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The appreciation of the Japanese yen does not explain why Japanese firms have doubled foreign direct investment (FDI) in Southeast Asia since the late 2000s. This study investigates the effects of exchange rates and other factors on new Japanese FDI in Indonesia, Malaysia, the Philippines, and Thailand, using industry-specific panel data from 2008 to 2015. It is found that new FDI was more strongly correlated with the expectation of consumption demand than with exchange rate movements for Indonesia and Thailand, while the agglomeration of Japanese firms predominantly affected new FDI in Malaysia and the Philippines.

Visual object tracking remains a challenging computer vision problem with numerous real-world applications. Discriminative correlation filter (DCF)-based methods are a recent state-of-the-art approach for dealing with this problem. The learning rate when applying a DCF is typically fixed, regardless of the situation. However, this rate is important for robust tracking, insofar as real-world video sequences include a variety of dynamical changes, such as occlusions, motion blur, and deformations. In this study, we propose Meta-Q-learning Correlation Filter (MQCF), a method for dynamically determining the learning rate of a baseline DCF-based tracker based on hand-crafted features of Histogram of Oriented Gradient (HOG), by means of reinforcement learning. The incorporation of reinforcement learning enables us to train a function for an image patch that outputs a situation-dependent learning rate of the baseline tracker in an autonomous fashion. We evaluated this method using two open benchmarks, namely, OTB-2015 and VOT-2105, and found our MQCF tracker outperformed a baseline state-of-the-art tracker by 1.8% in Area Under Curve on OTB-2015, and 8.4% relative gain in Expected Average Overlap in the VOT-2015 challenge. Our results demonstrate the advantages of the so-called meta-learning with DCF-based visual object tracking.

Here, we present an update on the next level of experiments studying the impact of the gamma radiation environment, created post-March, 2011 nuclear accident at Fukushima Daiichi nuclear power plant, on rice plant and its next generation-the seed. Japonica-type rice (Oryza sativa L. cv. Koshihikari) plant was exposed to low-level gamma radiation (~4 μSv/h) in the contaminated Iitate Farm field in Iitate village (Fukushima). Seeds were harvested from these plants at maturity, and serve as the treated group. For control group, seeds (cv. Koshihikari) were harvested from rice grown in clean soil in Soma city, adjacent to Iitate village, in Fukushima. Focusing on the multi-omics approach, we have investigated the dry mature rice seed transcriptome, proteome, and metabolome following cultivation of rice in the radionuclide contaminated soil and compared it with the control group seed (non-radioactive field-soil environment). This update article presents an overview of both the multi-omics approach/technologies and the first findings on how rice seed has changed or adapted its biology to the low-level radioactive environment.

This paper focuses on empirically investigating the efficacy of foreign exchange intervention in the Thai economy by simulation analyses. We find that foreign reserves are determinants of exchange rate dynamics, whereas the uncovered interest parity condition does not hold. We also find that foreign exchange intervention influences the inflation rate via the exchange rate, although by a lesser degree, whereas such an intervention for an extended period is likely to incur higher costs of the macro economy.

An ozone-sensitive mutant was isolated from T-DNA-tagged lines of Arabidopsis thaliana. The T-DNA was inserted at a locus on chromosome 3, where two genes encoding glycolate oxidases, GOX1 and GOX2, peroxisomal enzymes involved in photorespiration, reside contiguously. The amounts of the mutant's foliar transcripts for these genes were reduced, and glycolate oxidase activity was approximately 60% of that of the wild-type plants. No difference in growth and appearance was observed between the mutant and the wild-type plants under normal conditions with ambient air under a light intensity of 100 µmol photons m-2 s-1. However, signs of severe damage, such as chlorosis and ion leakage from the tissue, rapidly appeared in mutant leaves in response to ozone treatment at a concentration of 0.2 µl l-1 under a higher light intensity of 350 µmol photons m-2 s-1 that caused no such symptoms in the wild-type plant. The mutant also exhibited sensitivity to sulfur dioxide and long-term high-intensity light. Arabidopsis mutants with deficiencies in other photorespiratory enzymes such as glutamate:glyoxylate aminotransferase and hydroxypyruvate reductase also exhibited ozone sensitivities. Therefore, photorespiration appears to be involved in protection against photooxidative stress caused by ozone and other abiotic factors under high-intensity light.

