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Earthworm activities not only increase nitrogen (N) uptake by crops, but also lead to N losing to environment. Thus, the present study examined the transformation of

Coal bio-transformation technologies can convert the neglected low rank coal reserves to alternative fuels and non-fuel organic chemicals. Fungal mediated coal degradation can serve as biological beneficiation for alternative substances. For enhanced biodegradation, fungi can be isolated from coal environments and optimized for coal degradation processes. In this study, an indigenous fungal isolate AD-1, identified as Rhizopus oryzae by fungal ITS sequences, was isolated from a low rank coal sample collected from Darra Adam Khel, Frontier Region Kohat, Pakistan. The AD-1 mediated coal degradation was optimized and showed a substantial release of organics at 1.5% glucose and 0.5% coal loading ratio within 11 days. The GC–MS analysis of the black liquid from coal solubilization showed a variety of organic compounds including aromatic acids, fatty acids, alkanes, amines, and amides. The depolymerized liquid extract was analyzed by High-Performance Size-Exclusion Chromatography (HPSEC) for determining the molecular mass distribution of the released organics. The release of the organic fragments had molecular mass distribution ranging between 1.5 kDa and 26.7 kDa. In addition, the released fraction and residual coal sample after solubilization were analyzed with Fourier transform infrared spectroscopy (FT-IR) to explore the structural changes aroused by the AD-1 fungal treatment. The AD-1 fungal strain has the abilities of decarboxylation and deamination, as well as breaking the side chains of the aromatic rings. This demonstrates that indigenous fungi from coal environments can enhance the biodegradation and shows a potential for producing chemical feedstock or alternative fuels from low rank coal.

Chemosensory proteins (CSPs) have great contributions in performing diverse functions in insects. However, physiological appraisal of chemosensory protein genes still remains elusive in insects. We studied expression patterns and binding affinities of MsepCSP14, a chemosensory protein, in Mythimna separata. The distinct functions of MsepCSP14 were validated by employing different molecular techniques. The MsepCSP14 had high resemblance of sequence with chemosensory proteins of other insect family members. The MsepCSP14 expression was higher in antennal tissues of females than other tissues. Fluorescence binding assay validated that binding of nine out of 21 ligands to MsepCSP14 was higher at pH 7.4 than at pH 5.0. Three dimensional modeling (3D) and docking analysis predicted that amino acid residues of MsepCSP14 were involved in binding of compounds, and behavior assay displayed that adults of M. separata considerably responded to four volatiles from compounds demonstrating strong binding ability to MsepCSP14. Results of the present study suggest that MsepCSP14 is likely to mediate chemosensory functions in M. separata.

Skin darkening results as a consequence of the accumulation of skin pigment melanin. To combat this, the amplitude of skin lightening agents are commercially available, most of which inhibit melanin synthesis. Decolorization of melanin is an alternative method of skin lightening. In this study, we show that lignin peroxidase (LiP), an extracellular enzyme purified from Phanerochaete chrysosporium NK-1 isolated from a forest soil can effectively degrade and decolorize melanin in vitro. Decolorization conditions including pH, temperature, incubation time, enzyme concentration, and mediator addition were investigated to optimize the reaction conditions. The results indicate that pH 3, 40 °C, 15 IU/ml, and 10 h incubation were the optimal conditions for the decolorization of the melanin. The use of the mediator, veratryl alcohol was also found effective to enhance the efficacy of the melanin decolonization, with up to 92% decolorization. The scanning electron microscopy results showed void spaces on the treated melanin granules as compared to the untreated sample, indicating the degradation of melanin. Changes in the fingerprint region of the melanin were observed. Between wavenumbers 1500–500 cm−1, for example, the presence of new peaks in the treated melanin at 1513, 1464, and 1139 cm−1 CH2, CH3 bend and C–O–C stretch represented structural changes. A new peak at 2144 cm−1 (alkynyl C≡C stretch) was also detected in the decolorized melanin. The cytotoxicity study has shown that the treated melanin and LiP have low cytotoxic effects; however, the mediator of veratryl alcohol could result in high mortality which suggests that its use should be meticulously tested in formulating health and skincare products. The findings of the study suggest that LiP produced by Phanerochaete chrysosporium has the potential to be used in the medical and cosmetic industries, particularly for the development of biobased cosmetic whitening agents.

