Molecular dissection of an intronic enhancer governing cold-induced expression of the vacuolar invertase gene in potato.

Potato (Solanum tuberosum) is the third most important food crop in the world. Potato tubers must be stored at cold temperatures to minimize sprouting and losses due to disease. However, cold temperatures strongly induce the expression of the potato vacuolar invertase gene (VInv) and cause reducing sugar accumulation. This process, referred to as "cold-induced sweetening," is a major postharvest problem for the potato industry. We discovered that the cold-induced expression of VInv is controlled by a 200 bp enhancer, VInvIn2En, located in its second intron. We identified several DNA motifs in VInvIn2En that bind transcription factors involved in the plant cold stress response. Mutation of these DNA motifs abolished VInvIn2En function as a transcriptional enhancer. We developed VInvIn2En deletion lines in both diploid and tetraploid potato using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9)-mediated gene editing. VInv transcription in cold-stored tubers was significantly reduced in the deletion lines. Interestingly, the VInvIn2En sequence is highly conserved among distantly related Solanum species, including tomato (Solanum lycopersicum) and other non-tuber-bearing species. We conclude that the VInv gene and the VInvIn2En enhancer have adopted distinct roles in the cold stress response in tubers of tuber-bearing Solanum species.

Global Supply Chain Drivers of Agricultural Antibiotic Emissions in China.

Antibiotic pollution causes serious environmental and social issues. China is the largest antibiotic producer and user in the world, with a large share of antibiotics used in agriculture. This study quantified agricultural antibiotic emissions of mainland China in 2014 as well as critical drivers in global supply chains. Results show that China's agriculture discharged 4131 tons of antibiotics. Critical domestic supply chain drivers are mainly located in Central China, North China, and East China. Foreign final demand contributes 9% of agricultural antibiotic emissions in mainland China and leads to 5-40% of emissions in each province. Foreign primary inputs (e.g., labor and capital) contribute 5% of agricultural antibiotic emissions in mainland China and lead to 2-63% of emissions in each province. Critical international drivers include the final demand of the United States and Japan for foods and textile products, as well as the primary inputs of the oil seeds sector in Brazil. The results indicate the uniqueness of supply chain drivers for antibiotic emissions compared with other emissions. Our findings reveal supply chain hotspots for multiple-perspective policy decisions to control China's agricultural antibiotic emissions as well as for international cooperation.

Etiolated Stem Branching Is a Result of Systemic Signaling Associated with Sucrose Level.

The potato (Solanum tuberosum) tuber is a swollen stem. Sprouts growing from the tuber nodes represent loss of apical dominance and branching. Long cold storage induces loss of tuber apical dominance and results in secondary branching. Here, we show that a similar branching pattern can be induced by short heat treatment of the tubers. Detached sprouts were induced to branch by the heat treatment only when attached to a parenchyma cylinder. Grafting experiments showed that the scion branches only when grafted onto heat- or cold-treated tuber parenchyma, suggesting that the branching signal is transmitted systemically from the bud-base parenchyma to the grafted stem. Exogenous supply of sucrose (Suc), glucose, or fructose solution to detached sprouts induced branching in a dose-responsive manner, and an increase in Suc level was observed in tuber parenchyma upon branching induction, suggesting a role for elevated parenchyma sugars in the regulation of branching. However, sugar analysis of the apex and node after grafting showed no distinct differences in sugar levels between branching and nonbranching stems. Vacuolar invertase is a key enzyme in determining the level of Suc and its cleavage products, glucose and fructose, in potato parenchyma. Silencing of the vacuolar invertase-encoding gene led to increased tuber branching in combination with branching-inducing treatments. These results suggest that Suc in the parenchyma induces branching through signaling and not by excess mobilization from the parenchyma to the stem.

Long-term effects of multi-walled carbon nanotubes on the performance and microbial community structures of an anaerobic granular sludge system.

