HCN Production Research Articles

Phytochemical Pharmacological and Growth Promoting Study of Endophytic Fungi Rhizopus delemer Isolated from Abutilon indicum (L.) Sweet.

Endophytic fungi have been perceived as a potential source of bioactive secondary metabolites. Whereas Rhizopus as the endophytic fungi were isolated from the medicinal plant Abutilon indicum (L) sweet under the family Malvaceae found in tropical areas. This work aims to isolate and identify the fungal endophyte Rhizopus sp. from the medicinal plant Abutilon indicum at the molecular level, and the second objective is to test the Plant Growth Promoting activity like Indole Acidic Acid A, Hydrogen cyanide, Phosphate solubility (PO4), Catalase test, and it is the proof for plant endophyte association, lastly to analyse the extract of Rhizopus in phytochemical and pharmacology activity using Gas chromatography and Mass spectrometry study. The isolated fungal plant growthpromoting activity like Indole acetic acid production, Phosphate solubility assay, HCN production and Catalase enzyme synthesis was tested. As compared to IAA, PO4 solubility assay, HCN and Catalase synthesis test culture gave positive results. It described endophytic fungal within-hosts that are actively responsible against pathogens and stress tolerance conditions. Isolated fungal culture was able to synthesize catalase enzymes and HCN compounds. Rhizopus can act as a Phyto stimulator, broad-spectrum antimicrobial agent, increased stress tolerance capacity and also can increase the water availability in the nutrition supply to the host plant. In the qualitative screening test, there are about 12 phytochemicals that showed positive results. It gives the effort of the plant to act as a defensive mechanism against pathogens. Based on morphological and phylogenetic analyses, fungal species were identified: as Rhizopus delemer UICC 26. Accession no: LC514308. It is an unfamiliar mutant strain. Further GC-MS analysis revealed that the Identification of 20 bioactive compounds in the ethanolic extract of fungus Rhizopus delemer was Butane, 1,1-diethoxy-2-methyl-. 3,3-diethoxy-2-butanone. identified. The identification of bioactive chemical compounds is based on a Database in the National Institute of Standard Technology. The compound's retention time, peak area, molecular weight and molecular formula were matched with WILEY and NIST Library. It could be concluded that the isolated endophytic fungus Rhizopus delemer UICC 26 extract was prominent endophytic fungi in Abutilon indicum plant and also it enhances the plant growth-promoting activity. It was confirmed by the positive results of HCN and Catalase production test. In 12 different phytochemical compounds, and among 20 bioactive agent’s presence 9 shown our fungi has active form inside of plant part against the pathogen.

Evolution in response to climate in the native and introduced ranges of a globally distributed plant.

The extent to which species can adapt to spatiotemporal climatic variation in their native and introduced ranges remains unresolved. To address this, we examined how clines in cyanogenesis (hydrogen cyanide [HCN] production-an antiherbivore defense associated with decreased tolerance to freezing) have shifted in response to climatic variation in space and time over a 60-year period in both the native and introduced ranges of Trifolium repens. HCN production is a polymorphic trait controlled by variation at two Mendelian loci (Ac and Li). Using phenotypic assays, we estimated within-population frequencies of HCN production and dominant alleles at both loci (i.e., Ac and Li) from 10,575 plants sampled from 131 populations on five continents, and then compared these frequencies to those from historical data collected in the 1950s. There were no clear relationships between changes in the frequency of HCN production, Ac, or Li and changes in temperature between contemporary and historical samples. We did detect evidence of continued evolution to temperature gradients in the introduced range, whereby the slope of contemporary clines for HCN and Ac in relation to winter temperature became steeper than historical clines and more similar to native clines. These results suggest that cyanogenesis clines show no clear changes through time in response to global warming, but introduced populations continue to adapt to their contemporary environments.

