Stress signaling in response to polycyclic aromatic hydrocarbon exposure in Arabidopsis thaliana involves a nucleoside diphosphate kinase, NDPK-3.

MAIN CONCLUSION: The study is the first to reveal the proteomic response in plants to a single PAH stress, and indicates that NDPK3 is a positive regulator in the Arabidopsis response to phenanthrene stress. Polycyclic aromatic hydrocarbons (PAHs) are highly carcinogenic pollutants that are byproducts of carbon-based fuel combustion, and tend to persist in the environment for long periods of time. PAHs elicit complex, damaging responses in plants, and prior research at the physiological, biochemical, and transcriptional levels has indicated that reactive oxygen species (ROS) and oxidative stress play major roles in the PAH response. However, the proteomic response has remained largely unexplored. This study hypothesized that the proteomic response in Arabidopsis thaliana to phenanthrene, a model PAH, would include a strong oxidative stress signature, and would provide leads to potential signaling molecules involved. To explore that proteomic signature, we performed 2D-PAGE experiments and identified 30 proteins levels that were significantly altered including catalases (CAT), ascorbate peroxidase (APX), peroxiredoxins (POD), glutathione-S-transferase, and glutathione reductase. Also upregulated was nucleoside diphosphate kinase 3 (NDPK-3), a protein known to have metabolic and stress signaling functions. To address whether NDPK-3 functions upstream of the oxidative stress response, we measured levels of stress-responsive enzymes in NDPK-3 overexpressor, loss-of-function knockout, and wild-type plant lines. In the NDPK-3 overexpressor, the enzyme activities of APX, CAT, POD, as well as superoxide dismutase were all increased compared to wild type; in the NDPK-3 knockout line, these enzymes had reduced activity. This pattern occurred in untreated as well as phenanthrene-treated plants. These data support a model in which NDPK-3 is a positive regulator of the Arabidopsis stress response to PAHs.

Specific ion effects on the soil shear strength and clay surface properties of collapsing wall in Benggang.

Benggangs are a special type of soil erosion in the hilly granite regions of the tropical and subtropical areas of Southern China. They cause severe soil and water loss, which can severely deteriorate soil quality and threat to the local ecological environment. Soils (red soil, sandy soil and detritus soil) were collected from collapsing wall of a typical Benggang in Changting County of Fujian Province, and their physicochemical and mineralogical properties were analyzed. Five different monovalent cations were used to saturate the soil samples to examine the specific ion effects on the shear strength and clay surface properties. Red soil had a higher clay content, plastic limit, liquid limit and shear strength than sandy soil and detritus soil. The studied soils mainly consisted of kaolinite, hydroxy-interlayer vermiculite, illite and gibbsite clay minerals. The soils saturated with K+, NH4 +and Cs+ had greater cohesion than the Li+- and Na+-saturated soils, e.g., the cohesion of the red soil saturated with Li+, K+, NH4 + and Cs+ cations were 1.05, 1.23, 1.45 and 1.20 times larger than that of the Na+-saturated soil, respectively. While the internal friction angle was slightly different, which indicated that different monovalent cations affected the shear strength differently. K+-, NH4 +- and Cs+-saturated clay particles had higher zeta potentials and thinner shear plane thicknesses than Li+- and Na+-saturated clay particles and showed strong specific ion effects on the clay surface properties. The changes in clay surface properties strongly affected the soil mechanical properties. Soils saturated with K+, NH4 + and Cs+ could increase the shear strength, and then increase the stability of the collapsing wall, thus might decrease the erosion intensity of Benggang. The results provide a scientific basis for the interpretation of and practical treatment of Benggang.

Impact of soil surface properties on soil swelling of different soil layers in collapsing wall of Benggang.

