Yoshihisa Kohno

Ozone stress-induced proteomic changes in leaf total soluble and chloroplast proteins of soybean reveal that carbon allocation is involved in adaptation in the early developmental stage

Considerable soybean yield losses caused by ozone (O3) stress have been demonstrated by large‐scale meta‐analyses of free‐gas concentration enrichment systems. In this study, comparative proteomic approach was employed to explore the differential changes of proteins in O3 target structures such as leaf and chloroplasts of soybean seedlings. Acute O3 exposure (120 parts‐per‐billion) for 3 days did not cause any visible symptoms in developing leaves. However, higher amounts of ROS and lipid peroxidation indicated that severe oxidative burst occurred. Immunoblot analysis of O3‐induced known proteins revealed that proteins were modulated before symptoms became visible. Proteomic analysis identified a total of 20 and 32 differentially expressed proteins from O3‐treated leaf and chloroplast, respectively. Proteins associated with photosynthesis, including photosystem I/II and carbon assimilation decreased following exposure to O3. In contrast, proteins involved in antioxidant defense and carbon metabolism increased. The activity of enzymes involved in carbohydrate metabolism increased following exposure to O3, which is consistent with the decrease in starch and increase in sucrose concentrations. Taken together, these results suggest that carbon allocation is tightly programmed, and starch degradation probably feeds the tricarboxylic acid cycle while the photosynthesis pathway is severely affected during O3 stress.

Establishing critical levels of air pollutants for protecting East Asian vegetation — A challenge

Critical levels of ozone (O3) and sulfur dioxide (SO2) for protecting European forests are not evaluated to apply to East Asian vegetation. Based on the results obtained from the long-term experimental studies on the effects of chronic exposure to O3 or SO2 on 30 young potted grown tree species using open-top chambers, we analyzed dose-response relationships between the whole-plant dry mass increment during the experiment and concentration of air pollutants to establish Asian critical levels.

In Vitro and In Vivo Activities of Novel Fluoroquinolones Alone and in Combination with Clarithromycin against Clinically Isolated Mycobacterium avium Complex Strains in Japan

ABSTRACT The recommended treatments for Mycobacterium avium complex (MAC) infectious disease are combination regimens of clarithromycin (CLR) or azithromycin with ethambutol and rifamycin. However, these chemotherapy regimens are sometimes unsuccessful. Recently developed antimicrobial agents, such as newer fluoroquinolones (FQs) containing C-8 methoxy quinolone (moxifloxacin [MXF] and gatifloxacin [GAT]), are expected to be novel antimycobacterial agents. Here, we evaluated the in vitro and in vivo antimycobacterial activities of three FQs (MXF, GAT, and levofloxacin) and CLR against clinically isolated MAC strains. Subsequently, the in vitro and in vivo synergic activities of FQ-CLR combinations against MAC strains were investigated. CLR and the individual FQs alone showed promising activity against MAC strains in vitro, and the bacterial counts in organs (lungs, liver, and spleen) of MAC-infected mice treated with single agents were significantly reduced compared to control mice. CLR showed the best anti-MAC effect in vivo. When the three FQs were individually combined with CLR in vitro, mild antagonism was observed for 53 to 57% of the tested isolates. Moreover, mice were infected with MAC strains showing mild antagonism for FQ-CLR combinations in vitro, and the anti-MAC effects of the FQ-CLR combinations were evaluated by counting the viable bacteria in their organs and by histopathological examination after 28 days of treatment. Several FQ-CLR combinations exhibited bacterial counts in organs significantly higher than those in mice treated with CLR alone. Our results indicate that the activity of CLR is occasionally attenuated by combination with an FQ both in vitro and in vivo and that this effect seems to be MAC strain dependent. Careful combination chemotherapy using these agents against MAC infectious disease may be required.