N migration and transformation during the co-combustion of sewage sludge and coal slime

The co-combustion of sewage sludge and coal slime is considered a promising technique for reducing the volume of sewage sludge, adding value, and decreasing the risks associated with these wastes. This work aimed to study N migration and transformation mechanisms and the related interactions during the co-combustion of sewage sludge (SS) and coal slime (CS) by thermogravimetric-mass spectrometry combined with X-ray photoelectron spectroscopy. The results revealed that the main N-containing gases produced during the combustion of SS and CS were NH3 generated from Amino-N at 200–400 °C and HCN generated from heterocyclic nitrogen at 400–600 °C, respectively. The increase of CS ratio led to a decrease in the release of NH3 and NO, but an increase in the release of HCN. Distinct interactions were observed during the co-combustion process, which promoted the production of NH3 and inhibited the production of HCN and NO. Co-combustion inhibited the release of NO by 36.9% when the CS ratio was 50%. The interaction mechanism suggested that H radicals from SS promoted the premature decomposition of N species in CS, and increased the selectivity of N species for NH3 formation by promoting the conversion of heterocyclic-N to Amino-N. In addition, the interaction of char (in SS) and char (in CS) enhanced the reduction of NO. Above 600 °C, co-combustion promoted the retention of N species in the ash.

Measurement of CO, HCN, and NO productions in atmospheric reaction induced by femtosecond laser filament

It is proved that the chemical reaction induced by femtosecond laser filament in the atmosphere produces CO, HCN, and NO, and the production CO and HCN are observed for the first time. The concentrations of the products are measured by mid-infrared tunable laser absorption spectroscopy. In the reduced pressure air, the decomposition of CO2 is enhanced by vibration excitation induced by laser filament, resulting in the enhanced production of CO and HCN. At the same time, the CO and HCN generated from the atmosphere suffer rotation excitation induced by laser filament, enhancing their absorption spectra. It is found that NO, CO, and HCN accumulate to 134 ppm, 80 ppm, and 1.6 ppm in sealed air after sufficient reaction time. The atmospheric chemical reaction induced by laser filament opens the way to changing the air composition while maintaining environmental benefits.

Whole Genome, Functional Annotation and Comparative Genomics of Plant Growth-Promoting Bacteria Pseudomonas aeruginosa (NG61) with Potential Application in Agro-Industry.

A plant growth-promoting Rhizobacteria (PGPR) Pseudomonas aeruginosa (NG61) isolated from rhizosphere of Sunflower plant. The isolate was identified by 16S rRNA gene sequencing (Accession no. MK455763). NG61 showed various plant growth promotion and biocontrol activities like, Phosphate solubilisation, Nitrogen fixation, Ammonia production, IAA production, siderophore production, HCN production. The whole genome sequence of Pseudomonas aeruginosa (NG61) was reported and analysed. The estimated genome size was 6537180bp with 66.18% of G+C content. The genome encoded 6186 protein-coding genes, 6252 genes were predicted, 66RNA genes. Phylogenetic tree showed that the P. aeruginosa( NG61) was closely related to P.aeruginosa strain DSM 50071. The annotated draft genome has been deposited at the NCBI database under the accession number PRJNA707114 BioProject and BioSample: SAMN18174979. The analysis of genome sequence of P. aeruginosa (NG61) showed various genes encoding plant growth promotion and biocontrol activities.

Unveiling the mechanism of NOx precursor formation during sewage sludge pyrolysis: Effects of carbohydrate–protein interactions

Municipal sewage sludge poses environmental and health risks, and thus, requires proper disposal using pyrolytic techniques. However, these techniques are hindered by the production of NH3 and HCN (NOx precursors) by the sludge. Hence, a comprehensive understanding of NH3 and HCN formation during sludge pyrolysis is required to minimize its NOx footprint. As sludge N mainly occurs in proteins forms, its transformations can be modeled using amino acids. Here, we aimed to the study the mechanism by which carbohydrates influence the formation of NOx precursors during sludge protein pyrolysis at different temperatures using glutamic acid, tyrosine, and histidine as protein models and cellulose and lignin as carbohydrate models. During pyrolysis, the release of NH3 and HCN was promoted by high temperatures and inhibited by carbohydrates. Despite this inhibitory effect, the results suggested that the release of NH3 should be considered for samples rich in aliphatic amino acids. For glutamic acid and tyrosine, NOx precursor formation was inhibited by N fixation in coke under the action of volatiles produced during carbohydrate pyrolysis. For glutamic acid, adding cellulose and lignin increased the coke-N content by 25.32% and 44.73% at 700 ℃. For histidine, this effect was ascribed to the ring-opening reactions induced by the free radical products of carbohydrate decomposition and the enhanced transfer of N-containing compounds to tar after ring-opening recombination. Further, heterocyclic-N within tar increased to 69.12% due to lignin-histidine interactions at 700 ℃. The results of this study can assist in regulating sludge protein pyrolysis intended for minimizing the production of NOx precursors.

