scholarly journals ISOGENIC STRAINS OF PHYCOMYCES BLAKESLEEANUS SUITABLE FOR GENETIC ANALYSIS

Genetics ◽  
1983 ◽  
Vol 105 (4) ◽  
pp. 873-879
Author(s):  
M I Alvarez ◽  
A P Eslava
Keyword(s):  
Genetic Analysis ◽  
Mating Type ◽  
Type Strain ◽  
Wild Type ◽  
Tetrad Analysis ◽  
Phycomyces Blakesleeanus ◽  
Isogenic Strains ◽  
Type Strains

ABSTRACT The progeny of crosses between wild-type strains of Phycomyces usually do not exhibit all of the expected genotypes from meiosis. By backcrossing, we have isolated a new (+) mating-type strain, A56, which is nearly isogenic with the (-) wild-type NRRL1555 commonly used in Phycomyces research. Tetrad analysis of the backcrosses shows that meiosis becomes more regular as the parental (+) and (-) strains become more isogenic. In our two-factor crosses with unlinked markers, the regularity of meiosis is measured as the percent of reciprocal ditypes plus tetratypes in the progeny. We have shown that this percentage increases from about 15% for crosses between nonisogenic parents to 90% in the eighth backcross. The results indicate that routine, reliable recombination analyses are possible in P. blakesleeanus.

scholarly journals Influence of parental strains on the germination ofPhycomyces blakesleeanuszygospores

1988 ◽  
Vol 52 (2) ◽  
pp. 91-95 ◽  
Author(s):  
María José F. Sarabia ◽  
Arturo P. Eslava ◽  
María Isabel Alvarez
Keyword(s):  
Type Strain ◽  
Wild Type Strain ◽  
Wild Type ◽  
Standard Strain ◽  
Tetrad Analysis ◽  
Phycomyces Blakesleeanus ◽  
The Third ◽  
Dormancy Period ◽  
Sexual Crosses ◽  
Regular Meiosis

SummaryPhycomyces blakesleeanuswild-type NRRL1555( − ), the standard strain, when crossed with UBC21( + ), another wild type, gives zygospores that germinate in 50–60 days. By backcrossing to UBC21 and selecting for shorter dormancy we have isolated a ( − ) strain, A803, and a ( + ) strain A804, which when crossed give zygospores that germinate in 32 days, the shortest dormancy period found inPhycomyces.The same result was obtained when A803 was crossed with UBC21. Zygospore dormancy decreased as the parental strains became more isogenic with UBC21, but the number of zygospores giving germsporangia with viable germspores also decreased to zero in the third backcross. The existence of germspore-killer alleles in the strain UBC21 is postulated. The strains of shortest dormancy can be used as helper strains (Orejaset al.1985) in sexual crosses. Tetrad analysis of the cross NRRL1554 × S102, a two-factor cross, showed 90% of reciprocal ditypes plus tetratypes in the progeny, indicating that the ( + ) wild-type strain NRRL1554, when crossed with the standard strain, gives regular meiosis and, contrary to current beliefs, may be used inPhycomycesgenetic analysis.

Development of mutants of Gliocladium virens tolerant to benomyl

1990 ◽  
Vol 36 (7) ◽  
pp. 484-489 ◽  
Author(s):  
G. C. Papavizas ◽  
D. P. Roberts ◽  
K. K. Kim
Keyword(s):  
Carbon Source ◽  
Type Strain ◽  
Wild Type Strain ◽  
Carbon Sources ◽  
Sclerotium Rolfsii ◽  
Wild Type ◽  
Gliocladium Virens ◽  
Rhizosphere Competence ◽  
Filter Paper Cellulase ◽  
Type Strains