Multi-walled carbon nanotubes (MWCNTs) released into the sewage may cause negative and/or positive effects on the treatment system. The objective of this study was to explore over 110 days' effect of MWCNTs on the performance of anaerobic granular sludge and microbial community structures in an upflow anaerobic sludge blanket (UASB) reactor. The results showed that MWCNTs had no significant effect on the removal of chemical oxidation demand (COD) and ammonia in UASB reactor, but the total phosphorus (TP) removal efficiency increased by 29.34%. The biogas production of the reactor did not change. The anaerobic granular sludge tended to excrete more EPS to resist the effects of MWCNTs during the long-term impact. Illumina MiSeq sequencing of 16S rRNA gene revealed that MWCNTs did not affect the microbial diversity, but altered the composition and structure of microbial community in the reactor. In this process, Saccharibacteria replaced Proteobacteria as the highest abundant bacterial phylum. MWCNTs promoted the differentiation of methanogen structure, resulting in increase of Methanomassiliicoccus, Methanoculleus, and the uncultured WCHA1-57. These results indicated that MWCNTs impacted the performance of UASB reactor and the structures of the microbial community in anaerobic granular sludge.

Revealing the anaerobic acclimation of microbial community in a membrane bioreactor for coking wastewater treatment by Illumina Miseq sequencing.

The dynamic change of microbial community during sludge acclimation from aerobic to anaerobic in a MBR for coking wastewater treatment was revealed by Illumina Miseq sequencing in this study. The diversity of both Bacteria and Archaea showed an increase-decrease trajectory during acclimation, and exhibited the highest at the domestication interim. Ignavibacteria changed from a tiny minority (less than 1%) to the dominant bacterial group (54.0%) along with acclimation. The relative abundance of Betaproteobacteria kept relatively steady, as in this class some species increased coupled with some other species decreased during acclimation. The dominant Archaea shifted from Halobacteria in initial aerobic sludge to Methanobacteria in the acclimated anaerobic sludge. The dominant bacterial and archaeal groups in different acclimation stages were indigenous microorganisms in the initial sludge, though some of them were very rare. This study supported that the species in "rare biosphere" might eventually become dominant in response to environmental change.

Responses of Bacterial Communities to CuO Nanoparticles in Activated Sludge System.

The main objectives of this study were to investigate the influence of copper oxide nanoparticles (CuO NPs) on wastewater nutrient removal, bacterial community and molecular ecological network in activated sludge. The results showed that long-term exposure to 1 mg/L CuO NPs induced an increase of effluent concentrations of ammonia and total phosphorus, which was consistent with the inhibition of enzyme activities of ammonia monooxygenase, nitrite oxidoreductase, exopolyphosphatase, and polyphosphate in the presence of CuO NPs. MiSeq sequencing data indicated that CuO NPs significantly decreased the bacterial diversity and altered the overall bacterial community structure in activated sludge. Some genera involved in nitrogen and phosphorus removal, such as Nitrosomonas, Acinetobacter, and Pseudomonas decreased significantly. Molecular ecological network analysis showed that network interactions among different phylogenetic populations were markedly changed by CuO NPs. For example, ß-Proteobacteria, playing an important role in nutrients removal, had less complex interactions in the presence of CuO NPs. These shifts of the abundance of related genera, together with the network interactions may be associated with the deterioration of ammonia and phosphorus removal. This study provides insights into our understanding of shifts in the bacteria community and their molecular ecological network under CuO NPs in activated sludge systems.

Performance and microbial community of a membrane bioreactor system - Treating wastewater from ethanol fermentation of food waste.

In this study, a lab-scale biological anaerobic/anaerobic/anoxic/membrane bioreactor (A3-MBR) was designed to treat wastewater from the ethanol fermentation of food waste, a promising way for the disposal of food waste and reclamation of resources. The 454 pyrosequencing technique was used to investigate the composition of the microbial community in the treatment system. The system yielded a stable effluent concentration of chemical oxygen demand (202±23mg/L), total nitrogen (62.1±7.1mg/L), ammonia (0.3±0.13mg/L) and total phosphorus (8.3±0.9mg/L), and the reactors played different roles in specific pollutant removal. The exploration of the microbial community in the system revealed that: (1) the microbial diversity of anaerobic reactors A1 and A2, in which organic pollutants were massively degraded, was much higher than that in anoxic A3 and aerobic MBR; (2) although the community composition in each reactor was quite different, bacteria assigned to the classes Clostridia, Bacteroidia, and Synergistia were important and common microorganisms for organic pollutant degradation in the anaerobic units, and bacteria from Alphaproteobacteria and Betaproteobacteria were the dominant microbial population in A3 and MBR; (3) the taxon identification indicated that Arcobacter in the anaerobic reactors and Thauera in the anoxic reactor were two representative genera in the biological process. Our results proved that the biological A3-MBR process is an alternative technique for treating wastewater from food waste.

