Identification and characterization of thermophilic amylase producing bacterial isolates from the brick kiln soil

The present experiment was designed to isolate bacterial strains from the brick kiln soil and to check the activity and enzyme kinetics of amylase from these isolates. The bacterial colonies were isolated from soil samples through the serial dilution method. The bacterial isolates were identified through morphological, electron microscopic and molecular analysis. The 16S ribosomal RNA sequences of the isolates IR-1, IR-2, IR-3, IR-8, and IR-9 showed high similarities with Bacillus tequilensis, Bacillus paramycoides, Proteus alimentorum, Bacillus wiedmannii, and Pseudomonas aeruginosa, respectively. All of the bacterial isolates showed a positive catalase activity except IR-9. Furthermore, the isolates showed variable antagonistic effects against different bacterial pathogens. All of the strains produced indole acetic acid (IAA), and the concentrations increased in the presence of tryptophan application. The isolates showed the amylase enzyme activity and maximum activity of isolates was achieved in 4% starch concentration. The IR-9 isolate showed the highest amylase activity of 5.9 U/ml. The V_max values of the extracellular amylase from different bacterial isolates ranged between 12.90 and 50.00 IU ml⁻¹. The lowest K_m value of 6.33 mg starch was recorded for IR-8 and the maximum K_cat value of 2.50 min⁻¹ was observed for IR-3. The amylase activity of the isolates was significantly affected by a range of different incubation time, temperature, and pH values. Further tests are required before the potential utilization of these isolates for amylase production, and in the biopesticide and biofertilizer applications.     

Microbial plankton across Drake Passage

We determined biomass and activity of microbial plankton across the Polar Front (PF) in Drake Passage during January 1994. Temperature was around 0°C south and between 3 and 5°C north of the PF. Both biomass and activities of microorganisms were significantly lower in the Antarctic waters south of the PF than in the sub-Antarctic waters north of it. Thus, values of chlorophyll a, integrated between 0 and 200 m, reached 150 mgm⁻² north, but only 25 mg m⁻² south of the PF. Likewise, bacteria varied between 10¹⁴ and 4×10¹³ cells m⁻². However, the abundance of heterotrophic nanoflagellates was extremely low throughout Drake Passage (around 3×10¹⁰ cells m⁻²). Bacterial doubling times were long (mean of 25 days). Bacterivory was estimated from the abundance of predators and prey and from temperature. The grazing impact on bacterioplankton biomass was insignificant (less that 0.05% per day) and low on bacterial heterotrophic production (15% per day). Neither biomass nor the activities of microorganisms were found to increase at the PF. The microbial food web was uncoupled and the bacteria did not seem to be controlled by predation.     

Dinitrogen fixation in a unicellular chlorophyll d-containing cyanobacterium

Marine cyanobacteria of the genus Acaryochloris are the only known organisms that use chlorophyll d as a photosynthetic pigment. However, based on chemical sediment analyses, chlorophyll d has been recognized to be widespread in oceanic and lacustrine environments. Therefore it is highly relevant to understand the genetic basis for different physiologies and possible niche adaptation in this genus. Here we show that unlike all other known isolates of Acaryochloris, the strain HICR111A, isolated from waters around Heron Island, Great Barrier Reef, possesses a unique genomic region containing all the genes for the structural and enzymatically active proteins of nitrogen fixation and cofactor biosynthesis. Their phylogenetic analysis suggests a close relation to nitrogen fixation genes from certain other marine cyanobacteria. We show that nitrogen fixation in Acaryochloris sp. HICR111A is regulated in a light–dark-dependent fashion. We conclude that nitrogen fixation, one of the most complex physiological traits known in bacteria, might be transferred among oceanic microbes by horizontal gene transfer more often than anticipated so far. Our data show that the two powerful processes of oxygenic photosynthesis and nitrogen fixation co-occur in one and the same cell also in this branch of marine microbes and characterize Acaryochloris as a physiologically versatile inhabitant of an ecological niche, which is primarily driven by the absorption of far-red light.     

