Isolation and characterization of Azotobacter vinelandii mutant strains with potential as bacterial fertilizer

1983 ◽  
Vol 29 (8) ◽  
pp. 973-978 ◽  
Author(s):  
Joyce K. Gordon ◽  
Marty R. Jacobson
Keyword(s):  
Azotobacter Vinelandii ◽  
Nitrogenase Activity ◽  
Resistant Mutant ◽  
Free Medium ◽  
Wild Type ◽  
Isolation And Characterization ◽  
Mutant Strains ◽  
Resistant Strains ◽  
Resistant Mutants

Mutant strains of Azotobacter vinelandii which might have potential for use as bacterial fertilizer have been isolated and fall into two categories: constitutive mutants that synthesize nitrogenase in the presence of ammonium and mutants that overproduce nitrogenase when grown in nitrogen-free medium. The constitutive mutants described in this paper were isolated from the wild type as methylalanine-resistant strains and express up to 23% of the fully derepressed nitrogenase level when grown in medium containing excess ammonium. By contrast, ammonium-grown cultures of wild type have less than 0.003% of the fully derepressed level. Strains which fix more N2 than the wild type in nitrogen-free medium were isolated as mefhylammonium-resistant mutants. Although the methylammonium-resistant mutant strains fix more N2 than the wild type, they grow no faster. The excess nitrogen produced by these mutants is excreted into the medium, resulting in up to 60% more nitrogen than in the medium of the wild type. Higher nitrogenase activity in the methylammonium-resistant mutant strains was found to be a result of increased levels of nitrogenase protein, suggesting that regulation of nitrogenase synthesis may be altered.

scholarly journals Isolation and characterization of the SPT2 gene, a negative regulator of Ty-controlled yeast gene expression.

1985 ◽  
Vol 5 (7) ◽  
pp. 1543-1553 ◽  
Author(s):  
G S Roeder ◽  
C Beard ◽  
M Smith ◽  
S Keranen
Keyword(s):  
Gene Product ◽  
Regulatory Region ◽  
Negative Regulator ◽  
Transcriptional Level ◽  
Wild Type ◽  
Isolation And Characterization ◽  
Mutant Strains ◽  
Insertion Mutations ◽  
His4 Gene

The his4-917 mutation of Saccharomyces cerevisiae results from the insertion of the Ty element Ty917 into the regulatory region of the HIS4 gene and renders the cell His-. The hist4-912 delta mutant, which carries a solo delta in the 5'-noncoding region of HIS4, is His+ at 37 degrees C but His- at 23 degrees C. Both these mutations interfere with HIS4 expression at the transcriptional level. The His- phenotype of both insertion mutations is suppressed by mutations at the SPT2 locus. The product of the wild-type SPT2 gene apparently represses HIS4 transcription in these mutant strains; this repression is relieved when the SPT2 gene is destroyed by mutation. The repression of transcription by SPT2 presumably results from an interaction between the SPT2+ gene product and Ty or delta sequences. In this paper, we report the cloning and DNA sequence analysis of the wild-type SPT2 gene and show that the gene is capable of encoding a protein of 333 amino acids in length. In addition, we show that a dominant mutation of the SPT2 gene results from the generation of an ochre codon which is presumed to lead to a shortened SPT2 gene product.

scholarly journals In Vitro Isolation and Characterization of Oxazolidinone-Resistant Mycobacterium tuberculosis

2017 ◽  
Vol 61 (10) ◽  
Author(s):  
Matthew B. McNeil ◽  
Devon D. Dennison ◽  
Catherine D. Shelton ◽  
Tanya Parish
Keyword(s):  
Mycobacterium Tuberculosis ◽  
23S Rrna ◽  
Clinical Settings ◽  
Content Type ◽  
Isolation And Characterization ◽  
Mutant Strains ◽  
Ribosomal Protein L3 ◽  
Resistant Strains

ABSTRACT Oxazolidinones are promising candidates for the treatment of Mycobacterium tuberculosis infections. We isolated linezolid-resistant strains from H37Rv (Euro-American) and HN878 (East-Asian) strains; resistance frequencies were similar in the two strains. Mutations were identified in ribosomal protein L3 (RplC) and the 23S rRNA (rrl). All mutant strains were cross resistant to sutezolid; a subset was cross resistant to chloramphenicol. Mutations in rrl led to growth impairment and decreased fitness that may limit spread in clinical settings.