Stomatal movements regulate gas exchange, thus directly affecting the efficiency of photosynthesis and the sensitivity of plants to air pollutants such as ozone. The GARP family transcription factors GOLDEN 2-LIKE1 (GLK1) and GLK2 have known functions in chloroplast development. Here, we show that Arabidopsis thaliana (A. thaliana) plants expressing the chimeric repressors for GLK1 and -2 (GLK1/2-SRDX) exhibited a closed-stomata phenotype and strong tolerance to ozone. By contrast, plants that overexpress GLK1/2 exhibited an open-stomata phenotype and higher sensitivity to ozone. The plants expressing GLK1-SRDX had reduced expression of the genes for inwardly rectifying K(+) (K(+) in) channels and reduced K(+) in channel activity. Abscisic acid treatment did not affect the stomatal phenotype of 35S:GLK1/2-SRDX plants or the transcriptional activity for K(+) in channel gene, indicating that GLK1/2 act independently of abscisic acid signaling. Our results indicate that GLK1/2 positively regulate the expression of genes for K(+) in channels and promote stomatal opening. Because the chimeric GLK1-SRDX repressor driven by a guard cell-specific promoter induced a closed-stomata phenotype without affecting chloroplast development in mesophyll cells, modulating GLK1/2 activity may provide an effective tool to control stomatal movements and thus to confer resistance to air pollutants.

Previously, we conducted a roadside survey to reveal the occurrence of genetically modified (GM) oilseed rape along a Japanese roadside (Route 51). In this study, we performed successive and thorough fixed-route monitoring in 5 sections along another road (Route 23). Oilseed rape plants were detected on both sides of the road in each section between autumn 2009 and winter 2013, which included 3 flowering seasons. In four sections, more plants were found on the side of the road leading from the Yokkaichi port than on the opposite side. In the fifth section, the presence of clogged drains on the roadside, where juvenile plants concentrated, caused the opposite distribution: oilseed rape predominantly occurred along the inbound lanes (leading to the Yokkaichi port) in 2010 and 2012. Unlike in our previous survey, glyphosate- or glufosinate-resistant oilseed rape plants were abundant (>75% of analyzed plants over 3 years). Moreover, a few individuals bearing both herbicide resistance traits were also detected in some sections. The spillage of imported seeds may explain the occurrence of oilseed rape on the roadside. The abundance of herbicide-resistant oilseed rape plants may reflect the extent of contamination with GM oilseed rape seed within imports.

In the summer of 2012, 1 year after the nuclear accident in March 2011 at the Fukushima Daiichi nuclear power plant, we examined the effects of gamma radiation on rice at a highly contaminated field of Iitate village in Fukushima, Japan. We investigated the morphological and molecular changes on healthy rice seedlings exposed to continuous low-dose gamma radiation up to 4 µSv h(-1), about 80 times higher than natural background level. After exposure to gamma rays, expression profiles of selected genes involved in DNA replication/repair, oxidative stress, photosynthesis, and defense/stress functions were examined by RT-PCR, which revealed their differential expression in leaves in a time-dependent manner over 3 days (6, 12, 24, 48, and 72 h). For example, OsPCNA mRNA rapidly increased at 6, 12, and 24 h, suggesting that rice cells responded to radiation stress by activating a gene involved in DNA repair mechanisms. At 72 h, genes related to the phenylpropanoid pathway (OsPAL2) and cell death (OsPR1oa) were strongly induced, indicating activation of defense/stress responses. We next profiled the transcriptome using a customized rice whole-genome 4×44K DNA microarray at early (6h) and late (72 h) time periods. Low-level gamma radiation differentially regulated rice leaf gene expression (induced 4481 and suppressed 3740 at 6 h and induced 2291 and suppressed 1474 genes at 72 h) by at least 2-fold. Using the highly upregulated and downregulated gene list, MapMan bioinformatics tool generated diagrams of early and late pathways operating in cells responding to gamma ray exposure. An inventory of a large number of gamma radiation-responsive genes provides new information on novel regulatory processes in rice.