Benggang is one of the most serious soil erosion problems in tropical and subtropical areas in southern China. Little work has been reported on the surface properties of soil colloidal particle and its influence on soil swelling of different soil layers in collapsing wall of Benggang. In this present work, the effects of sodium concentration on soil swelling, and the correlations between soil swelling rates and soil colloidal surface properties were comprehensively evaluated by carefully examining soil physicochemical properties and soil colloidal surface properties of red, sandy and detritus soil layers from a collapsing wall. Our results showed that the soil swelling rates of red, sandy and detritus soil layers all exponentially decreased with increasing initial water contents. The relationship between soil swelling rate and the thickness of shear plane showed an extremely significant negative correlation for red soil layer and no correlation for sandy and detritus soil layers. Moreover, the elevating sodium concentrations reduced the thickness of shear plane from 39.69 to 0.76 nm for red soil layer, followed from 22.56 to 0.79 nm for sandy soil layer and from 18.61 to 0.64 nm for detritus soil layer. These findings indicated that the soil particle interactions played a crucial role in the development and occurrence of Benggang. This work will be helpful in understanding the mechanisms of soil mass loss on the gully head and collapsing wall of Benggang.

Verification of the detachment-transport coupling relationship of rill erosion using colluvium material in steep nonerodible slopes.

The detachment-transport coupling equation by Foster and Meyer is a classical equation that describes the relationship between detachment and transport. The equation quantifies the relationship between sediment loads and soil detachment rates, deepens the understanding of soil erosion and provides a reliable basis for the establishment of an erosion model. However, the applicability of this equation to slopes with gradients greater than 47% is limited. In this work, the detachment-transport coupling relationship is investigated using the colluvium material of Benggang. A nonerodible rill flume 4 m long and 0.12 m wide was adopted. The slope gradient ranged from 27% to 70%, the unit flow discharge ranged from 0.56 × 10-3 to 3.33 × 10-3 m2 s-1, and the sediment transport capacity (Tc ) was measured under each slope and discharge combination. The sediment was inputted into the flume according to the predetermined sediment addition rate (from 0% to 100% of Tc ), and the detachment rate (Dr ) under each combination of the slope and discharge was measured. Dr linearly decreased with increasing sediment loads, which is consistent with the detachment-transport coupling equation by Foster and Meyer. The linear equations can predict the detachment capacity (Dc ) and Tc well (Nash-Sutcliffe efficiency coefficient (NSE) = 0.98 for Dc , and NSE = 0.99 for Tc ). The detachment-transport coupling equation can adequately predict the Dr (NSE = 0.89). However, its applicability to slopes of <47% (NSE: 0.92-0.96) was greater than that to slopes of ≥47% (NSE: 0.81-0.89), and the predicted Dr under Tc levels of 20% and 40% were higher than the measured values, while the predicted value under a Tc level of 80% was lower than the measured value. In summary, the detachment-transport coupling equation by Foster and Meyer can accurately reflect the negative feedback relationship between detachments and transports along steep-slope fixed beds and is suitable for colluvial deposit research. The results provide a basis for the construction of steep-slope colluvial deposit erosion models. In the future, the study of the hydrodynamic characteristics of sediment transport processes should be strengthened to clarify the detachment-transport effect of flows through hydrodynamics.

The Distribution Characteristics and Potential Risk Assessment of Lead in the Soil of Tieguanyin Tea Plantations in Anxi County, China.

Assessing the distribution and risks associated with the soil lead content in the Tieguanyin tea plantations of Anxi County is critical, given the county's significance as the primary Tieguanyin tea production area in Fujian Province. This study examined the distribution characteristics of soil lead in Anxi County's tea plantations according to the Kriging spatial interpolation of the parameters of the semivariance function of the exponential model. Moreover, the sources of lead content were analyzed, considering geological backgrounds and anthropogenic influences. Ecological risks and the issuance of early warnings were also assessed. The soil lead content in the rocks of the Tieguanyin tea plantations in Anxi County followed the order: andesite > dacite > rhyolite > granite. The soil lead content gradually decreased from the center toward the east and west, forming four distinct north-south parallel zones. High-lead-content areas were identified at the border of Jiandou, Bailai, and Hushang; in the central part of Lutian; and in the southern part of Huqiu. The high levels of soil lead in the tea plantations possibly originated from industrial and mining activities, automobile exhaust, and agricultural activities. The distribution of single-factor pollution indices and potential risk evaluation based on the Soil Environmental Quality Standard, Environmental Technical Conditions for Tea Production Area, and Environmental Technical Conditions for Organic Tea Production Area indicated that the soil in Tieguanyin tea plantations in Anxi County was clean and safe for tea cultivation.