Aqueous suspensions of conidia of Gliocladium virens strains Gl-3 and Gl-21 were exposed to both ultraviolet radiation and ethyl methanesulfonate. Two mutants of Gl-3 and three of Gl-21 were selected for tolerance to benomyl at 10 μg∙mL−1, as indicated by growth and conidial germination on benomyl-amended potato dextrose agar. The mutants differed considerably from their respective wild-type strains in appearance, growth habit, sporulation, carbon-source utilization, and enzyme activity profiles. Of 10 carbon sources tested, cellobiose, xylose, and xylan were the best for growth, galactose and glucose were intermediate, and arabinose, ribose, and rhamnose were poor sources of carbon. The wild-type strains and the mutants did not utilize cellulose as the sole carbon source for growth. Two benomyl-tolerant mutants of Gl-3 produced less cellulase (β-1,4-glucosidase, carboxymethylcellulase, filter-paper cellulase) than Gl-3. In contrast, mutants of Gl-21 produced more cellulase than the wild-type strain. Only Gl-3 provided control of blight on snapbean caused by Sclerotium rolfsii. Wild-type strain Gl-21 and all mutants from both strains were ineffective biocontrol agents. Key words: Gliocladium, benomyl tolerance, Sclerotium, rhizosphere competence.

scholarly journals Genetic analysis of amidase mutants ofPseudomonas aeruginosa

1974 ◽  
Vol 23 (3) ◽  
pp. 335-359 ◽  
Author(s):  
Joan L. Betz ◽  
Jane E. Brown ◽  
Patricia H. Clarke ◽  
Martin Day
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Genetic Analysis ◽  
Structural Gene ◽  
Type Strain ◽  
Wild Type Strain ◽  
Relative Order ◽  
Regulator Gene ◽  
Wild Type ◽  
Growth Phenotype

SUMMARYMutants ofPseudomonas aeruginosa, which differed in amide growth phenotype from the wild-type strain, were subjected to genetic analysis using the generalized transducing phage F116. The map order of some mutational sites was determined by 3-factor crosses in which a mutation in the linked regulator geneamiRwas used as the outside marker to determine the relative order of mutations in the amidase structural geneamiE. Acetamide-positive transductants were recovered in crosses between amidase-negative strains and strains PhB3(PAC377), V2(PAC353) and V5(PAC356) producing mutant amidases which hydrolyse phenylacetamide and valeramide but not acetamide. Some recombinants carried the mutationamiE16 determining the properties of the mutant B amidase produced by strain B6(PAC351) from which both PhB and V class mutants were derived, while other recombinants produced A amidase determined by the wild-typeamiEgene.

Isolation and genetic analysis of self-sterility and perithecial pigmentation mutants in a homothallic isolate of Nectria haematococca

1982 ◽  
Vol 60 (1) ◽  
pp. 79-84 ◽  
Author(s):  
A. Babai-Ahary ◽  
M. J. Daboussi-Bareyre ◽  
D. Parisot
Keyword(s):  
Genetic Analysis ◽  
Sexual Reproduction ◽  
Type Strain ◽  
Wild Type Strain ◽  
Wild Type ◽  
Nectria Haematococca

Mutants with altered sexual reproduction were isolated from a homothallic wild-type strain of Nectria haematococca (Berk. & Br.) Wr. and genetically analysed. Most were self- and inter-sterile but some were interfertile and were used to improve genetic analysis by allowing an easy detection of hybrid perithecia. The study of these mutants also provides information about various aspects of the perithecial development in this organism.

scholarly journals A genetic analysis of resistance to nystatin inSaccharomyces cerevisiae

1967 ◽  
Vol 9 (2) ◽  
pp. 179-193 ◽  
Author(s):  
K. A. Ahmed ◽  
R. A. Woods
Keyword(s):  
Genetic Analysis ◽  
Resistance Genes ◽  
Type Strain ◽  
Wild Type Strain ◽  
Second Step ◽  
Wild Type ◽  
Recessive Resistance ◽  
Low Concentrations ◽  
Effects Of Temperature ◽  
Resistant Mutants

1. A number of stable nystatin-resistant mutants of the yeastSaccharomyces cerevisiaehave been isolated from platings of a sensitive wild-type strain on low concentrations of the antibiotic.2. These mutants were found to be resistant to 10, 15 or 60 units of drug/ml.3. Analysis of meiotic segregants from crosses of these mutants to wild-type indicate that resistance is determined by two types of genes; resistance genes and modifiers.4. Functional analysis of the mutants demonstrated the existence of three recessive resistance genes,nys-l,nys-2 andnys-3 and thatnys-1 andnys-2 were linked.5. Genetic analysis showed thatnys-1 was affected by two modifiers,Mnys-1 andMnys-2, but that onlyMnys-2 affectednys-2 andnys-3.6. The modifiersMnys-1 andMnys-2 are dominant.7. An investigation of the effects of temperature and medium on resistance demonstrated marked interactions between genotype and environment for both the resistance genes and the modifiers.8. Second-step mutants have been isolated by plating first-step mutants on higher concentrations of the drug. Some of these are resistant to 800 units/ml.9. Some possible mechanisms of nystatin resistance are discussed.

