Diversity and Antimicrobial Activity of Vietnamese Sponge-Associated Bacteria.

This study aimed to assess the diversity and antimicrobial activity of cultivable bacteria associated with Vietnamese sponges. In total, 460 bacterial isolates were obtained from 18 marine sponges. Of these, 58.3% belonged to Proteobacteria, 16.5% to Actinobacteria, 18.0% to Firmicutes, and 7.2% to Bacteroidetes. At the genus level, isolated strains belonged to 55 genera, of which several genera, such as Bacillus, Pseudovibrio, Ruegeria, Vibrio, and Streptomyces, were the most predominant. Culture media influenced the cultivable bacterial composition, whereas, from different sponge species, similar cultivable bacteria were recovered. Interestingly, there was little overlap of bacterial composition associated with sponges when the taxa isolated were compared to cultivation-independent data. Subsequent antimicrobial assays showed that 90 isolated strains exhibited antimicrobial activity against at least one of seven indicator microorganisms. From the culture broth of the isolated strain with the strongest activity (Bacillus sp. M1_CRV_171), four secondary metabolites were isolated and identified, including cyclo(L-Pro-L-Tyr) (1), macrolactin A (2), macrolactin H (3), and 15,17-epoxy-16-hydroxy macrolactin A (4). Of these, compounds 2-4 exhibited antimicrobial activity against a broad spectrum of reference microorganisms.

Simple stain-free screening method for pectinolytic microorganisms under alkalophilic conditions.

OBJECTIVES: To develop a simple pectin-degrading microorganism screening method. RESULTS: We developed a method utilizing the phenomenon whereby cooling an alkaline agar medium containing pectin causes the agar to become cloudy. This highly simplified method involves culturing the microorganisms on pectin-containing agar medium until colony formation is observed, and subsequent overnight cooling of the agar medium to 4 °C. Using this simple procedure, we successfully identified pectin-degrading microorganisms by observing colonies with halos on the clouded agar medium. We used alkaline pectinase and Bacillus halodurans, which is known to secrete alkaline pectinase, to establish the screening method. We demonstrated the screening of pectin-degrading microorganisms using the developed method and successfully isolated pectin-degrading microorganisms (Paenibacillus sp., Bacillus clausii, and Bacillus halodurans) from a soil sample. CONCLUSIONS: The developed method is useful for identifying pectin-degrading microorganisms.

Isolating, identifying and evaluating of oil degradation strains for the air-assisted microbial enhanced oil recovery process.

Due to the inefficient reproduction of microorganisms in oxygen-deprived environments of the reservoir, the applications of microbial enhanced oil recovery (MEOR) are restricted. To overcome this problem, a new type of air-assisted MEOR process was investigated. Three compounding oil degradation strains were screened using biochemical experiments. Their performances in bacterial suspensions with different amounts of dissolved oxygen were evaluated. Water flooding, microbial flooding and air-assisted microbial flooding core flow experiments were carried out. Carbon distribution curve of biodegraded oil with different oxygen concentration was determined by chromatographic analysis. The long-chain alkanes are degraded by microorganisms. A simulation model was established to take into account the change in oxygen concentration in the reservoir. The results showed that the optimal dissolved oxygen concentration for microbial growth was 4.5~5.5mg/L. The main oxygen consumption in the reservoir happened in the stationary and declining phases of the microbial growth systems. In order to reduce the oxygen concentration to a safe level, the minimum radius of oxygen consumption was found to be about 145m. These results demonstrate that the air-assisted MEOR process can overcome the shortcomings of traditional microbial flooding techniques. The findings of this study can help for better understanding of microbial enhanced oil recovery and improving the efficiency of microbial oil displacement.

Microbiology of secondary infections in Buruli ulcer lesions; implications for therapeutic interventions.