Study on soil-water characteristic curves in the profiles of collapsing walls of typical granite Benggang in southeast China.

Benggang with steep collapsing walls is one of the worst soil erosion problems in South China. The collapse of walls is the most critical process in Benggang development. This is mainly due to the soil water properties. The soil water characteristic curve (SWCC) is a key indicator for analyzing soil moisture, but the SWCC and its mechanism of influence in collapsing walls remain obscure. A pressure plate meter was used for drying experiments to research the SWCCs of undisturbed soils of five layers (from top to bottom: red soil layer, transition layer I, sand soil layer, transition layer II and detrital layer) of two typical collapsing walls. The van Genuchten (VG) model can be fitted to the SWCCs for different layers (NSE ≥ 0.90). With increasing soil depth, the parameters a and θ s first decreased and then increased, the parameters n first increased and then decreased, θ r declined as the soil depth increased. These findings illustrate that soil water holding capacity decreases with increasing soil depth. The bottom of the soil is weak in water retention and water can easily reach saturation, resulting in a decline in soil stability, thus promoting soil collapse and finally inducing upper soil collapse. Furthermore, gravel content and particle morphology are factors that should not be neglected for SWCCs. The results of this study provide a theoretical basis for understanding the process of wall collapse in Benggang landforms in South China.

[Benggang (collapsing hill) erosion hazard zoning based on the minimum cumulative resistance model: A case study of a small watershed in Anxi County, Fujian, China]. / 基于最小累积阻力模型的崩岗侵蚀危险性分区­­ä»¥å®‰æºªåŽ¿å°æµåŸŸä¸ºä¾‹.

In order to understand the possibility and spatial pattern of Benggang (collapsing hill) erosion in risk assessment and distinguish the primary and secondary areas in Benggang prevention, we took a small watershed in Anxi County (Fujian, China) as the study area and Benggang as the source, and constructed the Benggang expansion resistance surface using the minimum cumulative resistance (MCR) model and divided the risk zoning. The results showed that the area around Benggang displayed a low resistance value, while the northwest and southeast areas displayed a high resistance value. The Benggang expansion resistance surface had an island type form. Based on resistance surface, the research region was divided into very high-risk, high-risk, medium risk, low-risk and very low-risk zones. We proposed corresponding Benggang management suggestions for those zones. A total of 21 strategic saddle points were extracted based on the resistance surface morphology. The strategic points in the lower safety zone were considered as the priority areas for Benggang prevention. We compared the hazard results based on the MCR model and the information quantity method. These results were consistent in spatial distribution, indicating the reliability of the results of hazard zoning by the MCR model.

Regioselective 1-alkylation of 2'-deoxyguanosine.

A method has been found for the regioselective alkylation of the nitrogen at the 1-position of 2'-deoxyguanosine. This consists in the reaction, in tetrahydrofuran solution, of a fully protected form of dG, namely the 3'5'-O-bis(tert-butyldimethylsilyl)-N2-dimethylaminomethylene derivative, with an alkyl halide in the presence of cesium carbonate. The yields of these previously unavailable derivatives of 2'-deoxyguanosine range from good to excellent. Confirmation of the structure of these substances comes from a comparison of their spectroscopic properties with those of the known 1-methyl homologue. In particular, the UV spectra of these new derivatives and the known 1-methyl homologue are essentially identical.

[Responses of Arabidopsis thaliana to oxidative stress induced by polycyclic aromatic hydrocarbon fluoranthene].

With the seedlings of model plant Arabidopsis thaliana and the polycyclic aromatic hydrocarbon fluoranthene as test materials, this paper studied the physiological responses of plants to polycyclic aromatic hydrocarbons from the aspects of antioxidases and membrane protection system that are tightly related to abiotic stresses. The results showed that under fluoranthene stress, the seedlings experienced oxidative stress and membrane-lipid peroxidization. At 0.75 mmol x L(-1) of fluoranthene, the photosynthetic process was inhibited; at 1.00 mmol x L(-1) of fluoranthene, the malonaldehydic acid (MDA) content increased and the ascorbate peroxidase (APX) activity decreased significantly, indicating an accelerated membrane-lipid peroxidization; and at 1.25 mmol x L(-1) of fluoranthene, the peroxidase (POD) activity decreased significantly and H2O2 had a high accumulation, making the seedlings injured severely.

