Driving factors of community-level leaf stoichiometry patterns in a typical temperate mountain meadow ecosystem of northern China.

In ecological stoichiometry, the stoichiometry and spatial distribution of leaf carbon, nitrogen, and phosphorus are important research topics. Various studies have assessed leaf stoichiometry and its relationships with environmental factors at different scales. However, how the leaf carbon, nitrogen and phosphorus stoichiometric characteristics of the same vegetation type at the community level vary with environmental factors along a continuous altitudinal gradient remains poorly understood. In this paper, 13 sampling sites along an altitudinal gradient of 1,800-3,011 m in a typical temperate mountain meadow ecosystem on the southern slope of the Wutai Mountain in North China were sampled to explore the response of leaf carbon, nitrogen and phosphorus stoichiometric characteristics to altitude change using correlation analysis, and then quantified the contribution of driving factors using canonical correspondence analysis (CCA) and variation partitioning. We found that the community-level leaf stoichiometry of mountain meadows differed significantly at different altitudes, and an increase in altitude significantly decreased community-level leaf total nitrogen (LTN) and leaf total phosphorus (LTP); however, the leaf total carbon (LTC), C∶N, C∶P, and N∶P increased with an increase in altitude. Additionally, with increasing altitude, soil properties showed significant trends. Soil organic carbon (SOC), soil total nitrogen (STN), soil total phosphorus (STP), soil water content and soil electrical conductivity increased significantly, but soil temperature, soil bulk density and soil pH exhibited the opposite trend. Our results suggested that altitude, soil electrical conductivity and soil bulk density significantly influenced the changes in the leaf stoichiometric characteristics, explaining 75.5% of the total variation, and altitude had the greatest influence (36.6%). In the temperate mountains, altitude played a decisive role in affecting patterns of meadow plant nutrients and stoichiometry and was more important than soil in explaining leaf C∶N∶P stoichiometry variations. Our findings provide important references to understand the responses of plant stoichiometry to altitudinal gradients.

Three-dimensional single-photon imaging through realistic fog in an outdoor environment during the day.

Due to the strong scattering of fog and the strong background noise, the signal-to-background ratio (SBR) is extremely low, which severely limits the 3D imaging capability of single-photon detector array through fog. Here, we propose an outdoor three-dimensional imaging algorithm through fog, which can separate signal photons from non-signal photons (scattering and noise photons) with SBR as low as 0.003. This is achieved by using the observation model based on multinomial distribution to compensate for the pile-up, and using dual-Gamma estimation to eliminate non-signal photons. We show that the proposed algorithm enables accurate 3D imaging of 1.4 km in the visibility of 1.7 km. Compared with the traditional algorithms, the target recovery (TR) of the reconstructed image is improved by 20.5%, and the relative average ranging error (RARE) is reduced by 28.2%. It has been successfully demonstrated for targets at different distances and imaging times. This research successfully expands the fog scattering estimation model from indoor to outdoor environment, and improves the weather adaptability of the single-photon detector array.

Pathways and signatures of mutagenesis at targeted DNA nicks.

Nicks are the most frequent form of DNA damage and a potential source of mutagenesis in human cells. By deep sequencing, we have identified factors and pathways that promote and limit mutagenic repair at a targeted nick in human cells. Mutations were distributed asymmetrically around the nick site. BRCA2 inhibited all categories of mutational events, including indels, SNVs and HDR. DNA2 and RPA promoted resection. DNA2 inhibited 1 bp deletions but contributed to longer deletions, as did REV7. POLQ stimulated SNVs. Parallel analysis of DSBs targeted to the same site identified similar roles for DNA2 and POLQ (but not REV7) in promoting deletions and for POLQ in stimulating SNVs. Insertions were infrequent at nicks, and most were 1 bp in length, as at DSBs. The translesion polymerase REV1 stimulated +1 insertions at one nick site but not another, illustrating the potential importance of sequence context in determining the outcome of mutagenic repair. These results highlight the potential for nicks to promote mutagenesis, especially in BRCA-deficient cells, and identify mutagenic signatures of DNA2, REV1, REV3, REV7 and POLQ.

Treatment of human cells with 5-aza-dC induces formation of PARP1-DNA covalent adducts at genomic regions targeted by DNMT1.