Several studies have been carried out to examine nitrous oxide (N2O) emissions from agricultural soils in the past. However, the emissions of N2O particularly during amelioration of acidic soils have been rarely studied. We carried out the present study using a rice-rapeseed rotation soil (pH 5.44) that was amended with dolomite (0, 1 and 2 g kg−1 soil) under 60% water filled pore space (WFPS) and flooding. N2O emissions and several soil properties (pH, NH4+ N, NO3−-N, and nosZ gene transcripts) were measured throughout the study. The increase in soil pH with dolomite application triggered soil N transformation and transcripts of nosZ gene controlling N2O emissions under both water regimes (60% WFPS and flooding). The 60% WFPS produced higher soil N2O emissions than that of flooding, and dolomite largely reduced N2O emissions at higher pH under both water regimes through enhanced transcription of nosZ gene. The results suggest that ameliorating soil acidity with dolomite can substantially mitigate N2O emissions through promoting nosZ gene transcription.

Soil N2O emissions are generally larger from soil with straw incorporated, and adjusting soil pH can substantially affect the mineralization of incorporated straw. However, the interactive effects of straw return and adjustment of soil pH on N2O emissions have not been well studied. Therefore, we investigated the role of adjusting soil pH using dolomite with incorporated crop straw in changing soil properties and N2O emissions. We performed a 105‐day incubation study in which dolomite was added to a rice (Oryza sativa L.) (C/N: 36) and green bean (Phaseolus vulgaris L.) straw (C/N: 17) amended acidic soil (Ultisol). The pH of the original soil (4.92) steadily increased to 5.35 and 5.45 at day 105 in rice and green bean straw treatments, respectively. Application of dolomite increased pH to a maximum value of 6.35 in the rice straw + dolomite and 6.55 in the green bean straw + dolomite amended soil at day 105 following establishment of the trial. Application of dolomite and the crop straws significantly increased soil microbial biomass C (MBC), dissolved organic C (DOC) and nitrate (NO3−), whereas it had a short‐term effect on exchangeable ammonium (NH4+). The Ultisol amended with green bean straw emitted larger cumulative N2O emissions (7.25 mg N2O‐N kg−1) during the 105‐day study than soil amended with rice straw (4.23 mg N2O‐N kg−1) and the control soil (1.42 mg N2O‐N kg−1). Dolomite application significantly (P ≤ 0.001) decreased N2O emissions from both rice and green bean straw treatments by increasing soil pH. The results of the current study can help to mitigate N2O emissions and so improve field management.

Chemosensory proteins (CSPs) have been suggested to perform several functions in insects, including chemoreception. To find out whether MsepCSP5 identified from Mythimna separata shows potential physiological functions in olfaction, gene expression profiles, ligand-binding experiments, molecular docking, RNA interference, and behavioral test were performed. Results showed that MsepCSP5 was highly expressed in female antennae. MsepCSP5 showed high binding affinities to a wide range of host-related semiochemicals, and displayed that 26 out of 35 candidate volatiles were highly bound (Ki < 10 µM) at pH 5.0 rather than pH 7.4. The binding sites of MsepCSP5 to candidate volatiles were well predicted by three-dimensional structure modeling and molecular docking experiments. Pursuing further, biological activities of M. separata to highly bound compounds elicited strong behavioral responses, such as alcoholic compounds displayed strong attractiveness whereas terpenes showed repellency to M. separata. The transcript expression level of MsepCSP5 gene significantly decreased after injecting target dsRNAs, and resulted in non-significant preference responses of M. separata to semiochemicals, such as 3-pentanol and 1-octene-3-ol. In conclusion, MsepCSP5 may involve in semiochemical reception of M. separata.

