Synthesis of Short-chain-length/Medium-chain-length Polyhydroxyalkanoate (PHA) Copolymers in Peroxisome of the Transgenic Arabidopsis Thaliana Harboring the PHA Synthase Gene from Pseudomonas sp. 61-3

In this paper, the photosynthetic production of short-chain-length/medium-chain-length polyhydroxyalkanoate (PHA) copolymers is reported. The wild-type and highly active doubly mutated PHA synthase 1 (S325T/Q481K, abbreviated ST/QK) genes from Pseudomonas sp. 61-3 were introduced into Arabidopsis thaliana. Peroxisome targeting signal 1 (PTS1) was used to target PHA synthases into the peroxisome to synthesize PHA from the intermediates of the β-oxidation pathway. The transgenic Arabidopsis produced PHA copolymers consisting of 40–57 mol% 3-hydroxybutyrate, 21–49 mol% 3-hydroxyvalerate, 8–18 mol% 3-hydroxyhexanoate, and 2–8 mol% 3-hydroxyoctanoate. The maximum PHA contents were 220μ g/g cell dry weight (cdw) in leaves, and 36μ g/g cdw in stems, respectively. The expression of the ST/QK mutated PHA synthase in leaves gene did not lead to significant difference in PHA content and monomer composition of PHAs, compared to the wild-type PHA synthase gene, suggesting that the supply of monomers may be a rate-determining step of PHA biosynthesis in the peroxisome. However, in stems, there were significant differences dependent on whether the wild-type or ST/QK mutated PHA synthase was expressed. These results suggest that tissue-specific monomer availability is important in determining the final mol% composition of PHA copolymers produced by the peroxisome in plants.     

Ecological impacts of umbrella effects of radiation on the individual members

In order to study the interactions in a model aquatic microcosm, an individual-based computer simulation model was developed. The microcosm consists of Euglena gracilis as an autotroph algae, Tetrahymena thermophila as a heterotroph protozoa and Escherichia coli as a saprotroph bacteria. There exists a strong interaction between Tetrahymena and E. coli as the first is the predator of the second. Ecological toxicity tests were conducted to test the population level impacts of the biological effects of radiation and toxicants on the lethality and mobility factors that influence directly or indirectly growth and reproduction. Radiological effects on lethality of E. coli individuals were translated to the reduction of the equilibrium population of Tetrahymena. A synergistic effect at the community level was also observed by the simulation of a combined exposure of radiation and a toxicant which reduced the feeding efficiency of Tetrahymena.     

Impact of gamma irradiation on the transformation efficiency for extracellular plasmid DNA

Extracellular DNA is omnipresent in aquatic environments and is thought to be a genetic material for horizontal gene transformation between microorganisms. We studied the impact of gamma irradiation on the transformation efficiency (transformants number per ng of DNA per ml) of extracellular DNA. Plasmid pEGFP as a model extracellular DNA was irradiated by gamma rays. The transformation efficiency decreased with the increase in radiation dose. A total dose of 10Gy is normally not lethal for microorganisms but certainly affects the transformation efficiency of extracellular DNA. The decrease in the efficiency would be induced by strand breaks of extracellular DNA because the yield of both single-strand breaks (SSBs) and double-strand breaks (DSBs) increased with the increase in radiation dose. The relative transformation efficiency of SSBs and DSBs to that of covalently closed circles (CCCs) was 30.3% and 0.2%, respectively. This impact on natural transformation suggests an inability of microorganisms to acquire new characteristics which should be normally acquired.     

Improvement of an efficient separation method for chemicals in diesel exhaust particles: analysis for nitrophenols

GOAL, SCOPE, AND BACKGROUND: Diesel exhaust is believed to consist of thousands of organic constituents and is a major cause of urban pollution. We recently reported that a systematic separation procedure involving successive solvent extractions, followed by repeated column chromatography, resulted in the isolation of vasodilatory active nitrophenols. These findings indicated that the estimation of the amount of nitrophenols in the environment is important to evaluate their effect on human health. The isolation procedure, however, involved successive solvent extractions followed by tedious, repeated chromatography, resulting in poor fractionation and in a significant loss of accuracy and reliability. Therefore, it was crucial to develop an alternative, efficient, and reliable analytical method. Here, we describe a facile and efficient acid-base extraction procedure for the analysis of nitrophenols. MATERIALS AND METHODS: Diesel exhaust particles (DEP) were collected from the exhaust of a 4JB1-type engine (ISUZU Automobile Co., Tokyo, Japan). Gas chromatography-mass spectrometry (GC-MS) analysis was performed with a GCMS-QP2010 instrument (Shimadzu, Kyoto, Japan). RESULTS: A solution of DEP in 1-butanol was extracted with aqueous NaOH to afford a nitrophenol-rich oily extract. The resulting oil was methylated with trimethylsilyldiazomethane and subsequently subjected to GC-MS analysis, revealing that 4-nitrophenol, 3-methyl-4-nitrophenol, 2-methyl-4-nitrophenol, and 4-nitro-3-phenylphenol were present in significantly higher concentrations than those reported previously. DISCUSSION: Simple acid-base extraction followed by the direct analysis of the resulting extract by GC-MS gave only broad peaks of nitrophenols with a poor detection limit, while the GC-MS analysis of the sample pretreated with (trimethylsilyl)diazomethane gave satisfactorily clear chromatograms with sharp peaks and with a significantly lowered detection limit (0.5 ng/ml, approximately 100 times). CONCLUSION: The present method involving an acid-base extraction, in situ derivatization, and GC-MS analysis has shown to be a simple, efficient, and reliable method for the isolation and identification of the chemical substances in DEP.     