The nucleoside analog 5-aza-2'-deoxycytidine (5-aza-dC) is used to treat some hematopoietic malignancies. The mechanism of cell killing depends upon DNMT1, but is otherwise not clearly defined. Here we show that PARP1 forms covalent DNA adducts in human lymphoblast or fibroblasts treated with 5-aza-dC. Some adducts recovered from 5-aza-dC-treated cells have undergone cleavage by apoptotic caspases 3/7. Mapping of PARP1-DNA adducts, by a new method, "Adduct-Seq", demonstrates adduct enrichment at CpG-dense genomic locations that are targets of maintenance methylation by DNMT1. Covalent protein-DNA adducts can arrest replication and induce apoptosis, and these results raise the possibility that induction of PARP1-DNA adducts may contribute to cell killing in response to treatment with 5-aza-dC.

POLQ suppresses interhomolog recombination and loss of heterozygosity at targeted DNA breaks.

Interhomolog recombination (IHR) occurs spontaneously in somatic human cells at frequencies that are low but sufficient to ameliorate some genetic diseases caused by heterozygous mutations or autosomal dominant mutations. Here we demonstrate that DNA nicks or double-strand breaks (DSBs) targeted by CRISPR-Cas9 to both homologs can stimulate IHR and associated copy-neutral loss of heterozygosity (cnLOH) in human cells. The frequency of IHR is 10-fold lower at nicks than at DSBs, but cnLOH is evident in a greater fraction of recombinants. IHR at DSBs occurs predominantly via reciprocal end joining. At DSBs, depletion of POLQ caused a dramatic increase in IHR and in the fraction of recombinants exhibiting cnLOH, suggesting that POLQ promotes end joining in cis, which limits breaks available for recombination in trans These results define conditions that may produce cnLOH as a mutagenic signature in cancer and may, conversely, promote therapeutic correction of both compound heterozygous and dominant negative mutations associated with genetic disease.

[Value evaluation and ecological compensation of endangered species from the perspective of stakeholders: A cases study with Elaeagnus mollise]. / åˆ©ç›Šç›¸å ³è€ è§†è§’ä¸‹çš„æ¿’å±ç‰©ç§ä»·å€¼è¯„ä¼°ä¸Žç”Ÿæ€è¡¥å¿­­ä»¥ç¿ æžœæ²¹æ ‘ä¸ºä¾‹.

The value evaluation of endangered species can provide important supports for policy-making in biodiversity conservation. We evaluated the stakeholders' preference and willingness to pay (WTP) from the perspective of stakeholders by the choice experiment method (CEM) and discussed the ecological compensation mechanism, with Elaeagnus mollis, a national secondary key protected plant as an example. Based on the interest demands analysis of three stakeholders, the management departments, enterprises and farmers, we measured five species attributes of E. mollis, including planting area, product classification, seedling varieties, protection investment, and far-mers' income. Furthermore, the preference of stakeholders and WTP for different attribute variables were analyzed using random parameter model (RPL). The results showed that the preference of respondents for the variable of "increase investment in protection" was the strongest and WTP was the highest (331.00 yuan·a-1·household-1). The stakeholders had stronger preference for the variables of "simultaneous development of high and low-end products" and "development of high-end products", with WTP being 242.71 and 227.57 yuan·a-1·household-1, respectively. For the variables of "farmers' income", "unchanged investment in protection", "seedling varieties" and "planting area", stakeholders showed certain preference and WTP from strong to weak. The prefe-rence for "no investment in protection" and "development of low-end products" was weaker and the WTP was negative. Through direct and indirect approaches of compensation, ecological compensation could be implemented for the objects of ecological compensation (E. mollis and associated habitat). The value of compensating surplus (CS) calculated was 285.62 yuan·a-1·household-1.

Climate change-induced migration patterns and extinction risks of Theaceae species in China.

