Chelators effect on soil Cu extractability and uptake by Elsholtzia splendens

Phytoremediation is emerging as a potential cost-effective solution for remediation of contaminated soils, and bioavailability of metal in the soil for plant uptake is an important factor for successful phytoremediation.This study aimedat investigating the ability of EDTA and citric acid for enhancing soil bioavailability of Cu and phytoremediation by E1sholtzia splendens in two types of soils contaminated with heavy metals [i.e. mined soil from copper mining area (MS), and paddy soil (PS) polluted by copper refining].The results showed that addition of 2.5 mmol/kg EDTA significantly increased the H2O extractable Cu concentration from 1.20 to 15.78mg/kg in MS and from 0.26 to 15.72mg/kg in PS,and that shoot Cu concentration increased 4-fold and 8-fold as compared to the control.There was no significant difference between the treatment with 5.0mmol/kg EDTA and that with 2.5mmol/kg EDTA, probably because that 2.5mmol/kg EDTA was enough for elevating Cu bioavailability to the maximum level.As compared with the control, citric acid had no marked effect on both soil extractable Cu and shoot Cu concentration or accumulation. The results indicated that EDTA addition can increase the potential and efficiency of Cu phytoextraction by E.splendens in polluted soils.     

SRO1 regulates heavy metal mercury stress response in Arabidopsis thaliana

Heavy metals in the environment are harmful limiting factors for the normal growth and development of plants. Here, we isolated and identified an Arabidopsis thaliana T-DNA insertion mutant, named srol-1, which showed a hyper-sensitive response to HgCl2. The SRO1 protein contains a WWE domain that mediates proteinprotein interactions. Under HgCl2 treatment, when compared with the wild-type plants, the growth of srol-1 was repressed dramatically and the number of true leaves was reduced and etiolated. The electrolyte leakage rates showed that cell membrane integrity in srol-1 was damaged more severely than in the wild type. DAB （3,5-diaminobenzidine） staining and confocal microscopy showed that Hg2＋ stress induced more hydrogen peroxide accumulation in srol-1 than in the wild type. The qRT-PCR results indicated that the expression of some abiotic stress-induced genes, such as L-ascorbate peroxidase （APX1）, was reduced under oxidative or Hg2＋ stress. Transgenic plants containing a GFP：：SRO1 fusion protein showed that SRO1 was localized in the nucleus of the cells. SRO1 was shown to be expressed in various tissues, and was most highly expressed in the vigorous tissues. Our results suggest thatSRO1 may play an important role in the stress response of A. thaliana to heavy metals.     

Rapid genome evolution in Pms 1 region of rice revealed by comparative sequence analysis

Pms1, a locus for photoperiod sensitive genic male sterility in rice, was identified and mapped to chromosome 7 in previous studies. Here we report an effort to identify the candidate genes for Pms1 by comparative sequencing of BAC clones from two cultivars Minghui 63 and Nongken 58, the parents for the initial mapping population. Annotation and comparison of the sequences of the two clones resulted in a total of five potential candidates which should be functionally tested. We also conducted com-parative analysis of sequences of these two cultivars with two other cultivars, Nipponbare and 93-11, for which sequence data were available in public databases. The analysis revealed large differences in sequence composition among the four genotypes in the Pms1 region primarily due to retroelement activity leading to rapid recent growth and divergence of the genomes. High levels of polymorphism in the forms of indels and SNPs were found both in intra- and inter-subspecific comparisons. Dating analysis using LTRs of the retroelements in this region showed that the substitution rate of LTRs was much higher than reported in the literature. The results provided strong evidence for rapid genomic evolution of this region as a consequence of natural and artificial selection.     

