Bacteria assisted green synthesis of copper oxide nanoparticles and their potential applications as antimicrobial agents and plant growth stimulants.

Copper oxide nanoparticles (CuO-NPs) have piqued the interest of agricultural researchers due to their potential application as fungicides, insecticides, and fertilizers. The Serratia sp. ZTB29 strain, which has the NCBI accession number MK773873, was a novel isolate used in this investigation that produced CuO-NPs. This strain can survive concentrations of copper as high as 22.5 mM and can also remove copper by synthesizing pure CuO-NPs. UV-VIS spectroscopy, DLS, Zeta potential, FTIR, TEM, and XRD techniques were used to investigate the pure form of CuO-NPs. The synthesized CuO-NPs were crystalline in nature (average size of 22 nm) with a monoclinic phase according to the XRD pattern. CuO-NPs were found to be polydisperse, spherical, and agglomeration-free. According to TEM and DLS inspection, they ranged in size from 20 to 40 nm, with a typical particle size of 28 nm. CuO-NPs were extremely stable, as demonstrated by their zeta potential of -15.4 mV. The ester (C=O), carboxyl (C=O), amine (NH), thiol (S-H), hydroxyl (OH), alkyne (C-H), and aromatic amine (C-N) groups from bacterial secretion were primarily responsible for reduction and stabilization of CuO-NPs revealed in an FTIR analysis. CuO-NPs at concentrations of 50 µg mL-1 and 200 µg mL-1 displayed antibacterial and antifungal activity against the plant pathogenic bacteria Xanthomonas sp. and pathogenic fungus Alternaria sp., respectively. The results of this investigation support the claims that CuO-NPs can be used as an efficient antimicrobial agent and nano-fertilizer, since, compared to the control and higher concentrations of CuO-NPs (100 mg L-1) considerably improved the growth characteristics of maize plants.

Bioprospecting of novel ligninolytic bacteria for effective bioremediation of agricultural by-product and synthetic pollutant dyes.

Lignin is a significant renewable carbon source that needs to be exploited to manufacture bio-ethanol and chemical feedstocks. Lignin mimicking methylene blue (MB) dye is widely used in industries and causes water pollution. Using kraft lignin, methylene blue, and guaiacol as a full carbon source, 27 lignin-degrading bacteria (LDB) were isolated from 12 distinct traditional organic manures for the current investigation. The ligninolytic potential of 27 lignin-degrading bacteria was assessed by qualitative and quantitative assay. In a qualitative plate assay, the LDB-25 strain produced the largest zone, measuring 6.32 ± 0.297, on MSM-L-kraft lignin plates, while the LDB-23 strain produced the largest zone, measuring 3.44 ± 0.413, on MSM-L-Guaiacol plates. The LDB-9 strain in MSM-L-kraft lignin broth was able to decolorize lignin to a maximum of 38.327 ± 0.011% in a quantitative lignin degradation assay, which was later verified by FTIR assay. In contrast, LDB-20 produced the highest decolorization (49.633 ± 0.017%) in the MSM-L-Methylene blue broth. The highest manganese peroxidase enzyme activity, measuring 6322.314 ± 0.034 U L-1, was found in the LDB-25 strain, while the highest laccase enzyme activity, measuring 1.5105 ± 0.017 U L-1, was found in the LDB-23 strain. A preliminary examination into the biodegradation of rice straw using effective LDB was carried out, and efficient lignin-degrading bacteria were identified using 16SrDNA sequencing. SEM investigations also supported lignin degradation. LDB-8 strain had the highest percentage of lignin degradation (52.86%), followed by LDB-25, LDB-20, and LDB-9. These lignin-degrading bacteria have the ability to significantly reduce lignin and lignin-analog environmental contaminants, therefore they can be further researched for effective bio-waste management mediated breakdown.

Methylotroph bacteria and cellular metabolite carotenoid alleviate ultraviolet radiation-driven abiotic stress in plants.