BACKGROUND: Buruli ulcer (BU) is a skin disease caused by Mycobacterium ulcerans and is the second most common mycobacterial disease after tuberculosis in Ghana and Côte d'Ivoire. M. ulcerans produces mycolactone, an immunosuppressant macrolide toxin, responsible for the characteristic painless nature of the infection. Secondary infection of ulcers before, during and after treatment has been associated with delayed wound healing and resistance to streptomycin and rifampicin. However, not much is known of the bacteria causing these infections as well as antimicrobial drugs for treating the secondary microorganism. This study sought to identify secondary microbial infections in BU lesions and to determine their levels of antibiotic resistance due to the prolonged antibiotic therapy required for Buruli ulcer. RESULTS: Swabs from fifty-one suspected BU cases were sampled in the Amansie Central District from St. Peters Hospital (Jacobu) and through an active case surveillance. Forty of the samples were M. ulcerans (BU) positive. Secondary bacteria were identified in all sampled lesions (N = 51). The predominant bacteria identified in both BU and Non-BU groups were Staphylococci spp and Bacilli spp. The most diverse secondary bacteria were detected among BU patients who were not yet on antibiotic treatment. Fungal species identified were Candida spp, Penicillium spp and Trichodema spp. Selected secondary bacteria isolates were all susceptible to clarithromycin and amikacin among both BU and Non-BU patients. Majority, however, had high resistance to streptomycin. CONCLUSIONS: Microorganisms other than M. ulcerans colonize and proliferate on BU lesions. Secondary microorganisms of BU wounds were mainly Staphylococcus spp, Bacillus spp and Pseudomonas spp. These secondary microorganisms were less predominant in BU patients under treatment compared to those without treatment. The delay in healing that are experienced by some BU patients could be as a result of these bacteria and fungi colonizing and proliferating in BU lesions. Clarithromycin and amikacin are likely suitable drugs for clearance of secondary infection of Buruli ulcer.

Integrated use of phosphate-solubilizing Bacillus subtilis strain IA6 and zinc-solubilizing Bacillus sp. strain IA16: a promising approach for improving cotton growth.

Mineral nutrition of crop plants is one of the major challenges faced by modern agriculture, particularly in arid and semi-arid regions. In alkaline calcareous soils, the availability of phosphorus and zinc is critically less due to their fixation and precipitation as complexes. Farmers use fertilizers to fulfill crop requirements, but their efficacy is less, which increases production costs. Plant growth-promoting rhizobacteria (PGPR) can improve the availability of crop nutrients through solubilizing the insoluble compounds of phosphorus and zinc in soil. In the present study, a total of 40 rhizobacterial isolates were isolated from cotton rhizosphere and screened for improving cotton growth through the solubilization of phosphorus and zinc. Out of these 40 isolates, seven isolates (IA2, IA3, IA6, IA7, IA8, IA13, and IA14) efficiently solubilized insoluble rock phosphate while seven isolates (IA10, IA16, IA20, IA23, IA24, IA28, and IA30) were more efficient in solubilizing insoluble zinc oxide. In liquid media, strain IA7 (2.75 µg/mL) solubilized the highest amount of phosphate while the highest concentration of soluble zinc was observed in the broth inoculated with strain IA20 (3.94 µg/mL). Seven phosphate-solubilizing and seven zinc-solubilizing strains were evaluated using jar trial to improve the growth of cotton seedlings, and the results were quite promising. All the inoculated treatments showed improvement in growth parameters in comparison with control. Best results were shown by the combined application of IA6 and IA16, followed by the combination of strains IA7 and IA20. Based on the jar trial, the selected isolates were further characterized by plant growth-promoting characters such as siderophores production, HCN production, ammonia production, and exopolysaccharides production. These strains were identified through 16S rRNA sequencing as Bacillus subtilis IA6 (accession # MN005922), Paenibacillus polymyxa IA7 (accession # MN005923), Bacillus sp. IA16 (accession # MN005924), and Bacillus aryabhattai IA20 (accession # MN005925). It is hence concluded that the integrated use of phosphate-solubilizing and zinc-solubilizing strains as potential inoculants can be a promising approach for improving cotton growth under semi-arid conditions.