Heavy metals contents in soil profiles of typical agricultural lands in Yixing,Jiangsu Province,China

The effect of land use pattern on the heavy metals(HMs)distribution in soil profiles was investigated in Yixing City,China,for the four typical types of land utilization,i.e.vegetable fields on the plain,paddy fields,tea gardens on hilly lands and bamboo groves.The variance of HMs concentration in the depth of 0-15 cm,15-30 cm,30-60 cm of soil profiles was compared among the different tillage ages. Results showed there was a notable accumulation of Cd,Hg,Cu,Pb,Zn and Cr in the soil profiles of vegetable fields and paddy fields causing a pollution of Cu,Hg and Cd in some profiles,while no remarkable accumulation of HMs in the soil profiles of the tea gardens and bamboo groves was found. The accumulation extent of HMs in the soil profiles of the four kinds of land use followed a sequence of vegetable fields>paddy fields>tea gardens>bamboo groves.The accumulation of HMs was mainly due to the industry,transportation,other urban human activities,and intensive use of fertilizers and pesticides.The downward leaching of HMs was relatively remarkable in the soil profiles of vegetable fields. There was an accumulation in surface and subsurface layers for Cd,Hg,Pb,Zn in the soil profiles of paddy fields and tea gardens,and for Cd,Hg in those of bamboo groves.Serious pollution effects of Hg, Pb,and Zn have been shown in the soil of vegetable fields.All the four kinds of agricultural land are basically free from the pollution of the external sources of As and Ni.With the increase of tillage time, the soil of vegetable and paddy fields will face the potential risk of HMs pollution,which mainly comes from Hg,Cd and Cu,whereas that of tea gardens and bamboo groves faces a relatively small risk.     

Chk1 prevents abnormal mitosis of S-phase HeLa cells containing DNA damage

To explore effects of DNA damage on cell-cycle progression in p53-deficient tumor cells, synchronized HeLa cells at G1, S and G2/M phases were treated with methyl methanesulfnate （MMS）. The results showed that the MMS treatment resulted in the cell-cycle arrest or delay in all 3 phases, while the S-phase cells were the most sensitive to MMS. Further studies demonstrated that ATM-Chk2 and p38 MAPK signaling pathways were activated in all 3 phases when the cells were treated with MMS; whereas Chk1 was activated only in S phase under the drug treatment, indicating that Chk1 specifically participated in S-phase checkpoints. To analyze the role of Chk1 in S-phase checkpoints, we administered a specific Chk1 inhibitor, UCN-01, to the S-phase cells. The results showed that the S-phase cells treated with MMS＋UCN-01 could enter aberrant mitosis without finishing DNA replication, indicating that Chk1 mainly functions in the DNA damage checkpoint rather than in the replication checkpoint. In addition, MMS treatment alone inhibited the accumulation of cyclin B1, a key component of M-phase CDK-cyclin complex, in the S-phase cells, whereas the inhibition of Chk1 activation resulted in the accumulation of cyclin B1 in the MMS-treated S-phase cells. This observation further supports the view that DNA-damaged S-phase cells enter abnormal mitosis when Chk1 activation is inhibited. Our results demonstrate that Chk1 is a specific kinase that plays an important role in the MMS-induced S-phase DNA damage checkpoint. As p53 is not involved in this process, Chk1 may be a potential target for p53-deficient tumor therapy.     

Salt tolerance conferred by over-expression of OsNHX1 gene in Poplar 84K

OsNHX1 gene (Na^＋/H^＋ antiporter gene of Oryza sativa L.) was introduced into Poplar 84K with Agrobacterium tumefaciens-mediated transformation. PCR, Southern and Northern blot analysis showed that OsNHX1 gene was incorporated successfully into the genome of Poplar 84K and expressed in these transgenic plants. Salt tolerance test showed that three lines of transgenic plants grew normally in the presence of 2OO mmol/L NaCI, while the Na^＋ content in the leaves of the transgenic plants grown at 2OO mmol/L NaCl was significantly higher than that in plants grown at 0mmol/L NaCI. The osmotic potential in the transgenic plants with high salinity treatment was lower than that of control plants. Our results demonstrate the potential use of these transgenic plants for agricultural use in saline soils.     

Polaronic effect on a bound polaron

Feynman variational path-integral theory was used to obtain the groud-state energy of a polaron in a quantum well in the presence of a Coulomb potential for arbitray values of the electron-phonon couplng constant α.Numerical and analytical results showed that the energy shift was more sensitive to α than to the Coulomb binding parameter(β) and increased with the decrease of effective quantum well width lZ.it was intersting that due to the electronic confinement in the quasi-2D(quantum well)structures,the lower bound of the strong coupling regime was shifted to smaller values of α.Comparison of the polaron in the quantum well with that in the quantum wire or dot showed that the polaronic effect strengthened with decrease of the confinement dimension.     