Increasing UV radiation in the atmosphere due to the depletion of ozone layer is emerging abiotic stress for agriculture. Although plants have evolved to adapt to UV radiation through different mechanisms, but the role of phyllosphere microorganisms in counteracting UV radiation is not well studied. The current experiment was undertaken to evaluate the role of phyllosphere Methylobacteria and its metabolite in the alleviation of abiotic stress rendered by ultraviolet (UV) radiation. A potential pink pigmenting methylotroph bacterium was isolated from the phylloplane of the rice plant (oryzae sativa). The 16S rRNA gene sequence of the bacterium was homologous to the Methylobacter sp. The isolate referred to as Methylobacter sp N39, produced beta-carotene at a rate (µg ml-1 d-1) of 0.45-3.09. Biosynthesis of beta-carotene was stimulated by brief exposure to UV for 10 min per 2 days. Carotenoid biosynthesis was predicted as y = 3.09 × incubation period + 22.151 (r 2 = 0.90). The carotenoid extract of N39 protected E. coli from UV radiation by declining its death rate from 14.67% min-1 to 4.30% min-1 under UV radiation. Application of N39 cells and carotenoid extract also protected rhizobium (Bradyrhizobium japonicum) cells from UV radiation. Scanning electron microscopy indicated that the carotenoid extracts protected E. coli cells from UV radiation. Foliar application of either N39 cells or carotenoid extract enhanced the plant's (Pigeon pea) resistance to UV irradiation. This study highlight that Methylobacter sp N39 and its carotenoid extract can be explored to manage UV radiation stress in agriculture.

Boosting Sustainable Agriculture by Arbuscular Mycorrhiza under Stress Condition: Mechanism and Future Prospective.

Global agriculture is frequently subjected to stresses from increased salt content, drought, heavy metals, and other factors, which limit plant growth and production, deteriorate soil health, and constitute a severe danger to global food security. Development of environmentally acceptable mitigation techniques against stresses and restrictions on the use of chemical fertilizers in agricultural fields is essential. Therefore, eco-friendly practises must be kept to prevent the detrimental impacts of stress on agricultural regions. The advanced metabolic machinery needed to handle this issue is not now existent in plants to deal against the stresses. Research has shown that the key role and mechanisms of arbuscular mycorrhiza fungi (AMF) to enhance plant nutrient uptake, immobilisation and translocation of heavy metals, and plant growth-promoting attributes may be suitable agents for plant growth under diversed stressed condition. The successful symbiosis and the functional relationship between the plant and AMF may build the protective regulatory mechansm against the key challenge in particular stress. AMF's compatibility with hyperaccumulator plants has also been supported by studies on gene regulation and theoretical arguments. In order to address this account, the present review included reducing the impacts of biotic and abiotic stress through AMF, the mechanisms of AMF to improve the host plant's capacity to endure stress, and the strategies employed by AM fungus to support plant survival in stressful conditions.

Genomic Diversity of Pigeon Pea (Cajanus cajan L. Millsp.) Endosymbionts in India and Selection of Potential Strains for Use as Agricultural Inoculants.

Pigeon pea (Cajanus cajan L. Millsp. ) is a legume crop resilient to climate change due to its tolerance to drought. It is grown by millions of resource-poor farmers in semiarid and tropical subregions of Asia and Africa and is a major contributor to their nutritional food security. Pigeon pea is the sixth most important legume in the world, with India contributing more than 70% of the total production and harbouring a wide variety of cultivars. Nevertheless, the low yield of pigeon pea grown under dry land conditions and its yield instability need to be improved. This may be done by enhancing crop nodulation and, hence, biological nitrogen fixation (BNF) by supplying effective symbiotic rhizobia through the application of "elite" inoculants. Therefore, the main aim in this study was the isolation and genomic analysis of effective rhizobial strains potentially adapted to drought conditions. Accordingly, pigeon pea endosymbionts were isolated from different soil types in Southern, Central, and Northern India. After functional characterisation of the isolated strains in terms of their ability to nodulate and promote the growth of pigeon pea, 19 were selected for full genome sequencing, along with eight commercial inoculant strains obtained from the ICRISAT culture collection. The phylogenomic analysis [Average nucleotide identity MUMmer (ANIm)] revealed that the pigeon pea endosymbionts were members of the genera Bradyrhizobium and Ensifer. Based on nodC phylogeny and nod cluster synteny, Bradyrhizobium yuanmingense was revealed as the most common endosymbiont, harbouring nod genes similar to those of Bradyrhizobium cajani and Bradyrhizobium zhanjiangense. This symbiont type (e.g., strain BRP05 from Madhya Pradesh) also outperformed all other strains tested on pigeon pea, with the notable exception of an Ensifer alkalisoli strain from North India (NBAIM29). The results provide the basis for the development of pigeon pea inoculants to increase the yield of this legume through the use of effective nitrogen-fixing rhizobia, tailored for the different agroclimatic regions of India.

Nitrous oxide production from soybean and maize under the influence of weedicides and zero tillage conservation agriculture.