A non-invasive soil-based setup to study tomato root volatiles released by healthy and infected roots.

The role of root exudates in mediating plant-microbe interactions has been well documented. However, the function of volatile organic compounds (VOCs) emitted by plant roots has only recently begun to attract attention. This newly recognized relevance of belowground VOCs has so far mostly been tested using systems limited to a two-compartment Petri-dish design. Furthermore, many of the plant-microbe interaction studies have only investigated the effects of microbial VOCs on plant growth. Here, we go two steps further. First we investigated the volatile profile of healthy and pathogen (Fusarium oxysporum) infected tomato roots grown in soil. We then used a unique soil-based olfactometer-choice assay to compare the migration pattern of four beneficial bacteria (Bacillus spp.) towards the roots of the tomato plants. We demonstrate that the blend of root-emitted VOCs differs between healthy and diseased plants. Our results show that VOCs are involved in attracting bacteria to plant roots.

Isolation, selection and evaluation of Bacillus spp. as potential multi-mode probiotics for poultry.

Bacillus based probiotics are becoming relevant as alternatives to antibiotics used in poultry production and in other animal husbandry. This study describes the isolation of 48 Bacillus spp. candidates, from chickens and chicken environments, for use as potential probiotics in poultry production. These isolates, plus a further 18, were tested in a comprehensive in vitro screening regime that was specifically designed to select the best isolates that satisfied multiple modes of action desirable for commercial poultry probiotics. This screening programme involved the evaluation of the ability of the isolates to survive and grow in the limiting conditions of the chicken gastrointestinal tract. Only 11 of the isolates fulfilled these criteria; hence, they were further evaluated for the ability to adhere to epithelial cells, produce extracellular enzymes, and to demonstrate antagonistic activity against selected pathogens of significant importance in poultry production. Of these, a total of 6 isolates were selected, due to their all-round probiotic capability. Identification by 16S RNA sequencing confirmed these isolates as B. subtilis and B. velezensis, identities which are generally regarded as safe. The Bacillus isolates reported in our study exhibit strong all-inclusive probiotic effects and can potentially be formulated as a probiotic preparation for poultry production.

Characterisation of antagonistic Bacillus paralicheniformis (strain EAL) by LC-MS, antimicrobial peptide genes, and ISR determinants.

Plants have their own defense mechanisms such as induced systemic resistance (ISR) and systemic-acquired resistance. Bacillus spp. are familiar biocontrol agents that trigger ISR against various phytopathogens by eliciting various metabolites and producing defense enzyme in the host plant. In this study, B. paralicheniformis (strain EAL) was isolated from the medicinal plant Enicostema axillare. Butanol extract of B. paralicheniformis showed potential antagonism against Fusarium oxysporum compared to control well (sterile distilled water) A liquid chromatography mass spectrometry analysis showed 80 different compounds. Among the 80 compounds, we selected citrulline, carnitine, and indole-3-ethanol based on mass-to-charge ratio, database difference, and resolution of mass spectrum. The synthetic form of the above compounds showed biocontrol activity against F. oxysporum under in vitro condition in combination, not as individual compounds. However, the PCR amplification of 11 antimicrobial peptide genes showed that none of the genes amplified in the strain. B. paralicheniformis inoculation challenged with F. oxysporum on tomato plants enhanced production of defense enzymes such as peroxidase (POD), superoxide dismutase (SOD), phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and proline compared to control plants (without inoculation of B. paralicheniformis) at significant level (p < 0.005). Stem of tomato plants expressed higher POD (2.2-fold), SOD (2.2-fold), PPO (1.9-fold), and PAL (1.3-fold) contents followed by the leaf and root. Elevated proline accumulation was observed in the leaf (1.8-fold) of tomato plants. Thus, results clearly showed potentiality of B. paralicheniformis (EAL) in activation of antioxidant defense enzyme against F. oxysporum-infected tomato plants and prevention of oxidative damage though hydroxyl radicals scavenging activities that suppress the occurrence of wilt diseases.

