Peidong Tai

Variations in the bioavailability of polycyclic aromatic hydrocarbons in industrial and agricultural soils after bioremediation

The aim of this study was to demonstrate the variations in bioavailability remaining in industrial and agricultural soils contaminated by polycyclic aromatic hydrocarbons (PAHs) after bioremediation. After inoculation of Mycobacterium sp. and Mucor sp., PAH biodegradation was tested on a manufactured gas plant (MGP) soil and an agricultural soil. PAH bioavailability was assessed before and after biodegradation using solid-phase extraction (Tenax-TA extraction) and solid-phase micro-extraction (SPME) to represent bioaccessibility and chemical activity of PAHs, respectively. Only 3- and 4-ring PAHs were noticeably biodegradable in the MGP soil. PAH biodegradation in the agricultural soil was different from that in the MGP soil. The rapidly desorbing fractions (F(rap)) extracted by Tenax-TA and the freely dissolved concentrations of 3- and 4-ring PAHs determined by SPME from the MGP soil decreased after 30 days biodegradation; those values of the 5- and 6-ring PAHs changed to a lesser degree. For the agricultural soil, the F(rap) values of the 3- and 4-ring PAHs also decreased after the biodegradation experiment. The Tenax-TA extraction and the SPME have the potential to assess variations in the bioavailability of PAHs and the degree of biodegradation in contaminated MGP soils. In addition, Tenax-TA extraction is more sensitive than SPME when used in the agricultural soil.

Cadmium-induced DNA damage and mutations in Arabidopsis plantlet shoots identified by DNA fingerprinting

Random amplified polymorphic DNA (RAPD) test is a feasible method to evaluate the toxicity of environmental pollutants on vegetal organisms. Herein, Arabidopsis thaliana (Arabidopsis) plantlets following Cadmium (Cd) treatment for 26 d were screened for DNA genetic alterations by DNA fingerprinting. Four primers amplified 20-23 mutated RAPD fragments in 0.125-3.0 mg L(-1) Cd-treated Arabidopsis plantlets, respectively. Cloning and sequencing analysis of eight randomly selected mutated fragments revealed 99-100% homology with the genes of VARICOSE-Related, SLEEPY1 F-box, 40S ribosomal protein S3, phosphoglucomutase, and noncoding regions in Arabidopsis genome correspondingly. The results show the ability of RAPD analysis to detect significant genetic alterations in Cd-exposed seedlings. Although the exact functional importance of the other mutated bands is unknown, the presence of mutated loci in Cd-treated seedlings, prior to the onset of significant physiological effects, suggests that these altered loci are the early events in Cd-treated Arabidopsis seedlings and would greatly improve environmental risk assessment.

Cadmium-induced genomic instability in Arabidopsis: Molecular toxicological biomarkers for early diagnosis of cadmium stress.

Microsatellite instability (MSI) analysis, random-amplified polymorphic DNA (RAPD), and methylation-sensitive arbitrarily primed PCR (MSAP-PCR) are methods to evaluate the toxicity of environmental pollutants in stress-treated plants and human cancer cells. Here, we evaluate these techniques to screen for genetic and epigenetic alterations of Arabidopsis plantlets exposed to 0-5.0 mg L(-1) cadmium (Cd) for 15 d. There was a substantial increase in RAPD polymorphism of 24.5, and in genomic methylation polymorphism of 30.5-34.5 at CpG and of 14.5-20 at CHG sites under Cd stress of 5.0 mg L(-1) by RAPD and of 0.25-5.0 mg L(-1) by MSAP-PCR, respectively. However, only a tiny increase of 1.5 loci by RAPD occurred under Cd stress of 4.0 mg L(-1), and an additional high dose (8.0 mg L(-1)) resulted in one repeat by MSI analysis. MSAP-PCR detected the most significant epigenetic modifications in plantlets exposed to Cd stress, and the patterns of hypermethylation and polymorphisms were consistent with inverted U-shaped dose responses. The presence of genomic methylation polymorphism in Cd-treated seedlings, prior to the onset of RAPD polymorphism, MSI and obvious growth effects, suggests that these altered DNA methylation loci are the most sensitive biomarkers for early diagnosis and risk assessment of genotoxic effects of Cd pollution in ecotoxicology.