scholarly journals MUTATIONS AFFECTING SEXUAL CONJUGATION AND RELATED PROCESSES IN SACCHAROMYCES CEREVISIAE. II. GENETIC ANALYSIS OF NONMATING MUTANTS

Genetics ◽  
1974 ◽  
Vol 76 (2) ◽  
pp. 273-288
Author(s):  
Vivian Mackay ◽  
Thomas R Manney
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genetic Analysis ◽  
Mating Type ◽  
Tetrad Analysis ◽  
Genetic Loci ◽  
Type Locus ◽  
Mating Type Locus

ABSTRACT Rare diploids formed by sterile mutants have been studied by tetrad analysis. Sixteen classes of mutants representing at least five distinct genetic loci have been defined. One group of mutations, isolated only in α, maps at the mating-type locus, while none of the others shows any linkage to mating type. Some of the mutations are nonspecific for mating type, while others act only on a or α. In addition, mutations were found that prevent sporulation when heterozygous in diploids. These appear to be mutations of the mating-type alleles.

scholarly journals ISOLATION AND GENETIC ANALYSIS OF MUTANT STRAINS OF CHLAMYDOMONAS REINHARDI DEFECTIVE IN GAMETIC DIFFERENTIATION

Genetics ◽  
1976 ◽  
Vol 82 (2) ◽  
pp. 169-186
Author(s):  
Ursula W Goodenough ◽  
Carol Hwang ◽  
Howard Martin
Keyword(s):  
Genetic Analysis ◽  
Cell Fusion ◽  
Mating Type ◽  
Normal Growth ◽  
The Other ◽  
Genetic Dissection ◽  
Structural Studies ◽  
Tetrad Analysis ◽  
Mating Structure ◽  
Mutant Strains

ABSTRACT Impotent mutant strains of Chlamydomonas reinhardi, mating-type (mt) plus, are described that have normal growth and motility but fail to differentiate into normal gametes. Procedures for their isolation and their genetic analysis are described. Five of the imp strains (imp-2, imp-5, imp-6, imp-7, and imp-8) exhibit no flagellar agglutination when mixed with mt  - or mt  + gametes; these strains have been induced to form rare zygotes with mt  - gametes and the mutations are shown to be unlinked to the mt locus (with the possible exception of imp-7). Two of the strains (imp-3 and imp-4) carry leaky mutations that affect cell fusion; neither mutation is found by tetrad analysis to be linked to mt or to the other. Cells of the imp-1 strain agglutinate well with mt  - gametes and active agglutination continues for up to 48 hours, but cell fusion occurs only very rarely. Analysis of these rare zygotes indicates that imp-1 is closely linked to the mt  + locus, and fine-structural studies reveal that imp-1gametes produce a mutant mating structure involved in zygotic cell fusion. The development of sexuality in C. reinhardi therefore appears amenable to genetic dissection.

scholarly journals LINKAGE OF MUTATIONS AFFECTING MINUS FLAGELLAR MEMBRANE AGGLUTINABILITY TO THE mt  - MATING-TYPE LOCUS OF CHLAMYDOMONAS

Genetics ◽  
1981 ◽  
Vol 99 (1) ◽  
pp. 41-47 ◽  
Author(s):  
Carol J Hwang ◽  
Brian C Monk ◽  
Ursula W Goodenough
Keyword(s):  
Uv Irradiation ◽  
Mating Type ◽  
Zygote Formation ◽  
Wild Type ◽  
Tetrad Analysis ◽  
Type Locus ◽  
Mating Type Locus ◽  
Flagellar Membrane ◽  
Mutant Strains

ABSTRACT Two independently isolated mutant strains, imp-10 and imp-12, were obtained by UV irradiation of wild-type mating-type minus (wt-). Each fails to agglutinate sexually with gametes of either mating type, but mating and zygote formation can be elicited by agglutinating either strain to wt+ gametes by means of anti-flagellar antiserum. Tetrad analysis of the resultant zygotes shows that both imp-10 and imp-12 are very closely linked to mt  -, with no recombinants observed. Diploid strains constructed between imp-10 or imp-12 and wt+ gametes are wt-, that is, they agglutinate and fuse like normal minus cells. Tetrad analysis of triploids from imp-10 diploid x wt+ haploid crosses shows that only imp-10 and wt+ products are recovered. A model is proposed to account for these results.