Bacterial and heavy metal contamination in selected commonly sold herbal medicine in Blantyre, Malawi.

Background: There has been an increase in use of herbal medicine worldwide. It is either used as a stand-alone or complementary therapy to conventional medicine due to past good experience, poverty and family traditions. In Malawi, there are no regulations governing the supply, acquisition, marketing and quality enforcement of herbal medicine. This compromises its safety thereby exposing consumers to avoidable bacteria and heavy metals leading to various adverse health effects. Methods: Cross-sectional laboratory experiments were conducted to determine bacterial and heavy metal contamination of herbal medicine commonly sold in Blantyre, Malawi. A total of 47 samples which were in three formulations namely liquid, powder and tablet were used in the experiments. 29 samples were used for bacterial limit tests and 18 samples were used for heavy metal analysis. Bacterial contamination was determined by streak plate method and biochemical tests while heavy metals were determined by atomic absorption spectroscopy. Descriptive statistics and t-tests were calculated using Microsoft excel and SPSS software programs. Results: Twenty out of the 29 samples (68.9%) were contaminated with Bacillus, coagulase negative Staphylococcus, Klebsiella, Enterobacter, Citrobacter and other-Coliform bacterial species. Most isolated microorganism was Citrobacter spp. (30%), followed by Bacillus spp. (25%). Out of 20 contaminated samples, 75% were contaminated with coliforms. From these 75% which were contaminated with coliforms, 93.3% of them exceeded WHO regulatory limit (103 CFU/g for enterobacteria). Although liquid samples had the highest level of bacterial contaminants, the count was not statistically different from other formulations (P = 0.058). For heavy metals, lead and cadmium were detected and 67% of the samples had lead levels exceeding regulatory limits. Conclusion: Levels of bacterial and lead contamination in herbal medicine from Blantyre markets are far above acceptable limits set by WHO and Canadian guidelines. The use of these herbal medicines is a major risk to the health of consumers.

Diversity of sugarcane root-associated endophytic Bacillus and their activities in enhancing plant growth.

AIMS: Plant tissues are the reservoirs of beneficial and harmful microbes that regulates plant growth. In the present study, we investigated the diversity, function and colonization of sugarcane roots associated with Bacillus spp. METHODS AND RESULTS: A total of 20 Bacillus strains were isolated and identified by 16S rRNA gene sequencing, and their genetic diversity was examined by BOX, ERIC, REP, (GTG)5 PCR techniques. Among all Bacillus isolates, 65% showed indole acetic acid-like compounds production, 50% solubilized phosphorus and 25% of the isolates were able to secrete siderophore. Moreover, all 20 Bacillus isolates showed antifungal activity against eight fungal pathogens and 11 of them (55%) antagonized tomato grey mold. Based on the plant growth-promoting traits and antifungal potential, isolate Y8 was selected for root and plant tissue colonization assays and a greenhouse-level sugarcane growth promotion study. Fluorescence microscopy results confirmed that isolate Y8 has a strong ability to colonize in the sugarcane root and leaves, and the root surface association of Y8 was confirmed by scanning electron microscopy. Furthermore, greenhouse experimental results demonstrated that Y8 has a significant effect on enhancing sugarcane biomass and root length. CONCLUSIONS: Endophytic Bacillus strains have growth-promoting properties and anti-fungal ability that can enhance plant fitness in an eco-friendly manner. SIGNIFICANCE AND IMPACT OF THE STUDY: Endophytic Bacillus strains would be a potential alternative to chemical fertilizer as well as a biocontrol agent in the future.

The central bacterial community in Pericarpium Citri Reticulatae 'Chachiensis'.

