalists and others plants are soft targets for O₃. Ozone enters plants through stomata, where it disolves in the apoplastic fluid. O₃ has several potential effects on plants: direct reaction with cell membranes; conversion into ROS and H₂O₂ (which alters cellular function by causing cell death); induction of premature senescence; and induction of and up- or down-regulation of responsive components such as genes , proteins and metabolites. In this review we attempt to present an overview picture of plant O₃ interactions. We summarize the vast number of available reports on plant responses to O₃ at the morphological, physiological, cellular, biochemical levels, and address effects on crop yield, and on genes, proteins and metabolites. it is now clear that the machinery of photosynthesis, thereby decreasing the economic yield of most plants and inducing a common morphological symptom, called the "foliar injury". The "foliar injury" symptoms can be authentically utilized for biomonitoring of O₃ under natural conditions. Elevated O₃ stress has been convincingly demonstrated to trigger an antioxidative defense system in plants. The past several years have seen the development and application of high-throughput omics technologies (transcriptomics, proteomics, and metabolomics) that are capable of identifying and prolifiling the O₃-responsive components in model and nonmodel plants. Such studies have been carried out ans have generated an inventory of O₃-Responsive components--a great resource to the scientific community. Recently, it has been shown that certain organic chemicals ans elevated CO₂ levels are effective in ameliorating O₃-generated stress. Both targeted and highthroughput approaches have advanced our knowledge concerning what O₃-triggerred signaling and metabolic pathways exist in plants. Moreover, recently generated information, and several biomarkers for O₃, may, in the future, be exploited to better screen and develop O₃-tolerant plants.

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.

Sample preparation in plant proteomics is tedious, requiring modifications depending on the type of tissue involved. Here, we describe a protein extractlon protocoM for both monocotyledonous (monocot) and dicotyledonous (dicot) species, which significantly improves the soiubilization of total proteins~ For exam#e, we used the primary leaf tissue and seeds from rice, a cereai crop and genome model system. Total protein was first precipitated with trichRoroacetic acid/acetone extraction buffer (

The OsSIPK expression is transcriptionally regulated in time and space by diverse environmental stresses and phytohormones. Rice OsSIPK and its orthologs in other plants are highly conserved and appear to have overlapping physiological responses. Given our interest in understanding the signaling and metabolic pathways responsible for environmental factors, we briefly discuss the role of OsSIPK in ozone-triggered physiological responses, particularly in rice. We also provide evidence on tight correlation between ozone-induced OsSIPK expression and ethylene production.