Theaceae, an economically important angiosperm family, is widely distributed in tropical and subtropical forests in Asia. In China, Theaceae has particularly high abundances and endemism, comprising ~75% of the total genera and ~46% of the total species worldwide. Therefore, predicting the response of Theaceae species to climate change is vital. In this study, we collected distribution data for 200 wild Theaceae species in China, and predicted their distribution patterns under current and future climactic conditions by species distribution modeling (SDM). We revealed that Theaceae species richness is highest in southeastern China and on Hainan Island, reaching its highest value (137 species) in Fujian Province. According to the IUCN Red List criteria for assessing species threat levels under two dispersal assumptions (no dispersal and full dispersal), we evaluated the conservation status of all Theaceae species by calculating loss of suitable habitat under future climate scenarios. We predicted that nine additional species will become threatened due to climate change in the future; one species will be classified as critically endangered (CR), two as endangered (EN), and six as vulnerable (VU). Given their extinction risks associated with climate change, we recommended that these species be added to the Red List. Our investigation of migration patterns revealed regional differences in the number of emigrant, immigrant, and persistent species, indicating the need for targeted conservation strategies. Regions containing numerous emigrants are concentrated in Northern Taiwan and coastal regions of Zhejiang and Fujian provinces, while regions containing numerous immigrants include central Sichuan Province, the southeastern Tibet Autonomous Region, southwest Yunnan Province, northwest Sichuan Province, and the junction of Guangxi and Hunan provinces. Lastly, regions containing persistent species are widely distributed in southern China. Importantly, regions with high species turnover are located on the northern border of the entire Theaceae species distribution ranges owing to upwards migration; these regions are considered most sensitive to climate change and conservation planning should therefore be prioritized here. This study will contribute valuable information for reducing the negative impacts of climate change on Theaceae species, which will ultimately improve biodiversity conservation efficiency.

[Flora and geographic pattern of mountain forests at community level in Taihang Mountains: Results based on plant community survey]. / å¤ªè¡Œå±±å±±åœ°æ£®æž—ç¾¤è½æ¤ç‰©åŒºç³»ä¸Žåœ°ç†æ ¼å±€­­åŸºäºŽæ¤ç‰©ç¾¤è½æ¸ 查数据.

Taihang Mountains is located on the eastern edge of the second step of Chinese topography, which is the natural boundary between Loess Plateau and North China Plain and is considered as one of the important priority areas for biodiversity conservation in China. We took 108 counties involved in the generalized Taihang Mountains as the research area to systematically analyze the characteristics of family and genus characters, floristic composition, geographical pattern of plant diversity and species richness hotspots at community level of mountain forests in Taihang Mountains. A total of 963 species of seed plants belonging to 447 genera and 100 families were recorded in 778 forest plots in Taihang Mountains. Within all the species, 12 species of gymnosperms belonging to 7 genera of 3 families, 951 species of angiosperms belonging to 440 genera of 97 families. Herbaceous plants (71.1%) was the dominant life form. The distribution types of families were mainly tropical (38%) and temperate (24%), and the distribution types of genera were mainly temperate (68.7%). The horizontal distribution pattern of plant diversity showed a trend of increasing from southwest to northeast. Species richness was positively correlated with the latitude and longitude. However, the richness patterns of different life-form plants were different, in that herbaceous richness was positively correlated with the longitude and latitude but that of woody plants was not. In the vertical gradient, plant richness of Taihang Mountains presented a single-peak distribution, which was concentrated in the low and middle elevations of 400-1800 m and peaked at 1000-1200 m. Based on the community inventory data, we mapped plant richness of Taihang Mountain forest community. The mountain areas such as Xiaowutai Mountain, Yuntai Mountain, Taiyue Mountain, Wangwu Mountain and Zhongtiao Mountain were identified as hotspots of plant richness, which should be included in the key planning and management areas of the Taihang Mountains priority protection.

[Interactive Effects of Ozone and Drought on Antioxidant Enzyme Activities of Poplar Leaves].

Ground-level ozone (O3) pollution and drought have become major environmental factors that threaten the normal growth of plants. In this study, we determined the effects of three O3 concentrations (charcoal-filtered air, CF, non-filtered air, NF and non-filtered air+40 nmol·mol-1, NF40), two water treatments (well-watered, WW, and mild drought, MD, 60% of WW in volumetric soil water content), and their interactions on the light-saturated photosynthesis rate (Asat) and on the activity of antioxidant enzymes and soluble proteins of the poplar '546' (Populus deltoids cv. 55/56×P. deltoides cv. Imperial). Results showed that Asat significantly decreased with elevated O3, and with interactions between O3 and drought. Drought significantly reduced the activities of catalase (CAT) and superoxide dismutase (SOD) but did not affect the activities of peroxidase (POD) or ascorbate peroxidase (APX). Reduction ranges in CAT and SOD increased with drought duration. APX activity significantly decreased with increasing O3 and drought, but soluble protein content did not. Activities of antioxidant enzymes were not significantly affected by elevated O3 and their interactions. Results provide a scientific basis for protection of poplar plantations in response to increasing O3 concentrations and drought under environmental changes.

Activity and fidelity of human DNA polymerase α depend on primer structure.