The dried and aged pericarps of Citri Reticulatae are condiments and medicinal products in southeast and eastern Asia for hundreds of years, among which Pericarpium Citri Reticulatae 'Chachiensis' (PCR-C) is the premium one with obvious health benefits. In order to explore the microbiota in PCR-C and their relationship with the chemical components during aging, culture-independent methods were applied to investigate PCR-C microbiota for the first time. Here in different PCR-C samples, 16S rRNA gene amplicon sequencing revealed common central bacterial community, which were absent or only accounted for small proportion in fresh pericarps or jute bag controls. Bacillus and Lactococcus were the top two dominant genera in PCR-C with acidic pH (4.06-4.51) and low moisture (11.48%-19.13%). Several OTUs were found to closely relate with specific compositions in essential oils and phenolics, such as d-limonene and nobiletin, which contributed to PCR-C flavor and quality. As the first study to reveal the central bacterial communities in PCR-C, it provides new insights to improve the quality and aging process of traditional Pericarpium Citri Reticulatae, and lays foundation for functional characterization of the microbes within.

Characteristics and Complete Genome Analysis of Bacillus asahii OM18, a Bacterium in Relation to Soil Fertility in Alkaline Soils Under Long-Term Organic Manure Amendment.

Bacillus asahii strain OM18, a bacterium in relation to soil fertility, was isolated from alkaline soils under long-term organic manure application in the North China Plain. B. asahii species play a pivotal role in the promotion of both crop yield and soil fertility via accelerating carbon and phosphorus cycling. However, little is known about the characteristics of B. asahii and its underlying molecular mechanism involved in soil nutrient cycling as well as its potential in promoting crop growth. To this end, we report the characteristics and complete genome analysis of strain OM18, which is relevant to promoting plant growth in phosphorus-deficient alkaline soils. Our results provide a glimpse into the metabolic function of B. asahii OM18.

Antagonistic activity of an epibiotic Bacillus strain SG3 from Gorgonian coral, Junceella juncea (Pallas, 1766).

Marine environment continues to be a huge source of pharmacologically active compounds that cure deadly disease. This research investigates the bioactive efficacy of bacteria isolated from surface of the coral, Junceella juncea (Pallas, 1766). 128 bacterial strains were isolated from the coral Junceella juncea from Tuticorin coast, Gulf of Mannar region, south east coast of India. The strains were tested against selected five human pathogens. Initial screening shows that the strain SG3 was found to exhibit broad spectral activity inhibiting Staplylococcus aureus. Also, twenty other strains were found to be active against various pathogens. Based on 16S rRNA sequencing and phylogenetic identification, the stain SG3 was identified to fell under the genera Bacillus. The ethanol precipitated of the culture broth (SG3) was done and its activity was noted. Mass spectrophotometry (MALDI-TOF) analysis has shown that the mass of the molecules ranged from 1225 Da to 1927 Da. Thus the marine bacteria isolated from corals are a potential source of novel bioactive agents and other natural products. Epibiotic bacteria also direct future isolation of peptide anti-MRSA compounds from marine source.

Culturable endophytic bacteria of Camellia species endowed with plant growth promoting characteristics.