Insects have evolved highly sensitive olfactory sensory systems to detect plant hosts and mates, with plant volatiles playing an important role in informing insect behavior. Chemosensory proteins (CSPs) are thought to play a key role in this process, but in this respect, there is limited information on brown planthopper Nilaparvata lugens, one of the most destructive pests of rice. To expand our understanding of CSP function in N. lugens we explored expression profiles and binding characteristics of NlugCSP3. The ligands with higher binding affinity were also validated by molecular docking and behavioral assays. NlugCSP3 mRNA was expressed at relatively higher levels in antennae and abdomen of 3-day-old unmated macropterous males as well as in antennae of 3-day mated macropterous and brachypterous females. Fluorescence competitive binding assays revealed that 5 out of 25 candidate volatiles are strong binders (Ki

Heavy metals are believed to impact soil processes by influencing microbial communities, nutrient cycling or exchanging for essential plant nutrients. Soil pH adjustment highly influences the bio-availability of nutrients and microbial processes. We examined the effect of soil pH manipulation and copper (Cu as CuCl

Microbial solubilization of coal has been considered as a promising technology to convert raw coal into valuable products. In the present study, initially a total of 50 different aerobic bacterial and fungal isolates have been isolated from soil, coal and water samples of Dulmial Coal Mines, Chakwal, Pakistan, but on the basis of solubilization potential, only four isolates were selected for further study. The intensity of biosolubilization was measured by determining the weight loss of the coal pieces, which was observed to be about 25.93% by Pseudomonas sp. AY2, 36.36% by Bacillus sp. AY3 and 50% by Trichoderma sp. AY6, while Phanerochaete sp. AY5 showed maximum coal solubilization potential i.e. 66.67% in 30 days. UV/Vis spectrum revealed an increase in the pattern of absorbance of all treated samples compared to control referring to solubilization. Fourier transform infrared spectroscopy indicated alterations in the structure of treated coal in comparison to control coal suggesting breakdown i...

BACKGROUND A robust integrated pest management (IPM) programme is needed to reduce the use of insecticides in controlling Helicoverpa armigera. Therefore, a 2 year field study was conducted to evaluate the use of alternative control measures (biochemical use) for H. armigera relative to exclusively using chemical insecticides. The entomopathogenic fungus Beauveria bassiana, jasmonic acid and the insecticide chlorantraniliprole were each applied twice during the chickpea growing season. RESULTS All three applied materials (either alone or combined) significantly (P ≤ 0.05) reduced the larval population of H. armigera and pod infestation. Effects increased with time, and the maximum difference was observed 7 days after the second application in each year. The lowest numbers of larvae per plant and pod infestation were in the B. bassiana 3.21 × 106 + chlorantraniliprole treatment in both 2009/2010 and 2010/2011 year. The reduction in the larval population and pod infestation increased chickpea yield and the highest yield in both seasons, and the maximum yield was obtained in the B. bassiana 3.21 × 106 + chlorantraniliprole treatment. The populations of natural enemies were highest in the jasmonic acid treatment. CONCLUSION The results suggest that B. bassiana, jasmonic acid and chlorantraniliprole may be useful components for the H. armigera IPM strategy. © 2016 Society of Chemical Industry