Non-Rhizobial Endophytes Associated with Nodules of Vigna radiata L. and Their Combined Activity with Rhizobium sp.

Root nodules of legume plants are devoted for hosting endophytic symbiotic bacteria that fix atmospheric nitrogen but recently proved as a niche for various non-rhizobial endophytes (NRE) also. In the present investigation, one rhizobial and two NRE were isolated and characterized as Rhizobium sp. AAU B3, Bacillus sp. AAU B6 and Bacillus sp. AAU B12. These isolates were studied for in vitro biocontrol activity against two pathogenic fungi. NRE isolates exhibited antifungal activity against root rot causing Macrophomina phaseolina (ITCC-6749) isolated from Vigna radiata and wilt causing pathogen Fusarium udum Butler isolated from Cajanus cajan in liquid as well as on solid medium. Furthermore, their antagonism was increased markedly when combined with Rhizobium sp. Moreover, Bacillus sp. AAU B6 showed amplification of the zwittermicin A gene (~ 950bp) which is evident for the production of antibiotics. All three isolates showed HCN production in vitro also, Bacillus sp. AAU B12 exhibited amplification of its gene hcnC. Pathogenic fungal hyphae became thin, transparent, and bent as well as fungi lost their normal growth and branching patterns when exposed to volatile compounds produced by NRE. All the 3 isolates produced siderophores, however siderophore production was increased considerably when all three strains are mixed together. Furthermore, all the three isolates produced cell wall degrading enzymes (chitinase, protease, and cellulase) but lipolytic activity was exhibited only by Rhizobium sp. AAU B3. When NRE inoculated in combination of Rhizobium; overcomes the disease severity against M. phaseolina under pot study. Thus, from present study it is concluded that co-inoculation of NRE and Rhizobium sp. can be exploited as biocontrol bio-agents against M. phaseolina in green gram at field levels.

Multifarious Indigenous Diazotrophic Rhizobacteria of Rice (Oryza sativa L.) Rhizosphere and Their Effect on Plant Growth Promotion.

A diverse group of rhizobacteria persists in the rhizospheric soil, on the surface of roots, or in association with rice plants. These bacteria colonize plant root systems, enhance plant growth and crop yield. Indigenous rhizobacteria are known to promote soil health, grain production quality and serve as sustainable bioinoculant. The present study was aimed to isolate, identify and characterize indigenous plant growth promoting (PGP) diazotrophic bacteria associated with the rhizosphere of rice fields from different areas of Jammu and Kashmir, India. A total of 15 bacteria were isolated and evaluated for various PGP traits, antagonistic activity against phytopathogens, production of hydrolytic enzymes and biofilm formation under in-vitro conditions. The majority of the isolated bacteria were Gram-negative. Out of 15 bacterial isolates, nine isolates produced IAA (12.24 ± 2.86 to 250.3 ± 1.15 μg/ml), 6 isolates exhibited phosphate solubilization activity (36.69 ± 1.63 to 312.4 ± 1.15 μg/ml), 7 isolates exhibited rock phosphate solubilization while 5 isolates solubilized zinc (10–18 mm), 7 isolates showed siderophore production, 8 isolates exhibited HCN production, 6 isolates exhibited aminocyclopropane-1-carboxylate (ACC) deaminase activity, 13 isolates exhibited cellulase activity, nine isolates exhibited amylase and lipase activity and six isolates exhibited chitinase activity. In addition, 5 isolates showed amplification with the nifH gene and showed a significant amount of nitrogenase activity in a range of 0.127–4.39 μmol C2H4/mg protein/h. Five isolates viz., IHK-1, IHK-3, IHK-13, IHK-15 and IHK-25 exhibited most PGP attributes and successfully limited the mycelial growth of Rhizoctonia solani and Fusarium oxysporum in-vitro. All the five bacterial isolates were identified based on morphological, biochemical and 16S rDNA gene sequencing study, as Stenotrophomonas maltophilia, Enterobacter sp., Bacillus sp., Ochrobactrum haematophilum and Pseudomonas aeruginosa. Rice plants developed from seeds inoculated with these PGP strains individually had considerably higher germination percentage, seed vigor index and total dry biomass when compared to control. These findings strongly imply that the PGP diazotrophic bacteria identified in this work could be employed as plant growth stimulators in rice.