Current experiment envisages evaluating N2O production from nitrification and denitrification under the influence of weedicides, cropping systems and conservation agriculture (CA). The weed control treatments were conventional hand weeding (no weedicide), pre emergence weedicide pendimethalin and post emergence weedicide imazethapyr for soybean, atrazine for maize. Experiment was laid out in randomized block design with three replicates. Soils were collected from different depths and incubated at different moisture holding capacity (MHC). N2O production from nitrification varied from 2.77 to 6.04 ng N2O g-1 soil d-1 and from denitrification varied from 0.05 to 1.34 ng N2O g-1 soil d-1. Potential nitrification rate (0.16-0.39 mM NO3 produced g-1 soil d-1) was higher than potential denitrification rate (0.45-0.93 mM NO3 reduced g-1 soil d-1). N2O production, nitrification, denitrification, and microbial gene abundance were higher in maize than soybean. Both N2O production and nitrification decreased (p < 0.05) with soil depth, while denitrification increased (p < 0.05) with soil depth. Abundance of eubacteria and ammonia oxidizing bacteria (AOB) were high (p < 0.01) at upper soil layer and declined with depth. Abundance of ammonia oxidizing archaea (AOA) increased (p < 0.05) with soil depth. Study concludes that intensive use of weedicides in CA may stimulate N2O production.

Do methanotrophs drive phosphorus mineralization in soil ecosystem?

Experiments were carried out to elucidate linkage between methane consumption and mineralization of phosphorous (P) from different P sources. The treatments were (i) no CH4 + no P amendment (absolute control), (ii) with CH4 + no P amendment (control), (iii) with CH4 + inorganic P as Ca3(PO4)2, and (iv) with CH4 + organic P as sodium phytate. P sources were added at 25 µg P·(g soil)-1. Soils were incubated to undergo three repeated CH4 feeding cycles, referred to as feeding cycle I, feeding cycle II, and feeding cycle III. CH4 consumption rate k (µg CH4 consumed·(g soil)-1·day-1) was 0.297 ± 0.028 in no P amendment control, 0.457 ± 0.016 in Ca3(PO4)2, and 0.627 ± 0.013 in sodium phytate. Rate k was stimulated by 2 to 6 times over CH4 feeding cycles and followed the trend of sodium phytate > Ca3(PO4)2 > no P amendment control. CH4 consumption stimulated P solubilization from Ca3(PO4)2 by a factor of 2.86. Acid phosphatase (µg paranitrophenol released·(g soil)-1·h-1) was higher in sodium phytate than the no P amendment control. Abundance of 16S rRNA and pmoA genes increased with CH4 consumption rates. The results of the study suggested that CH4 consumption drives mineralization of unavailable inorganic and organic P sources in the soil ecosystem.

Microbial Fabrication of Zinc Oxide Nanoparticles and Evaluation of Their Antimicrobial and Photocatalytic Properties.

Zinc oxide (ZnO) nanoparticles have attracted significant interest in a number of applications ranging from electronics to biomedical sciences due to their large exaction binding energy (60 meV) and wide bandgap of 3.37 eV. In the present study, we report the low-cost bacterium based "eco-friendly" efficient synthesis of ZnO nanoparticles by using the zinc-tolerant bacteria Serratia nematodiphila. The physicochemical characterization of ZnO nanoparticles was performed by employing UV-vis spectroscopy, XRD, TEM, DLS, Zeta potential, and Raman spectroscopy. The antimicrobial and antifungal studies were investigated at different concentrations using the agar well-diffusion method, whereby the microbial growth rate decreases with the increase in nanoparticle concentration. Further, photocatalytic performance studies were conducted by taking methyl orange (MO) as a reference dye.

Zinc tolerant plant growth promoting bacteria alleviates phytotoxic effects of zinc on maize through zinc immobilization.

