Hai-Ming Zhao

Complete degradation of the endocrine disruptor di-(2-ethylhexyl) phthalate by a novel Agromyces sp. MT-O strain and its application to bioremediation of contaminated soil.

A newly isolated strain Agromyces sp. MT-O could utilize various phthalates and efficiently degraded di-(2-ethylhexyl) phthalate (DEHP). Response surface methodology was successfully employed for the optimization of culture conditions including pH (7.2), temperature (29.6), and inoculum size (OD600 of 0.2), resulting in almost complete degradation of DEHP (200mgL(-1)) within 7days. At different initial concentrations (50-1000mgL(-1)), DEHP degradation curves were fitted well with the first-order kinetic model, and the half-life of DEHP degradation ranged from 0.83 to 2.92days. Meanwhile, the substrate inhibition model was used to describe the special degradation rate with qmax, Ks, and Ki of 0.6298day(-1), 86.78mgL(-1), and 714.3mgL(-1), respectively. The GC-MS analysis indicated that DEHP was degraded into mono-ethylhexyl phthalate and phthalate acid before its complete mineralization. Bioaugmentation of DEHP-contaminated soils with strain MT-O has greatly enhanced DEHP disappearance rate in soils, providing great potential for efficiently remediating DEHP-contaminated environment.

Efficient phytoremediation of organic contaminants in soils using plant–endophyte partnerships

Soil pollution with organic contaminants is one of the most intractable environmental problems today, posing serious threats to humans and the environment. Innovative strategies for remediating organic-contaminated soils are critically needed. Phytoremediation, based on the synergistic actions of plants and their associated microorganisms, has been recognized as a powerful in situ approach to soil remediation. Suitable combinations of plants and their associated endophytes can improve plant growth and enhance the biodegradation of organic contaminants in the rhizosphere and/or endosphere, dramatically expediting the removal of organic pollutants from soils. However, for phytoremediation to become a more widely accepted and predictable alternative, a thorough understanding of plant-endophyte interactions is needed. Many studies have recently been conducted on the mechanisms of endophyte-assisted phytoremediation of organic contaminants in soils. In this review, we highlight the superiority of organic pollutant-degrading endophytes for practical applications in phytoremediation, summarize alternative strategies for improving phytoremediation, discuss the fundamental mechanisms of endophyte-assisted phytoremediation, and present updated information regarding the advances, challenges, and new directions in the field of endophyte-assisted phytoremediation technology.

Variation in accumulation and translocation of di-n-butyl phthalate (DBP) among rice (Oryza sativa L.) genotypes and selection of cultivars for low DBP exposure.

AbstractDi-n-butyl phthalate (DBP) is a typical endocrine-disrupting chemical with higher detection frequency and concentration in agricultural soil (particularly in paddy-field soil of Guangdong Province) of China. In this study, a greenhouse experiment was conducted to investigate variation in uptake and accumulation of DBP by 20 rice cultivars and to screen low DBP-accumulating cultivars. DBP concentrations in plants varied greatly with rice cultivars, growth stages, and tissues. The highest DBP concentrations in both roots and shoots were observed at the ripening stage, with concentrations 2–100-fold higher than those at tillering, jointing, and flowering stages. At the ripening stage, DBP concentrations decreased in the order of leaf > root > stem > grain, and significant differences of DBP concentrations were observed among various rice cultivars. Moreover, the magnitude of variation in DBP concentrations among various cultivars was greater in stems and grains than in roots and leaves. The translocation factors of DBP from roots to stems and from shoots to grains were <1.0, and those from stems to leaves were almost >1.0. Overall, cultivars Yuxiangyouzhan, Jinnongsimiao, Tianyou 122, and Wuyou 380 accumulated relatively lower DBP in grains, resulting in lower DBP exposure. The DBP uptake and translocation pathways in rice require further investigation. Graphical abstractᅟ

Genotypic variation in the uptake, accumulation, and translocation of di-(2-ethylhexyl) phthalate by twenty cultivars of rice (Oryza sativa L.)