The development of bio-processing low rank coal into fuels and non-fuel products has received growing interest in recent years, e.g., production of alternative chemical stock (i.e., humic acids) by the fungal transformation. Previous studies have demonstrated that fungal strains isolated from coal environments can play an important role in the coal-transformation processes with in vivo optimization and chemical pretreatments. In this study, the influence of chemical pretreatments (nitric acid and hydrogen peroxide) with a fungal isolate from coal on the depolymerization of Pakistani subbituminous coal was conducted. The chemical pretreatment exhibited enhanced production of humic acid from 13.5% to 54.2% for nitric acid and 45.7% for hydrogen peroxide. Nitric acid was more effective pretreatment in individual and combination with fungal-mediated depolymerization than hydrogen peroxide. The production of fungal-transformed liquid-derived humic acids were 36.4% and 31.1% for nitric acid and hydrogen peroxide pretreated coal. The results showed that the fungal-transformed humic acid has a high molecular weight, elevated aromatic condensation with mono and poly-substituted functional groups than the chemically treated and raw coal-derived humic acids. The fungal treatment also increased the nitrogen content in the resultant extract, which can be beneficial to the functionality of humic acids. By combining chemical and fungal treatment, it is possible to improve the quantity and quality of humic acids extracted from low rank coal. This is a promising approach in the development of more sustainable-cleaner products from Pakistan’s coal resources for the production of bio-active humic acid.

Nitrous oxide (N2O) is an overwhelming greenhouse gas and agricultural soils, particularly acidic soils, are the main source of its release to the atmosphere. To ameliorate acidic soil condition, liming materials are added as an amendment. However, the impact of liming materials has not been well addressed in terms of exploring the effect of soil pH change on N2O emissions. In the present study, a soil with pH 5.35 was amended with liming materials (CaMg(CO3)2, CaCO3, Ca(OH)2 and CaO) to investigate their effects on N2O emissions. The results indicate that application of liming materials reduced the magnitudes of N2O emissions. The maximum reduction of soil N2O emissions took place for Ca(OH)2 treatment when compared to the other liming materials, and was related to increasing soil pH. Mineral N, dissolved organic C, and microbial biomass C were also influenced by liming materials, but the trend was inconsistent to the soil pH change. The results suggest that N2O emission mitigation is more dependent on soil pH than C and N dynamics when comparing the different liming materials. Moreover, ameliorating soil acidity is a promising option to mitigate N2O emissions from acidic soils The authors would like to thank the funding bodies of the National Science Foundation of China (417501 10485), China Post-Doctoral Science Foundation (2017 M 622478), and National Key R&D Program (2017 YFD 0800102) for financially assisting the present research SI

The present study investigated the possibility of microbial transformations of coal to gas (biogasification) as an alternative to conventional coal mining because this approach has the potential to be less expensive, cleaner, and provide greater access to deeper coal resources. Biogasification is often associated with low rank coal such as lignite and subbituminous coal that has produced enough coalbed methane to be commercially viable in the United States and Australia. However, little work has been done to analyze the potential of biogasification in higher rank coal. For this purpose, bioassay using a wetland-derived consortium and a coal-derived consortium were used to analyze coal samples from Pakistan belonging to different ranks (lignite to semi-anthracite). Among all samples a low volatile bituminous coal produced the maximum methane 34.95 μmol CH4/g coal with the wetland-derived microbial consortium, followed by subbituminous coal (30.18 μmol CH4/g coal). Lower methane levels were recorded with the coal-derived consortium, with subbituminous coal yielding the highest concentration (25.1 μmol CH4/g coal). Methane levels appeared to be increasing on the last measurement indicating the coal-derived consortium was slower than the wetland-derived consortium but could still catalyze biogasification in higher rank coals. Quantitative polymerase chain reaction analysis for mcrA functional genes indicated that the microbial community members that produce methane (methanogens) varied during the incubations. Energy conversion efficiency of different strategies (other biological and underground coal gasification processes) was also compared and discussed. This study was the first to compare bioassay using consortia of microbes non-indigenous and indigenous to coal and indicate the potential of biogasification from many different coalbeds across Pakistan.