Rapid TaqMan-Based Quantification of Chlorophyll d-Containing Cyanobacteria in the Genus Acaryochloris

Reports of the chlorophyll (Chl) d-containing cyanobacterium Acaryochloris have accumulated since its initial discovery in 1996. The majority of this evidence is based on amplification of the gene coding for the 16S rRNA, and due to the wide geographical distribution of these sequences, a global distribution of Acaryochloris species was suggested. Here, we present a rapid, reliable, and cost-effective TaqMan-based quantitative PCR (qPCR) assay that was developed for the specific detection of Acaryochloris species in complex environmental samples. The TaqMan probe showed detection limits of ∼10 16S rRNA gene copy numbers based on standard curves consisting of plasmid inserts. DNA from five Acaryochloris strains, i.e., MBIC11017, CCMEE5410, HICR111A, CRS, and Awaji-1, exhibited amplification efficiencies of >94% when tested in the TaqMan assay. When used on complex natural communities, the TaqMan assay detected the presence of Acaryochloris species in four out of eight samples of crustose coralline algae (CCA), collected from temperate and tropical regions. In three out of these TaqMan-positive samples, the presence of Chl d was confirmed via high-performance liquid chromatography (HPLC), and corresponding cell estimates of Acaryochloris species amounted to 7.6 × 10¹ to 3.0 × 10³ per mg of CCA. These numbers indicate a substantial contribution of Chl d-containing cyanobacteria to primary productivity in endolithic niches. The new TaqMan assay allows quick and easy screening of environmental samples for the presence of Acaryochloris species and is an important tool to further resolve the global distribution and significance of this unique oxyphototroph.     

A <Emphasis Type="Italic">Bacillus subtilis</Emphasis> strain HPC248 from an effluent treatment plant with antimicrobial activity

This study reports the identification and demonstration of an organism with antimicrobial activity isolated from activated biomass of an effluent treatment plant (ETP) treating wastewater containing pesticides. While assessing the heterotrophic diversity of biomass collected from ETP, clear zones were observed on Luria Bertani plates. The bacterial isolate producing the zone as well as the bacterial cells surrounding the zone were isolated and purified by sub-culturing. Both isolates were identified by partial sequencing of the 16S rDNA clone. Presence of antimicrobial activity was demonstrated against various laboratory strains, isolated from different treatment plants and also against waterborne pathogens. The isolate that produced antimicrobial activity was identified as Bacillus subtilis strain HPC248 and the sensitive strain was identified as Bacillus sphaericus strain HPC249.     

Isolation and molecular characterization of Bacillus thuringiensis found in soils of the Cerrado region of Brazil,and their toxicity to Aedes aegypti larvae

This study investigated the potential of Bacillus thuringiensis isolates obtained in the Cerrado region of the Brazilian state of Maranhão for the biological control of Aedes aegypti larvae. The isolates were obtained from soil samples and the identification of the B. thuringiensis colonies was based on morphological characteristics. Bioassays were run to assess the pathogenicity and toxicity of the different strains of the B. thuringiensis against third-instar larvae of A. aegypti. Protein profiles were obtained by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Polymerase chain reaction assays were used to detect the toxin genes found in the bacterial isolates. Overall, 12 (4.0%) of the 300 isolates obtained from 45 soil samples were found to present larvicidal activity, with the BtMA-104, BtMA-401 and BtMA-560 isolates causing 100% of mortality. The BtMA-401 isolate was the most virulent, with the lowest median lethal concentration (LC₅₀) (0.004 × 10⁷ spores/mL), followed by the Bacillus thuringiensis var. israelensis standard (0.32 × 10⁷ spores/mL). The protein profiles of BtMA-25 and BtMA-401 isolates indicated the presence of molecular mass consistent with the presence of the proteins Cry4Aa, Cry11Aa and Cyt1, similar to the profile of Bacillus thuringiensis var. israelensis IPS-82. Surprisingly, however, none of the cry and cyt genes analyzed were amplified in the isolate BtMA-401. The results of the present study revealed the larvicidal potential of B. thuringiensis isolates found in the soils of the Cerrado region from Maranhão, although further research will be necessary to better elucidate and describe other genes associated with the production of insecticidal toxins in these isolates.     

Chlorophyll d and Acaryochloris marina: current status

The discovery of the chlorophyll d-containing cyanobacterium Acaryochloris marina in 1996 precipitated a shift in our understanding of oxygenic photosynthesis. The presence of the red-shifted chlorophyll d in the reaction centre of the photosystems of Acaryochloris has opened up new avenues of research on photosystem energetics and challenged the unique status of chlorophyll a in oxygenic photosynthesis. In this review, we detail the chemistry and role of chlorophyll d in photosynthesis and summarise the unique adaptations that have allowed the proliferation of Acaryochloris in diverse ecological niches around the world.     