The increasing heavy metal contamination in agricultural soils has become a serious concern across the globe. The present study envisages developing microbial inoculant approach for agriculture in Zn contaminated soils. Potential zinc tolerant bacteria (ZTB) were isolated from zinc (Zn) contaminated soils of southern Rajasthan, India. Isolates were further screened based on their efficiency towards Zn tolerance and plant growth promoting activities. Four strains viz. ZTB15, ZTB24, ZTB28 and ZTB29 exhibited high degree of tolerance to Zn up to 62.5 mM. The Zn accumulation by these bacterial strains was also evidenced by AAS and SEM-EDS studies. Assessment of various plant growth promotion traits viz., IAA, GA3, NH3, HCN, siderophores, ACC deaminase, phytase production and P, K, Si solubilization studies revealed that these ZTB strains may serve as an efficient plant growth promoter under in vitro conditions. Gluconic acid secreted by ZTB strains owing to mineral solubilization was therefore confirmed using high performance liquid chromatography. A pot experiment under Zn stress conditions was performed using maize (Zea mays) variety (FEM-2) as a test crop. Zn toxicity reduced various plant growth parameters; however, inoculation of ZTB strains alleviated the Zn toxicity and enhanced the plant growth parameters. The effects of Zn stress on antioxidant enzyme activities in maize under in vitro conditions were also investigated. An increase in superoxide dismutase, peroxidase, phenylalanine ammonia lyase, catalase and polyphenol oxidase activity was observed on inoculation of ZTB strains. Further, ZIP gene expression studies revealed high expression in the ZIP metal transporter genes which were declined in the ZTB treated maize plantlets. The findings from the present study revealed that ZTB could play an important role in bioremediation in Zn contaminated soils.

Zinc biosorption, biochemical and molecular characterization of plant growth-promoting zinc-tolerant bacteria.

Zinc plays a key role in plant nutrition at low levels; however, at higher concentrations Zn ions can be highly phytotoxic and plant growth-promoting rhizobacteria can be used to reduce such metal toxicity. In the present investigation we had reported the zinc biosorption and molecular characterization of plant growth-promoting zinc-tolerant bacteria. Initially, thirty bacteria having zinc solubilizing ability were screened for MIC against zinc ion and displayed high value of MIC ranging from 2.5 to 62.5 mM. Biochemically, all the 30 isolates showed significant difference in the 6 biochemical tests performed. The molecular diversity studies based on the repetitive DNA PCR viz, REP, ERIC and BOX elements showed significant genetic diversity among these 30 zinc-tolerant bacteria. These ZTB strains also showed multiple PGP activities and all ZTB strains were found positive for production of IAA, GA3 and ammonia, whereas 24 were found positive for ACC deaminase activity, 8 showed siderophore production and 9 ZTB isolates were positive for HCN production. Out of 30 isolates, 24 showed phosphorus solubilization activity, 30 showed potash solubilization, 15 showed silica solubilization and 27 showed phytase production activities. All the 30 ZTB stains showed zinc solubilization up to 0.25% insoluble ZnO in the medium, whereas at 2% ZnO in MSM only 12 isolates showed solubilization which were further selected for zinc biosorption and pot studies. The heavy metal removal studies revealed that ZTB stains were able to remove zinc ions effectively from the medium efficiently and the highest zinc biosorption (< 90%) was recorded with the bacterial strain Z-15. Further, the inoculation of ZTB strains under zinc stress conditions (pot containing 1000 mg/kg Zn) resulted in significant increase of shoot length, root length and total chlorophyll content in maize seedlings compared with the uninoculated control. The partial 16S rDNA sequence of the potential ZTB isolates viz. Z-15, Z-24, Z-28 and Z-29 revealed their identity as Serratia sp. The ability of these zinc-tolerant bacteria to tolerate the toxic level of zinc may serve as suitable candidates for developing microbial formulations for the growth of crop plants in Zn-contaminated areas.

Nitrification Rates Are Affected by Biogenic Nitrate and Volatile Organic Compounds in Agricultural Soils.

The processes regulating nitrification in soils are not entirely understood. Here we provide evidence that nitrification rates in soil may be affected by complexed nitrate molecules and microbial volatile organic compounds (mVOCs) produced during nitrification. Experiments were carried out to elucidate the overall nature of mVOCs and biogenic nitrates produced by nitrifiers, and their effects on nitrification and redox metabolism. Soils were incubated at three levels of biogenic nitrate. Soils containing biogenic nitrate were compared with soils containing inorganic fertilizer nitrate (KNO3) in terms of redox metabolism potential. Repeated NH4-N addition increased nitrification rates (mM NO3 1- produced g-1 soil d-1) from 0.49 to 0.65. Soils with higher nitrification rates stimulated (p < 0.01) abundances of 16S rRNA genes by about eight times, amoA genes of nitrifying bacteria by about 25 times, and amoA genes of nitrifying archaea by about 15 times. Soils with biogenic nitrate and KNO3 were incubated under anoxic conditions to undergo anaerobic respiration. The maximum rates of different redox metabolisms (mM electron acceptors reduced g-1 soil d-1) in soil containing biogenic nitrate followed as: NO3 1- reduction 4.01 ± 0.22, Fe3+ reduction 5.37 ± 0.12, SO4 2- reduction 9.56 ± 0.16, and CH4 production (µg g-1 soil) 0.46 ± 0.05. Biogenic nitrate inhibited denitrificaton 1.4 times more strongly compared to mineral KNO3. Raman spectra indicated that aliphatic hydrocarbons increased in soil during nitrification, and these compounds probably bind to NO3 to form biogenic nitrate. The mVOCs produced by nitrifiers enhanced (p < 0.05) nitrification rates and abundances of nitrifying bacteria. Experiments suggest that biogenic nitrate and mVOCs affect nitrification and redox metabolism in soil.

