Sole Nitrogen Source For Growth Research Articles

Factors Influencing the Nitrogen-Source Dependent Flucytosine Resistance in Cryptococcus Species.

Flucytosine (5-FC) is an antifungal agent commonly used for treatment of cryptococcosis and several other systemic mycoses. In fungi, cytosine permease and cytosine deaminase are known major players in flucytosine resistance by regulating uptake and deamination of 5-FC, respectively. Cryptococcus species have three paralogs each of cytosine permease (FCY2, FCY3, and FCY4) and cytosine deaminase (FCY1, FCY5 and FCY6). As in other fungi, we found FCY1 and FCY2 to be the primary cytosine deaminase and permease gene, respectively, in C. neoformans H99 (VNI), C. gattii R265 (VGIIa) and WM276 (VGI). However, when various amino acids were used as the sole nitrogen source, C. neoformans and C. gattii diverged in the function of FCY3 and FCY6. Though there was some lineage-dependent variability, the two genes functioned as the secondary permease and deaminase, respectively, only in C. gattii when the nitrogen source was arginine, asparagine, or proline. Additionally, the expression of FCY genes, excluding FCY1, was under nitrogen catabolic repression in the presence of NH4. Functional analysis of GAT1 and CIR1 gene deletion constructs demonstrated that these two genes regulate the expression of each permease and deaminase genes individually. Furthermore, the expression levels of FCY3 and FCY6 under different amino acids corroborated the 5-FC susceptibility in fcy2Δ or fcy1Δ background. Thus, the mechanism of 5-FC resistance in C. gattii under diverse nitrogen conditions is orchestrated by two transcription factors of GATA family, cytosine permease and deaminase genes. IMPORTANCE 5-FC is a commonly used antifungal drug for treatment of cryptococcosis caused by Cryptococcus neoformans and C. gattii species complexes. When various amino acids were used as the sole nitrogen source for growth, we found lineage dependent differences in 5-FC susceptibility. Deletion of the classical cytosine permease (FCY2) and deaminase (FCY1) genes caused increased 5-FC resistance in all tested nitrogen sources in C. neoformans but not in C. gattii. Furthermore, we demonstrate that the two GATA family transcription factor genes GAT1 and CIR1 are involved in the nitrogen-source dependent 5-FC resistance by regulating the expression of the paralogs of cytosine permease and deaminase genes. Our study not only identifies the new function of paralogs of the cytosine permease and deaminase and the role of their regulatory transcription factors but also denotes the differences in the mechanism of 5-FC resistance among the two etiologic agents of cryptococcosis under different nitrogen sources.

Characterization of nitrilases from Variovorax boronicumulans that functions in insecticide flonicamid degradation and β-cyano-L-alanine detoxification.

To characterize the functions of nitrilases of Variovorax boronicumulans CGMCC 4969 and evaluate flonicamid (FLO) degradation and β-cyano-L-alanine (Ala(CN)) detoxification by this bacterium. Variovorax boronicumulans CGMCC 4969 nitrilases (NitA and NitB) were purified, and substrate specificity assay indicated that both of them degraded insecticide FLO to N-(4-trifluoromethylnicotinoyl)glycinamide (TFNG-AM) and 4-(trifluoromethyl)nicotinol glycine (TFNG). Ala(CN), a plant detoxification intermediate, was hydrolysed by NitB. Escherichia coli overexpressing NitA and NitB degraded 41.2 and 93.8% of FLO (0.87 mmol·L-1 ) within 1h, with half-lives of 1.30 and 0.25 h, respectively. NitB exhibited the highest nitrilase activity towards FLO. FLO was used as a substrate to compare their enzymatic properties. NitB was more tolerant to acidic conditions and organic solvents than NitA. Conversely, NitA was more tolerant to metal ions than NitB. CGMCC 4969 facilitated FLO degradation in soil and surface water and utilized Ala(CN) as a sole nitrogen source for growth. CGMCC 4969 efficiently degraded FLO mediated by NitA and NitB; NitB was involved in Ala(CN) detoxification. This study promotes our understanding of versatile functions of nitrilases from CGMCC 4969 that is promising for environmental remediation.