Risk assessment of ozone impact on Fagus crenata in Japan: consideration of atmospheric nitrogen deposition

Tropospheric ozone (O3) is considered to be the air pollutant relating to the decline of Fagus crenata forest in Japan. In the present study, we assessed a risk of O3 impact on the growth of F. crenata in Japan, giving consideration to the effects associated with atmospheric nitrogen (N) deposition based on the experimental study, national monitoring data for oxidant concentration and atmospheric N deposition, and a national vegetation survey. The average and maximum O3-induced relative growth reduction (RGred) of F. crenata across Japan were estimated to be 3.2 and 9.7%, respectively. Current levels of atmospheric N deposition were found to significantly affect the sensitivity of F. crenata to O3. When the N deposition was assumed as zero, the estimated average and maximum RGred were 2.3% and 5.7%, respectively. The inclusion of atmospheric N deposition data thus increased the estimated values for average and maximum RGred (by 38% and 71%, respectively). Our results demonstrate that a change in the sensitivity to O3 associated with atmospheric N deposition is an important consideration in the risk assessment of O3 impact on the growth of F. crenata in Japan.

Risk assessment of ozone impact on the carbon absorption of Japanese representative conifers.

A risk assessment of ozone (O3) impact on the annual carbon absorption (ACA) of Japanese representative conifers was conducted based on the results of an experimental study, monitoring data of oxidant concentrations and vegetation surveys. The areas with high O3-induced reduction in ACA did not necessarily correspond to the areas with relatively high O3-exposure. Widespread distribution of O3-sensitive tree species such as Pinus densiflora and Larix kaempferi, and high ACA were important factors that induced a high risk of O3 impact on the ACA. Therefore, we concluded that not only the accumulated O3-exposure but also the variety of tree habitat, the tree sensitivity to O3 and the ACA among the tree species must be taken into account to assess the risk of O3 impact on the ACA of Japanese conifers. The O3-induced reduction in the total ACA of the three tree species in Japan was estimated to be 0.8%.

Epidemiology and Clinical Features of Pulmonary Nontuberculous Mycobacteriosis in Nagasaki, Japan

Background and Objectives Recent reports indicate that the incidence of nontuberculous mycobacterial-lung disease (NTM-LD) is increasing. This study aimed to investigate the epidemiology and clinical features of NTM-LD patients in Nagasaki prefecture, Japan to identify the negative prognostic factors for NTM-LD in Japan. Methods The medical records of patients newly diagnosed with NTM-LD in eleven hospitals in Nagasaki prefecture between January 2001 and February 2010 were reviewed. Data regarding the annual population of each region and the incidence of all forms of tuberculosis were collected to assess geographic variations in NTM-LD incidence, isolates, and radiological features. Results A total 975 patients were diagnosed with NTM-LD. The incidence increased over the study period and reached 11.0 and 10.1 per 100,000 population in 2008 and 2009, respectively. M. intracellulare was the most common pathogen in the southern region, and M. avium most common in other regions. The most common radiographic pattern was the nodular-bronchiectatic pattern. Age >60 years, body mass index <18.5 kg/m2, underlying lung disease, and cavitary pattern were the negative prognostic factors at the 1-year follow-up. Conclusions The incidence of NTM-LD has been increasing in Nagasaki prefecture. The isolates and radiographic features of patients vary markedly by region.

Ozone-Induced Rice Grain Yield Loss Is Triggered via a Change in Panicle Morphology That Is Controlled by ABERRANT PANICLE ORGANIZATION 1 Gene