Synthesis and characterization of enantiomeric anti-2-fluorobenzo[a]pyrene-7,8-dihydrodiol-9,10-epoxides and their 2'-deoxyguanosine and oligodeoxynucleotide adducts.

Benzo[a]pyrene diol epoxides (BPDEs) are the ultimate carcinogenic species of benzo[a]pyrene, a prototype polycyclic aromatic hydrocarbon (PAH). BPDE-modified DNA duplexes can adopt multiple conformations depending on the nature of the modified bases, the stereochemistry at the location of the covalent linkage, and the sequence context surrounding the lesion site. In this paper, we describe the preparation of enantiomeric 2-fluoro-BPDEs, trans-(7R,8S)-dihydroxy-(9S,10R)- and trans-(7S,8R)-(9R,10S)-epoxy-7,8,9,10-tetrtahydro-2-fluorobenzo[a]pyrene (22 and 23, respectively), as models for probing the BPDE-induced conformational heterogeneity. The multistep synthesis of the target diol epoxides described herein entails regiospecific succinoylation of 2-fluoropyrene, followed by a ring closure, regio- and stereospecific construction of the 7,8-dihydrodiol functionality, and a subsequent meta-chloroperbenzoic acid-mediated epoxidation. Stereoselectivity was achieved by using Jacobsen chiral catalysts, which produced greater than approximately 90% enantiomeric excess. Absolute configurations at the C(7,8) carbons of the FBP derivatives were determined by comparison of the circular dichroism (CD) spectra with those reported for the BP analogues. Analysis of the 3J(7,8) vicinal coupling constants, CD shape, and charge density calculations all indicated that the prepared anti-FBPDEs preferentially adopt the pseudo-diequatorial C(7,8) diol conformation. Hydrolysis of anti-FBPDEs produced a 9:1 ratio of trans- to cis-opened tetraols. Reactions of each of the anti-FBPDEs with deoxyguanosine 5'-monophosphate produced predominantly trans-anti-N2-dG as the major adducts. Analogous reactions with two 11-mer oligodeoxynucleotides (5'-CCATXGCTACC-3' where X = dT, dC) gave FBP-modified oligodeoxynucleotides with structures that were characterized by enzyme digest/HPLC and electrospray ionization time-of-flight mass spectrometry data. The oligonucleotide adducts were annealed with the appropriate sequences to form fully complementary duplexes [(5'-CCATXGCTACC-3')(5'-GGTAGCYATGG-3'), G = FBP-N2-dG adduct, X = dT, Y = dA in duplex I; X = dC, Y = dG in duplex II] for CD and UV melting studies. The results of the present study were consistent with those reported previously for BPDE-modified duplexes in the same sequence contexts and support the utility of FBPDEs as useful structural probes.

Regioisomeric synthesis and characteristics of the alpha-hydroxy-1,N(2)-propanodeoxyguanosine.

Acrolein, a known mutagen, undergoes reaction in vitro under physiological conditions with both 2'-deoxyguanosine and native DNA to give rise to exocyclic adducts of the 5,6,7,8-tetrahydropyrimido[1,2-a]purine-10(3H)-one class having a hydroxyl group at either the 6 or the 8 position (these positions are respectively designated alpha and gamma when referring to the 1,N(2)-(propano-bridge). Previously, we have shown that the 8-hydroxy derivative has very low mutagenicity probably because, in double-stranded DNA, this residue exists in the open-chain aldehydic form [N(2)-(3-oxopropyl)-2'-deoxyguanosine] (5). To continue our investigation in this area, we needed ample supplies of the 6-hydroxy isomers. This current paper describes high-yield simple methods for the synthesis in bulk of the 6-hydroxy and the 6-methoxy exocyclic adducts 1 and 3 and a new efficient synthesis of 1,N(2)()-(prop-1,3-diyl)-2'-deoxyguanosine (4), previously used as a chemically stable model in studying the physico-biological implications of 1,N(2) exocyclic adduction to dG.