Microbial validation of organic preparations used in natural farming

In this study, the scientific validation of different organic preparations like amritpani, panchagavya, vermiwash and Jeevamrita in terms of microbial population and their activity has been performed. Microbes isolated from these preparations were tested for plant growth promoting (PGP) properties and finally characterized by biochemical and molecular tools. Strains of Bacilluspumilus and other bacillus sp. isolated form panchagavya showed positive results for HCN and ammonia production while bacteria isolated from vermiwash (Bacilluspumilus and Brevibacteriumlaterosporus) showed positive results for HCN production and P-solubilization. Plant growth promoting strain Brevibacillusborstelensis (isolated from amritpani) showed positive results for all PGP properties (including HCN, siderophore, ammonia, catalase and IAA production) except

Plant Growth-promoting and Bio-control Activity of Micrococcus luteus Strain AKAD 3-5 Isolated from the Soybean (Glycine max (L.) Merr.) Rhizosphere

Background: Applications of bioinoculants for improving crop productivity may be an eco-friendly alternative to chemical fertilizers. Rhizosphere or soil-inhabiting beneficial microbes can enhance plant growth and productivity through direct and indirect mechanisms, i.e., phosphate solubilization, nutrient acquisition, phytohormone production, etc. Objective: This study is based on the hypothesis that diseases resistant plants can act as a source of potential microbes that can have good plant growth-promoting traits and bio-control potential. Methods: In this study, we have isolated the rhizobacterial strains (AKAD 2-1, AKAD 2-10, AKAD 3-5, AKAD 3-9) from the rhizosphere of a disease-resistant variety of soybean (JS-20-34) (Glycine max (L.) Merr.). These bacterial strains were further screened for various plant growth-promoting traits (phosphate solubilization, indole acetic acid (IAA), ammonia, biofilm, HCN, Exopolysaccharide (EPS), and enzyme production activity (catalase, cellulase, and chitinase)). Results: Among four, only bacterial strain AKAD 3-5 has shown plant-growth-promoting and biocontrol (98%) activity against Fusarium oxysporum. Morphological, biochemical, and molecular characterization (16S rRNA) revealed that this rhizobacterial isolate AKAD 3-5 closely resembles Micrococcus luteus (Gene bank accession: MH304279). Conclusion: Here, we conclude that this strain can be utilized to promote soybean growth under varied soil stress conditions.

Rhizospheric PGPR Strains of Wheat, Barley and Trefoil Grown in Ağrı Province

The present study aimed to isolate potential plant growth rhizobacteria (PGPR) from various crops in agricultural areas in Ağrı-Turkey and their characterization. For this goal, rhizospheric soil samples of wheat, barley and trefoil were collected from 12 different locations in Ağrı province. Bacteria isolation studies were carried out with these rhizospheric samples. Then, to determine the PGPR properties of each isolate; nitrogen fixation, phosphate dissolution, siderophore, ammonia and HCN production tests were performed. Molecular identification of active isolates determined as suitable for development of biofertilizers, biostimulants and/or bioprotectants was done by PCR and sequencing applications performed with universal 16S rRNA primers. According to the results, 29 potential PGPR isolates were determined and their molecular characterization was done. These isolates were distributed in Arthrobacter, Bacillus, Brevibacterium, Enterobacter, Pantoea, Pseudomonas, Microbacterium and Stenotrophomonas according to their 16S rRNA sequence similarities.

Exploration of Klebsiella pneumoniae M6 for paclobutrazol degradation, plant growth attributes, and biocontrol action under subtropical ecosystem.