Background: We have previously demonstrated that long-term inhibition of Rho-kinase ameliorates pulmonary arterial hypertension (PAH) in animal models. In the present study, we examined the clinical effects of mid-term oral treatment with an extended release formulation of AT-877 (fasudil hydrochloride), a specific Rho-kinase inhibitor (AT-877ER) on PAH. Methods and Results: 23 PAH patients were treated with either placebo (10/2 females/males, 51±16 years, idiopathic PAH (IPAH) in 6, PAH associated with connective tissue disease (CTD-PAH) in 3, PAH with congenital heart disease (CHD-PAH) in 2, and portal PAH in 1) or AT-877ER (6/5 females/males, 47±14 years, IPAH in 2, CTD-PAH in 5, and CHD-PAH in 4); 3 patients were excluded. We performed a 6-min walk test and right heart catheterization in the remaining 20 patients, before and 3 months after the treatment (placebo n=11, AT-877ER n=9). Although there were no significant differences between the 2 groups for the 6-min walk distance, pulmonary hemodynamics tended to be improved in the AT-877ER group, especially the prevalence of improved cardiac index from baseline, which was significantly higher in the AT-877ER than in the placebo group. In the AT-877ER group, serum levels of hydroxyfasudil, an active metabolite of AT-877ER tended to correlate with improvements in the cardiac index and mean pulmonary artery pressure. Conclusions: Mid-term treatment with oral AT-877ER showed additional improvement in pulmonary hemodynamics in patients with PAH. (Circ J 2013; 77: 2619–2625)

The ‘ozone (O3)-responsive transcriptome’ behavior in the panicles and grains of rice plant was studied individually through high-throughput oligo-DNA microarray technique. O3 differentially and separately regulated 620 and 130 genes in the panicles and grains. Among the O3-responsive genes, 176 and 444 genes were up- and down-regulated in panicle compared to 24 and 106 genes in grain, respectively. Further mapping revealed that the majority of differentially expressed genes were mainly involved in signaling, hormonal, cell wall, transcription, proteolysis, and defense events. Many previously unknown O3-responsive novel genes were identified. Inventory of 745 O3-responsive genes and their mapping will expand our knowledge on novel regulatory processes in both panicles and grains of rice; and, serve as a resource towards the designing of rice crops for future high-O3world. Tropospheric ozone (O3) severely affects agricultural production worldwide. Present study aims to reveal a detailed O3 responsive gene network in panicle and grains of rice plants through transcriptomics approach. Our results provide an insight into the basis of O3-response in rice plants, and will help to develop suitable rice genotype for future high O3- world.

This paper empirically investigates the economic relationship between the US and Asian economies after the Asian currency crisis in Indonesia, Korea, the Philippines, Singapore, and Thailand, employing a cointegration methodology. Based on the empirical results, we conclude that the interdependence between the US and these Asian economies has intensified especially in information technology industries, and that their stock markets are integrated. On the other hand, the relationship between the domestic stock and foreign exchange markets is found to have a negative sign, interpreted by portfolio balance approach, in Indonesia, Korea, and Thailand. This result implies that the exchange rates of these countries are relatively vulnerable to fluctuation in international portfolio investments.

A combined approach of evaluating ozone (O 3 )-caused foliar injury symptom and global gene expression profiling was used to identify potential genes associated with severity of injury on leaves of O 3 (200 ppb)-fumigated (1, 12, 24, 48, and 72 h) two-week-old rice (cv. Nipponbare) seedling. Foliar injuries were evaluated up to 72 h using both qualitative visual scale and quantitative RGB (red-green-blue) image analysis methods. The (R-G)/(R+G) was an optimal quantitative RGB parameter to assess foliar injury. Large-scale transcript profiling of leaves identified 270 genes linked with foliar injury. Genes were subjected to Pearson’s correlation test between their expression changes and O 3 -induced foliar injury changes (relative (R-G)/(R+G) parameters). Expression of 146 genes was found to increase with increased foliar severity up to 72 h, showing a positive, tight correlation between a subset of gene expression and commonly observed O 3 -triggerred symptom of foliar injury with correlation coefficient below -0.80. Of 146 genes, genes involved in metabolism (25%) formed a major functional category. Metabolic networks with identified metabolic genes provided insight into cellular responses such as photorespiration, biosynthesis of secondary metabolites, and detoxification. The O 3 effect on these cellular responses has been previously reported based on physiological and biochemical studies, validating our approach used in this study to globally identify O 3 -responsive biomarkers tightly linked with foliar injury symptom. This study provides evidence for presence of large number of genes associated with foliar injury symptom than thought before, serving as a resource of potential biomarkers to study mechanisms of visible injury development by O 3 . DOI: http://dx.doi.org/10.3126/ijls.v6i1.6059