DNA polymerase α (Polα) plays an important role in genome replication. In a complex with primase, Polα synthesizes chimeric RNA-DNA primers necessary for replication of both chromosomal DNA strands. During RNA primer extension with deoxyribonucleotides, Polα needs to use double-stranded helical substrates having different structures. Here, we provide a detailed structure-function analysis of human Polα's interaction with dNTPs and DNA templates primed with RNA, chimeric RNA-DNA, or DNA. We report the crystal structures of two ternary complexes of the Polα catalytic domain containing dCTP, a DNA template, and either a DNA or an RNA primer. Unexpectedly, in the ternary complex with a DNA:DNA duplex and dCTP, the "fingers" subdomain of Polα is in the open conformation. Polα induces conformational changes in the DNA and hybrid duplexes to produce the universal double helix form. Pre-steady-state kinetic studies indicated for both duplex types that chemical catalysis rather than product release is the rate-limiting step. Moreover, human Polα extended DNA primers with higher efficiency but lower processivity than it did with RNA and chimeric primers. Polα has a substantial propensity to make errors during DNA synthesis, and we observed that its fidelity depends on the type of sugar at the primer 3'-end. A detailed structural comparison of Polα with other replicative DNA polymerases disclosed common features and some differences, which may reflect the specialization of each polymerase in genome replication.

Assaying Repair at DNA Nicks.

Nicks are the most common form of DNA damage, but they have only recently been shown to initiate damage that requires repair. Analysis of the pathways of nick repair in human cells has benefited from the development of enzymes that target nicks to specific sites in the genome and of reporters that enable rapid analysis of homology-directed repair and mutagenic end joining. Nicks undergo efficient repair by single-stranded oligonucleotide donors complementary to either the nicked or intact DNA strand, via pathways that are normally suppressed by RAD51. Here we discuss the details of reporter assays that take advantage of the convenience and sensitivity of flow cytometry to analyze pathways of repair at targeted DNA nicks. These assays are readily carried out in 96-well format cell culture plates, enabling mechanistic questions to be addressed by determining the contributions of specific factors by depletion and/or ectopic expression.

Comment on "The [4Fe4S] cluster of human DNA primase functions as a redox switch using DNA charge transport".

O'Brien et al (Research Article, 24 February 2017, eaag1789) proposed a novel mechanism of primase function based on redox activity of the iron-sulfur cluster buried inside the C-terminal domain of the large primase subunit (p58C). Serious problems in the experimental design and data interpretation raise concerns about the validity of the conclusions.

Occurrence, profiles, and ecological risks of polybrominated diphenyl ethers in mangrove sediments of Shantou, China.

Surface sediments were collected from three mangrove wetlands (Yifeng Xi, Shuanghan, and Su'ai Wan) in Shantou coastal zone of South China to investigate spatial distributions of polybrominated diphenyl ethers (PBDEs). The results demonstrate that PBDEs were detected in all the samples, indicating their widespread occurrence in coastal sediments of the studied area. Σ9PBDEs (defined as the sum of nine targeted PBDE congeners except BDE-209) and BDE-209 are in the range of 2.3 to 11.5 and 16.7 to 58.2 ng/g, respectively. BDE-209 is the dominant PBDE congener in all sediment samples. The sediment concentrations of ∑9PBDEs and BDE-209 among the three wetlands decrease in the order of Su'ai Wan > Shuanghan > Yifeng Xi. The concentrations of ∑9PBDEs are higher in mangrove sediments than in mudflats, but no obvious regularity can be found on the correlation between mangrove species and PBDE levels in sediments. The contents of total organic carbon are moderately correlated with BDE-209 concentrations in sediments but not with ∑9PBDE concentrations. The samples collected from different locations show slightly different composition profiles except BDE-209, with BDE-100 and BDE-47 being the pre-dominated congeners. The mudflats exhibit higher abundances of tri- to hexa-substituted congeners than the mangrove sediments. Ecological risk assessment demonstrates that the surface sediments from Shantou may pose a potential ecological risk of exposure to sediment-dwelling organisms.

Divalent ions attenuate DNA synthesis by human DNA polymerase α by changing the structure of the template/primer or by perturbing the polymerase reaction.