AIM: Tea (Camellia sinensis (L.) O. Kuntze) is an economically important caffeine-containing beverage crop with massive plantation in the Northeast corner of the agroclimatic belt of India. The main aim of the work was to isolate, identify and characterize the native plant growth promoting endophytes associated with tea for future microbe based bioformulation. METHODS AND RESULTS: A total of 129 endophytic bacteria were isolated and characterized for plant growth promoting traits such as indole-3-acetic acid (IAA), phosphate solubilization, ammonia production, biocontrol traits like siderophore and extracellular enzyme production. BOX-PCR fingerprinting was used to differentiate the various bacterial isolates obtained from six different tea species. 16S rRNA sequencing and blast analysis showed that these isolates belonged to different genera, that is, Bacillus, Brevibacterium, Paenibacillus and Lysinibacillus. Lysinibacillus sp. S24 showed the highest phosphate solubilization and IAA acid production efficiency of 268·4 ± 14·3 and 13·5 ± 0·5 µg ml-1 , respectively. Brevibacterium sp. S91 showed the highest ammonia production of 6·2 ± 0·5 µmol ml-1 . Chitinase, cellulase, protease and pectinase activities were shown by 4·6, 34·1, 27·13 and 13·14% of the total isolates, respectively. Similarly, 41% of the total isolates were positive for 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. Further, the potent PGP isolates, S24 and S91 were able to enhance the vegetative parameters such as dry/fresh weight of root and shoot of tea plants in nursery conditions. CONCLUSION: Our findings corroborate that tea endophytic bacteria possess the potential to demonstrate multiple PGP traits both, in vivo and in vitro and have the potential for further large-scale trials. SIGNIFICANCE AND IMPACT OF THE STUDY: The exploration of tea endophytic bacterial community is suitable for the development of bioformulations for an integrated nutrient management and thus sustainable crop production and decreasing the hazardous effects of chemical fertilizers on the environment and human health.

GC-MS analysis of volatile organic compounds from Bambara groundnut rhizobacteria and their antibacterial properties.

Bacterial metabolites have been observed to be important in new drug formulation for both plant, animals and human beings. The aim of this study was to identify the different bioactive compounds found in three rhizobacterial isolates (B. amyloliquefaciens, B. thuringiensis and Bacillus sp.) from the rhizosphere of Bambara groundnut and to assay for their antibacterial properties. Gas chromatography mass spectrometry (GC-MS) was used to carry out the analysis using seven extraction solvents. In the GC-MS analysis, 68 compounds were identified based on peak area percentage, retention time and structure. From the bioactive compounds in B. amyloliquefaciens and B. thuringiensis, the peak area percentage shows that dimethylfuvene from ethyl acetate extraction had the highest relative abundance with 89.11% while Formic acid 2-methylpropyl ester from hexane extraction had the lowest with 6.25%. Others are tridecane, acetic acid butyl ester, paraldehyde, s-(+)-1,2 propanediol, tropone, phthalan and p-xylene with relative abundance of 61.72%, 60.41%, 83.79%, 71.53%, 24.06%, 86.72% and 64.33% respectively. These extracts inhibited the growth of the four test organisms, Bacillus cereus, Pseudomonas aeruginosa, Micrococcus cryophilus and Enterococcus feacalis. Butanol extract from B. amyloliquefaciens had 28 mm zone of inhibition against B. cereus compared to 18 mm and 16 mm by Bacillus sp. and B. thuringiensis respectively. Its zone of inhibition was 24 mm zone against M. cryophilus compared to 12 mm and 19 mm by Bacillus sp. and B. thuringiensis respectively. Butanol extract from B. thuringiensis suppressed E. feacalis and P. aeruginosa having 23 mm and 26 mm zones of inhibition respectively. This was higher compared to Bacillus sp. and B. amyloliquefaciens having 18 mm/15 mm and 21 mm/15 mm against E. feacalis and P. aeruginosa respectively. Hexane and ethyl acetate extract from Bacillus sp. suppressed P. aeruginosa with 12 mm and 17 mm inhibition zones respectively compared to no inhibition zones from hexane extract of B. amyloliquefaciens and B. thuringiensis. Zones of inhibition of 2 mm and 6 mm were observed against P. aeruginosa from ethyl acetate extract of B. amyloliquefaciens and B. thuringiensis respectively. These results suggest that the three isolates are quite rich in the production of bioactive compounds that are also very effective antibacterial agents. These volatile organic compounds are promising compounds for more antibacterial bioactivity development.

Bacterial communities associated with anthracnose symptomatic and asymptomatic leaves of guarana, an endogenous tropical crop, and their pathogen antagonistic effects.

