共查询到20条相似文献,搜索用时 15 毫秒
1.
Luis M. S. Magalhães Carlos A. Neyra Johanna Döbereiner 《Archives of microbiology》1978,117(3):247-252
Chlorate resistant spontaneous mutants ofAzospirillum spp. (syn.Spirillum lipoferum) were selected in oxygen limited, deep agar tubes with chlorate. Among 20 mutants fromA. brasilense and 13 fromA. lipoferum all retained their functional nitrogenase and 11 from each species were nitrate reductase negative (nr–). Most of the mutants were also nitrite reductase negative (nir–), only 3 remaining nir+. Two mutants from nr+ nir+ parent strains lost only nir and became like the nr+ nir– parent strain ofA. brasilense. No parent strain or nr+ mutant showed any nitrogenase activity with 10 mM NO
3
–
. In all nr– mutants, nitrogenase was unaffected by 10 mM NO
3
–
. Nitrite inhibited nitrogenase activity of all parent strains and mutants including those which were nir–. It seems therefore, that inhibition of nitrogenase by nitrate is dependent on nitrate reduction. Under aerobic conditions, where nitrogenase activity is inhibited by oxygen, nitrate could be used as sole nitrogen source for growth of the parent strains and one mutant (nr– nir–) and nitritite of the parent strains and 10 mutants (all types). This indicates the loss of both assimilatory and dissimilatory nitrate reduction but only dissimilatory nitrite reduction in the mutants selected with chlorate. 相似文献
2.
PCR Detection of Genes Encoding Nitrite Reductase in Denitrifying Bacteria 总被引:11,自引:6,他引:5
下载免费PDF全文
![点击此处可从《Applied microbiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Using consensus regions in gene sequences encoding the two forms of nitrite reductase (Nir), a key enzyme in the denitrification pathway, we designed two sets of PCR primers to amplify cd1- and Cu-nir. The primers were evaluated by screening defined denitrifying strains, denitrifying isolates from wastewater treatment plants, and extracts from activated sludge. Sequence relationships of nir genes were also established. The cd1 primers were designed to amplify a 778 to 799-bp region of cd1-nir in the six published sequences. Likewise, the Cu primers amplified a 473-bp region in seven of the eight published Cu-nir sequences. Together, the two sets of PCR primers amplified nir genes in nine species within four genera, as well as in four of the seven sludge isolates. The primers did not amplify genes of nondenitrifying strains. The Cu primers amplified the expected fragment in all 13 sludge samples, but cd1-nir fragments were only obtained in five samples. PCR products of the expected sizes were verified as nir genes after hybridization to DNA probes, except in one case. The sequenced nir fragments were related to other nir sequences, demonstrating that the primers amplified the correct gene. The selected primer sites for Cu-nir were conserved, while broad-range primers targeting conserved regions of cd1-nir seem to be difficult to find. We also report on the existence of Cu-nir in Paracoccus denitrificans Pd1222. 相似文献
3.
温度和水分变化对冻土区泥炭地土壤氮循环功能基因丰度的影响 总被引:2,自引:0,他引:2
以大兴安岭多年冻土区泥炭地为研究对象,通过室内模拟增温实验,研究温度升高对不同深度(0-150 cm)土壤氮循环功能基因丰度的影响。同时针对0-20 cm和20-40 cm土壤设置两个水分处理,分别为土壤原始含水量和淹水状态,研究水分变化对表层土壤氮循环功能基因丰度的影响。结果表明温度升高显著提高了活动层(0-60 cm)、过渡层(60-80 cm)、永冻层(80-100 cm)中nifH、nirK基因丰度,温度升高显著提高了活动层(0-40 cm)和过渡层(60-80 cm)中nirS基因丰度。温度升高显著提高了过渡层(60-80 cm)NH4+-N和较深永冻层(140-150 cm)NO3--N的含量,但降低了过渡层(60-80 cm)NO3--N和较深永冻层(120-150 cm)NH4+-N的含量,相关性分析表明,NH4+-N含量与nifH和nirS基因丰度呈显著正相关,NO3--N含量与nirK基因丰度呈显著正相关,说明温度升高能够通过改变微生物丰度促进过渡层固氮作用和反硝化作用。在增温条件下,淹水处理使表层土壤nirS和nirK基因丰度及NH4+-N含量降低,但提高了NO3--N含量,说明淹水造成了过度还原的条件使反硝化底物浓度降低,降低反硝化微生物活性进而抑制了土壤反硝化作用。该结果对于明确未来气候变化影响下冻土区泥炭地土壤氮循环过程具有重要意义。 相似文献
4.