Vibrio nitrifigilis sp. nov., a marine nitrogen-fixing bacterium isolated from the lagoon sediment of an islet inside an atoll.

A nitrogen-fixing isolate of facultatively anaerobic, marine bacterium, designated strain NFV-1T, was recovered from the lagoon sediment of Dongsha Island, Taiwan. It was a Gram-negative rod which exhibited motility with monotrichous flagellation in broth cultures. The strain required NaCl for growth and grew optimally at about 25-35°C, 3% NaCl and pH 7-8. It grew aerobically and could achieve anaerobic growth by fermenting D-glucose or other carbohydrates as substrates. NH4Cl could serve as a sole nitrogen source for growth aerobically and anaerobically, whereas growth with N2 as the sole nitrogen source was observed only under anaerobic conditions. Cellular fatty acids were predominated by C16:1 ω7c, C16:0, and C18:1 ω7c. The major polar lipids consisted of phosphatidylethanolamine and phosphatidylserine. Strain NFV-1T had a DNA G + C content of 42.5mol%, as evaluated according to the chromosomal DNA sequencing data. Analyses of sequence similarities and phylogeny based on the 16S rRNA genes, together with the housekeeping genes, gyrB, ftsZ, mreB, topA and gapA, indicated that the strain formed a distinct species-level lineage in the genus Vibrio of the family Vibrionaceae. These phylogenetic data and those from genomic and phenotypic characterisations support the establishment of a novel Vibrio species, for which the name Vibrio nitrifigilis sp. nov. (type strain NFV-1T = BCRC 81211T = JCM 33628T) is proposed.

Simultaneous sulfamethoxazole biodegradation and nitrogen conversion by Achromobacter sp. JL9 using with different carbon and nitrogen sources

This study investigated sulfamethoxazole (SMX) biodegradation and nitrogen conversion by Achromobacter sp. JL9 using different carbon and nitrogen sources. Results showed that SMX and sodium acetate could be co-metabolized as carbon sources for bacterial growth and nitrogen conversion with highest removal efficiencies of 82.44%, 80.2%, and 79.45% for NH4+-N, NO3−-N, and SMX, respectively. Strain JL9 was able to utilize SMX as its sole nitrogen source for growth, with an SMX biodegradation efficiency of 63.10%. In addition, carbon and nitrogen balance analyses showed that approximately 35.31% and 63.22% of carbon and nitrogen, respectively, were lost as gaseous products. Finally, medium toxicity gradually decreased during the carbon and nitrogen dependence experiments. This study, thus, suggests that carbon and nitrogen play vital roles in SMX biodegradation and biotoxicity reduction.

Enhanced biodegradation of atrazine by Arthrobacter sp. DNS10 during co-culture with a phosphorus solubilizing bacteria: Enterobacter sp. P1