Plants are colonized by diverse microorganisms that can substantially impact their health and growth. Understanding bacterial diversity and the relationships between bacteria and phytopathogens may be key to finding effective biocontrol agents. We evaluated the bacterial community associated with anthracnose symptomatic and asymptomatic leaves of guarana, a typical tropical crop. Bacterial communities were assessed through culture-independent techniques based on extensive 16S rRNA sequencing, and cultured bacterial strains were evaluated for their ability to inhibit the growth of Colletotrichum sp. as well as for enzyme and siderophore production. The culture-independent method revealed that Proteobacteria was the most abundant phylum, but many sequences were unclassified. The emergence of anthracnose disease did not significantly affect the bacterial community, but the abundance of the genera Acinetobacter, Pseudomonas and Klebsiella were significantly higher in the symptomatic leaves. In vitro growth of Colletotrichum sp. was inhibited by 11.38% of the cultured bacterial strains, and bacteria with the highest inhibition rates were isolated from symptomatic leaves, while asymptomatic leaves hosted significantly more bacteria that produced amylase and polygalacturonase. The bacterial isolate Bacillus sp. EpD2-5 demonstrated the highest inhibition rate against Colletotrichum sp., whereas the isolates EpD2-12 and FD5-12 from the same genus also had high inhibition rates. These isolates were also able to produce several hydrolytic enzymes and siderophores, indicating that they may be good candidates for the biocontrol of anthracnose. Our work demonstrated the importance of using a polyphasic approach to study microbial communities from plant diseases, and future work should focus on elucidating the roles of culture-independent bacterial communities in guarana anthracnose disease.

The changes of microbial community and flavor compound in the fermentation process of Chinese rice wine using Fagopyrum tataricum grain as feedstock.

Chinese rice wine (CRW), a unique wine species, has a long history in China. Fagopyrum tataricum grain is a kind of high-quality grain with function in health care. The production of CRW wine with F. tataricum grain is beneficial to the development of new rice wine products. The flavor compounds and microorganisms in F. tataricum grain rice wine were studied. One hundred and seven volatile compounds (including 11 kinds of pyrazines that were rarely detected in wine) were detected and eight organic acids were measured. The microecological diversity in the fermentation process of F. tataricum rice wine was studied. It was found that Bacillus was the main bacterial genus, and the unclassfied_O_Saccharomycetales was the main fungi. Correlation analysis between microorganism and flavor compound shown there are 838 correlations. A total of 108 microbial genera maybe participate in the formation of flavor compounds. In addition, fourteen genera included unclassified_O_Saccharomycetales, Lactococcus, Pediococcus, Aspergillus, Cladosporium, Cochliobolus, Sporidiobolus, Pichia and Saccharomycopsis et al. were screened as functional significant microbiota and built correlation with flavor compounds. This work provides a perspective for bridging the gap between flavor compound and microbial community, and advances our understanding of mechanisms in F. tataricum rice wine fermentation.

An indigenous strain of potassium-solubilizing bacteria Bacillus pseudomycoides enhanced potassium uptake in tea plants by increasing potassium availability in the mica waste-treated soil of North-east India.