Polyene antibiotic affinities for the sterols of resistant and sensitive strains of Neurospora crassa

1977 ◽  
Vol 23 (1) ◽  
pp. 113-115 ◽  
Author(s):  
D. Johnson ◽  
R. Subden
Keyword(s):  
Candida Albicans ◽  
Neurospora Crassa ◽  
Resistant Mutant ◽  
Wild Type ◽  
Polyene Antibiotic ◽  
Polyene Antibiotics ◽  
Mutant Strains ◽  
Resistant Strains

Ergosterol, the principle sterol of many wild-type Neurospora and other Ascomycetes, had a greater affinity for polyene antibiotics than did lichesterol or eburicol, the sterols of some resistant mutant strains. The affinity was demonstrated by comparing the sterols extracted from sensitive and resistant strains of Neurospora crassa and Candida albicans for protection against polyene inhibition of sensitive N. crassa and for their ability to alter specific polyene absorption maxima.

Isolation and characterization of catabolite repression-resistant mutants of Escherichia coli

1977 ◽  
Vol 23 (10) ◽  
pp. 1384-1393 ◽  
Author(s):  
Glen D. Armstrong ◽  
Hiroshi Yamazaki
Keyword(s):  
Escherichia Coli ◽  
Catabolite Repression ◽  
Coli Strain ◽  
Wild Type ◽  
Phosphotransferase System ◽  
E Coli ◽  
Isolation And Characterization ◽  
In The Wild ◽  
Resistant Mutants

A method has been developed for the isolation of Escherichia coli mutants which are resistant to catabolite repression. The method is based on the fact that a mixture of glucose and gluconate inhibits the development of chemotactic motility in the wild type, but not in the mutants. A motile E. coli strain was mutagenized and grown in glucose and gluconate. Mutants which were able to swim into a tube containing a chemotactic attractant (aspartic acid) were isolated. Most of these mutants were able to produce β-galactosidase in the presence of glucose and gluconate and were normal in their ability to degrade adenosine 3′,5′-cyclic monophosphate. Some of these mutants were defective in the glucose phosphotransferase system.

scholarly journals ISOLATION AND CHARACTERIZATION OF NEUROSPORA MUTANTS AFFECTED IN INVERTASE SYNTHESIS

Genetics ◽  
1984 ◽  
Vol 106 (4) ◽  
pp. 591-599
Author(s):  
Deborah B Lee ◽  
Stephen J Free
Keyword(s):  
Neurospora Crassa ◽  
Large Scale ◽  
Invertase Activity ◽  
Free Medium ◽  
Wild Type ◽  
Isolation And Characterization ◽  
Normal Enzyme ◽  
Enzyme Molecules ◽  
Large Scale Screening

ABSTRACT We have outlined a procedure that allows the large-scale screening of mutagenized Neurospora crassa populations for invertaseless mutants. We have isolated and characterized three mutations, inv(DBL1), inv(DBL9) and inv(DBL14), which have been mapped at or near the invertase structural gene. One of these, inv(DBL1), is particularly interesting. Our experiments indicate that the reduced level of invertase activity in the inv(DBL1)-containing cell can be explained as the result of a reduced number of normal enzyme molecules. We also show that wild-type Neurospora is able to respond rapidly to a change of medium and can dramatically increase its production of invertase within 20 min after a transfer to a carbon-free medium.