In recent times, injudicious use of paclobutrazol (PBZ) in mango orchards deteriorates the soil quality and fertility by persistence nature and causes a serious ecosystem imbalance. In this study, a new Klebsiella pneumoniae strain M6 (MW228061) was isolated from mango rhizosphere and characterized as a potent plant growth promoter, biocontrol, and PBZ degrading agent. The strain M6 efficiently utilizes PBZ as carbon, energy and nitrogen source and degrades up to 98.28% (50 mgL-1 initial conc.) of PBZ at 15th day of incubation in MS medium. In the soil system first order degradation kinetics and linear model suggested 4.5 days was the theoretical half-life (t1/2 value) of PBZ with strain M6. Box Behnken design (BBD) model of Response surface methodology (RSM) showed pH 7.0, 31°C temperature, and 2.0 ml inoculum size (8 x 109 CFU mL-1) was optimized condition for maximum PBZ degradation with strain M6. Plant growth promoting attributes such as Zn, K, PO4 solubilization IAA, HCN and NH3 production of strain M6 showed positive results and were assessed quantitatively. The relation between plant growth promotion and PBZ degradation was analyzed by heat map, principal component analysis (PCA) and, clustal correlation analysis (CCA). Strain M6 was also showing a significant biocontrol activity against pathogenic fungi such as Fusarium oxysporum (MTCC–284), Colletotrichum gloeosporioides (MTCC– 2190), Pythium aphanidermatum (MTCC– 1024), Tropical race 1 (TR -1), and Tropical race 4 (TR -4). Hence, results of the study suggested that strain M6 can be utilized as an effective bio-agent to restore degraded land affected by persistent use of paclobutrazol.

Impact of Pseudomonas spp. on Plant Growth, Lytic Enzymes and Secondary Metabolites Production

Seven strains of Pseudomonas spp. were isolated from the south Gujarat region of India. Antifungal and bacterial activities of bacterial strains were evaluated against important plant pathogens in vitro, among them, PaRS was found most effective. The indole acetic acid production was recorded in all isolated Pseudomonas spp. Seed treatment with PaRS at 6 g/kg was significantly superior over other treatments for plant growth promotion (germination, shoot and root length, shoot and root weight, vigor index, and both shoot and root colonization) under standard roll towel method and pot conditions. The maximum siderophore production was observed in PaRS and medium production in PfRB, PaNS, and PfNC. PaNS and PaRS strains recorded strong HCN production but moderate production recorded in PaWP, PaWS, and PfNC. The maximum phosphate solubalization zone (22 mm) was found in PaRS. PaRS recorded maximum chitinase, ß-1,3-glucanase activity, hydrogen cyanide, and salicylic acid production as compared to other strains.

The Effect of Accelerated Ageing on Reaction-to-Fire Properties–Composite Materials

As material age, the durability, strength, and other mechanical properties are impacted. The lifespan of a material generally decreases when exposed to weathering conditions such as wind, temperature, humidity, and light. It is important to have knowledge of how materials age and how the material properties are affected. Regarding materials´ fire behaviour and the effect of ageing on these properties, the knowledge is limited. The research questions of the current work are: Are the fire properties of composite materials affected by ageing? And if so, how is it affected? The study is on material at Technology Readiness Level 9 (TRL). In this study, three composite fibre laminates developed for marine applications were exposed to accelerated ageing. Two different ageing conditions were selected, thermal ageing with an increased temperature of 90°C and moisture ageing in a moderately increased temperature of 40°C and a relative humidity of 90%. Samples were collected after one, two and four weeks of ageing. The reaction-to-fire properties after ageing was evaluated using the ISO 5660–1 cone calorimeter and the EN ISO 5659–2 smoke chamber with FTIR gas analysis. The test results showed that the fire behaviour was affected. Two of the composite laminates, both phenolic/basalt composites, showed a deteriorated fire behaviour from the thermal ageing and the third composite laminate, a PFA/glass fibre composite, showed an improved fire behaviour both for thermal and moisture ageing. The smoke toxicity was affected by the accelerated ageing, especially for the PFA/glass fibre composite that showed a higher production of CO and HCN, both for the thermal aged and the moisture aged samples.