Trace element characteristics of a Gorham-Stout Syndrome sufferer

Gorham-Stout Syndrome （GSS） is an infrequent and mysterious bone disease characterized by massive bone dissolving or even disappearing due to an unknown pathogeny. It is quite different from the Itai-itai disease and osteoporosis. In 2001, an 8-year-old boy from a small town in Xinjiang was found to get GSS disease. Some parts of his bones vanished without any external force. Results showed that the concentrations of Cd and Cu, especially Na and K in his hair were far higher than those of the healthy people, and the concentration of Cd was 2 times that of the reference while those of Cr and Zn were insufficient for health. The ratio of K/Na was also higher than that of healthy group. Four different ratios revealed weak bone growth potential and strong bone breakage occurred simultaneously in the GSS sufferer＇s skeletal system. It might be the synergistic effect of the trace elements leading to the baffling syndrome. Further investigation demonstrated that the trace elements can cause a series of diseases, including GSS disease and a typical rheumatoid arthritis （ARA）. Epidemiological investigation also proved that there were about 25% of the inhabitants in the town suffered from an ARA. All of these data implied there was significant relationship among Gorham-Stout Syndrome, ARA and prolapse of lumbar intervertebral disc.     

Molecular cloning and polymorphism of major histocompatibility complex class Ⅰ genes from grass carp (Ctenophayngodon idellus)

In order to clarify the molecular sequences, allelic polymorphism and the tertiary structure of grass carp(Ctenophayngodon idellus) MHC class Ⅰ, and to further study their relationship with disease resistances, grass carp MHC class Ⅰ gene (Ctid-MHC I) was cloned from a cDNA library and the allelic polymorphism in the population was investigated. The results showed that most of the variations exist in the peptide-binding domain (PBD) and high polymorphism was identified in the Ctid-MHC I allelic genes from 12 individuals. Based on the genetic distance, Ctid-MHC class Ⅰ can be classified into 6 types (from Ctid-MHC I-UA to Ctid-MHC I-UF) which were subdivided into 9 lineages (from A to I). Comparison of the Ctid-MHC I among animals and humans showed that the key amino acids of the peptide binding sites are conserved. Analysis of the tertiary structure of the PBD between Grass carp and human crystallographic data of HLA-A2, the variation with insertion or deletion was found in eight regions (A-H). The p     

Subcellular distribution and compartmentalization of arsenic in <Emphasis Type="Italic">Pteris vittata</Emphasis> L.

The subcellular distribution of arsenic （As） in Pteris vittata L., an As-hyperaccumniator, was studied to determine As compartmentalization and to explore the mechanisms that confer As tolerance. When the plant was grown in a nutrient solution without additional As, most of the accumulated As was isolated to the cell wall. However, in plants growing in a nutrient solution containing 0.1 or 0.2 mmol/L As, approximately 78% of the total As accumulated within the pinna. The proportions of As accumulation in the cyto- plasmic supernatant fraction were 78 % of that in the pinna and 61% of that in the plant. In either treatment group （0.1 or 0.2 mmol/L As）, the fraction containing the lowest level of As was the organelle fraction. These results suggest that As accumulates in the pinna where it is primarily distributed in the cytoplasmic supernatant fraction. The role of As compartmentalization may be intricately linked with As detoxification in P. vittata L.     

Edaphic factors determining variation of two ecotypes of Leymus chinensis in North China

To identify ecotypic variation of L. chinensis, the physiological characteristics and environmental factors were measured and assessed for grey-green and yellow-green ecotypes of L. chinensis grown in the salt-alkalized and aeolian sandy soils. Although both ecotypes presented a similar salt-alkaline tolerance under the experimental conditions, they differed in several aspects. The L. chinensis grown in the aeolian sandy soils had higher water potential and chlorophyll content and lower proline content in their leaves than those grown in the salt-alkalized habitat, and the grey-green ecotype of L. chinensis in both types of soils had higher water potential, chlorophyll and proline contents than the yellow-green ecotype. Besides, grey-green ecotypes had higher salt-alkaline tolerance than the yellow-green ecotype. The stepwise regression analysis showed that soil conductance and pH value were the main ecological factors affecting the physiology of L. chinensis. From the natural distribution of the two ecotypes, we considered that salt-alkaline in the soils is the leading factor in determining ecotypic variation of L. chinensis. The two ecotypes were first named grey-green and yellow-green ecotypes respectively.     