The interaction between pure culture microorganisms has been evaluated allowing for the enhanced biodegradation of various kinds of pollutants. Arthrobacter sp. DNS10 previously enriched in an atrazine-containing soil was capable of utilizing atrazine as the sole nitrogen source for growth, and Enterobacter sp. P1 is a phosphorus-solubilizing bacterium that releases various kinds of organic acids but lacks the ability to degrade atrazine. Whether strain P1 could enhance atrazine biodegradation by the degrader strain DNS10 was investigated in this experiment. Gas chromatography and high-performance liquid chromatography results showed that co-culture of both strains degraded 99.18 ± 1.00% of the atrazine (initial concentration was 100 mg L−1), while the single strain DNS10 only degraded 38.57 ± 7.39% after a 48 h culture, and the resulting concentration of the atrazine final metabolite cyanuric acid were 63.91 ± 3.34 mg L−1 and 26.60 ± 3.87 mg L−1, respectively. In addition, the expression of the atrazine degradation-related genes trzN, atzB and atzC in co-culture treatments was 6.61, 1.81 and 3.09 times that of the single strain DNS10 culture treatment. A substrates utilization test showed that the atrazine-degrading metabolites ethylamine and isopropylamine could serve as the nitrogen source to support strain P1 growth, although strain P1 cannot degrade atrazine or utilize atrazine for growth. Furthermore, the pH of the medium was significantly decreased when strain P1 utilized ethylamine and isopropylamine as the nitrogen source for growth. The results suggest that nondegrader strain P1 could promote the atrazine biodegradation when co-cultured with strain DNS10. This phenomenon is due to metabolite exchange between the two strains. Culturing these two strains together is a new biostimulation strategy to enhance the biodegradation of atrazine by culturing these two strains together.

Nitrogen transformation under different dissolved oxygen levels by the anoxygenic phototrophic bacterium Marichromatium gracile.

Marichromatium gracile: YL28 (M. gracile YL28) is an anoxygenic phototrophic bacterial strain that utilizes ammonia, nitrate, or nitrite as its sole nitrogen source during growth. In this study, we investigated the removal and transformation of ammonium, nitrate, and nitrite by M. gracile YL28 grown in a combinatorial culture system of sodium acetate-ammonium, sodium acetate-nitrate and sodium acetate-nitrite in response to different initial dissolved oxygen (DO) levels. In the sodium acetate-ammonium system under aerobic conditions (initial DO = 7.20-7.25mg/L), we detected a continuous accumulation of nitrate and nitrite. However, under semi-anaerobic conditions (initial DO = 4.08-4.26mg/L), we observed a temporary accumulation of nitrate and nitrite. Interestingly, under anaerobic conditions (initial DO = 0.36-0.67mg/L), there was little accumulation of nitrate and nitrite, but an increase in nitrous oxide production. In the sodium acetate-nitrite system, nitrite levels declined slightly under aerobic conditions, and nitrite was completely removed under semi-anaerobic and anaerobic conditions. In addition, M. gracile YL28 was able to grow using nitrite as the sole nitrogen source in situations when nitrogen gas produced by denitrification was eliminated. Taken together, the data indicate that M. gracile YL28 performs simultaneous heterotrophic nitrification and denitrification at low-DO levels and uses nitrite as the sole nitrogen source for growth. Our study is the first to demonstrate that anoxygenic phototrophic bacteria perform heterotrophic ammonia-oxidization and denitrification under anaerobic conditions.

Biodegradation kinetics of nitriles with easily degradable substrate by Klebsiella oxytoca GS-4-08

Nitrile compounds are common toxic and refractory organic wastes in industrial wastewater, and can be efficiently treated through biodegradation. The strain Klebsiella oxytoca GS-4-08 could utilize nitriles as the sole nitrogen source for growth, and the nitrile biodegradation kinetics showed good linear correlation with cell growth and glucose consumption. Degradation kinetics showed that the reaction order was dependent on initial cell concentration, and the degradation rates were increased when more easily degradable substrate was available. Higher acetonitrile (ACN) concentration inhibited not only the cell growth but also the ACN degradation. Haldane model constants indicated a high tolerance ability of K. oxytoca to acetonitrile. The formation of acetate during High Performance Liquid Chromatography (HPLC) wastewater treatment showed that the strain has a great potential for treating real nitriles-containing wastewater, and for organic acid production.

Isolation and characterization of atrazine-degrading strain Shewanella sp. YJY4 from cornfield soil.