DNA polymerases (pols) are sophisticated protein machines operating in the replication, repair and recombination of genetic material in the complex environment of the cell. DNA pol reactions require at least two divalent metal ions for the phosphodiester bond formation. We explore two understudied roles of metals in pol transactions with emphasis on polα, a crucial enzyme in the initiation of DNA synthesis. We present evidence that the combination of many factors, including the structure of the template/primer, the identity of the metal, the metal turnover in the pol active site, and the influence of the concentration of nucleoside triphosphates, affect DNA pol synthesis. On the poly-dT70 template, the increase of Mg(2+) concentration within the range typically used for pol reactions led to the severe loss of the ability of pol to extend DNA primers and led to a decline in DNA product sizes when extending RNA primers, simulating the effect of "counting" of the number of nucleotides in nascent primers by polα. We suggest that a high Mg(2+) concentration promotes the dynamic formation of unconventional DNA structure(s), thus limiting the apparent processivity of the enzyme. Next, we found that Zn(2+) supported robust polα reactions when the concentration of nucleotides was above the concentration of ions; however, there was only one nucleotide incorporation by the Klenow fragment of DNA pol I. Zn(2+) drastically inhibited polα, but had no effect on Klenow, when Mg(2+) was also present. It is possible that Zn(2+) perturbs metal-mediated transactions in pol active site, for example affecting the step of pyrophosphate removal at the end of each pol cycle necessary for continuation of polymerization.

Mechanism of Concerted RNA-DNA Primer Synthesis by the Human Primosome.

The human primosome, a 340-kilodalton complex of primase and DNA polymerase α (Polα), synthesizes chimeric RNA-DNA primers to be extended by replicative DNA polymerases δ and ϵ. The intricate mechanism of concerted primer synthesis by two catalytic centers was an enigma for over three decades. Here we report the crystal structures of two key complexes, the human primosome and the C-terminal domain of the primase large subunit (p58C) with bound DNA/RNA duplex. These structures, along with analysis of primase/polymerase activities, provide a plausible mechanism for all transactions of the primosome including initiation, elongation, accurate counting of RNA primer length, primer transfer to Polα, and concerted autoregulation of alternate activation/inhibition of the catalytic centers. Our findings reveal a central role of p58C in the coordinated actions of two catalytic domains in the primosome and ultimately could impact the design of anticancer drugs.

Insight into the Human DNA Primase Interaction with Template-Primer.

DNA replication in almost all organisms depends on the activity of DNA primase, a DNA-dependent RNA polymerase that synthesizes short RNA primers of defined size for DNA polymerases. Eukaryotic and archaeal primases are heterodimers consisting of small catalytic and large accessory subunits, both of which are necessary for the activity. The mode of interaction of primase subunits with substrates during the various steps of primer synthesis that results in the counting of primer length is not clear. Here we show that the C-terminal domain of the large subunit (p58C) plays a major role in template-primer binding and also defines the elements of the DNA template and the RNA primer that interact with p58C. The specific mode of interaction with a template-primer involving the terminal 5'-triphosphate of RNA and the 3'-overhang of DNA results in a stable complex between p58C and the DNA/RNA duplex. Our results explain how p58C participates in RNA synthesis and primer length counting and also indicate that the binding site for initiating NTP is located on p58C. These findings provide notable insight into the mechanism of primase function and are applicable for DNA primases from other species.

Molecular Characterization of Two Lysophospholipid:acyl-CoA Acyltransferases Belonging to the MBOAT Family in Nicotiana benthamiana.

In the remodeling pathway for the synthesis of phosphatidylcholine (PC), acyl-CoA-dependent lysophosphatidylcholine (lysoPC) acyltransferase (LPCAT) catalyzes the reacylation of lysoPC. A number of genes encoding LPCATs have been cloned and characterized from several plants in recent years. Using Arabidopsis and other plant LPCAT sequences to screen the genome database of Nicotiana benthamiana, we identified two cDNAs encoding the putative tobacco LPCATs (NbLPCAT1 and NbLPCAT2). Both of them were predicted to encode a protein of 463 amino acids with high similarity to LPCATs from other plants. Protein sequence features such as the presence of at least eight putative transmembrane regions, four highly conserved signature motifs and several invariant residues indicate that NbLPCATs belong to the membrane bound O-acyltransferase family. Lysophospholipid acyltransferase activity of NbLPCATs was confirmed by testing lyso-platelet-activating factor (lysoPAF) sensitivity through heterologous expression of each full-length cDNA in a yeast mutant Y02431 (lca1△) disrupted in endogenous LPCAT enzyme activity. Analysis of fatty acid profiles of phospholipids from the NbLPCAT-expressing yeast mutant Y02431 cultures supplemented with polyunsaturated fatty acids suggested more incorporation of linoleic acid (18:2n6, LA) and α-linolenic acid (18:3n3, ALA) into PC compared to yeast mutant harbouring empty vector. In vitro enzymatic assay demonstrated that NbLPCAT1had high lysoPC acyltransferase activity with a clear preference for α-linolenoyl-CoA (18:3), while NbLPCAT2 showed a high lysophosphatidic acid (lysoPA) acyltransferase activity towards α-linolenoyl-CoA and a weak lysoPC acyltransferase activity. Tissue-specific expression analysis showed a ubiquitous expression of NbLPCAT1 and NbLPCAT2 in roots, stems, leaves, flowers and seeds, and a strong expression in developing flowers. This is the first report on the cloning and characterization of lysophospholipid acyltransferases from N. benthamiana.