L-type amino-acid transporter 1 (LAT1) is the first identified light chain of CD98 molecule, disulfide-linked to a heavy chain of CD98. Following cDNA cloning of chicken full-length LAT1, we have constructed targeting vectors for the disruption of chicken LAT1 gene from genomic DNA of chicken LAT1 consisting of 5.4kb. We established five homozygous LAT1-disrupted (LAT1(-/-)) cell clones, derived from a heterozygous LAT1(+/-) clone of DT40 chicken B cell line. Reactivity of anti-chicken CD98hc monoclonal antibody (mAb) with LAT1(-/-) DT40 cells was markedly decreased compared with that of wild-type DT40 cells. All LAT1(-/-) cells were deficient in L-type amino-acid transporting activity, although alternative-splice variant but not full-length mRNA of LAT1 was detected in these cells. LAT1(-/-) DT40 clones showed outstandingly slow growth in liquid culture and decreased colony-formation capacity in soft agar compared with wild-type DT40 cells. Cell-cycle analyses indicated that LAT1(-/-) DT40 clones have prolonged cell-cycle phases compared with wild-type or LAT1(+/-) DT40 cells. Knockdown of human LAT1 by small interfering RNAs resulted in marked in vitro cell-growth inhibition of human cancer cells, and in vivo tumor growth of HeLa cells in athymic mice was significantly inhibited by anti-human LAT1 mAb. All these results indicate essential roles of LAT1 in the cell proliferation and occurrence of malignant phenotypes and that LAT1 is a promising candidate as a molecular target of human cancer therapy.

Using relatively recent data, this paper examines the causal relation between trade and production in some Asian developing countries. We find that causality analyses provide no evidence of export-led growth. Export-oriented growth has not been the primary strategy.

The 12-oxo-phytodienoic acid reductase (OPR) is a key enzyme in jasmonic acid (JA) biosynthesis. Previously, we reported the presence of 13 OPR isogenes (OsOPR1-13) in rice. OsOPRs phylogenetically belong to two subgroups, OPRI and OPRII. OsOPR13 is assigned to the second subgroup, which is involved in JA biosynthesis, while the others are found in the first subgroup. Here, we systematically investigated transcript levels of OsOPRs in various tissues and against diverse environmental stresses. Each gene was differentially involved in flower maturation, showing a tissue-specific response. OsOPR1, OsOPR2, and OsOPR13 were also active in responses to wounding, a fungal elicitor (chitosan), salt, UV-C irradiation, H2O2, and ozone exposure. In the case of JA-responsive OsOPRs (OsOPR1, OsOPR2, OsOPR6, OsOPR10, and OsOPR13), co-application of JA and SA suppressed jasmonate-induced transcript levels and delayed OsOPR10 expression. We also investigated the biochemical properties of OsOPR1 and found a flavin cofactor with optimal activity at pH 7.8 and values of 0.048 min−1 kcat and 8.33 μM Km for (9S,13R)-12-oxo-phytodienoic acid. Here, we discuss the role of OsOPRs in stress responses and floral development.

This paper empirically investigates the transmission of the Reserve Bank of India (RBI)'s monetary policy at a time when multiple indicator approaches are popular, using a structural VAR approach. We find that non-recursive zero restrictions must be imposed on the contemporaneous structural parameters to identify a monetary policy shock and that the RBI regards the current value of money as the centered information variable of the monetary policy, though exercising leverage over the real interest rate.