Atrazine has been used worldwide for over 50years as a chemical herbicide. A strain of bacteria, YJY4 which utilizes atrazine as its sole nitrogen source for growth was isolated from agricultural black soil in northeastern China. 16S rDNA sequencing identified YJY4 as a Shewanella sp. PCR analysis and sequencing confirmed that YJY4 contained atrazine-degrading atzA, atzB and atzC genes. These genes revealed high similarity with those in Pseudomonas sp. ADP and Arthrobacter sp. TC1. The strain YJY4 was observed to degrade atrazine (100mgl(-1) ) to cyanuric acid completely after 36h. To the best of our knowledge, YJY4 was the first reported Shewanella sp. to grow in pure culture with atrazine serving as a sole source of nitrogen. Therefore, YJY4 may help with atrazine biodegradation and may become an abundant resource of atrazine degradation strains. A new isolated strain, YJY4 showed high atrazine-degrading ability, being able to degrade 100mgl(-1) atrazine completely in 36h. Strain YJY4 was identified as Shewanella sp. and contained atrazine-degrading atzA, atzB and atzC genes. This study examined the degradation mechanism and metabolic ability of this strain and for the bioremediation of contaminated environments, provides more strain selection and determines the strain of atrazine bioremediation potential.

Nitrogen removal characteristics of heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis C16

Alcaligenes faecalis C16 was found to have the ability to heterotrophically nitrify and aerobically denitrify. In order to further understand its nitrogen removal ability and mechanism, the growth and ammonium removal response were investigated at different C/N ratios and ammonium concentrations in the medium with citrate and acetate as carbon source separately. Furthermore, experiments of nitrogen sources, production of nitrogen gas and enzyme assay were conducted. Results show that the bacterium converts NH4+-N and produces NH2OH during the growing phase and nitrite accumulation is its distinct metabolic feature. A. faecalis C16 is able to tolerate not only high ammonium concentration but also high C/N ratio, and the ammonium tolerance is associated with carbon source and C/N ratio. The nitrogen balance under different conditions shows that approximately 28%–45% of the initial ammonium is assimilated into the cells, 44%–60% is denitrified and several percent is converted to nitrification products. A. faecalis C16 cannot utilize hydroxylamine, nitrite or nitrate as the sole nitrogen source for growth. However, nitrate can be used when ammonium is simultaneously present in the medium. A possible pathway for nitrogen removal by C16 is suggested. The preliminary enzyme assay provides more evidence for this nitrogen removal pathway.

Identification and characterization of the nitrate assimilation genes in the isolate of Streptomyces griseorubens JSD-1.

BackgroundStreptomyces griseorubens JSD-1 is a novel actinomycete isolated from soil that can utilize nitrate as its sole nitrogen source for growth and these nitrate assimilation genes active in this biotransformation are expected to be crucial. However, little is known about its genomic or genetic background related to nitrogen metabolism in this isolate. Thus, this study concentrates on identification and characterization of genes involved in nitrate assimilation.ResultsTo investigate the molecular mechanism of nitrate metabolism, genome sequencing was performed by Illumina Miseq platform. Then the draft genome of a single linear chromosome with 8,463,223 bp and an average G+C content of 72.42% was obtained, which has been deposited at GenBank under the accession number JJMG00000000. Sequences of nitrate assimilation proteins such as nitrate reductase (EC 1.7.99.4), nitrite reductase (EC 1.7.1.4), glutamine synthetase (EC 6.3.1.2), glutamate synthase (EC 1.4.1.13) and glutamate dehydrogenase (EC 1.4.1.2) were acquired. All proteins were predicted to be intracellular enzymes and their sequences were highly identical to those from their similar species owing to the conservative character. Putative 3D structures of these proteins were also modeled based on the templates with the most identities in the PDB database. Through KEGG annotated map, these proteins proved to be located on the key positions of nitrogen metabolic signaling pathway. Finally, quantitative RT-PCR indicated that expression responses of all genes were up-regulated generally and significantly when stimulated with nitrate.ConclusionIn this manuscript, we describe the genome features of an isolate of S. griseorubens JSD-1 following with identification and characterization of these nitrate assimilation proteins such as nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase and glutamate dehydrogenase accounts for the ability to utilize nitrate as its sole nitrogen source for growth through cellular localization, multiple sequence alignment, putative 3D modeling and quantitative RT-PCR. In summary, our findings provide the genomic and genetic background of utilizing nitrate of this strain.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-014-0174-4) contains supplementary material, which is available to authorized users.