Molecular identification and in vitro screening of antagonistic bacteria from agricultural byproduct compost: Effect of compost on development and photosynthetic efficiency of tomato plant

The dynamics of mesophilic and thermophilic bacterial population of compost was studied. The bacteria population in the compost ranged from 10⁹ to 10⁵ CFU g^?1 and was found to be maximum during mesophilic phase, and then decreased during the thermophilic, the cooling and maturation phases. Assessment of culturable bacteria by 16S rDNA revealed phylogenetic lineage of different polymorphic class bacilli, γ, β-proteobacteria and actinobacteria. Bacterial isolates produced extracellular enzymes: proteases, cellulase, xylanase, pectinase, tannase and amylase. Among them, mesophilic bacteria exhibited xylanolytic (81.25 %) and cellulolytic (63 %) activity. Thermophilic bacteria showed cellulolytic (75 %) and xylanolytic (66.6 %) activity, but a few isolates also produced tannase and pectinase. All bacterial isolates were observed to cause inhibition of three isolates of Bacillus pumilus and one isolate each of Staphylococcus sciuri and Kocuria sp. The physiological effect of compost on shoot length, leaf size and fruit maturation of tomato have been evaluated; the compost (75 g/pot) improved these parameters as compared to known compost (SOM). The efficacy of compost and SOM on photochemistry of tomato leaves was studied, based on imaging-PAM of the chlorophyll fluorescence parameters. F_v/F_m and electron transport rate (ETR) were increased significantly in compost (75 g) amended pot within 30 days of growth. Likewise, highest Y (II) of photosystem II (PS II) yield was found in compost (75 g) pot in 15 days. The findings of this study proved that the compost comprising of various bacteria involved in degradation of substrates was found to be beneficial for enhancement of tomato growth and development.     

Characterization of Bacillus thuringiensis isolated from soils in the Jazan region of Saudi Arabia,and their efficacy against Spodoptera littoralis and Aedes aegypti larvae

Pest control in Saudi Arabia depends on applying chemical insecticides, which have many undesirable considerations and impacts on the environment. Therefore, the aim of this study was to isolate Bacillus thuringiensis from different rhizosphere soil samples in the Jazan region for the biological control of Spodoptera littoralis and Aedes aegypti larvae. The samples were collected from the rhizosphere of different plants located in eight agricultural areas in Jazan, Saudi Arabia. Out of 100 bacterial isolates, four bacterial isolates belonging to Bacillus species were selected namely JZ1, JZ2, JZ3, and JZ4, and identified using classical bacteriological and molecular identification using 16S rRNA. JZ1 and JZ2 isolates were identified as Bacillus thuringiensis. SDS-PAGE analysis and the detection of the Cry1 gene were used to describe the two isolates JZ1 and JZ2 in comparison to Bacillus thuringiensis reference strain Kurstaki HD1 (BTSK) were revealed that slightly different from each other due to the place of their isolation and namely Khlab JZ1 and Ayash JZ2. The EC₅₀ of JZ1 and JZ2 isolates, BTSK, and the commercial biopesticide DiPEL 6.4 DF against the second-instar larvae of Aedes aegypti were 207, 932, 400, and 500 ppm respectively, while EC₅₀ against first-instar larvae of Spodoptera littoralis were 193.93, 589.7, 265.108, and 342.9, ppm respectively. Isolate JZ1 recorded the highest mortality while JZ2 isolate gave the lowest mortality. It can be concluded that the local isolate of JZ1 and JZ2 can be developed for bio formulations to be used in Spodoptera littoralis and Aedes aegypti biological control programs.     

Culture-Dependent and Culture-Independent Analysis of Hydrocarbonoclastic Microorganisms Indigenous to Hypersaline Environments in Kuwait

The halophilic, hydrocarbonoclastic bacteria and archaea inhabiting two hypersaline coastal areas in Kuwait, one in the north and the other in the south, were counted and characterized. Environmental parameters in both areas were similar, with the exception of the soil organic carbon content, which was in the north higher than in the south. The hydrocarbonoclastic bacterial and haloarchaeal numbers and identities as analyzed using nutrient media of various salinities were similar in soil and pond water samples from both areas. The bacterial species recorded by this culture-dependent method belonged to the genera Halomonas, Chromohalobacter, Marinobacter, Exiguobacterium, Stenotrophomonas, Pseudomonas, Salinivibrio, and Bacillus. The haloarchaeal species belonged to the genera Haloferax and Halobacterium. When analyzed by fingerprinting of their amplified genomic DNA followed by sequencing of the electrophoresis-resolved bands, the same environmental samples revealed a different microbial composition. Bacterial phylotypes recorded by this culture-independent method were affiliated with the genera Ochrobactrum, Stenotrophomonas, Rhodococcus, and “Halomicrobium,” whereas the archaeal phylotypes were affiliated with Halorussus, Halomicrobium, and Halorientalis. The observed diversity and composition similarity of the hydrocarbonocalastic microflora in both hypersaline areas suggest an effective potential for oil mineralization therein. This potential has been confirmed experimentally.     