Silencing of vacuolar invertase and asparagine synthetase genes and its impact on acrylamide formation of fried potato products.

Acrylamide is produced in a wide variety of carbohydrate-rich foods during high-temperature cooking. Dietary acrylamide is a suspected human carcinogen, and health concerns related to dietary acrylamide have been raised worldwide. French fries and potato chips contribute a significant proportion to the average daily intake of acrylamide, especially in developed countries. One way to mitigate health concerns related to acrylamide is to develop potato cultivars that have reduced contents of the acrylamide precursors asparagine, glucose and fructose in tubers. We generated a large number of silencing lines of potato cultivar Russet Burbank by targeting the vacuolar invertase gene VInv and the asparagine synthetase genes StAS1 and StAS2 with a single RNA interference construct. The transcription levels of these three genes were correlated with reducing sugar (glucose and fructose) and asparagine content in tubers. Fried potato products from the best VInv/StAS1/StAS2-triple silencing lines contained only one-fifteenth of the acrylamide content of the controls. Interestingly, the extent of acrylamide reduction of the best triple silencing lines was similar to that of the best VInv-single silencing lines developed previously from the same potato cultivar Russet Burbank. These results show that an acrylamide mitigation strategy focused on developing potato cultivars with low reducing sugars is likely to be an effective and sufficient approach for minimizing the acrylamide-forming potential of French fry processing potatoes.

Bioaugmentation with isolated strains for the removal of toxic and refractory organics from coking wastewater in a membrane bioreactor.

The bioaugmentation strains for phenol, pyridine, quinoline, carbazole, and naphthalene degradation were employed to treat coking wastewater in a membrane bioreactor (MBR). The results showed that the bioaugmented MBR was much better in pollutant removal than that of the control MBR with conventional activated sludge. Compared to the control MBR, the bioaugmented MBR displayed an additional 3.2 mg/L of phenol, pyridine, quinoline, naphthalene and carbazole in total by the addition of the degrading strains. Also, about 10 % of the chemical oxygen demand in the effluent was further removed by the bioaugmentation. The pyrosequencing analysis of the sludge in the MBRs revealed that the microbial community shifted in response to the addition of the degrading strains. The diversity of the microbial community increased during the bioaugmentation, and some bacterial taxa favorable to the removal of toxic and refractory pollutants appeared in the bioaugmented MBR. The results indicated that the use of high-efficiency bacteria was a feasible method for industrial coking wastewater treatment.

Drought priming at seedling stage improves photosynthetic performance and yield of potato exposed to a short-term drought stress.

Potato (Solanum tuberosum L.) is an important food and vegetable crop worldwide. In recent years, the arid environment resulting from climate change has caused a sharp decline in potato yield. To clarify the effect of drought priming at the seedling stage on the tolerance of potato plants to drought stress during tuber expansion, we conducted a pot experiment to investigate the physiological response of the plants generated from seed potatoes of the variety 'Favorita' to varied water supply conditions: normal water supply at the seedling stage (control), normal water supply at the seedling stage and drought stress at the mid-tuber-expansion stage (non-primed), and drought priming at the seedling stage plus drought stress at the mid-tuber-expansion stage (primed). Drought priming resulted in an increase in the number of small vascular bundles in potato plants compared to non-primed plants. It also altered the shape and density of stomata, enhancing water use efficiency and reducing whole-plant transpiration. The primed plants maintained the basal stem cambium for a longer time under drought stress, which gained an extended differentiation ability to generate a greater number of small vascular bundles compared to non-primed plants. Drought priming increased the amount and rate of dry matter translocation, and so reduced the adverse effects on tubers of potato under drought stress. Therefore, drought priming at the seedling stage improved the photosynthetic performance and yield, and probably enhanced the drought tolerance of potato.

Composition and dynamics of microbial community in a zeolite biofilter-membrane bioreactor treating coking wastewater.