A Stable 44S Intermediate in the Autodigestion of 50S Ribosomal Subunits

1971 ◽  
Vol 29 ◽  
pp. 430-431
Author(s):  
John H. Nisbet ◽  
Henry S. Slayter
Keyword(s):  
Escherichia Coli ◽  
Molecular Weight ◽  
Ribosomal Rna ◽  
Gel Electrophoresis ◽  
Type Strain ◽  
Wild Type Strain ◽  
Ribosomal Subunit ◽  
Wild Type ◽  
Ribosomal Subunits ◽  
Type Strains

Wild - type strains of Escherichia coli are known to contain as many as four endogenous nucleases (Ref. 1). These are commonly found associated with the ribosomes after extraction from the cell, but may be removed, with the exception of RNase IV, by washing the ribosomes in NH4Cl (at 0.2 M and higher concentrations). We have examined the effect of these nucleases on the 50S ribosomal subunit of one wild-type strain, K12 (Hfr 3000), by incubating the unwashed particles at 37° in the presence of varying magnesium concentrations.At 10-4 molar magnesium (slower at 10-3 molar), the 50S particle is converted to a species sedimenting at about 44S. About 20% of the total O.D260 is liberated at the same time. Continued incubation leads to the release of more O.D260 material while the RNA remaining in the 44S (Fig. 1) particle is progressively cleaved, eventually to the point where it consists of one principal fragment of molecular weight 0.42 x 106 daltons and several lesser fragments. The ribosomal RNA and proteins have been characterized by acrylamide gel electrophoresis.

scholarly journals The Secreted Esterase of Propionibacterium freudenreichii Has a Major Role in Cheese Lipolysis

2013 ◽  
Vol 80 (2) ◽  
pp. 751-756 ◽  
Author(s):  
María Claudia Abeijón Mukdsi ◽  
Hélène Falentin ◽  
Marie-Bernadette Maillard ◽  
Victoria Chuat ◽  
Roxana Beatriz Medina ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Type Strain ◽  
Wild Type Strain ◽  
Lipolytic Activity ◽  
Premature Stop Codon ◽  
Propionibacterium Freudenreichii ◽  
Wild Type ◽  
Swiss Cheese ◽  
Content Type ◽  
Type Strains

ABSTRACTFree fatty acids are important flavor compounds in cheese.Propionibacterium freudenreichiiis the main agent of their release through lipolysis in Swiss cheese. Our aim was to identify the esterase(s) involved in lipolysis byP. freudenreichii. We targeted two previously identified esterases: one secreted esterase, PF#279, and one putative cell wall-anchored esterase, PF#774. To evaluate their role in lipolysis, we constructed overexpression and knockout mutants ofP. freudenreichiiCIRM-BIA1Tfor each corresponding gene. The sequences of both genes were also compared in 21 wild-type strains. All strains were assessed for their lipolytic activity on milk fat. The lipolytic activity observed matched data previously reported in cheese, thus validating the relevance of the method used. The mutants overexpressing PF#279 or PF#774 released four times more fatty acids than the wild-type strain, demonstrating that both enzymes are lipolytic esterases. However, inactivation of thepf279gene induced a 75% reduction in the lipolytic activity compared to that of the wild-type strain, whereas inactivation of thepf774gene did not modify the phenotype. Two of the 21 wild-type strains tested did not display any detectable lipolytic activity. Interestingly, these two strains exhibited the same single-nucleotide deletion at the beginning of thepf279gene sequence, leading to a premature stop codon, whereas they harbored apf774gene highly similar to that of the other strains. Taken together, these results clearly demonstrate that PF#279 is the main lipolytic esterase inP. freudenreichiiand a key agent of Swiss cheese lipolysis.

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