Interactive effect of climate factors, biochar and insecticide chlorpyrifos on methane consumption and microbial abundance in a tropical Vertisol.

Climate change may increase the pest infestation leading to intensive use of insecticides. However, the effect of insecticide and climate factors on soil methane (CH4) consumption is less understood. A laboratory experiment was carried out to evaluate the effect of temperature (15 °C, 35 °C, and 45 °C), moisture holding capacity (MHC) (60%, 100%), biochar (0%, 1%) and chlorpyrifos (0 ppm, 10 ppm) on CH4 consumption and microbial abundance in a tropical Vertisol of central India. Methane consumption rate k (ng CH4 consumed g-1 soil d-1) varied from 0.065 ±â€¯0.005 to 0.608 ±â€¯0.018. Lowest k was in 15 °C-60% moisture holding capacity (MHC)-no biochar and with 10 ppm chlorpyrifos. Highest k was in 35 °C-100% MHC-1% biochar and without (0 ppm) chlorpyrifos. Cumulative CO2 production (ng CO2 produced g-1 soil d-1) varied from 446 ±â€¯15 to 1989 ±â€¯116. Both CH4 consumption and CO2 production peaked in the treatment of 35 °C-100% MHC-1% biochar. Chlorpyrifos inhibited CH4 consumption irrespective of treatments. Abundance of 16S rRNA of eubacteria (× 106 g-1 soil) varied from 2.33 ±â€¯0.58 to 85.67 ±â€¯7.00. Abundance of 16S rRNA genes representing Actinomycetes (× 104 g-1 soil) varied from 7.67 ±â€¯1.53 and pmoA gene (Methanotrophs) (× 105 g-1 soil) varied from 1.23 ±â€¯0.59 to 34.33 ±â€¯6.51. Chlorpyrifos inhibited abundance of heterotrophic bacteria and methanotrophs but stimulated actinomycetes. Biochar stimulated the CH4 consumption, CO2 production and microbial abundance. Study highlighted that use of chlorpyrifos under climate change factors may inhibit CH4 consumption but the use of biochar may alleviate the negative effect of the chlorpyrifos.

Phylloplane bacteria of Jatropha curcas: diversity, metabolic characteristics, and growth-promoting attributes towards vigor of maize seedling.

The complex role of phylloplane microorganisms is less understood than that of rhizospheric microorganisms in lieu of their pivotal role in plant's sustainability. This experiment aims to study the diversity of the culturable phylloplane bacteria of Jatropha curcas and evaluate their growth-promoting activities towards maize seedling vigor. Heterotrophic bacteria were isolated from the phylloplane of J. curcas and their 16S rRNA genes were sequenced. Sequences of the 16S rRNA gene were very similar to those of species belonging to the classes Bacillales (50%), Gammaproteobacteria (21.8%), Betaproteobacteria (15.6%), and Alphaproteobacteria (12.5%). The phylloplane bacteria preferred to utilize alcohol rather than monosaccharides and polysaccharides as a carbon source. Isolates exhibited ACC (1-aminocyclopropane-1-carboxylic acid) deaminase, phosphatase, potassium solubilization, and indole acetic acid (IAA) production activities. The phosphate-solubilizing capacity (mg of PO4 solubilized by 108 cells) varied from 0.04 to 0.21. The IAA production potential (µg IAA produced by 108 cells in 48 h) of the isolates varied from 0.41 to 9.29. Inoculation of the isolates to maize seed significantly increased shoot and root lengths of maize seedlings. A linear regression model of the plant-growth-promoting activities significantly correlated (p < 0.01) with the growth parameters. Similarly, a correspondence analysis categorized ACC deaminase and IAA production as the major factors contributing 41% and 13.8% variation, respectively, to the growth of maize seedlings.