In this study, a lab-scale anaerobic/anoxic/zeolite biofilter-membrane bioreactor (A1/A2/ZB-MBR) was designed to treat coking wastewater. The 454 pyrosequencing was used to obtain the composition and dynamics of microbial community about the treatment system. The results showed that the system yielded stable effluent chemical oxidation demand (158.5 ± 21.8 mg/L) and ammonia (8.56 ± 7.30 mg/L), but fluctuant total nitrogen (31.4-165.1 mg/L) concentrations. In addition, 66,256 16S rRNA gene sequences were obtained from A2 and ZB-MBR, and the microbial diversity and richness for five samples were determined. Although community compositions in the five samples were quite different, bacteria assigned to phylum Proteobacteria and class Flavobacteria commonly existed and dominated the microbial populations. The pyrosequencing analysis revealed that the microbial community shifted in the ZB-MBR with the presence of zeolite. Some taxa began to appear in ZB-MBR and contributed to the system performance. Additionally, Nitrosomonas and Nitrobacter gradually became the dominant ammonia-oxidizing bacteria and nitrite-oxidizing bacteria during the operation, respectively, which are favorable for the stabilized ammonia removal. Our results proved that the ZB-MBR is an alternative technique for treating coking wastewater.

Enhanced gaseous acetone adsorption on montmorillonite by ball milling generated Si-OH and interlayer under synergistic modification with H2O2 and tetramethylammonium bromide.

Montmorillonite (Mt) is a potential adsorbent for volatile organic vapor removal from contaminated soils because of its rich reserves and porous nature, but its inertia surface property has limited its application for polar compounds. In this study, modifications of Mt were carried out by high energy ball milling with H2O2 and tetramethylammonium bromide (TMAB) to obtain adsorbents with both high porosity and abundant Si-OH groups (BHTMt). The microporous structure produced by TMAB insertion as well as the silanol (Si-OH) groups formed by H2O2 oxidation improved the adsorption of acetone by the modified material. The adsorption capacity of BHTMt for acetone was increased by 80% compared to the original Mt. The effect of H2O2 dosage on the adsorption performance for gaseous acetone was investigated by dynamic adsorption experiments. The adsorption kinetic results demonstrated that the adsorption of acetone by the modified material was subject to both physical and chemical adsorption. Density functional theory calculations indicated that there was no obvious interaction between TMAB and acetone, and the materials adsorbed acetone mainly through hydrogen bonding interaction of Si-OH as well as pore filling effects.

Sodium hydroxide or tetramethylammonium hydroxide modified corncob combined with biodegradable polymers to prepare slow-release carbon source for wastewater denitrification.

This study proposed a method to improve the bioavailability of artificially prepared carbon sources for the purpose of wastewater denitrification. This carbon source (named SPC) was prepared by mixing corncobs with poly(3-hydroxybutyrate-3-hydroxyvalerate) (PHBV), where the corncobs were pretreated by NaOH or TMAOH. The results of compositional analysis and FTIR showed that both NaOH and TMAOH degraded lignin, hemicellulose and their connection bonds in corncob, thus increased the cellulose content from 39% to 53% and 55%, respectively. The cumulative carbon release from SPC was about 9.3 mg/g and was consistent with both the first-order kinetic and Ritger-Peppas equation. The released organic matters contained low concentration of refractory components. Correspondingly, it showed excellent denitrification performance in simulated wastewater, and the total nitrogen (TN) removal rate was above 95% (influent NO3--N was 40 mg/L) and effluent residual chemical oxygen demand (COD) was less than 50 mg/L.

pH adjustable MgAl@LDH-coated MOFs-derived Co2.25Mn0.75O4 for SMX degradation in PMS activated system.

Sulfate radical-based advanced oxidation processes (SR-AOPs) is considered as one of the most promising technologies for antibiotic pollution. In this study, a core-shell catalyst of cobalt-manganese oxide derived from CoMn-MOFs coating by MgAl-LDH (Co/Mn@LDH) was synthesized for peroxymonosulfate (PMS) activation to degrade sulfamethoxazole (SMX). Degradation efficiency of nearly 100% and a mineralization efficiency of 68.3% for SMX were achieved in Co/Mn@LDH/PMS system. Mn species and out shell MgAl-LDH greatly suppressed the cobalt ions leaching, which only 23 µg/L Co ions were detected by ICP after the reaction. SO4·- was identified as dominant reactive species in the system. Furthermore, the possible reactive sites of SMX were predicted by the density functional theory (DFT) calculations. And the intermediates of SMX were detected by LC-MS and the degradation pathway was proposed based on the results above. The ECOSAR results suggested the intermediates of SMX showed a relatively low toxicity compared to SMX, indicating huge potential of utilization of Co/Mn@LDH in SR-AOPs system.

Biodegradation mechanism of chlortetracycline by a novel fungal Aspergillus sp. LS-1.