Edaphic factors determining variation of two ecotypes of Leymus chinensis in North China

To identify ecotypic variation of L. chinensis, the physiological characteristics and environmental factors were measured and assessed for grey-green and yellow-green ecotypes of L. chinensis grown in the salt-alkalized and aeolian sandy soils. Although both ecotypes presented a similar salt-alkaline tolerance under the experimental conditions, they differed in several aspects. The L. chinensis grown in the aeolian sandy soils had higher water potential and chlorophyll content and lower proline content in their leaves than those grown in the salt-alkalized habitat, and the grey-green ecotype of L. chinensis in both types of soils had higher water potential, chlorophyll and proline contents than the yellow-green ecotype. Besides, grey-green ecotypes had higher salt-alkaline tolerance than the yellow-green ecotype. The stepwise regression analysis showed that soil conductance and pH value were the main ecological factors affecting the physiology of L. chinensis. From the natural distribution of the two ecotypes, we considered that salt-alkaline in the soils is the leading factor in determining ecotypic variation of L. chinensis. The two ecotypes were first named grey-green and yellow-green ecotypes respectively.     

Screening of the interacting proteins with NifA in <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> Sp7 by the yeast two-hybrid system

NifA in AzospiriUum brasilense plays a key role in regulating the synthesis and activity of nitrogenase in response to ammonia and oxygen available. In this work we used the yeast two-hybrid system to identify the proteins that interact with NifA. The nifA gene was fused to the yeast two-hybrid vector pGBD-C2, and three A. brasilense Sp7 genomic libraries for use in yeast two-hybrid studies were constructed. Screening of the libraries identified four clones encoding proteins that interact with NifA. The confirmation of the interactions of each gene product of the four clones and NifA were carried out by exchanging the vectors for nifA and the four clones and by mutageneses of the four clones with shift reading frame experiments in yeast two-hybrid studies. DNA sequence analyses showed that two clones encode proteins containing PAS domains that play an important role in signal transduction. One clone has high similarity with the fhuE gene of Escherichia coli, whose gene product is involved in iron uptake and transportation, and the other clone encodes an unknown protein.     

Bioaccumulation of heavy metal in wild growing mushrooms from Liangshan Yi nationality autonomous prefecture,China

The heavy metal bioaccumulation levels of 13 species of mushrooms were studied. 13 different species of wild mushrooms growing in China were analyzed for Cu, Zn, Pb, Cd, and As. Contents of Cu, Zn, Cd and Pb in mushroom and Cu, Zn in Soil were tested by atomic absorption spectrometer （AAS）, and As in samples by atomic fluorescent light （AFL）. Contents of Cd and Pb in soil were analyzed by graphite furnace AAS. The results showed that contents of Cu, Pb, Cd, and As in Termitomyces microcarpus were the highest in the tested mushroom samples, which were 135.00, 13.28, 65.30, and 1.60 mg·kg^-1 （dry biomass） respectively, and the corresponding bioconcentration factor （BCF） for Cu and Cd was 57 and 1 674 separately, which was the highest data in 13 mushroom species. The capability ofBoletus griseus for Cd accumulation was very strong with BCF of 300. when people consume the wild edible mushrooms, contents of Cd and Pb should be greatly considered.     

Application of triple potential step amperometry method for quantitative electroanalysis

A novel quantitative electroanalysis method, triple potential step amperometry (TPSA), was developed and explained with an example of nitrobenzene analyzing in water. The selectivity of TPSA was im- proved by controlling the potential step within a narrow interval and using enzyme-modified electrode, the narrow potential step makes the method avoid most interferents, and enzyme-modified electrode can enhance the response of target substance selectively. The peak area was investigated for quanti- tative calibration, such as nitrobenzene concentration showing a linear relation with the peak area, with the correlation coefficients being 0.9995. The t-test and F-test were applied to evaluating the reliability of TPSA, the results showed that there was no evidence of systematic error for TPSA, and the method was of no significant difference from CV. The merit of fast detecting and few potential changing times make the TPSA suitably applicable to low-cost automatic monitoring equipments.