Diversity of bacteria and archaea in the rhizosphere of bioenergy crop Jatropha curcas.

Plant-microbial interaction in rhizosphere plays vital role in shaping plant's growth and ecosystem function. Most of the rhizospheric microbial diversity studies are restricted to bacteria. In natural ecosystem, archaea also constitutes a major component of the microbial population. However, their diversity is less known compared to bacteria. Experiments were carried out to examine diversity of bacteria and archaea in the rhizosphere of bioenergy crop Jatropha curcas (J. curcas). Samples were collected from three locations varying widely in the soil physico-chemical properties. Diversity was estimated by terminal restriction fragment length polymorphism (TRFLP) targeting 16S rRNA gene of bacteria and archaea. Fifteen bacterial and 17 archaeal terminal restriction fragments (TRFs) were retrieved from J. curcas rhizosphere. Bacterial indicative TRFs were Actinobacteria, Firmicutes, Acidobacteria, Verrumicrobiaceae, and Chlroflexi. Major archaeal TRFs were crenarchaeota, and euryarchaeota. In case of bacteria, relative fluorescence was low for TRF160 and high for TRF51, TRF 420. Similarly, for archaea relative fluorescence of TRF 218, and TRF 282 was low and high for TRF 278, TRF468 and TRF93. Principal component analysis (PCA) of bacterial TRFs designated PC 1 with 46.83% of variation and PC2 with 31.07% variation. Archaeal TRFs designated 90.94% of variation by PC1 and 9.05% by PC2. Simpson index varied from 0.530 to 0.880 and Shannon index from 1.462 to 3.139 for bacteria. For archaea, Simpson index varied from 0.855 to 0.897 and Shannon index varied from 3.027 to 3.155. Study concluded that rhizosphere of J. curcas constituted of diverse set of both bacteria and archaea, which might have promising plant growth promoting activities.

Aquatic microphylla Azolla: a perspective paradigm for sustainable agriculture, environment and global climate change.

This review addresses the perspectives of Azolla as a multifaceted aquatic resource to ensure ecosystem sustainability. Nitrogen fixing potential of cyanobacterial symbiont varies between 30 and 60 kg N ha(-1) which designates Azolla as an important biological N source for agriculture and animal industry. Azolla exhibits high bioremediation potential for Cd, Cr, Cu, and Zn. Azolla mitigates greenhouse gas emission from agriculture. In flooded rice ecosystem, Azolla dual cropping decreased CH4 emission by 40 % than did urea alone and also stimulated CH4 oxidation. This review highlighted integrated approach using Azolla that offers enormous public health, environmental, and cost benefits.

Methane oxidation and abundance of methane oxidizers in tropical agricultural soil (vertisol) in response to CuO and ZnO nanoparticles contamination.

There is worldwide concern over the increase use of nanoparticles (NPs) and their ecotoxicological effect. It is not known if the annual production of tons of industrial nanoparticles (NPs) has the potential to impact terrestrial microbial communities, which are so necessary for ecosystem functioning. Here, we have examined the consequences of adding the NPs particularly the metal oxide (CuO, ZnO) on CH4 oxidation activity in vertisol and the abundance of heterotrophs, methane oxidizers, and ammonium oxidizers. Soil samples collected from the agricultural field located at Madhya Pradesh, India, were incubated with either CuO and ZnO NPs or ionic heavy metals (CuCl2, ZnCl2) separately at 0, 10, and 20 µg g(-1) soil. CH4 oxidation activity in the soil samples was estimated at 60 and 100 % moisture holding capacity (MHC) in order to link soil moisture regime with impact of NPs. NPs amended to soil were highly toxic for the microbial-mediated CH4 oxidation, compared with the ionic form. The trend of inhibition was Zn 20 > Zn 10 > Cu 20 > Cu 10. NPs delayed the lag phase of CH4 oxidation to a maximum of 4-fold and also decreased the apparent rate constant k up to 50 % over control. ANOVA and Pearson correlation analysis (α = 0.01) revealed significant impact of NPs on the CH4 oxidation activity and microbial abundance (p < 0.0001, and high F statistics). Principal component analysis (PCA) revealed that PC1 (metal concentration) rendered 76.06 % of the total variance, while 18.17 % of variance accounted by second component (MHC). Biplot indicated negative impact of NPs on CH4 oxidation and microbial abundance. Our result also confirmed that higher soil moisture regime alleviates toxicity of NPs and opens new avenues of research to manage ecotoxicity and environmental hazard of NPs.