5.
M. Chippaux D. Giudici A. Abou-Jaoudé F. Casse M. C. Pascal 《Molecular & general genetics : MGG》1978,160(2):225-229
Summary Mutants of E. coli, completely devoid of nitrite reductase activity with glucose or formate as donor were studied. Biochemical analysis indicates that they are simultaneously affected in nitrate reductase, nitrite reductase, fumarate reductase and hydrogenase activities as well as in cytochrome c552 biosynthesis. The use of an antiserum specific for nitrate reductase shows that the nitrate reductase protein is probably missing. A single mutation is responsible for this phenotype: the gene affected, nir R, is located close to tyr R i.e. at 29 min on the chromosomal map.Abbreviations BV
Benzyl-Viologen
- NTG
N-methyl-N-nitro-N-nitrosoguanidine
- NR
nitrate reductase
- NIR
nitrite reductase
- FR
fumarate reductase
- HYD
hydrogenase
- CYT c552
cytochrome c552 相似文献
6.
7.
Amélie A. M. Cantarel Juliette M. G. Bloor Thomas Pommier Nadine Guillaumaud Caroline Moirot Jean‐François Soussana Franck Poly 《Global Change Biology》2012,18(8):2520-2531
Emissions of the trace gas nitrous oxide (N2O) play an important role for the greenhouse effect and stratospheric ozone depletion, but the impacts of climate change on N2O fluxes and the underlying microbial drivers remain unclear. The aim of this study was to determine the effects of sustained climate change on field N2O fluxes and associated microbial enzymatic activities, microbial population abundance and community diversity in an extensively managed, upland grassland. We recorded N2O fluxes, nitrification and denitrification, microbial population size involved in these processes and community structure of nitrite reducers (nirK) in a grassland exposed for 4 years to elevated atmospheric CO2 (+200 ppm), elevated temperature (+3.5 °C) and reduction of summer precipitations (?20%) as part of a long‐term, multifactor climate change experiment. Our results showed that both warming and simultaneous application of warming, summer drought and elevated CO2 had a positive effect on N2O fluxes, nitrification, N2O release by denitrification and the population size of N2O reducers and NH4 oxidizers. In situ N2O fluxes showed a stronger correlation with microbial population size under warmed conditions compared with the control site. Specific lineages of nirK denitrifier communities responded significantly to temperature. In addition, nirK community composition showed significant changes in response to drought. Path analysis explained more than 85% of in situ N2O fluxes variance by soil temperature, denitrification activity and specific denitrifying lineages. Overall, our study underlines that climate‐induced changes in grassland N2O emissions reflect climate‐induced changes in microbial community structure, which in turn modify microbial processes. 相似文献
8.
Anthonius P. N. de Boer Willem N. M. Reijnders J. Gijs Kuenen Adriaan H. Stouthamer Rob J. M. van Spanning 《Antonie van Leeuwenhoek》1994,66(1-3):111-127
By using the gene encoding the C-terminal part of thecd
1-type nitrite reductase ofPseudomonas stutzeri JM300 as a heterologous probe, the corresponding gene fromParacoccus denitrificans was isolated. This gene,nirS, codes for a mature protein of 63144 Da having high homology withcd
1-type nitrite reductases from other bacteria. Directly downstream fromnirS, three othernir genes were found in the ordernirECF. The organization of thenir gene cluster inPa. denitrificans is different from the organization ofnir clusters in some Pseudomonads.nirE has high homology with a S-adenosyl-L-methionine:uroporphyrinogen III methyltransferase (uro'gen III methylase). This methylase is most likely involved in the hemed
1 biosynthesis inPa. denitrificans. The third gene,nirC, codes for a small cytochromec of 9.3 kDa having high homology with cytochromec
55X ofPs. stutzeri ZoBell. The 4th gene,nirF, has no homology with other genes in the sequence databases and has no relevant motifs. Inactivation of either of these 4 genes resulted in the loss of nitrite and nitric oxide reductase activities but not of nitrous oxide reductase activity.nirS mutants lack thecd
1-type nitrite reductase whilenirE, nirC andnirF mutants produce a small amount ofcd
1-type nitrite reductase, inactive due to the absence of hemed
1. Upstream from thenirS gene the start of a gene was identified which has limited homology withnosR, a putative regulatory gene involved in nitrous oxide reduction. A potential FNR box was identified between this gene andnirS.Abbreviations SDS
sodium dodecyl sulfate
- NBT
nitroblue tetrazolium
- PAGE
polyacrylamide gel electrophoresis 相似文献
9.