Isolation and characterization of atrazine mineralizing Bacillus subtilis strain HB-6.

Atrazine is a widely used herbicide with great environmental concern due to its high potential to contaminate soil and waters. An atrazine-degrading bacterial strain HB-6 was isolated from industrial wastewater and the 16S rRNA gene sequencing identified HB-6 as a Bacillus subtilis. PCR assays indicated that HB-6 contained atrazine-degrading genes trzN, atzB and atzC. The strain HB-6 was capable of utilizing atrazine and cyanuric acid as a sole nitrogen source for growth and even cleaved the s-triazine ring and mineralized atrazine. The strain demonstrated a very high efficiency of atrazine biodegradation with a broad optimum pH and temperature ranges and could be enhanced by cooperating with other bacteria, suggesting its huge potential for remediation of atrazine-contaminated sites. To our knowledge, there are few Bacillus subtilis strains reported that can mineralize atrazine, therefore, the present work might provide some new insights on atrazine remediation.

Identification and characterization of the nitrate assimilation genes in the isolate of Streptomyces griseorubens JSD-1

Streptomyces griseorubens JSD-1 is a novel actinomycete isolated from soil that can utilize nitrate as its sole nitrogen source for growth and these nitrate assimilation genes active in this biotransformation are expected to be crucial. However, little is known about its genomic or genetic background related to nitrogen metabolism in this isolate. Thus, this study concentrates on identification and characterization of genes involved in nitrate assimilation. To investigate the molecular mechanism of nitrate metabolism, genome sequencing was performed by Illumina Miseq platform. Then the draft genome of a single linear chromosome with 8,463,223 bp and an average G+C content of 72.42% was obtained, which has been deposited at GenBank under the accession number JJMG00000000. Sequences of nitrate assimilation proteins such as nitrate reductase (EC 1.7.99.4), nitrite reductase (EC 1.7.1.4), glutamine synthetase (EC 6.3.1.2), glutamate synthase (EC 1.4.1.13) and glutamate dehydrogenase (EC 1.4.1.2) were acquired. All proteins were predicted to be intracellular enzymes and their sequences were highly identical to those from their similar species owing to the conservative character. Putative 3D structures of these proteins were also modeled based on the templates with the most identities in the PDB database. Through KEGG annotated map, these proteins proved to be located on the key positions of nitrogen metabolic signaling pathway. Finally, quantitative RT-PCR indicated that expression responses of all genes were up-regulated generally and significantly when stimulated with nitrate. In this manuscript, we describe the genome features of an isolate of S. griseorubens JSD-1 following with identification and characterization of these nitrate assimilation proteins such as nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase and glutamate dehydrogenase accounts for the ability to utilize nitrate as its sole nitrogen source for growth through cellular localization, multiple sequence alignment, putative 3D modeling and quantitative RT-PCR. In summary, our findings provide the genomic and genetic background of utilizing nitrate of this strain.

Nitrogen Oxyanion-dependent Dissociation of a Two-component Complex That Regulates Bacterial Nitrate Assimilation