Chemosensory proteins (CSPs) play important roles in chemosensation in insects, but their exact physiological functions remain elusive. In order to investigate the functions of CSPs in the oriental armyworm Mythimna separata, in the present study we explored expression patterns and binding characteristics of the CSP, MsepCSP8. The distinctive functions of MsepCSP8 were also validated by RNAi. The results showed that MsepCSP8 shares high sequence similarity with CSPs of other insect family members, including the characteristic four-cysteine signature motif. MsepCSP8 mRNA was specifically expressed in antennae of females at levels well above those in other tissues. Competitive binding assays confirmed that 20 out of 56 ligands bound more strongly to MsepCSP8 at pH 7.4 than at pH 5.0. Protein structure modeling and molecular docking analyses identified amino acid residues involved in binding volatile compounds, and behavioral response experiments showed that M. separata elicited significant responses to five volatiles from compounds displaying high binding affinity to MsepCSP8. MsepCSP8 transcript abundance was decreased by dsMsepCSP8 injection, which affected the behavioral responses of M. separata to representative semiochemicals. Our findings demonstrate that MsepCSP8 likely contributes to mediating responses of M. separata adults to plant volatiles.

Chemosensory proteins (CSPs) play important roles in insects' chemoreception, although their specific functional roles have not been fully elucidated. In this study, we conducted the developmental expression patterns and competitive binding assay as well as knock-down assay by RNA interference both in vitro and in vivo to reveal the function of NlugCSP10 from the brown planthopper (BPH), Nilaparvata lugens (Stal), a major pest in rice plants. The results showed that NlugCSP10 messenger RNA was significantly higher in males than in females and correlated to gender, development and wing forms. The fluorescence binding assays revealed that NlugCSP10 exhibited the highest binding affinity with cis-3-hexenyl acetate, eicosane, and (+)-β-pinene. Behavioral assay revealed that eicosane displayed attractant activity, while cis-3-hexenyl acetate, similar to (+)-β-pinene significantly repelled N. lugens adults. Silencing of NlugCSP10, which is responsible for cis-3-hexenyl acetate binding, significantly disrupted cis-3-hexenyl acetate communication. Overall, findings of the present study showed that NlugCSP10 could selectively interrelate with numerous volatiles emitted from host plants and these ligands could be designated to develop slow-release mediators that attract/repel N. lugens and subsequently improve the exploration of plans to control this insect pest.

Application of biochar to soil can play a significant role in the alteration of nutrients dynamics, soil contaminants as well as microbial functions. Therefore, strategic biochar application to soil may provide agronomic, environmental and economic benefits. Key environmental outcomes may include reduced availability of toxic metals and organic pollutants, reduced soil N losses and longer-term storage of carbon in soil. The use of biochar can certainly address key soil agronomic constraints to crop production including Al toxicity, low soil pH and may improve nutrient use efficiency. Biochar application has also demerits to soil properties and attention should be paid when using a specific biochar for a specific soil property improvement. This review provides a concise assessment and addresses impacts of biochar on soil properties.

Several studies have been carried out to examine nitrous oxide (N

Nitrous oxide (N2O) is a devastating greenhouse gas mainly released from soils to the atmosphere. Pasture soils, particularly acidic in nature, are large contributors of atmospheric N2O through deposition of urine-N. Devising strategies for reducing N2O emissions in acidic soils are the utmost need of the time. Therefore, the present study was carried out to investigate the possible efficacy of dolomite application to reduce N2O emissions from urine treated acidic soil. Application of urine to soil enlarged the production of NH4+-N, NO3−-N, microbial biomass C (MBC) and dissolved organic C (DOC), resulting in higher N2O emissions as compared to the control (soil only). The highest N2O emission rate (1.35 μg N2O-N kg−1 h−1) and cumulative flux (408 μg N2O-N kg−1) occurred in urine only treated soil. Dolomite addition, especially higher application dose, greatly reduced N2O emissions through improved soil pH. The results suggest that increasing pH of acidic soils is a good applicable approach for reducing N2O emissions from urine-treated soils.