Using cointegration and structural vector autoregression (SVAR) techniques this paper investigates the effect of Bank Indonesia's (BI) monetary policy on inflation during the post-1997 crisis monetary-targeting period. Our analysis suggests that BI's monetary policy does not have systematic impact on the price level, apparently because of unstable money demand. Unreliable effects of BI's monetary policy are reflected in frequent and substantial deviations of the actual inflation rate from its targeted ranges.

Monitoring for escape of genetically modified (GM) oilseed rape (Brassica napus) during transport can be performed by means of roadside evaluations in areas where cultivation of this GM crop is not conducted, such as in Japan. We performed a survey of oilseed rape plants growing along a 20-km section of Japan's Route 51, one of the main land transportation routes in central Japan for imports of GM oilseed rape from the Port of Kashima into Keiyo District. Oilseed rape plants were found each year, but the number of plants varied substantially during the three years of our study: 2162 plants in 2005, 4066 in 2006, and only 278 in 2007. The low number in 2007 was probably caused by roadwork. Herbicide-resistant individuals were detected in the three consecutive years (26, 8, and 5 individuals with glyphosate resistance), but glufosinate-resistant plants (9 individuals) were detected only in 2005. The roadside plants occurred mainly along the inbound lane from Kashima to Narita. These plants are likely to have their origin in seeds spilled during transportation of cargo from the port, since there are no potential natural seed source plants for B. napus near Route 51. This is the first detailed report on the transition and distribution of herbicide-resistant oilseed rape plants following loss and spillage along Japanese roads.

Cyclobutane pyrimidine dimers (CPDs) constitute a majority of DNA lesions caused by ultraviolet-B (UVB). CPD photolyase, which rapidly repairs CPDs, is essential for plant survival under sunlight containing UVB. Our earlier results that the transcription of the cucumber CPD photolyase gene (CsPHR) was activated by light have prompted us to propose that this light-driven transcriptional activation would allow plants to meet the need of the photolyase activity upon challenges of UVB from sunlight. However, molecular mechanisms underlying the light-dependent transcriptional activation of CsPHR were unknown. In order to understand spectroscopic aspects of the plant response, we investigated the wavelength-dependence (action spectra) of the light-dependent transcriptional activation of CsPHR. In both cucumber seedlings and transgenic Arabidopsis seedlings expressing reporter genes under the control of the CsPHR promoter, the action spectra exhibited the most predominant peak in the long-wavelength UVB waveband (around 310 nm). In addition, a 95-bp cis-acting region in the CsPHR promoter was identified to be essential for the UVB-driven transcriptional activation of CsPHR. Thus, we concluded that the photoperception of long-wavelength UVB by UVB photoreceptor(s) led to the induction of the CsPHR transcription via a conserved cis-acting element.

This paper investigates the monetary transmission mechanism in Thailand, employing a VAR approach. It is found that the Bank of Thailand has leverage over the real interest rate in the short run due to inflation inertia. It is also found that the Thai monetary transmission mechanism has important international dimensions. More specifically, monetary contraction has stronger negative effects on import demand in the short run even though import prices fall.

To understand better the plant response to ozone, we isolated and characterized an ozone-sensitive (ozs1) mutant strain from a set of T-DNA-tagged Arabidopsis thaliana ecotype Columbia. The mutant plants show enhanced sensitivity to ozone, desiccation and sulfur dioxide, but have normal sensitivity to hydrogen peroxide, low temperature and high light levels. The T-DNA was inserted at a single locus which is linked to ozone sensitivity. Identification of the genomic sequences flanking the T-DNA insertion revealed disruption of a gene encoding a transporter-like protein of the tellurite resistance/C(4)-dicarboxylate transporter family. Plants with either of two different T-DNA insertions in this gene were also sensitive to ozone, and these plants failed to complement ozs1. Transpiration levels, stomatal conductance levels and the size of stomatal apertures were greater in ozs1 mutant plants than in the wild type. The stomatal apertures of ozs1 mutant plants responded to light fluctuations but were always larger than those of the wild-type plants under the same conditions. The stomata of the mutant and wild-type plants responded similarly to stimuli such as light, abscisic acid, high concentrations of carbon dioxide and ozone. These results suggest that OZS1 helps to close stomata, being not involved in the responses to these signals.