AIM: Potassium (K) is one of the three major nutrients required of plant growth and muriate of potash (MoP) is the only recognized chemical fertilizer used in agriculture. In many countries, 100% of the applied MoP is imported costing huge revenue. Application of suitable potassium-solubilizing bacteria (KSB) as biofertilizer could be an integral part of K management in arable soil. The object of this study was to evaluate K-solubilizing ability of a ubiquitous micro-organism as KSB to supplement K in soil. METHODS AND RESULTS: Strain (O-5) was isolated from tea-growing soil and identified as Bacillus pseudomycoides. Phylogenetic analysis revealed that the nearest neighbours of B. pseudomycoides strain O-5 were Bacillus cereus, Bacillus thuringiensis and Bacillus toyonensis. Though the species was first identified in 1998 and is ubiquitous in soil, the role of this group of micro-organisms in nutrient cycling in soil has not been studied before. Strain solubilized 33·32 ± 2·40 µg K ml-1 in mica waste (MW; muscovite type mineral)-amended broth after 7 days incubation at 30 ± 1°C. In a soil microcosm study under laboratory condition, B. pseudomycoides strain O-5 increased K availability by 47·0 ± 7·1 mg kg-1 after 105 days incubation, while the strain released 104·9 ± 15·3 mg K kg-1 in MW-treated soil. In this study, application of isolated B. pseudomycoides with MW significantly increased K availability in soil, and that in turn facilitated K uptake by tea plants. CONCLUSION: Based on the data, it could be inferred that B. pseudomycoides could mobilize K from bound form in soil and can be utilized as K-solubilizing biofertilizer especially in combination with MW for supplementing K in soil. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacillus pseudomycoides strain O-5 has potential to be used as K-solubilizing biofertilizer in agriculture.

Characterization of Hydrocarbon-Degrading Bacteria in Constructed Wetland Microcosms Used to Treat Crude Oil Polluted Water.

Ten plant species were grown in constructed wetlands (CWs) to remediate water containing 2% (w/v) crude oil. The plant species with better growth and biomass production were Typha latifolia and Cyperus laevigatus, and they were significantly correlated (R2 = 0.91) with hydrocarbon degradation. From T. latifolia and C. laevigatus, 33 hydrocarbon-degrading bacterial strains were isolated from the rhizosphere, and root and shoot interiors. More diversified bacteria were found in the rhizosphere and endosphere of C. laevigatus than those of T. latifolia. The predominant cultural hydrocarbon-degrading bacteria were shown to belong to the genera Pseudomonas, Acinetobacter and Bacillus. In addition to genes involved in hydrocarbon degradation, most of the bacteria displayed multiple plant growth promoting (PGP) activities. This study suggests the importance of selecting suitable bacterial strains with hydrocarbon degradation and PGP activities for improving the efficacy of CWs used in remediating water contaminated with crude oil.

Biosurfactant production and hydrocarbon degradation activity of endophytic bacteria isolated from Chelidonium majus L.

BACKGROUND: The process of plant growth in the contaminated environment is often inhibited and entails the neutralization of harmful compounds. To reduce the negative impact of harmful compounds microorganisms produce unique compounds called biosurfactants. This paper describes the potential of culturable endophytic microorganisms from synanthropic plant-Chelidonium majus L. for the production of biosurfactants, as indirect plant promoting factors as well as their degradation activity. Emulsifying activity and degradation potential of tested strains were assessed by cultivation of isolates in the presence of diesel oil and waste engine oil. RESULTS: Ten bacterial strain were isolated. Analysis of emulsifying activity revealed that all isolates possessed the ability for biosurfactant production. However, one of the isolated endophytes-2A, identified as Bacillus pumilus, exhibited the highest emulsifying activity (OD500 1.96). The same strain has shown very high degradation potential, both for diesel oil and waste engine oil hydrocarbons. Results obtained with the Phytotoxkit tests revealed that the addition of biosurfactant isolated from B. pumilus 2A strain resulted in stimulation of seed germination in soil contaminated with diesel oil (137%) and waste engine oil (120%). Positive impact of the biosurfactant produced by B. pumilus 2A on the growth of Sinapis alba in hydrocarbons contaminated soil was demonstrated. CONCLUSIONS: The endophytic strain identified as Bacillus pumilus 2A produce biosurfactant that is able to act as plant-growth promoting agent. Endophytic bacteria isolated from Chelidonium majus L. exhibit potential for hydrocarbons degradation and biosurfactant production. These properties provide promising perspectives for application of biosurfactants as potential agents for bioremediation of environment contaminated with hydrocarbons.