Agricultural soil in China contains high levels of di-(2-ethylhexyl) phthalate (DEHP), especially in paddy-field soil of Guangdong province of China, but the accumulation and translocation of DEHP by rice (Oryza sativa L.) remains unknown. In the present study, twenty rice cultivars were cultivated in paddy soil spiked with DEHP, and variations in DEHP accumulation and translocation among various cultivars were investigated. Our results showed that DEHP concentrations in roots and shoots of different rice cultivars at four growth stages (i.e., ripening, tillering, jointing, and flowering stages) varied greatly from 0.26 to 11.8 mg/kg (dry weight, dw) and 0.40 to 7.58 mg/kg (dw), respectively. No obvious change over time was observed. The greatest variation in DEHP concentrations among the rice cultivars occurred at ripening stage, whereas the lowest variation at flowering stage. During ripening stage, the largest variation in DEHP concentrations among cultivars were observed in stems (varying from 0.35 to 13.2 mg/kg), whereas the least one was observed in roots (ranging from 1.01 to 5.72 mg/kg). Significant differences in DEHP concentrations in the roots, stems, leaves and grains of most rice cultivars were found. The translocation factors of DEHP from roots to stems or stems to leaves were higher than those from shoots to grains. Overall, cultivars Tianfengyou 316, Wuyou 308, and Peizataifeng, which contained low levels of DEHP in grains but high levels in shoots, were ideal cultivars for simultaneous production of safe food and phytoremediation of contaminated soil.

Biodegradation of di-n-butyl phthalate (DBP) by a novel endophytic Bacillus megaterium strain YJB3

Phthalic acid esters (PAEs) are a group of recalcitrant and hazardous organic compounds that pose a great threat to both ecosystem and human beings. A novel endophytic strain YJB3 that could utilize a wide range of PAEs as the sole carbon and energy sources for cell growth was isolated from Canna indica root tissue. It was identified as Bacillus megaterium based on morphological characteristics and 16S rDNA sequence homology analysis. The degradation capability of the strain YJB3 was investigated by incubation in mineral salt medium containing di-n-butyl-phthalate (DBP), one of important PAEs under different environmental conditions, showing 82.5% of the DBP removal in 5days of incubation under the optimum conditions (acetate 1.2g·L-1, inocula 1.8%, and temperature 34.2°C) achieved by two-step sequential optimization technologies. The DBP metabolites including mono-butyl phthalate (MBP), phthalic acid (PA), protocatechuic acid (PCA), etc. were determined by GC-MS. The PCA catabolic genes responsible for the aromatic ring cleavage of PCA in the strain YJB3 were excavated by whole-genome sequencing. Thus, a degradation pathway of DBP by the strain YJB3 was proposed that MBP was formed, followed by PA, and then the intermediates were further utilized till complete degradation. To our knowledge, this is the first study to show the biodegradation of PAEs using endophyte. The results in the present study suggest that the strain YJB3 is greatly promising to act as a competent inoculum in removal of PAEs in both soils and crops.

Analysis of Trace Quaternary Ammonium Compounds (QACs) in Vegetables Using Ultrasonic-Assisted Extraction and Gas Chromatography–Mass Spectrometry

A reliable, sensitive, and cost-effective method was developed for determining three quaternary ammonium compounds (QACs) including dodecyltrimethylammonium chloride, cetyltrimethylammonium chloride, and didodecyldimethylammonium chloride in various vegetables using ultrasonic-assisted extraction and gas chromatography-mass spectrometry. The variety and acidity of extraction solvents, extraction times, and cleanup efficiency of sorbents were estimated to obtain an optimized procedure for extraction of the QACs in nine vegetable matrices. Excellent linearities (R(2) > 0.992) were obtained for the analytes in the nine matrices. The limits of detection and quantitation were 0.7-6.0 and 2.3-20.0 μg/kg (dry weight, dw) in various matrices, respectively. The recoveries in the nine matrices ranged from 70.5% to 108.0% with relative standard deviations below 18.0%. The developed method was applied to determine the QACs in 27 vegetable samples collected from Guangzhou in southern China, showing very high detection frequency with a concentration of 23-180 μg/kg (dw).

Soil contamination and sources of phthalates and its health risk in China: A review

&NA; Phthalates (PAEs) are extensively used as plasticizers and constitute one of the most frequently detected organic contaminants in the environment. With the deterioration of eco‐environment in China during the past three decades, many studies on PAE occurrence in soils and their risk assessments have been conducted which allow us to carry out a fairly comprehensive assessment of soil PAE contamination on a nation‐wide scale. This review combines the updated information available associated with PAE current levels, distribution patterns (including urban soil, rural or agricultural soil, seasonal and vertical variations), potential sources, and human health exposure. The levels of PAEs in soils of China are generally at the high end of the global range, and higher than the grade II limits of the Environmental Quality Standard for soil in China. The most abundant compounds, di‐n‐butyl phthalate (DBP) and di‐(2‐ethylhexyl) phthalate (DEHP), display obvious spatial distribution in different provinces. It is noted that urbanization and industrialization, application of plastic film (especially plastic film mulching in agricultural soil) and fertilizer are the major sources of PAEs in soil. Uptake of PAEs by crops, and human exposure to PAEs via ingestion of soil and vegetables are reviewed, with scientific gaps highlighted. Graphical abstract Figure. No Caption available. HighlightsPAE levels in soils of China were generally higher than the other countries.DBP and DEHP are the dominant compounds in soil with obvious spatial distribution.Urbanization and industrialization, application of plastic film and fertilizer are the major sources of PAEs in soil.Uptake of PAEs by crops and human exposure to PAEs are reviewed.