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.

The present study continues our previous research on investigating the biological effects of low-level gamma radiation in rice at the heavily contaminated Iitate village in Fukushima, by extending the experiments to unraveling the leaf proteome. 14-days-old plants of Japonica rice (Oryza sativa L. cv. Nipponbare) were subjected to gamma radiation level of upto 4 µSv/h, for 72 h. Following exposure, leaf samples were taken from the around 190 µSv/3 d exposed seedling and total proteins were extracted. The gamma irradiated leaf and control leaf (harvested at the start of the experiment) protein lysates were used in a 2-D differential gel electrophoresis (2D-DIGE) experiment using CyDye labeling in order to asses which spots were differentially represented, a novelty of the study. 2D-DIGE analysis revealed 91 spots with significantly different expression between samples (60 positive, 31 negative). MALDI-TOF and TOF/TOF mass spectrometry analyses revealed those as comprising of 59 different proteins (50 up-accumulated, 9 down-accumulated). The identified proteins were subdivided into 10 categories, according to their biological function, which indicated that the majority of the differentially expressed proteins consisted of the general (non-energy) metabolism and stress response categories. Proteome-wide data point to some effects of low-level gamma radiation exposure on the metabolism of rice leaves.

Ozone (O 3 ), a major photochemical oxidant, induces leaf injury concomitant with salicylic acid (SA) synthesis. In pathogen-infected leaves, SA is synthesized via two pathways, involving phenylalanine or isochorismate. SA biosynthesis under O 3 fumigation is not well understood. When we applied 14 C-labeled benzoic acid (a precursor of SA in the pathway via phenylalanine) to O3-exposed tobacco leaves, it was effectively metabolized to SA. However, the activity and mRNA level of isochorismate synthase (ICS) were not increased. In contrast, ICS activity was increased in O3-exposed Arabidopsis thaliana L. These results suggest that SA is synthesized via benzoic acid from phenylalanine in O 3 -exposed tobacco leaves but via isochorismate in Arabidopsis. Ethylene is a plant hormone that promotes leaf damage in O3-exposed plants. During O3 exposure, transgenic plants with a phenotype of reduced O 3 -induced ethylene production accumulated less SA than did wild-type plants. O3 increased the activity of phenylalanine ammonia-lyase (PAL) and the transcript levels of the chorismate mutase (CM) and PAL genes in wild-type tobacco, but their induction was suppressed in the transgenic plants. These results indicate that ethylene promotes SA accumulation by regulating the expression of the CM and PAL genes in O3-exposed tobacco.

Plants in the field are subjected to numerous environmental stresses. Lengthy continuation of such environmental stresses or a rapid increase in their intensity is harmful to vegetation. Assessments of the phytotoxicity of various stresses have been performed in many countries, although they have largely been based on estimates of leaf injury. We developed a novel method of detecting plant stresses that is more sensitive and specific than those previously available. This method is based on the detection of mRNA expression changes in 205 ozone-responsive Arabidopsis expressed sequence tags (ESTs) by cDNA macroarray analysis. By using this method, we illustrated shifts in gene expression in response to stressors such as drought, salinity, UV-B, low temperature, high temperature, and acid rain, as distinct from those in response to ozone. We also made a mini-scale macroarray with 12 ESTs for diagnosis of the above environmental stresses in plants. These results illustrate the potential of our cDNA macroarray for diagnosis of various stresses in plants.

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This report examines for the first time, the response of rice ( Oryza sativa japonica-type cv. Nipponbare) seedlings exposed to the air pollutant sulfur dioxide (SO 2 ). Distinctive reddish-brown necrotic spots and interveinal browning appeared on the leaf surface after exposure to SO 2 , over control, partly reminiscent of the hypersensitive reaction lesions. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis and immunoblotting analysis revealed strong induction of ascorbate peroxidase(s), and changes in cysteine proteinase inhibitors (‘phytocystatins’)-like proteins. Employing classical two-dimensional electrophoresis (2-DE) followed by amino acid sequencing, we identified several changes in the 2-DE protein profiles of SO 2 -fumigated leaves. Most prominent changes in leaves were the induced accumulation of a pathogenesis-related (PR) class 5 (OsPR5) protein, three PR 10 class proteins (OsPR10s), a novel ATP-dependent CLP protease, and an unknown protein. Subsequent Northern analysis showed potent accumulation of OsPR5 and OsPR10 transcripts in leaves. Finally, mass spectrometry analysis revealed a strong production of phytoalexins, sakuranetin and momilactone A in SO 2 stressed leaves. Our results not only demonstrate the highly damaging effect of SO 2 , but also identify SO 2 triggered multiple events linked with defense/stress response in the leaves of rice seedlings.