Bacillus aryabhattai FACU: A promising bacterial strain capable of manipulate the glyphosate herbicide residues

Glyphosate is a commonly used organophosphate herbicide that has an adverse impact on humans, mammals and soil microbial ecosystems. The redundant utilize of glyphosate to control weed growth cause the pollution of the soil environment by this chemical. The discharge of glyphosate in the agricultural drainage can also cause serious environmental damage and water pollution problems. Therefore, it is important to develop methods for enhancing glyphosate degradation in the soil through bioremediation. In this study, thirty bacterial isolates were selected from an agro-industrial zone located in Sadat City of Monufia Governorate, Egypt. The isolates were able to grow in LB medium supplemented with 7.2 mg/ml glyphosate. Ten isolates only had the ability to grow in a medium containing different concentrations of glyphosate (50, 100, 150, 200 and 250 mg/ml). The FACU3 bacterial isolate showed the highest CFU in the different concentrations of glyphosate. The FACU3 isolate was Gram-positive, spore-forming and rod-shape bacteria. Based on API 50 CHB/E medium kit, biochemical properties and 16S rRNA gene sequencing, the FACU3 isolate was identified as Bacillus aryabhattai. Different bioinformatics tools, including multiple sequence alignment (MSA), basic local alignment search tool (BLAST) and primer alignment, were used to design specific primers for goxB gene amplification and isolation. The goxB gene encodes FAD-dependent glyphosate oxidase enzyme that responsible for biodegradation process. The selected primers were successfully used to amplify the goxB gene from Bacillus aryabhattai FACU3. The results indicated that the Bacillus aryabhattai FACU3 can be utilized in glyphosate-contaminated environments for bioremediation. According to our knowledge, this is the first time to isolate of FAD-dependent glyphosate oxidase (goxB) gene from Bacillus aryabhattai.     

Toxic-Metabolite-Producing Bacteria and Fungus in an Indoor Environment

Toxic-metabolite-emitting microbes were isolated from the indoor environment of a building where the occupant was suffering serious building-related ill-health symptoms. Toxic substances soluble in methanol and inhibitory to spermatozoa at <10 μg (dry weight) ml⁻¹ were found from six bacterial isolates and one fungus. The substances from isolates of Bacillus simplex and from isolates belonging to the actinobacterial genera Streptomyces and Nocardiopsis were mitochondriotoxic. These substances dissipated the mitochondrial membrane potential (Δψ) of boar spermatozoa. The substances from the Streptomyces isolates also swelled the mitochondria. The substances from isolates of Trichoderma harzianum Rifai and Bacillus pumilus damaged the cell membrane barrier function of sperm cells.     

Alleviation of Drought Stress in Turfgrass by the Combined Application of Nano-compost and Microbes from Compost

Drought stress is a key environmental factor limiting the growth and productivity of plants. Turfgrasses are often affected by drought in north China due to water shortage. In the present study, the impact of nanosized compost either alone or in combination with drought tolerant isolates from compost on turfgrass response to drought was investigated. Municipal solid waste (MSW) compost was processed into nanosized particles and added in turfgrass soil. Microorganisms in the MSW compost were screened for drought stress tolerance using increasing concentrations of polyethylene glycol (PEG 6000). Festuca arundinacea Schreb. plants were inoculated with this mixture and exposed to drought stress by reducing the amount of water added at vegetative growth stage. The drought-tolerant isolates from compost were identified as Bacillus cereus, Lysinibacillus sp. and Rhodotorula glutinis. Our results revealed that nanocompost and microbial inoculation minimized the drought stress-imposed effects significantly increasing shoot biomass, root biomass, and chlorophyll content. Similarly, nanocompost-treated and inoculated seedlings showed higher levels of antioxidant enzymes and lower MDA content compared to nontreated control under drought stress. The combination of nano-sized compost and microbial inoculation were more efficient than nanocompost alone in terms of influencing growth and physiological status of the seedlings under drought stress. Our data suggest that nanocompost combined with drought-tolerant isolates may enhance drought tolerance in turfgrass by promoting plant growth and increasing the capacity to eliminate toxic reactive oxygen species (ROS).     