Effects of acute γ-irradiation on the aquatic microbial microcosm in comparison with chemicals

Effects of acute γ-irradiation were investigated in the aquatic microcosm consisting of green algae (Chlorella sp. and Scenedesmus sp.) and a blue–green alga (Tolypothrix sp.) as producers; an oligochaete (Aeolosoma hemprichi), rotifers (Lecane sp. and Philodina sp.) and a ciliate protozoan (Cyclidium glaucoma) as consumers; and more than four species of bacteria as decomposers. At 100 Gy, populations were not affected in any taxa. At 500–5000 Gy, one or three taxa died out and populations of two or three taxa decreased over time, while that of Tolypothrix sp. increased. This Tolypothrix sp. increase was likely an indirect effect due to interspecies interactions. The principal response curve analysis revealed that the main trend of the effects was a dose-dependent population decrease. For a better understanding of radiation risks in aquatic microbial communities, effect doses of γ-rays compared with copper, herbicides and detergents were evaluated using the radiochemoecological conceptual model and the effect index for microcosm.     

Challenging the current strategy of radiological protection of the environment: arguments for an ecosystem approach

The system of radiological protection of the environment that is currently under development is one contribution to the general need to adequately protect the environment against stress. Dominated by operational goals, it emphasizes conceptual and methodological approaches that are readily accessible today: reference organisms supported by individual-based traditional ecotoxicological data. Whilst there are immediate advantages to this approach (pragmatism, consistency with other approaches in use for man and biota), there are also clear limitations, especially in a longer run perspective, that need to be acknowledged and further considered. One can mention a few: uncertainties generated by the need for various extrapolations (from lower to higher levels of biological organisation, …), various features missed such as potential ecological impact through impairment of ecosystem processes, trans-generational impacts as mediated through genomic instability, indirect effects mediated through trophic interactions or disruption of ecological balances,… Such limitations have already been faced in other fields of environmental protection against other stressors, pushing a number of environment professionals to assign stronger emphasis on more systemic approaches. This review discusses the advantages and limitations of the current approach designed for the radiological protection of non-human biota in the broader context of environment protection as a whole, with especial reference to upcoming trends and evolutions. This leads in particular to advocating the need to boost scientific and methodological approaches featuring the ecosystem concept as a mean to access a unified goal of protection: preserving life sustainability through protection of ecosystem structure and functioning.     

Semiquantitative color profiling of soils over a land degradation gradient in Sakaerat, Thailand

In this study, we attempted multivariate color profiling of soils over a land degradation gradient represented by dry evergreen forest (original vegetation), dry deciduous forest (moderately disturbed by fire), and bare ground (severely degraded) in Sakaerat, Thailand. The soils were sampled in a dry-to-wet seasonal transition. Values of the red–green–blue (RGB), cyan–magenta–yellow–key black (CMYK), L*a*b*, and hue–intensity–saturation (HIS) color models were determined using the digital software Adobe Photoshop^TM. Land degradation produced significant variations (p?R, C, Y, L*, a*, b*, S, and I values (p?p?R, G, B, C, M, K, L*, b*, and I values. In discriminating the soils, the color models did not differ in discriminatory power, while discriminatory power was affected by seasonal changes. Most color variation patterns had nonlinear relationships with the intensity of the land degradation gradient, due to effects of fire that darkened the deciduous forest soil, masking the nature of the soil as the intermediate between the evergreen forest and the bare ground soils. Taking these findings into account, the utilization of color profiling of soils in land conservation and rehabilitation is discussed.     

Assessing the probability of land submergence for lowland rice cultivation in Africa using satellite imagery and geospatial data

Sub-Saharan African countries are being strongly urged to enhance their rice production, because their rice consumption and importation rates have been rapidly increasing in recent years. Areas planted to rice in Africa are classified agro-ecologically into rainfed upland, rainfed lowland, and irrigated. Rainfed lowland includes extensive areas of unexploited land that has great potential for the promotion of rice growing. For the unexploited rainfed lowlands of Ghana, we have been studying the development of low-cost rice-farming systems that require no large-scale irrigation or land reclamation. For such systems, it is important to select suitable areas where water for rice farming can be obtained naturally; floodwaters offer promise for this purpose. Delineation and mapping of floodwater prone areas suitable for rice production is important for successful utilization of this land resource. Here, we propose a method of assessing flood probability from submergence frequency, as estimated from satellite imagery and geospatial data. ALOS/PALSAR images acquired in May, June, August, and September 2010 were used to classify land and water, and then a submerged-area map was produced. From the results, we were able to accurately detect non-submerged areas and submerged areas with water depths of at least 3?cm. The number of times classified into submerged area was defined as submergence frequency, and it was approximated by distance from reservoirs representing White Volta River, ponds, and swamps. In addition, flood extent derived from reservoirs was simulated using digital elevation model (DEM). Finally, a flood probability assessment map was produced by integration of the estimated submergence frequency and flood extent simulation. The results of a comparison of soil moisture data measured at 69 points in the field and the NDVIs computed by ALOS/AVNIR showed that areas with high potential for flooding retained high levels of soil moisture and were more likely to show less deterioration of vegetation in the dry season. The validation of these results confirmed the adequacy of the flood probability assessment method.