Development of native Pseudomonas aeruginosa bioformulations with plant growth promoting potential for apple crop in Himachal Pradesh

This study was conducted to characterize different plant growth promoting Pseudomonas strains isolated from wet temperate zones of Himachal Pradesh and two best effective plant growth promoting P. aeruginosa strains An-14-Mg and An-15-Mg were tested under field conditions to evaluate their growth promotional effect on apple crop. Maximum production of all the plant growth promoting traits were shown by two Pseudomonas isolate An-14-Mg and An-15-Mg. Maximum phosphate solubilization was shown by An-15-Mg (46 mm) whereas highest siderophore units (SU) were recorded in An-14-Mg (71.23%SU) followed by An-15-Mg (70.12%SU) which were statistically at par with each other. Highest IAA production (95 µg/ml), HCN (++++) and ammonia (++++) production was observed again in Pseudomonas isolate An-15-Mg. Both the strains exhibited 99% level of similarity with P. aeruginosa. The presence of An-14-Mg, An-15-Mg alone and consortium were tested under field conditions to study their effect on percent survival of apple rootstocks, plant height and number of shoots up to 3 years period. Maximum increase in percent survival, plant height, number of shoots, chlorophyll content and available NPK content of soil were observed in consortial bioformulation after 3 years growth observed in apple plants. Both the P. aeruginosa strains were reported first time as plant growth promoter with apple tree, therefore can be used as biofertilizer in apple crop of Himachal Pradesh.

Potential Bacterial Antagonists for the Control of Charcoal Rot (Macrophomina phaseolina) in Strawberry

The effect of antagonistic bacteria to control Macrophomina phaseolina was evaluated under in vitro, growth chamber, greenhouse and field conditions. A total of 177 bacteria, isolated from Athrocaulon macrostachyum rhizosphere of the Lebrija marsh, were screened for their potential against M. phaseolina (causes charcoal rot in strawberry) by dual culture assay. Of these isolates, 14 most promising strains were molecularly identified by the 16S rDNA sequencing method using the EzBioCloud database. These strains were tested for in vitro hydrolytic enzymes, HCN production, and biocontrol against M. phaseolina in strawberry plants. All the 14 strains produced, at least, one hydrolytic enzymatic activity and one of them, which belongs to Brevibacterium genus (Hvs8), showed the lowest records of disease incidence (20%) and severity (0.4). With these results, greenhouse and field trials were carried out with the Hvs8 strain, compared to non-treated control. In the greenhouse assays, Hvs8 strain increased root dry mass by 30%, over the control. In the field trials, production and fruit quality were not significantly different between Hvs8 treatment and non-treated control, but plant mortality and plant mortality associated to M. phaseolina decreased by more than 24% and 65% respectively, in Hvs8 treatment. This study suggests that Brevibacterium sp. Hvs8 strain could be a candidate for controlling charcoal rot in strawberry.

Rhizosphere competent Pseudomonas indoloxydans (F3-47) as a plant growth promoter and enhancer of Zea mays L. under greenhouse and field trials

A rhizospheric bacterium belonging to the genus Pseudomonas isolated from the rhizosphere of maize was evaluated for the plant growth promoting and biocontrol traits. The strain was subjected to the different PGP and biocontrol traits such as ammonia, IAA, siderophores, phosphate solubilisation, ACC deaminase, HCN production, cellulase, pectinase and chitinase activity. 16S rRNA sequencing showed maximum homology with the strain Pseudomonas indoloxydans IPL-1(NR115922). In vivo plant growth promotion assay with the strains on maize seedlings showed a significant increase in the biometric parameters with respect to the number of shoots (32%) and roots (27%) and its fresh weight (20%) and dry weight (54%). Different formulations of the strain tested in fields revealed that the maize plants treated with the talc and lignite formulations showed a considerable increase in the growth of maize when compared to the plants without any treatment. The study concluded that the rhizosphere strains Pseudomonas indoloxydans (F3- 47) could be a potential plant growth promoter for future exploration in agriculture.