Components of a Rice-Oilseed Rape Production System Augmented with Trichoderma sp. Tri-1 Control Sclerotinia sclerotiorum on Oilseed Rape.

Sclerotinia sclerotiorum causes serious yield losses on many crops throughout the world. A multicomponent treatment that consisted of the residual rice straw remaining after rice harvest and Trichoderma sp. Tri-1 (Tri-1) formulated with the oilseed rape seedcake fertilizer was used in field soil infested with S. sclerotiorum. This treatment resulted in oilseed rape seed yield that was significantly greater than the nontreated control or when the fungicide carbendizem was used in the presence of this pathogen in field trials. Yield data suggested that the rice straw, oilseed rape seedcake, and Tri-1 components of this treatment all contributed incrementally. Similar treatment results were obtained regarding reduction in disease incidence. Slight improvements in yield and disease incidence were detected when this multicomponent treatment was combined with a fungicide spray. Inhibition of sclerotial germination by this multicomponent treatment trended greater than the nontreated control at 90, 120, and 150 days in field studies but was not significantly different from this control. This multicomponent treatment resulted in increased yield relative to the nontreated control in the absence of pathogen in a greenhouse pot study, while the straw alone and the straw plus oilseed rape seedcake treatments did not; suggesting that Tri-1 was capable of promoting growth. Experiments reported here indicate that a treatment containing components of a rice-oilseed rape production system augmented with Tri-1 can control S. sclerotiorum on oilseed rape, be used in integrated strategies containing fungicide sprays for control of this pathogen, and promote plant growth.

Crystal structure of the human primase.

DNA replication in bacteria and eukaryotes requires the activity of DNA primase, a DNA-dependent RNA polymerase that lays short RNA primers for DNA polymerases. Eukaryotic and archaeal primases are heterodimers consisting of small catalytic and large accessory subunits, both of which are necessary for RNA primer synthesis. Understanding of RNA synthesis priming in eukaryotes is currently limited due to the lack of crystal structures of the full-length primase and its complexes with substrates in initiation and elongation states. Here we report the crystal structure of the full-length human primase, revealing the precise overall organization of the enzyme, the relative positions of its functional domains, and the mode of its interaction with modeled DNA and RNA. The structure indicates that the dramatic conformational changes in primase are necessary to accomplish the initiation and then elongation of RNA synthesis. The presence of a long linker between the N- and C-terminal domains of p58 provides the structural basis for the bulk of enzyme's conformational flexibility. Deletion of most of this linker affected the initiation and elongation steps of the primer synthesis.

Column test-based optimization of the permeable reactive barrier (PRB) technique for remediating groundwater contaminated by landfill leachates.

We investigated the optimum composition of permeable reactive barrier (PRB) materials for remediating groundwater heavily contaminated by landfill leachate, in column tests using various mixtures of zero-valent iron (ZVI), zeolite (Zeo) and activated carbon (AC) with 0.01-0.25, 3.0-5.0 and 0.7-1.0mm grain sizes, respectively. The main contributors to the removal of organic/inorganic contaminants were ZVI and AC, and the optimum weight ratio of the three PRB materials for removing the contaminants and maintaining adequate hydraulic conductivity was found to be 5:1:4. Average reductions in chemical oxygen demand (COD) and contents of total nitrogen (TN), ammonium, Ni, Pb and 16 polycyclic aromatic hydrocarbons (PAHs) from test samples using this mixture were 55.8%, 70.8%, 89.2%, 70.7%, 92.7% and 94.2%, respectively. We also developed a systematic method for estimating the minimum required thickness and longevity of the PRB materials. A ≥ 309.6 cm layer with the optimum composition is needed for satisfactory longevity, defined here as meeting the Grade III criteria (the Chinese National Bureau of Standards: GB/T14848/93) for in situ treatment of the sampled groundwater for ≥ 10 years.