Isolation and characterization of the SPT2 gene, a negative regulator of Ty-controlled yeast gene expression

1985 ◽  
Vol 5 (7) ◽  
pp. 1543-1553
Author(s):  
G S Roeder ◽  
C Beard ◽  
M Smith ◽  
S Keranen
Keyword(s):  
Gene Product ◽  
Regulatory Region ◽  
Negative Regulator ◽  
Transcriptional Level ◽  
Wild Type ◽  
Isolation And Characterization ◽  
Mutant Strains ◽  
Insertion Mutations ◽  
His4 Gene

The his4-917 mutation of Saccharomyces cerevisiae results from the insertion of the Ty element Ty917 into the regulatory region of the HIS4 gene and renders the cell His-. The hist4-912 delta mutant, which carries a solo delta in the 5'-noncoding region of HIS4, is His+ at 37 degrees C but His- at 23 degrees C. Both these mutations interfere with HIS4 expression at the transcriptional level. The His- phenotype of both insertion mutations is suppressed by mutations at the SPT2 locus. The product of the wild-type SPT2 gene apparently represses HIS4 transcription in these mutant strains; this repression is relieved when the SPT2 gene is destroyed by mutation. The repression of transcription by SPT2 presumably results from an interaction between the SPT2+ gene product and Ty or delta sequences. In this paper, we report the cloning and DNA sequence analysis of the wild-type SPT2 gene and show that the gene is capable of encoding a protein of 333 amino acids in length. In addition, we show that a dominant mutation of the SPT2 gene results from the generation of an ochre codon which is presumed to lead to a shortened SPT2 gene product.

Interaction between finger millet (Eleusine coracana) genotypes and drug-resistant mutants ofAzospirillum brasilensein calcareous soil

1984 ◽  
Vol 102 (3) ◽  
pp. 521-529 ◽  
Author(s):  
R. Rai ◽  
V. Prasad ◽  
I. C. Shukla
Keyword(s):  
Azospirillum Brasilense ◽  
Finger Millet ◽  
Nitrogenase Activity ◽  
Resistant Mutant ◽  
Eleusine Coracana ◽  
Drug Resistant ◽  
Mutant Strains ◽  
Drug Resistant Mutant ◽  
Uninoculated Control ◽  
Resistant Mutants

SummaryAzospirillum brasilensewas treated with nitrosoguanidine and five drug-resistant mutant strains isolated. The effects of acriflavin on pre- and post-irradiation with u.v. light and the level of antibiotic resistance were studied. Variations in factors were found between the strains. Inoculation of finger millet withA. brasilenseand mutant strains led to significant increases in grain yield and nitrogenase activity compared with the uninoculated control, with significant strain x genotype interactions. Differential response of genotype and strain was noted on the protein and amino acid concentration of seeds.

Transformation of Azotobacter vinelandii strains unable to fix nitrogen with Rhizobium spp. DNA

1978 ◽  
Vol 24 (3) ◽  
pp. 209-214 ◽  
Author(s):  
William J. Page
Keyword(s):  
Transcriptional Control ◽  
Deoxyribonucleic Acid ◽  
Azotobacter Vinelandii ◽  
Nitrogenase Activity ◽  
Nitrogen Fixing ◽  
Wild Type ◽  
Spontaneous Reversion ◽  
Mutant Strains ◽  
Rhizobium Spp ◽  
Rhizobium Sp

The phenotypes of Azotobacter vinelandii ATCC 12837 strains defective in nitrogen fixation (Nif−) were characterized by intrageneric transformation with known Nif− strains of A. vinelandii OP. These former mutant strains were used as recipients for intergeneric transformation by deoxyribonucleic acid (DNA) prepared from Rhizobium spp. to determine if the rhizobia would transform the Azotobacter Nif− phenotypes to Nif+. The frequency of Nif+ transformants using Rhizobium DNA was always less than the frequency using Azotobacter wild-type DNA but was greater than the spontaneous reversion frequency. The Azotobacter Nif+ recombinants also were stable. DNA from all of the Rhizobium spp. transformed to Nif+Azotobacter mutants defective in the nitrogenase component I (molybdoferredoxin); however, some recombinants had a lower nitrogenase activity and a delayed nitrogenase depression time. Mutants defective in the pleiotrophic transcriptional control of both nitrogenase components were transformed to Nif+ by the asymbiotic nitrogen fixing Rhizobium sp. 32H1 and 41A1, but not the symbiotic nitrogen-fixing species. The significance of these results and the possible future applications of this system are discussed.