Heterodera glycines in Indiana: II. Morphology of Geographical Isolates

Although much morphometric overlap occurs among five geographical isolates of Heterodera glycines in Indiana, significant differences in means exist among the isolates for various comparisons of second-stage juveniles. By using combinations of means, most of the isolates can be distinguished from the rest: e.g., the Vanderburgh County isolate (southern Indiana) has the longest esophagus, tail, and tail terminus; the Vigo County isolate (also from the south) has the shortest esophagus; the White County isolate (northern Indiana) has the shortest tail and tail terminus and the greatest total length; the Benton County isolate (north) is the shortest. Morphological similarities and differences do not appear to be coordinated with reproductive behavioral patterns we observed in the northern versus the southern isolates.     

Isolation of facultatively anaerobic soil bacteria from Ny-Ålesund, Svalbard

Anaerobic conditions in soil commonly occur even in upland environments. Physiological and biogeochemical properties of individual anaerobic bacteria, however, have been poorly understood due to difficulties in culture. This study aimed to isolate anaerobic bacteria in the Arctic tundra soil and to identify their physiological characteristics. Anaerobic culture and 16S rRNA gene sequence-based phylogenetic analysis showed that total 33 bacterial strains were affiliated with 15 species from the following 8 genera: Bacillus, Carnobacterium, Clostridium, Paenibacillus, and Trichococcus (Firmicutes), Pseudomonas and Rahnella (Gamma-proteobacteria), and Cellulomonas (Actinobacteria). All isolates were identified as facultatively anaerobic bacteria; this finding might be partially attributed to the characteristics of sampling sites, which temporarily developed anaerobic conditions because of the presence of stagnant melting snow. Six of the 33 bacterial strains were revived subsequently from glycerol stocks held ?80 °C, and these were used for the physiological study: four isolates from Firmicutes, one isolate from Gamma-proteobacteria, and one isolate from Actinobacteria. Five isolates except KOPRI 80146 (Bacillus sp.) could grow at either 4 or 10 °C within a week. All six isolates showed cellulase or protease activities at 10 or 15 °C. Endospores were observed from four isolates belonging to Firmicutes. These physiological characteristics may contribute to the survival of these organisms at low temperatures and to their involvement in biogeochemical cycles in the tundra soil. These isolates may be used for further detailed studies for identifying their cold adaptation mechanisms and ecological roles in the Arctic.     

蕙兰病株根部内生细菌种群变化

为了研究褐斑病与蕙兰根部内生细菌群落结构和多样性的关联,从野生蕙兰健株和褐斑病株根部分离出内生细菌112株,采用核糖体DNA扩增片段限制性酶切分析(ARDRA),研究了健株和病株内生细菌多样性与群落结构。将内生细菌纯培养物扩增近全长的16S rDNA,并用ARDRA (Amplified Ribosomal DNA Restriction Analysis) 对所分离的菌株进行分型,根据酶切图谱的差异,将健株中的内生细菌分成8个ARDRA型,病株分成13个ARDRA型。并选取代表性菌株进行16S rDNA序列测定。结果表明,健株分离出内生细菌6个属,优势菌群为Bacillus;病株分离出11个属,优势菌群为 Mitsuaria和Flavobacterium。通过回接兰花植物和初步拮抗实验发现,从病株分离出的H5号菌株 (Flavobacterium resistens)使兰花产生病症,而健株中的B02 (Bacillus cereus) 和B22号菌株 (Burkholderia stabilis) 对菌株H5有拮抗作用。     

Two new subspecies of Bacillus thuringiensis isolated in Japan: Bacillus thuringiensis subsp. kumamotoensis (serotype 18) and Bacillus thuringiensis subsp. tochigiensis (serotype 19)

Bacteriological and serological characteristics of three Bacillus thuringiensis isolates obtained in Japan were investigated. They formed typical rhomboidal parasporal inclusions but flagellar (H) antigens of these isolates were different from those of the known 17 H serotypes of B. thuringiensis. The three isolates were divided into two new serotypes (serotypes 18 and 19). The serotype 18 isolate (3–71) produced thermostable exotoxin and the inclusions of this isolate were toxic to larvae of the silkworm, Bombyx mori, but nontoxic to larvae of the mosquito, Aedes aegypti. The other isolate (119-72) belonging to serotype 18 produced inclusions nontoxic to larvae of B. mori and A. aegypti and did not produce thermostable exotoxin. However, other bacteriological properties of the isolate 119-72 were similar to those of the isolate 3–71. The serotype 19 isolate (117-72) produced inclusions nontoxic to larvae of B. mori and A. aegypti and did not produce thermostable exotoxin. Acid production from saccharose and the production of brownish purple pigment were observed in the two serotype 18 isolates, while neither of them was observed in the serotype 19 isolate. In other 29 biochemical properties tested, there was no difference among the three isolates. Based on these characteristics, the following two subspecies names are proposed: Bacillus thuringiensis subsp. kumamotoensis (serotype 18) for the type strain 3–71 and Bacillus thuringiensis subsp. tochigiensis (serotype 19) for the type strain 117-72.     