Chlortetracycline (CTC), a widely used typical tetracycline antibiotic, has raised increasing concerns due to its potential health and environmental risks. Biodegradation is considered an effective method to reduce CTC in environment. In this study, a strain Aspergillus sp. LS-1, which can efficiently degrade CTC, was isolated from CTC-rich activated sludge. Under optimal conditions, the maximum removal efficiency of CTC could reach 95.41%. Temperature was the most significant factor affecting the degradation efficiency of LS-1. The 19 products were identified in the CTC degradation by strain LS-1, and three degradation pathways were proposed. All the degradation pathways for CTC exhibited ring-cleaving, which may accelerate the mineralization of CTC. To gain more comprehensive insights into this strain, we obtained the genome of LS-1, which had high GC content (50.1%) and completeness (99.3%). The gene annotation revealed that LS-1 contains some vital enzymes and resistance genes that may carry functional genes involved in the CTC degradation. In addition, other antibiotic resistance genes were found in the genome of LS-1, indicating that LS-1 has the potential to degrade other antibiotics. This study provides a more theoretical basis for the investigation of CTC degradation by fungi and new insights into the biodegradation of CTC.

Aging behavior of biodegradable polylactic acid microplastics accelerated by UV/H2O2 processes.

The usage of biodegradable plastics is expanding annually due to worldwide plastic limits, resulting in a substantial number of microplastics (MPs) particles formed from biodegradable plastic products entering the aquatic environment. Until now, the environmental behaviors of these plastic product-derived MPs (PPDMPs) have remained unclear. In this work, commercially available polylactic acid (PLA) straws and PLA food bags were used to evaluate the dynamic aging process and environmental behavior of PLA PPDMPs under UV/H2O2 conditions. By combining scanning electron microscopy, two-dimensional (2D) Fourier transform infrared correlation spectroscopy (COS) and X-ray photoelectron spectroscopy, it was determined that the aging process of the PLA PPDMPs was slower than that of pure MPs. The 2D-COS analysis revealed that the response orders for the functional groups on the PLA MPs differed during the aging process. The results demonstrated that the oxygen-containing functional groups of the PLA PPDMPs were the first to react. Subsequently, the -C-H and -C-C- structural responses began, and the polymer backbone was ruptured by the aging process. However, the aging of the pure-PLA MPs started with a brief oxidation process and then breakage of the polymer backbones, followed by continuous oxidation. Moreover, compared to the PLA PPDMPs, the pure-PLA MPs exhibited a greater adsorption capacity, which was increased by 88% after aging, whereas those of the two PPDMPs only increased by 64% and 56%, respectively. This work provides new insights into the behaviors of biodegradable PLA MPs in aquatic environments, which is critical for assessing the environmental risks and management policies for degradable MPs.

Systematic Analysis of Galactinol Synthase and Raffinose Synthase Gene Families in Potato and Their Expression Patterns in Development and Abiotic Stress Responses.

Raffinose family oligosaccharides (RFOs) are very important for plant growth, development, and abiotic stress tolerance. Galactinol synthase (GolS) and raffinose synthase (RFS) are critical enzymes involved in RFO biosynthesis. However, the whole-genome identification and stress responses of their coding genes in potato remain unexplored. In this study, four StGolS and nine StRFS genes were identified and classified into three and five subgroups, respectively. Remarkably, a total of two StGolS and four StRFS genes in potato were identified to form collinear pairs with those in both Arabidopsis and tomato, respectively. Subsequent analysis revealed that StGolS4 exhibited significantly high expression levels in transport-related tissues, PEG-6000, and ABA treatments, with remarkable upregulation under salt stress. Additionally, StRFS5 showed similar responses to StGolS4, but StRFS4 and StRFS8 gene expression increased significantly under salt treatment and decreased in PEG-6000 and ABA treatments. Overall, these results lay a foundation for further research on the functional characteristics and molecular mechanisms of these two gene families in response to ABA, salt, and drought stresses, and provide a theoretical foundation and new gene resources for the abiotic-stress-tolerant breeding of potato.

Mapping and validation of quantitative trait loci associated with wheat yellow mosaic bymovirus resistance in bread wheat.