Study on the pyrolysis behavior of coal-water slurry and coal-oil-water slurry

In this work, different concentrations of coal-water slurry and coal-oil-water slurry were prepared and pyrolyzed at various temperatures of 800–1000 °C. The chemical characteristics and combustion reactivities of chars and the formation mechanism of main gas components of CO, CO2, CH4, and H2 were investigated. With the water content of the coal-water slurry increasing, the formation of CO and CH4 were hindered, while CO2 and H2 were promoted. For the coal-oil-water slurry, oil promoted CH4 and H2 production but inhibited CO and CO2. Moreover, the variation of the nitrogen-containing pyrolytic gas during pyrolysis was studied for various concentrations of coal-water and coal-oil-water slurries at different temperatures. The relative content of HCN and NH3 gradually increased with the increase of pyrolysis temperature and water content. For coal-oil-water slurry, the waste lubricating oil promoted the formation of HCN and NH3, with the relative content of HCN being much higher than that of NH3. To study the mechanism of nitrogen migration during pyrolysis, Lammps was used to simulate pyrolytic behavior of pyrrole and pyridine under N2 and steam atmospheres. The increase in temperature promoted the decomposition of pyrrole and pyridine, thereby promoting the production of HCN and NH3. Finally, the properties and the combustion reactivities of the chars were analyzed. The water and oil content in the coal-water slurry increased the structural ordering degree of chars but decreased the combustion reactivities.

Growth Promotion of Rapeseed (Brassica napus L.) and Blackleg Disease (Leptosphaeria maculans) Suppression Mediated by Endophytic Bacteria

Rapeseed is an important oil crop strongly dependent on high agrochemical inputs. Some pathogens, including Leptosphaeria maculans, cause blackleg disease and can drastically decrease yields. Microbial inoculants seem to be a promising solution to these problems. However, a selection of potent bacterial strains able to improve growth and/or suppress disease is needed. Endophytic bacteria (n = 38) isolated from rapeseed plants with exceptionally good growth were screened for plant growth promoting (PGP) traits and L. maculans antifungal activity. A majority of isolates (35) showed the ability to produce siderophores, 17 isolates solubilized phosphate, and 28 isolates inhibited the growth of L. maculans. The six most promising isolates belonging to Bacillus genera were characterized in detail and compared to two previously published PGP strains. Plant growth measured as total weight and root length of rapeseed seedlings was stimulated by all isolates in comparison to control. The best isolate, 1L6, preliminary identified as Bacillus pumilus showed the highest phosphate solubilization, IAA and HCN production, and growth promotion of plants. Isolates with high antifungal activity in screening showed good potential to suppress disease on plants, with 87% reduction of lesions caused by L. maculans. These strains are good candidates to be explored under field use either solely or in combination.

Characterization and Plant Growth Promoting Properties of a Bacillus sp. Isolated from Maize Roots

Aim: To isolate plant growth promoting Bacillus strains from maize roots and to characterize using molecular methods, the strain with greatest potential for plant growth promotion.
 Place and Duration of Study: Department of Applied Microbiology and Brewing, Nnamdi Azikiwe University, Awka, between February 2019 and March 2020.
 Methodology: The isolation of plant growth promoting rhizobacteria (PGPR) from maize roots was made using Nitrogen Free Bromothymol Blue (NFB) broth. They were screened for Phosphate solubilizing activities on Pikovskaya (PVK) agar. Quantitative determination and solubilization of different types of Phosphates was carried out using Pikovskaya broth. Optimization of factors affecting phosphate such as NaCl concentration, initial pH of the medium, size of inoculum, was done using pvk broth. Evaluation of other plant growth promoting properties were carried out such as IAA, Ammonia, cellulase and HCN production. 
 Results: Eleven Nitrogen fixing bacteria were isolated using NFB broth based on colour change of the medium from green to blue. Test for phosphate solubilization abilities of the organisms revealed that nine of the isolates solubilized phosphate on PVK agar. Organism coded with IS52 gave the least solubilization index of 1.14 while isolate IS19 gave the highest index of 3.4. Isolate IS19 yielded the highest amount of 73.5µg/ml P, while isolate IS30 was the weakest solubilizer in PVK broth, yielding 19.4µg/ml P. The best isolate IS19, produced the plant growth hormone Indole Acetic acid at a concentration of 105.4μg/ml. The organism also gave a positive result for ammonia and cellulase production but did not produce Hydrogen cyanide. It was identified as Bacillus subtilis using the 16S rRNA gene sequencing.
 Conclusion: Bacillus subtilis fixed Nitrogen qualitatively and solubilized insoluble phosphates in addition to other plant growth promoting properties, thus Bacillus subtilis has potential for plant growth promotion, making it an efficient strain for biofertilizer production