Isolation and characterization of Azospirillum mutants excreting high amounts of indoleacetic acid

1983 ◽  
Vol 29 (8) ◽  
pp. 916-923 ◽  
Author(s):  
A. Hartmann ◽  
Mahavir Singh ◽  
W. Klingmüller
Keyword(s):  
Azospirillum Brasilense ◽  
Indoleacetic Acid ◽  
Acid Metabolism ◽  
Feedback Inhibition ◽  
Growth Patterns ◽  
Wild Type ◽  
Azospirillum Lipoferum ◽  
Isolation And Characterization ◽  
Resistant Mutants

Mutants of Azospirillum brasilense Sp Cd, resistant to 5-fluorotryptophan (FT) excreted 3-indoleacetic acid (IAA), i.e., auxin, producing up to 16 μg/mL which was 30 times greater than the wild-type level. Under conditions of nitrogen fixation, the mutants excreted IAA up to 1 μg/mL, 10 times more than the wild type. However, none of the FT-resistant mutants of Azospirillum lipoferum Sp RG 20a excreted high levels of IAA. This was probably due to differences in the tryptophan and IAA biosynthetic steps between A. brasilense and A. lipoferum strains. Some of the FT-resistant mutants of A. brasilense Sp Cd showed a reduced feedback inhibition of anthranilate synthetase by tryptophan. The increased synthesis of tryptophan could explain the observed excretion of tryptophan and related metabolites. In addition, the IAA-overproducing mutants excreted other amino acids, probably owing to pleiotropic effects of deregulated tryptophan biosynthesis on amino acid metabolism. The growth patterns of some mutants excreting large amounts of IAA were almost identical to those of the wild type.

scholarly journals The Na+-translocating NADH : ubiquinone oxidoreductase of Azotobacter vinelandii negatively regulates alginate synthesis

Microbiology ◽  
2009 ◽  
Vol 155 (1) ◽  
pp. 249-256 ◽  
Author(s):  
Cinthia Núñez ◽  
Alexander V. Bogachev ◽  
Gabriel Guzmán ◽  
Isaac Tello ◽  
Josefina Guzmán ◽  
...  
Keyword(s):  
Sodium Pump ◽  
Azotobacter Vinelandii ◽  
Soil Bacterium ◽  
Wild Type ◽  
Ubiquinone Oxidoreductase ◽  
Alginate Production ◽  
Isolation And Characterization ◽  
Nadh Ubiquinone Oxidoreductase ◽  
A Subunit

Azotobacter vinelandii is a nitrogen-fixing soil bacterium that produces the exopolysaccharide alginate. In this report we describe the isolation and characterization of A. vinelandii strain GG4, which carries an nqrE : : Tn5 mutation resulting in alginate overproduction. The nqrE gene encodes a subunit of the Na+-translocating NADH : ubiquinone oxidoreductase (Na+-NQR). As expected, Na+-NQR activity was abolished in mutant GG4. When this strain was complemented with the nqrEF genes this activity was restored and alginate production was reduced to wild-type levels. Na+-NQR may be the main sodium pump of A. vinelandii under the conditions tested (∼2 mM Na+) since no Na+/H+-antiporter activity was detected. Collectively our results indicate that in A. vinelandii the lack of Na+-NQR activity caused the absence of a transmembrane Na+ gradient and an increase in alginate production.

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