Wheat yellow mosaic (WYM) caused by wheat yellow mosaic bymovirus (WYMV) has been growing as one of the most serious diseases affecting wheat production in China. In this study, the association of quantitative trait loci (QTLs) governing WYMV resistance with molecular markers was established using 164 recombinant inbred lines (RILs) derived from 'Xifeng Wheat' (highly resistant) × 'Zhen 9523' (highly susceptible). Phenotypic data of WYMV resistance of the RILs were collected from 4-year, two-location replicated field trials. A molecular marker-based linkage map, which was comprised of 273 non-redundant loci and represented all the 21 wheat chromosomes, was constructed with the JoinMap 4.0 software. Using the Windows QTL Cartographer V2.5 software, three QTLs associated with WYMV resistance, QYm.njau-3B.1, QYm.njau-5A.1 and QYm.njau-7B.1, were detected on chromosomes 3BS, 5AL, and 7BS, respectively. The favorable allele effects were all contributed by 'Xifeng Wheat'. Among the three QTLs, QYm.njau-3B.1 and QYm.njau-5A.1 were detected in all the four trials and the overall mean, and could explain 3.3-10.2% and 25.9-53.7% of the phenotypic variation, respectively, while QYm.njau-7B.1 was detected in one trial and the overall mean and explained 4.9 and 3.3% of the phenotypic variation, respectively. A large portion of the variability for WYMV response was explained by a major QTL, QYm.njau-5A.1. The relationship of the molecular markers linked with QYm.njau-5A.1 and the WYMV resistance was further validated using a secondary F(2) population. The results showed that three markers, i.e., Xwmc415.1, CINAU152, and CINAU153, were closely linked to QYm.njau-5A.1 with the genetic distances of 0.0, 0.0, and 0.1 cM, respectively, indicating they should be useful in marker-assisted selection (MAS) wheat breeding for WYMV resistance. A panel of germplasm collection consisting of 46 wheat varieties with known WYMV response phenotypes was further used to validate the presence and effects of QYm.njau-5A.1 and the above three markers. It was found that QYm.njau-5A.1 was present in 12 of the 34 WYMV-resistant varieties.

Fe-Cu binary oxide loaded zeolite as heterogeneous Fenton catalyst for degradation of carbamazepine at near-neutral pH.

In this study, Fe-Cu binary oxide was loaded on zeolite (Fe/Cu/zeolite) to be used as heterogeneous Fenton catalyst, and the catalytic degradation of carbamazepine (CBZ) were optimized at near-neutral pH. The results showed that the Fe and Cu oxide, mainly Fe2O3, Fe3O4, and CuO nanoparticles, were uniformly distributed on the surface of zeolite particles. Under the optimized conditions, Fe/Cu/zeolite could completely degrade CBZ when initial pH ranged from 3 to 7, and the removal efficiency of CBZ still remained above 74% even though the initial pH increased to near 10. After 8 times' repeated use, the Fe/Cu/zeolite exhibited an over 95% removal efficiency of CBZ. The hydroxyl radicals (•OH) were verified to be the main active oxidants by quenching experiments and ESR testing. The XPS of the materials revealed that the high catalytic efficiency was attributed to the synergistic effect of Fe(III)/Fe(II) and Cu(II)/Cu(I) redox cycles. This catalyst can be used for the efficient degradation of organic pollutants in heterogeneous Fenton systems.

Mechanistic insights into ball milling enhanced montmorillonite modification with tetramethylammonium for adsorption of gaseous toluene.

Montmorillonite is widely used for pollutants adsorption due to its porous structure and low price. However, the low specific surface area and small porosity limit its application in gas adsorption field. In this study, montmorillonite was organically modified using a facile dry ball milling method by tetramethylammonium bromide. The adsorption behaviour of toluene as a model VOC compound on organic montmorillonite was systematically investigated through adsorption breakthrough curves, adsorption kinetics and isotherms. After modification by ball milling, the specific surface area of ball milling with tetramethylammonium bromide for montmorillonite modification (BMTMt) was increased from 20.6 m2/g to 186.4 m2/g, and the microporosity proportion was up to 47%. Dynamic adsorption experiments showed that the best performance of BMTMt for toluene (55.9 mg/g) was 6 times higher than that of original montmorillonite (8.8 mg/g). Compared with the water bath preparation method, ball milling method promoted the intercalation of tetramethylammonium bromide into the layers of montmorillonite, resulting in a higher proportion of micropores. Density functional theory calculations indicated that the interaction between tetramethylammonium bromide and montmorillonite was mainly electrostatic forces, and the enhanced adsorption performance for toluene was mainly through microporous filling. BMTMt was proved to be a promising adsorbent for VOCs removal.