The cDNA clones for l -galactono-γ-lactone dehydrogenase (EC 1.3.2.3; GLDHase) that catalyzes the final step in the ascorbic acid (AsA) synthetic pathway, were isolated from Arabidopsis thaliana. The gene (AtGLDH) encodes a polypeptide of 610 amino acids and the predicted amino acid sequence was highly homologous to GLDHases from other plant species. The AtGLDH mRNA level of seedlings exhibited a diurnal change; the level was low in the morning and increased during the day. Its level in the evening (at 18:00) was about 2-fold that in the morning (at 6:00). Then the level decreased during the night until dawn of the next day. Similar diurnal changes were also observed in GLDHase activity and AsA content, suggesting that the AsA pool size is, at least partly, determined by the transcription rate of AtGLDH. Moreover, the diurnal changes of mRNA level, enzyme activity, and AsA content were not observed if plants were kept in continuous darkness. These results suggest that the diurnal change in expression of AtGLDH, which may play an important role in the regulation of AsA pool size, is regulated not by a circadian rhythm but by light.

Bisphenol A (BPA, 4,4'-isopropylidenediphenol), an endocrine disrupter with estrogenic properties, was supplied to tobacco BY-2 cells in suspension culture and the chemical nature of its metabolites was investigated. The concentration of BPA in the culture medium decreased rapidly and became undetectable at 2.5 h after the application. Four metabolites of BPA were observed in a methanol extract of the cells when the culture was supplemented with [(14)C]BPA. The most abundant metabolite was determined to be 4,4'-isopropylidenediphenol-O-beta-D-glucopyranoside (BPAG) by mass spectrometry, nuclear magnetic resonance spectroscopy and by hydrolysis with beta-glucosidase. This identification was confirmed by synthesis. When [(14)C]BPA was administrated to tobacco seedlings from their roots, radioactivity was incorporated in BPAG and three unidentified metabolites. These metabolites were accumulated in the leaves after 4 h exposure, indicating that tobacco seedlings absorbed BPA through their root systems, metabolized to its beta-glucoside and translocated the metabolites to their leaves.

Antisense DNA for an ozone-inducible 1-aminocyclopropane-1-carboxylate (ACC) synthase (EC 4.4.1.14; LE-ACS6) from tomato, under the control of the cauliflower mosaic virus 35S promoter, was introduced into tobacco to generate transgenic plants. Lower rates of ozone-induced ethylene production were observed in three of seven transgenic plants than in the wild-type plants. Ozone-induced visible damage was attenuated in these three lines, and the extent of damage was positively related to the level of ozone-induced ethylene production. In the most ozone-resistant line, ozone-induced accumulation of ACC and levels of transcripts for ozone-inducible endogenous ACC synthases were suppressed compared with those in wild-type plants, demonstrating that ozone-inducible ACC synthases have a key role in the expression of leaf damage by ozone exposure. No significant differences in growth and morphology were observed between transgenic and wild-type plants. Stomatal conductance of transgenic plants during ozone exposure was higher than that of wild-type plants. These findings indicate that the introduction of antisense DNA for an ozone-inducible ACC synthase can improve the ozone tolerance of plants without reducing their gas absorption and productivity.

Various detrimental factors in the environment damage plants, resulting in growth inhibition or withering. However, it is not easy to identify causal factors by visually inspecting the damaged plants. Therefore, we have developed a sensitive and reliable method for plant diagnosis, based on measuring changes in expression of a set of genes in a DNA microarray. With this method, we have been able to detect and discriminate between plants stressed by ozone, drought, or wounding.