The results of the cross reactions of the 27 strains of Azospirillum spp. with 4 fluorescent antibodies (FA) show a neat differentiation between the two species. A. lipoferum represents a more homogenous group in respect to FA reactions and highly fluorescent preparations were obtained with strains from a large scope origin against Sp59 FA, the type strain. In contrast A. brasilense contains at least three sub groups in respect to FA reactions. The first includes all denitrifing strains (nir+) which react with FA from Sp7 the type strain. None of the nir- strains reacted strongly with Sp7 FA. One part of the A. brasilense nir- group which includes the strains isolated from well sterilized rice and wheat roots (Sp 107, 107 st, 106 and 109 st) reacts with FA of their reference strain Sp107 but not with that of Sp28 FA. The strains isolated from unsterilized roots and soils reacted with SP28 FA and not with that of Sp107 FA. In addition there were 3 strains (Sp A4, 34 and 67) which reacted with neither of the FAs.Abbreviations Fa
fluorescent antibody
- FITC
fluorescein isothiocyanate
- Rh ITC
gelatin-rhodamine isothiocyanate
- nir+
nitrite reductase positive
- nir-
nitrite reductase negative 相似文献
10.
To date, the association of coral–bacteria and the ecological roles of bacterial symbionts in corals remain largely unknown. In particular, little is known about the community components of bacterial symbionts of corals involved in the process of denitrification and ammonia oxidation. In this study, the nitrite reductase (nirS and nirK) and ammonia monooxygenase subunit A (amoA) genes were used as functional markers. Diverse bacteria with the potential to be active as denitrifiers and ammonia-oxidizing bacteria (AOB) were found in two East China Sea corals: stony coral Alcyonium gracillimum and soft coral Tubastraea coccinea. The 16S rRNA gene library analysis demonstrated different communities of bacterial symbionts in these two corals of the same location. Nitrite reductase nirK gene was found only in T. coccinea, while both nirK and nirS genes were detected in A. gracillimum, which might be the result of the presence of different bacterial symbionts in these two corals. AOB rather than ammonia-oxidizing archaea were detected in both corals, suggesting that AOB might play an important role in the ammonia oxidation process of the corals. This study indicates that the coral bacterial symbionts with the potential for nitrite reduction and ammonia oxidation might have multiple ecological roles in the coral holobiont, which promotes our understanding of bacteria-mediated nitrogen cycling in corals. To our knowledge, this study is the first assessment of the community structure and phylogenetic diversity of denitrifying bacteria and AOB in corals based on nirK, nirS, and amoA gene library analysis. 相似文献
11.