Rice grain yield is predicted to decrease in the future because of an increase in tropospheric ozone concentration. However, the underlying mechanisms are unclear. Here, we investigated the responses to ozone of two rice (Oryza Sativa L.) cultivars, Sasanishiki and Habataki. Sasanishiki showed ozone-induced leaf injury, but no grain yield loss. By contrast, Habataki showed grain yield loss with minimal leaf injury. A QTL associated with grain yield loss caused by ozone was identified in Sasanishiki/Habataki chromosome segment substitution lines and included the ABERRANT PANICLE ORGANIZATION 1 (APO1) gene. The Habataki allele of the APO1 locus in a near-isogenic line also resulted in grain yield loss upon ozone exposure, suggesting APO1 involvement in ozone-induced yield loss. Only a few differences in the APO1 amino acid sequences were detected between the cultivars, but the APO1 transcript level was oppositely regulated by ozone exposure: i.e., it increased in Sasanishiki and decreased in Habataki. Interestingly, the levels of some phytohormones (jasmonic acid, jasmonoyl-L-isoleucine, and abscisic acid) known to be involved in attenuation of ozone-induced leaf injury tended to decrease in Sasanishiki but to increase in Habataki upon ozone exposure. These data indicate that ozone-induced grain yield loss in Habataki is caused by a reduction in the APO1 transcript level through an increase in the levels of phytohormones that reduce leaf damage.

Effects of Nitrogen Supply on the Sensitivity to O3 of Growth and Photosynthesis of Japanese Beech (Fagus crenata) Seedlings

To obtain basic information for evaluating critical levels of O3 under different nitrogen loads for protecting Japanese beech forests, two-year-old seedlings of Fagus crenata Blume were grown in potted andisol supplied with N as NH4NO3 solution at 0, 20 or 50 kg ha year and exposed to charcoal-filtered air or O3 at 1.0, 1.5 and 2.0 times the ambient concentration from 16 April to 22 September 2004. The O3 induced significant reductions in the wholeplant dry mass, net photosynthetic rate at 380 μmol mol CO2 (A380), carboxylation efficiency (CE) and concentrations of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and total soluble protein (TSP) in the leaves. The concentrations of Rubisco and TSP were negatively correlated with the concentration of leaf acidic amino acid, suggesting that O3 enhanced the degradation of protein such as Rubisco. The N supply to the soil did not significantly change the whole-plant dry mass and A380, whereas it significantly increased the CE and concentrations of Rubisco and total amino acid. No significant interactive effects of O3 and N supply to the soil were detected on the growth, photosynthetic parameters and concentrations of protein and amino acid in the leaves. In conclusion, N supply to the soil at ≤50 kg ha year does not significantly change the sensitivity to O3 of growth and net photosynthesis of Fagus crenata seedlings.

EFFECT OF SIMULATED ACID RAIN ON THE GROWTH OF JAPANESE CONIFERS GROWN WITH OR WITHOUT FERTILIZER

Three-year old cuttings of Japanese cedar (Cryptomeria japonica), Japanese cypress (Chamaecyparis obtusa) and Sawara cypress (Chamaecyparis pisifera) were grown in pots with andosol and were exposed to simulated acid rain (SAR) at pH 2.0, 3.0, and 4.0 for 23 months. Total precipitation was 2460, 3960, or 5450 mm and SAR contained sulfuric, nitric and chloric acid at equivalent ratio of 5∶2∶3. Deionized water of pH 5.6 was a control. Exposure to SAR at pH 2.0 induced visible foliar injuries, but not any visible symptoms at pH 3.0 or higher. Total dry weights of 3 conifer cuttings grown with fertilizer were about two fold of those in plants grown without fertilizer and they reduced significantly by the exposure to SAR at pH 2.0. However, Cryptomeria japonica without fertilizer increased dry weights even though plants developed reddish-brown necrosis in large parts of tops. These 3 coniferous cuttings did not show any significant growth reduction at pH 3.0 and 4.0. Soil pH after receiving 5450 mm of SAR at pH 2.0 was 4.0 and molar ratio of (K+Ca+Mg)/Al in water and 1M ammonium acetate soluble fraction of the soil was about 0.3 and 0.4. respectively. However, dry weight of root in plants grown without fertilizer did not decrease. This suggested that growth reduction in plants fertilized and exposed to pH 2.0 was due to a reduction in photosynthetic organs associated with visible injuries, but not direct linkage with root growth inhibition due to soil acidification stress.

Genome-wide mapping of the ozone-responsive transcriptomes in rice panicle and seed tissues reveals novel insight into their regulatory events

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.Purpose of workTropospheric 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.