Endolithic chlorophyll d-containing phototrophs

Cyanobacteria in the genus Acaryochloris are the only known oxyphototrophs that have exchanged chlorophyll a (Chl a) with Chl d as their primary photopigment, facilitating oxygenic photosynthesis with near infrared (NIR) light. Yet their ecology and natural habitats are largely unknown. We used hyperspectral and variable chlorophyll fluorescence imaging, scanning electron microscopy, photopigment analysis and DNA sequencing to show that Acaryochloris-like cyanobacteria thrive underneath crustose coralline algae in a widespread endolithic habitat on coral reefs. This finding suggests an important role of Chl d-containing cyanobacteria in a range of hitherto unexplored endolithic habitats, where NIR light-driven oxygenic photosynthesis may be significant.     

Impact of Bacterial Biomass on Contaminant Sorption and Transport in a Subsurface Soil

The objective of this study was to investigate the impact of bacterial biomass on the sorption and transport of three solutes (quinoline, naphthalene, and ⁴⁵Ca) in a subsurface soil. Miscible displacement techniques were employed to measure sorption and transport of the above compounds during steady, saturated water flow in sterile and/or bacterium-inoculated soil columns. The soil was inoculated with either a quinoline-degrading bacterium, Pseudomonas sp. 3N3A isolate, or its mutant isolate, B53, which does not degrade quinoline. In soil columns inoculated with the B53 and 3N3A isolates, quinoline sorption was reduced by about 60 and 20%, respectively. In contrast, ⁴⁵Ca sorption was minimally reduced, which indicated that biomass did not significantly alter the cation-exchange capacity of the soil. Biomass impacts on sorption were solute specific, even when the sorption mechanism for both quinoline and ⁴⁵Ca was similar. Thus, the differential response is attributed to biomass-induced changes in quinoline speciation; an increase in pH at the sorbent-water interface would result in a larger proportion of the neutral species and a decrease in sorption. Sorption of naphthalene was reduced by about 30%, which was attributed to accessibility of hydrophobic regions. Minimal biosorption of all solutes indicated negligible biofacilitated transport. Alteration of the soil surfaces upon addition of bacterial biomass reduced sorption of quinoline and naphthalene, thereby enhancing transport.     

The invasive Sorghum halepense harbors endophytic N2-fixing bacteria and alters soil biogeochemistry

Exotic plants invading new habitats frequently initiate broad changes in ecosystem functioning. Sorghum halepense is an invasive grass capable of growing in nitrogen (N)-poor tallgrass prairie soils that creates near monocultures in once phylogenetically diverse-communities. The biogeochemistry of soils invaded by S. halepense was compared to that of un-invaded native prairie soils. Invaded soils contained two to four times greater concentrations of alkaline metals, micronutrients, and essential plant nutrients than native prairie soils. The notable exception was Ca⁺², which was always significantly lower in invaded soils. The N-content of S. halepense above-ground biomass was 6.4 mg g^?1 (320 mg N plant^?1) and suggested a supplemental N source supporting plant growth. Altered soil biogeochemistry in invaded areas coupled with high above-ground biomass in N-poor soils suggested N₂-fixing activity associated with S. halepense. Nitrogenase activity of plant tissues indicated that N₂-fixation was occurring in, and largely restricted to, S. halepense rhizomes and roots. A culture approach was used to isolate these N₂-fixing bacteria from plant tissues, and 16S rRNA gene sequencing was used to identify these bacterial isolates. Nitrogenase activity of bacterial isolates indicated several were capable of N₂-fixation. In addition to N₂-fixation, other roles involved in promoting plant growth, namely mobilizing phosphorus and iron chelation, are known for closest matching relatives of the bacterial isolates identified in this work. Our results indicate that these plant growth-promoting bacteria may enhance the ability of S. halepense to invade and persist by altering fundamental ecosystem properties via significant changes in soil biogeochemistry.