Effect of freeze-thawing cycles on soil aging behavior of individually spiked phenanthrene and pyrene at different concentrations

This work was initiated to study the concentration effects on phenanthrene and pyrene extraction during process of aging in phaeozem and burozem with or without freeze-thawing cycles. 1, 10, and 100 μg g(-1) phenanthrene and pyrene contaminated soils were extracted by 10,000 mg L(-1) surfactant SDBS at various times. The extraction amount decreased with increasing contact time. Aging process could be divided into two stages: an initially rapid and then a slow sorption period. The amount extraction of 1 μg g(-1) pyrene in phaeozem was higher at the initial sorption stage. The time for 100 μg g(-1) phenanthrene and pyrene extraction efficiency to reach equilibrium in both soils were shorter than 1 and 10 μg g(-1). Changes in extraction efficiency rate caused by slow sorption were affected by contact time, physicochemical properties of soils and chemicals and the applied concentration of chemicals. For 1 and 10 μg g(-1) phenanthrene and pyrene contaminated soils, freeze-thawing cycle increased the extraction efficiency at the initial 1st and 8th days. The extraction efficiency with no freeze-thawing cycle was higher than that with freeze-thawing cycles at 30 and 120 days. A conceptual model was proposed to account for this process.

Characterization of cadmium (108Cd) distribution and accumulation in Tagetes erecta L. seedlings: Effect of split-root and of remove-xylem/phloem

Tagetes erecta has a high potential for cadmium (Cd) phytoremediation. Through several hydroponic experiments, characteristics of (108)Cd distribution and accumulation were investigated in T. erecta with split -roots or removed xylem/phloem. The results showed that (108)Cd transport from roots to aboveground tissues showed the homolateral transport phenomenon in split-root seedlings. (108)Cd content of leaves on the +(108)Cd side and the -(108)Cd side was not significantly different, which implied that there was horizontal transport of (108)Cd from the +(108)Cd side to the -(108)Cd side in cut-root seedlings. Like (108)Cd transport, the transport of (70)Zn was homolateral. Reduction of water consumption in the removed xylem treatment significantly decreased (108)Cd accumulation; whereas, the removed phloem treatment had no significant effect on water consumption, but did decrease (108)Cd accumulation in leaves of the seedlings. The removal of phloem significantly reduced distal leaf (108)Cd content, which was significantly lower than that in the basal leaves in both the split-root and unsplit-root seedlings. Overall, the results presented in this study revealed that the root to aboveground cadmium translocation via phloem is as an important and common physiological process as xylem determination of the cadmium accumulation in stems and leaves of marigold seedlings.

Role of sulfur in cadmium accumulation of Tagetes erecta L.

ABSTRACT The role of sulfur in cadmium (Cd) accumulation was investigated in Tagetes erecta L. Shoot Cd concentrations were significantly increased with increasing of sulfur levels in the culture solution. In another experiment, leaf application of sodium sulfate (Na2SO4) or cysteine to sulfur (S)-starved plants significantly increased Cd accumulation in shoots up to three folds or twice that of those S-starved plants. When plants were cultured at 35°C, shoot Cd accumulation was significantly lower than in those cultured at 25°C, however, leaf application of glutathione or cysteine significantly increased Cd accumulations in shoots than that of no application. It is suggested that elevated temperature (35°C) might disturb sulfur assimilation, for example, cysteine and glutathione synthesis, thus reducing Cd accumulation. This phenomenon can be alleviated by adding thiol-containing compounds, such as cysteine and glutathione, but not by adding Na2SO4. These results indicate that thiol-containing compounds play an important role in Cd accumulation of T. erecta.

Short-term toxic effects of chlorobenzenes on broadbean (Vicia faba) seedlings

The root growth, changes in Superoxide dismutase (SOD, EC 1.15.1.1) activity, malonyldialdehyde (MDA) and total soluble protein level of broadbean (Vicia faba) seedlings were researched at different soil concentrations of chlorobenzene (CB), 1,2,4-trichlorobenzene (TCB) and hexachlorobenzene (HCB). The results showed that root growth of seedlings was interrupted after 5d of 50–200 μg · g−1 TCB treatment. During a 3 d of recovery period, root growth was, however, restored to some extent although there was a delay in returning to the control level. The total soluble protein content in seedlings increased with TCB concentration and duration of exposure. Effect of TCB stress on SOD activity in seedlings displayed a significant dose-effect relationship for 1–5 d of 50–200 μg · g−1 treatment. When broadbean seedlings were placed in clean tap water for 3 d following exposure to 5 d of TCB stress to clear tap water for 3 d, SOD activity at 50 μg · g−1 TCB recovered towards control level (P> 0.05) while a significant increase in SOD activity was observed at 100 and 200 μg · g−1 TCB compared to control (P< 0.05). The experiments also revealed that a significant increase of MDA level in seedlings occurred after 3 and 5 d of 100 and 200 μg · g−1 TCB treatment (P< 0.05 andP< 0.01), and there was a positive correlation between TCB concentration and MDA level. All the above results showed that SOD activity and MDA level of broadbean seedlings might be proposed as the biomarkers for short-term TCB contamination in soil. Compared to TCB, the toxicity of 50−1000 μg · g−1 CB or HCB in soil to broadbean seedlings was not observed after a 3 d exposure.