Chlorate toxicity in Aspergillus nidulans. Studies of mutants altered in nitrate assimilation. 总被引:20,自引:0,他引:20
D. J. Cove 《Molecular & general genetics : MGG》1976,146(2):147-159
Summary It had previously been held that chlorate is not itself toxic, but is rendered toxic as a result of nitrate reductase-catalysed conversion to chlorite. This however cannot be the explanation of chlorate toxicity in Aspergillus nidulans, even though nitrate reductase is known to have chlorate reductase activity. Among other evidence against the classical theory for the mechanism of chlorate toxicity, is the finding that not all mutants lacking nitrate reductase are clorate resistant. Both chlorate-sensitive and resistant mutants lacking nitrate reductase, also lack chlorate reductase. Data is presented which implicates not only nitrate reductase but also the product of the nirA gene, a positive regulator gene for nitrate assimilation, in the mediation of chlorate toxicity. Alternative mechanisms for chlorate toxicity are considered. It is unlikely that chlorate toxicity results from the involvement of nitrate reductase and the nirA gene product in the regulation either of nitrite reductase, or of the pentose phosphate pathway. Although low pH has an effect similar to chlorate, chlorate is not likely to be toxic because it lowers the pH; low pH and chlorate may instead have similar effects. A possible explanation for chlorate toxicity is that it mimics nitrate in mediating, via nitrate reductase and the nirA gene product, a shut-down of nitrogen catabolism. As chlorate cannot act as a nitrogen source, nitrogen starvation ensures. 相似文献
12.
13.
Lipopolysaccharides (LPS) were extracted by hot phenol-water from five strains each of Azospirillum lipoferum and Azospirillum brasilense. Rhamnose, glucose, glucosamine and 3-deoxy-d-mannooctulosonic acid were comon sugar constituents of all LPS preparations. 2-O-Mefucose, 3-O-Me-fucose, 3-O-Me-rhamnose and 2-O-Megalactose were found in LPSs of some A. brasilense strains. Fatty acid spectra from all LPSs studied were almost identical with predominance of 3-hydroxymyristic and 3-hydroxypalmitic acids. 3-Hydroxypalmitic acid was the only amide-linked fatty acid. Lipopolysaccharides isolated from A. brasilense showed higher heterogeneity in sugar composition than those from A. lipoferum.Abbreviations glc
gas liquid chromatography
- ms
mass spectrometry
- LPS
lipopolysaccharide
- dOclA
3-deoxy-d-mannooctulosonic acid
- 3-OH-16:0
3-hydroxypalmitic acid
- nir-
nitrite reductase negative
- nir+
nitrite reductase positive 相似文献
14.
The gene encoding nitrite reductase (nir) from the cyanobacterium Synechococcus sp. PCC 7942 has been identified and sequenced. This gene comprises 1536 nucleotides and would encode a polypeptide of 56506 Da that shows similarity to nitrite reductase from higher plants and to the sulfite reductase hemoprotein from enteric bacteria. Identities found at positions corresponding to those amino acids which in the above-mentioned proteins hold the Fe4S4-siroheme active center suggest that nitrite reductase from Synechococcus bears an active site much alike that present in those reductases. The fact that the Synechococcus and higher-plant nitrite reductases are homologous proteins gives support to the endosymbiont theory for the origin of chloroplasts. 相似文献
15.
16.
17.
Hee-Sung Bae Wan-Taek Im Yuichi Suwa James M. Lee Sung-Taik Lee Young-Keun Chang 《Archives of microbiology》2009,191(4):329-340
Although, there have been many published bacterial strains aerobically degrading the heterocyclic amine compounds, only one
strain to date has been reported to degrade pyrrolidine under denitrifying conditions. In this study, denitrifying bacteria
degrading pyrrolidine and piperidine were isolated from diverse geological and ecological origins through selective enrichment
procedures. Based on the comparative sequence results of 16S rRNA genes, 30 heterocyclic amine-degrading isolates were grouped
into ten distinct phylotypes belonging to the genera Thauera, Castellaniella, Rhizobium, or Paracoccus of the phylum Proteobacteria. The representative isolates of individual phylotypes were characterized by phylogenetic, phenotypic and chemotaxonomical
traits, and dissimilatory nitrite reductase gene (nirK and nirS). All isolates completely degraded pyrrolidine and piperidine under both aerobic and anaerobic conditions. The anaerobic
degradations were coupled to nitrate reduction. A metabolic pathway for the anaerobic degradation of pyrrolidine was proposed
on the basis of enzyme activities implicated in pyrrolidine metabolism from three isolates. The three key pyrrolidine-metabolizing
enzymes pyrrolidine dehydrogenase, γ-aminobutyrate/α-ketoglutarate aminotransferase, and succinic semialdehyde dehydrogenase,
were induced by heterocyclic amines under denitrifying conditions. They were also induced in cells grown aerobically on heterocyclic
amines, suggesting that the anaerobic degradation of pyrrolidine shares the pathway with aerobic degradation.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
18.