Nitrogen is an essential nutrient for growth and is readily available to microbes in many environments in the form of ammonium and nitrate. Both ions are of environmental significance due to sustained use of inorganic fertilizers on agricultural soils. Diverse species of bacteria that have an assimilatory nitrate/nitrite reductase system (NAS) can use nitrate or nitrite as the sole nitrogen source for growth when ammonium is limited. In Paracoccus denitrificans, the pathway-specific two-component regulator for NAS expression is encoded by the nasT and nasS genes. Here, we show that the putative RNA-binding protein NasT is a positive regulator essential for expression of the nas gene cluster (i.e. nasABGHC). By contrast, a nitrogen oxyanion-binding sensor (NasS) is required for nitrate/nitrite-responsive control of nas gene expression. The NasS and NasT proteins co-purify as a stable heterotetrameric regulatory complex, NasS-NasT. This protein-protein interaction is sensitive to nitrate and nitrite, which cause dissociation of the NasS-NasT complex into monomeric NasS and an oligomeric form of NasT. NasT has been shown to bind the leader RNA for nasA. Thus, upon liberation from the complex, the positive regulator NasT is free to up-regulate nas gene expression.

Isolation and characterization of a novel cinosulfuron degrading Kurthia sp. from a methanogenic microbial consortium

A novel bacterial strain LAM0713 was isolated from a methanogenic bacterial complexes and identified as Kurthia sp. based on morphological, cultural, physio-biochemical characteristics and analysis of 16S rDNA sequence. Strain LAM0713 was found to be capable of utilizing cinosulfuron as sole nitrogen source for growth over a wide range of temperature (20-40 °C) and pH (5.0-9.0). Response surface methodology was used to optimize the degradation conditions. Strain LAM0713 could efficiently degrade 92.4% of initially supplemented 50 mg·L(1) cinosulfuron under the optimum conditions (pH 6.9, 31.8 °C) within 5 days. Five intermediates formed during cinosulfuron degradation were detected by liquid chromatography mass spectrometry (LC-MS), and a metabolic pathway for cinosulfuron degradation was proposed via cleavage of the sulfonylurea bridge. It is the first report showing that Kurthia sp. strain could degrade sulfonylurea herbicides, suggesting that strain LAM0713 may provide new insight into microbial degradation of herbicides.

Biodegradation of nicosulfuron by a Talaromyces flavus LZM1

The fungal strain LZM1 was isolated from activated sludge and found to be capable of utilizing nicosulfuron as the sole nitrogen source for growth. Based on morphological and internal transcribed spacer evaluations, LZM1 was identified as a Talaromyces flavus strain. Under optimum conditions (pH 6.1, 29°C), T. flavus LZM1 degraded 100% of the initially added nicosulfuron (100 mg L(-1)) within 5d. T. flavus LZM1 was also found to be highly efficient in degrading tribenuron methyl, chlorsulfuron, bensulfuron methyl, ethametsulfuron methyl, cinosulfuron, and rimsulfuron. Metabolites from nicosulfuron degradation were identified by liquid chromatography mass spectrometry, and a possible degradation pathway was deduced. These results show that T. flavus LZM1 may possess potential to be used in bioremediation of nicosulfuron-contaminated environments.

Inadequate root uptake may represent a major component limiting rice to use urea as sole nitrogen source for growth

Background and aims Urea represents over 50 % of nitrogen fertilizers applied worldwide to crop production, however one-third of nitrogen fertilized could be recovered by crops. Previous studies have provided useful knowledge of urea-related plant nitrogen-nutrition, whereas information about crop growth-response to urea without its external degradation seems limiting. We thus assess the ability of rice seedlings to use urea at a physiological level.

Isolation and characterization of a high-efficiency soil atrazine-degrading Arthrobacter sp. strain

Atrazine has been a pollutant of great environmental concern due to its high potential to contaminate soil and waters. In north China, atrazine has widely applied in corn fields and is still in use. Isolation of a high-efficiency atrazine degrader whose degradation ability is not inhibited by organic matters and fertilziers in the soils is of practical importance for bioremediation of contaminated soils. A bacterial strain (DAT1) capable of utilizing atrazine as a sole nitrogen source for growth was isolated from an agricultural soil in Hebei Province, China. The strain DAT1 was identified as an Arthrobacter species and harbored degradation genes trzN, atzB and atzC on plasmid DNA. The strain demonstrated a very high efficiency of atrazine biodegradation with a broad optimum pH and temperature ranges and could be enhanced by addition of both carbon and nitrogen sources, suggesting its huge potential for remediation of atrazine-contaminated sites. This work might provide some new insights on atrazine remediation and contributes to the limited pool of knowledge on the function of Arthrobacter species.