Heavy metals are believed to impact soil processes by influencing microbial communities, nutrient cycling or exchanging for essential plant nutrients. Soil pH adjustment highly influences the bio-availability of nutrients and microbial processes. We examined the effect of soil pH manipulation and copper (Cu as CuCl2.2H2O) application on nitrogen (N) cycling and nitrous oxide (N2O) emissions from an acid soil. Increasing amounts of Cu (0, 250, 500 and 1000 mg kg−1) were added to an acidic soil (pH = 5.44) that was further amended with increasing amounts of dolomite [CaMg(CO3)2] to increase soil pH. Dolomite increased soil pH values, which reached a maximum without Cu application (-Cu) at day 42 of the experiment. The soil pH values decreased with increasing dose of Cu, and remained low as compared with both control and dolomite amended soil. Ammonium (NH4+-N) concentrations were higher in Cu contaminated soil as compared with the control and dolomite treated soil. Nitrate (NO3−-N) concentrations increased in dolomite treated soil when compared with the +Cu alone treatments and control. Microbial biomass carbon (MBC) and nitrogen (MBN) contents were higher in dolomite treated soil as compared with the +Cu treatments and control. The application of increasing amounts of Cu progressively decreased soil MBC and MBN. Nitrous oxide emissions were higher (p ≤ 0.01) in +Cu soil as compared with the control, and increased with increasing Cu concentration in soil. Application of dolomite highly suppressed soil N2O emissions in both +Cu and –Cu soils. The results indicate that the effects of heavy metal contamination (specifically Cu contamination) can increase N2O emissions, but this can be effectively mitigated through increasing soil pH, also decreasing potential toxic effects on soil microorganisms.

Tomato (Solanum lycopersicum L.) is an important food crop extensively grown all over the world and is susceptible to various abiotic and biotic stresses. Tomato leaf curl virus (TLCV) is a devastating disease of tomato, transmitted by whitefly, ultimately results in marked reduction of tomato production. In current experiment, four tomato cultivars i.e., Reo Grande, Roma, Moneymaker and Nagina were grown under field conditions to screen against TLCV. Correlation studies revealed a positive and significant relationship among TLCV, whitefly population and environmental factors (temperature and relative humidity). Highest whitefly infestation with TLCV severity was observed on moneymaker compared to other tested cultivars. Subsequently, moneymaker was further subjected to check efficacy of botanicals (neem leaf extract and garlic clove extract), detergent and Imidacloprid against disease vector i.e., whitefly and TLCV at different concentrations under glasshouse conditions. These botanicals and detergent elucidated promising results in reducing whitefly population followed by low severity of TLCV. Whereas, Imidacloprid was most effective in reducing whitefly population up to 1.00 adult/ plant followed by neem leaf extract, garlic clove extract and detergent that reduced up to 1.33, 1.67 and 2.33 adults/ plant, respectively. It is recommended that TLCV severity could be managed by using neem leaf and garlic clove extract which ultimately reduced the whitefly population and serves as a good alternative than hazardous pesticides on food crops.

A robust integrated pest management (IPM) programme is needed to reduce the use of insecticides in controlling Helicoverpa armigera. Therefore, a 2 year field study was conducted to evaluate the use of alternative control measures (biochemical use) for H. armigera relative to exclusively using chemical insecticides. The entomopathogenic fungus Beauveria bassiana, jasmonic acid and the insecticide chlorantraniliprole were each applied twice during the chickpea growing season.All three applied materials (either alone or combined) significantly (P ≤ 0.05) reduced the larval population of H. armigera and pod infestation. Effects increased with time, and the maximum difference was observed 7 days after the second application in each year. The lowest numbers of larvae per plant and pod infestation were in the B. bassiana 3.21 × 10The results suggest that B. bassiana, jasmonic acid and chlorantraniliprole may be useful components for the H. armigera IPM strategy. © 2016 Society of Chemical Industry.