Physiological differences in response to di-n-butyl phthalate (DBP) exposure between low- and high-DBP accumulating cultivars of Chinese flowering cabbage (Brassica parachinensis L.)

To increase understanding on the mechanisms of cultivar difference in contaminant accumulation in crops, this study was designed to compare the physiological responses to di-n-butyl phthalate (DBP) exposure between low (Lvbao70) and high (Huaguan) DBP cultivars of Chinese flowering cabbage (Brassica parachinensis L.). Under high DBP exposure, significant differences in various physiological responses were observed between the two cultivars, which might account for the variation in DBP accumulation. Ultrastructure observation also showed different alterations or damages in the mesophyll cell structures between both cultivars, especially for the chloroplast disintegration, starch grain quantity, and plastoglobuli accumulation. Compared with Huaguan, Lvbao70 suffered greater decreases in biomass, chlorophyll content, carbon assimilation, gas exchange parameters, photosynthetic electron transport capacity, and antioxidase activities, which would have resulted in a great reduction of photosynthetic capacity. Although Lvbao70 enhanced energy dissipation and activities of some antioxidant enzymes, they did not provide sufficient protection against oxidative damage caused by DBP. The result suggested that the lower DBP tolerance of Lvbao70 might be associated with its poor physiological performances, which was responsible for its lower DBP accumulation to protect itself from toxicity. Additionally, Lvbao70 had a significantly lower transpiration rate and stomatal conductance than Huaguan, which might be the factors regulating DBP-accumulation variation.

Low-molecular-weight organic acids correlate with cultivar variation in ciprofloxacin accumulation in Brassica parachinensis L.

To understand the mechanism controlling cultivar differences in the accumulation of ciprofloxacin (CIP) in Chinese flowering cabbage (Brassica parachinensis L.), low-molecular-weight organic acids (LMWOAs) secreted from the roots of high- and low-CIP cultivars (Sijiu and Cutai, respectively) and their effects on the bioavailability of CIP in soil were investigated. Significant differences in the content of LMWOAs (especially maleic acid) between the two cultivars played a key role in the variation in CIP accumulation. Based on the Freundlich sorption coefficient (Kf) and distribution coefficient (Kd), the presence of LMWOAs reduced the CIP sorption onto soil particles, and higher concentrations of LMWOAs led to less CIP sorption onto soil. On the other hand, LMWOAs enhanced CIP desorption by lowering the solution pH, which changed the surface charge of soil particles and the degree of CIP ionization. LMWOAs promoted CIP desorption from soil by breaking cation bridges and dissolving metal cations, particularly Cu2+. These results implied that the LMWOAs (mainly maleic acid) secreted from Sijiu inhibited CIP sorption onto soil and improved CIP desorption from soil to a greater extent than those secreted from Cutai, resulting in higher bioavailability of CIP and more uptake and accumulation of CIP in the former.

Variations in microbial community and ciprofloxacin removal in rhizospheric soils between two cultivars of Brassica parachinensis L.

Ciprofloxacin (CIP) is one of most used quinolone antibiotics detected frequently in agricultural soils and vegetables. In the present study, variations in microbial community and CIP removal in rhizospheric soils between two cultivars of Brassica parachinensis L. that accumulate higher and lower CIP (Sijiu and Cutai, respectively) were investigated under CIP stress (0mg/kg in CK, 2.94mg/kg in T1, and 67.11mg/kg in T2). The removal rates of CIP in rhizospheric soils of cultivar Sijiu were higher than those of cultivar Cutai, with a significant difference in T2 (48.7%>39.4%, P<0.05). The pyrosequencing of 16S rRNA and ITS gene indicated that the microbial diversity and community structure in rhizospheric soils of the two cultivars varied significantly. Spirochaeta and Trichosporon might be associated with CIP degradation, and higher relative abundances of Trichosporon in rhizoshperic soils of cultivar Sijiu might be responsible for higher CIP removal. Fourteen bacterial genera and ten fungal genera were screened as potential biomarkers for CIP removal process. The community level physiological profiling in rhizospheric soils of the two cultivars under CIP stress differed significantly, and more C substrates that favored CIP removal were observed in rhizoshperic soils of cultivar Sijiu. Our results demonstrate that variations in microbial community and the utilization of C substrates played important roles in differring the CIP removal in rhizospheric soils between the two cultivars.