Joana Falcão Salles Michele C. Pereira e Silva Francisco Dini-Andreote Armando C. F. Dias Nadine Guillaumaud Franck Poly Jan Dirk van Elsas 《Biogeochemistry》2017,132(1-2):185-201
Here, we investigated the patterns of microbial nitrogen cycling communities along a chronosequence of soil development in a salt marsh. The focus was on the abundance and structure of genes involved in N fixation (nifH), bacterial and archaeal ammonium oxidation (amoA; AOB and AOA), and the abundances of genes involved in denitrification (nirS, nirK, nosZ). Potential nitrification and denitrification activities were also measured, and increases in nitrification were found in soils towards the end of succession, whereas denitrification became maximal in soils at the intermediate stages. The nifH, nirK and nirS gene markers revealed increases in the sizes of the respective functional groups towards the intermediate stage (35 years), remaining either constant (for nifH) or slightly declining towards the latest stage of succession (for nirK and nirS). Moreover, whereas the AOB abundance peaked in soils at the intermediate stage, that of AOA increased linearly along the chronosequence. The abundance of nosZ was roughly constant, with no significant regression. The drivers of changes in abundance and structure were identified using path analysis; whereas the ammonia oxidizers (AOA and AOB) showed patterns that followed mainly N availability, those of the nitrogen fixers followed plant diversity and soil structure. The patterns of denitrifiers were group-dependent, following the patterns of plant diversity (nirK and nirS) and belowground shifts (nosZ). The variation observed for the microbial groups associated with the same function highlights their differential contribution at different stages of soil development, revealing an interplay of changes in terms of niche complementarity and adaptation to the local environment. 相似文献
19.
Rocio J. Alcántara-Hernández Carla M. Centeno Alejandro Ponce-Mendoza Silvia Batista Martin Merino-Ibarra Julio Campo Luisa I. Falcón 《Polar Biology》2014,37(3):403-416
Cyanobacterial microbial mats are highly structured communities commonly found in Antarctic inland waters including melt streams. These benthic microbial associations comprise a large number of microorganisms with different metabolic capacities, impacting nutrient dynamics where established. The denitrification process is a feasible nitrogen loss pathway and a biological source of nitrous oxide, a potent greenhouse gas that also promotes ozone depletion. Potential denitrifiers from five microbial mats were characterized using a PCR-DGGE (denaturing gradient gel electrophoresis) approach. Molecular markers encoding for key enzymes in the denitrification process (nirK, nirS and nosZ) were used. Fingerprints were obtained for the five sampled mats and compared for two successive years. Distance analysis showed that despite the sampled year, the denitrifying genetic potential was similar between most of the sites when represented in Euclidean space. The number of dominant denitrifiers detected for each sample ranged between 6 and 18 for nirK, 4–10 for nirS and 6–17 for nosZ. The seventy-two sequenced phylotypes showed 80–98 % identity to previously reported environmental sequences from water column, sediments and soil samples. These results suggest that Antarctic microbial mats have a large denitrification potential, previously uncharacterized and composed by both site-specific and common phylotypes belonging mainly to Alpha-, Beta- and Gammaproteobacteria. 相似文献
20.
Operon fusions in the nitrate reductase operon and study of the control gene nir R in Escherichia coli 总被引:24,自引:0,他引:24
Marc Chippaux Violaine Bonnefoy-Orth Jeannine Ratouchniak Marie-Claire Pascal 《Molecular & general genetics : MGG》1981,182(3):477-479
Summary Strains carrying operon fusions between the promotor of the chl I gene and the lac structural genes were constructed. From these strains in which the expression of the lac genes is under the control of both nitrate and oxygen, spontaneous regulatory mutants were selected: (i) mutants which synthesize -galactosidase constitutively in anaerobiosis; (ii) mutants in which -galactosidase synthesis is no longer repressed by oxygen.Introduction of the nir R mutated allele into strains carrying these fusions resulted in the total loss of -galactosidase synthesis, confirming that nir R is a regulatory gene controlling the expression of the biosynthesis of the nitrate reductase. 相似文献