Metabolic ability and gene characteristics of Arthrobacter sp. strain DNS10, the sole atrazine-degrading strain in a consortium isolated from black soil

Strain DNS10 was the only member that could utilize atrazine as the sole nitrogen source for growth in an atrazine-degrading consortium which was isolated from black soil previously in our laboratory. It belongs to the genus Arthrobacter according to the sequence of 16S rRNA gene and is designated as Arthrobacter sp. DNS10. 16S rRNA gene phylogenetic analysis showed that strain DNS10 was located in a different evolutionary branch comparing with other Arthrobacter sp. atrazine-degrading strains. The degrading genes such as trzN, atzB and atzC harbored in strain DNS10 revealed high sequence similarity with those in Arthrobacter aurescens TC1 and Pseudomonas sp. ADP. These genes enabled the strain DNS10 to decompose atrazine to cyanuric acid. This was further proved by the results that the strain DNS10 (10 8 CFU mL −1) could degrade the whole atrazine (100 mg L −1) in the medium within 24 h at 30 °C and there was 66.13 ± 2.11 mg L −1 cyanuric acid accumulated at 24 h. These results imply that the strain DNS10 seems to be an excellent atrazine-degrading strain. Furthermore, this paper helps us in the better understanding of the strain evolution by comparing the metabolic ability and gene characteristics of strain DNS10 with other geographically distinct atrazine-degrading strains.

Arthrobacter sp. strain KU001 isolated from a Thai soil degrades atrazine in the presence of inorganic nitrogen sources.

An atrazine degrading bacterium, strain KU001, was obtained from a sugarcane field at the Cane and Sugar Research and Development Center at the Kasetsart University, Kamphaeng Saen campus, Thailand. Strain KU001 had a rod-to-coccus morphological cycle during growth. Biolog carbon source analysis indicated that the isolated bacterium was Arthrobacter histidinolovorans. Sequence analysis of PCR product indicated the 16S rRNA gene in strain KU001 was 99% identical to the same region in Arthrobacter sp.. The atrazine degradation pathway in strain KU001 consists of the catabolic genes, trzN, atzB and, atzC. Strain KU001 was able to use atrazine as a sole nitrogen source for growth, and surprisingly, atrazine degradation was not inhibited in cells grown on ammonium, nitrate or urea, as compared to cells cultivated on growth-limiting nitrogen sources. During the atrazine degradation process, the supplementation of nitrate completely inhibits atrazine degradation activity in strain KU001, whereas ammonium and urea had no effect on atrazine degradation activity. The addition of strain KU001 to sterile or nonsterile soils resulted in the disappearance of atrazine at a rate that was 4 to 5-fold more than that achieved by the indigenous microbial community. The addition of citrate to soils resulted in enhanced atrazine degradation, 80% of atrazine disappeared within one day following nutrient supplementation.

Rapid and Complete Degradation of the Herbicide Picloram by Lipomyces kononenkoae

An enrichment culture approach was used to isolate a pure culture of the yeast Lipomyces kononenkoae, which had the ability to grow on the herbicide picloram. The yeast rapidly and completely degraded 50 microg mL(-1) picloram by 48 h of growth. While L. kononenkoae was found to use both N atoms of picloram as a sole nitrogen source for growth, it failed to mineralize the herbicide or use it as a sole C source. Product analysis done using LC-ESI-MS indicated that biodegradation of picloram by L. kononenkoae proceeds via a didechlorinated, dihydroxylated, pyridinecarboxylic acid derivative. Our results are consistent with the hypothesis that the majority of picloram degradation in the soil is likely due to microbial catabolic processes.