Selective adsorption of Mycobacterium Phlei on pyrite and sphalerite

The adsorption of Mycobacterium Phlei cells on the surfaces of pyrite and sphalerite was studied as functions of time and pH. The results indicated that a higher amount of cells adsorbing onto pyrite compared with that onto sphalerite under neutral and alkaline conditions, and it was also observed from photographs of scanning electron micrograph. To gain a better insight into the mechanisms of differential adsorption, the functional groups on cell surfaces and the chemical states of each element on mineral surfaces before and after interaction with bacterial cells were investigated. The results showed that many groups presented on cells surface, such as C-O-H, C-O-C, C=O, C-N, N-H and P=O. The change in state of each element on pyrite and sphalerite surfaces after interaction with bacterial cells revealed that there were chemical reactions between metal ions and S on mineral surface and atoms like N, O, P, etc. on cell surface, and the shifts in binding energy of each element on pyrite surface is larger than that of sphalerite. Possible mechanisms for selective adsorption of bacterial cells onto pyrite and sphalerite were discussed in the latter part of this paper.

CADMIUM ACCUMULATION OF TAGETES ERECTA L. AFFECTED BY PLANT GROWTH INHIBITORS AND GLUTATHIONE

The effect of two plant growth inhibitors maleic hydrazide (MH) and cyeloheximide (CHI) on cadmium (Cd) accumulation was investigated in Tagetes erecta L. The relative growth rate (RGR) as well as Cd content in shoots of Tagetes erecta L. was obviously decreased by the two inhibitors, indicating that there was close relationship between Cd accumulation in shoots and the plant growth. When the sulfur (S)-containing compound glutathione (GSH) was applied to the leaves of Tagetes erecta L. under the MH or CHI treatment, the decreased Cd content in shoots affected by MH was alleviated, but there was no effect of the GSH on Cd content in shoots under CHI treatment, suggesting that GSH has an counter-effect on Cd accumulation in shoots of Tagetes erecta L. under the MH treatment. All the results suggested that plant growth and S-containing compounds play an important role in Cd accumulation in Tagetes erecta L.

Cell cycle arrest mediated by Cd-induced DNA damage in Arabidopsis root tips

Accumulating evidence demonstrates that the aberrant expression of cell cycle regulation and DNA repair genes can result in abnormal cell proliferation and genomic instability in eukaryotic cells under different stresses. Herein, Arabidopsis thaliana (Arabidopsis) seedlings were grown hydroponically on 0.5 × MS media containing cadmium (Cd) at 0-2.5mgL-1 for 5d of treatment. Real time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis revealed that expression of DNA damage repair and cell cycle regulation genes, including BRCA1, MRE11, WEE1, CDKA;1 and PCNA1, showed an inverted U-shaped dose-response. In contrast, notably reduced expression was observed for G1-to-S transition-related genes, Histone H4, E2Fa and PCNA2; DSB end processing, GR1; G2-to-M transition-related gene, CYCB1;1; and DNA mismatch repair, MSH2, MSH6 and MLH1 genes in root tips exposed to 0.125-2.5mg/L Cd for 5d. Flow cytometry (FCM) analysis revealed significant increases of cells with a 2C nuclear content and with a 4C and 8C nuclear content under Cd stresses of 0.125 and 1-2.5mgL-1, respectively. Our results suggest that 0.125mgL-1 Cd-induced DNA damage induced the marked G1/S arrest, leading to accelerated growth in root tips, while 1.0-2.5mgL-1 Cd-induced DNA damage caused a notable G2/M arrest in root tips, leading to reduced growth in root tips. This may be a protective mechanism that prevents cells with damaged DNA from dividing under Cd stress.