Synthesis of the minor acrolein adducts of 2(')-deoxyguanosine and their generation in oligomeric DNA.

Acrolein, a known mutagen, undergoes reaction in vitro under physiological conditions with both 2(')-deoxyguanosine and native DNA to give rise to exocyclic adducts of the 5,6,7,8-tetrahydropyrimido[1,2-a]purine-10(3H)-one class having an hydroxy group at either the 6 or the 8 position. Previously we have shown that the 8-hydroxy derivative in a bacterial system has very low mutagenicity probably because in double-stranded DNA this residue exists in the open-chain aldehydic form [N(2)-(3-oxopropyl)-2(')-deoxyguanosine] (3). To continue our investigation in this area, we needed ample supplies of the 6-hydroxy isomers. This current paper describes high-yield simple methods for the synthesis in bulk of the 6-hydroxy adduct 1 and its incorporation into DNA oligomers. The basic methods for the synthesis of the adduct 1, involve 1-substitution of dG derivatives with a 3-butenyl group, dihydroxylation of the olefin with osmium tetroxide and N-methylmorpholine N-oxide, then diol cleavage with periodate ion after incorporation of the 1-(3,4-diacetoxybutyl)-2(')-deoxyguanosine into oligomeric DNA.

Regioselective synthesis of 1,N(2)-etheno-2'-deoxyguanosine and its generation in oligomeric DNA.

Chloroethylene oxide and chloroacetaldehyde, reactive intermediates derived from vinyl chloride, and the epoxy-hydroxy-alkanals, produced endogenously in the metabolism of polyunsaturated fatty acids, react with nucleic acid bases in DNA to form exocyclic etheno derivatives of 2'-deoxyadenosine, 2'-deoxyguanosine, and 2'-deoxycytidine. This paper describes an efficient method for the synthesis of the exocyclic 1,N(2)-etheno adduct of 2'-deoxyguanosine and its incorporation into DNA oligomers using automated synthesis techniques. The synthesis was initiated by a high-yield alkylation of N(2)-protected 2'-deoxyguanosine at the 1-position with 1,2-diacetoxy-3-bromopropane. The product was converted to the 5'-O-dimethoxytrityl-3'-O-phosphoramidite using published techniques and incorporated site specifically into DNA oligomers with 99% coupling efficiency. Ring closure to yield the 6-hydroxyethano derivative was accomplished by oxidation with sodium periodate, and facile dehydration then afforded DNA oligomers containing 1,N(2)-etheno-2'-deoxyguanosine. All oligomers were characterized fully by physicochemical methods.

Mutagenesis by acrolein-derived propanodeoxyguanosine adducts in human cells.

Acrolein, which is widely spread in the environment and is produced by lipid peroxidation in cells, reacts with DNA to form two exocyclic 1,N2-propanodeoxyguanosine (PdG) adducts. To establish their relative contribution to the acrolein mutagenicity, the genotoxic properties of alpha-OH-PdG and gamma-OH-PdG together with their model DNA adduct, PdG, were studied in human cells. DNA adducts were incorporated site-specifically into a SV40/BK virus origin-based shuttle vector and replicated in xeroderma pigmentosum complementation group A (XPA) cells. Analysis of progeny plasmid revealed that alpha-OH-PdG and PdG strongly block DNA synthesis and that both adducts induced base substitutions with G --> T transversions predominating. Primer extension studies, catalyzed by the 3'-->5' exonuclease-deficient Klenow fragment of Escherichia coli pol I, revealed limited extension from the 3' primer termini opposite these two adducts. In contrast, gamma-OH-PdG did not strongly block DNA synthesis or miscode in XPA cells. Primer extension from a dC terminus opposite gamma-OH-PdG was much more efficient than that opposite alpha-OH-PdG or PdG. These results indicate that the minor alpha-OH-PdG adduct is more genotoxic than the major gamma-OH-PdG. Furthermore, experiments using a HeLa whole cell extract indicate that all three DNA adducts are not efficiently removed from DNA by base excision repair.