scholarly journals CHROMOSOME FRAGMENTS IN DICTYOSTELIUM DISCOIDEUM OBTAINED FROM PARASEXUAL CROSSES BETWEEN STRAINS OF DIFFERENT GENETIC BACKGROUND

Genetics ◽  
1980 ◽  
Vol 95 (2) ◽  
pp. 289-304
Author(s):  
Keith L Williams ◽  
Gillian E Robson ◽  
Dennis L Welker
Keyword(s):  
Linkage Group ◽  
Genetic Analysis ◽  
Dictyostelium Discoideum ◽  
Genetic Background ◽  
Deleterious Effect ◽  
Fruiting Body Formation ◽  
Body Formation ◽  
Chromosome Fragments ◽  
Group Ii ◽  
Different Genetic Background

ABSTRACT The first aneuploid strains of Dictyostelium discoideum have been unambiguously characterized, using cytological and genetic analysis. Three independently isolated, but genetically similar, fragment chromosomes have been observed in segregants from diploids formed between haploid strains derived from the NC4 and VI2 isolates of D. discoideum. Once generated, the fragment chromosomes, all of which have Vl2-derived centromeres, can be maintained in a NC4 genetic background. Genetic evidence is consistent with the view that all three fragment chromosomes studied encompass the region from the centromere to the whiA locus of linkage group II and terminate in the interval between whiA and acrA. From cytological studies, one of the fragment chromosomes consists of approximately half of linkage group II.——We observed no deleterious effect on viability or asexual fruiting-body formation in either haploid or diploid strains carrying an additional incomplete chromosome and hence are disomic or trisomic, respectively, for part of linkage group II. The incomplete chromosome is lost at a frequency of 2 to 3% from disomic and trisomic strains, but surprisingly this loss is not increased in the presence of the haploidizing agent, benlate. A new locus (clyA), whose phenotype is altered colony morphology, is assigned to the region of linkage group II encompassed by the fragment chromosome.

Culmination in the slime mould Dictyostelium discoideum studied with a scanning electron microscope

Development ◽  
1976 ◽  
Vol 35 (2) ◽  
pp. 323-333
Author(s):  
D. J. Watts ◽  
T. E. Treffry
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Dictyostelium Discoideum ◽  
Fruiting Body ◽  
Maximum Height ◽  
Slow Process ◽  
Fruiting Body Formation ◽  
Basal Disc ◽  
Body Formation ◽  
Scanning Electron

Myxamoebae of Dictyostelium discoideum were allowed to develop on cellulose acetate filters, and specimens taken at various stages of fruiting body formation were prepared for study by scanning electron microscopy. In the immature fruiting body where the mass of pre-spore cells has just been lifted off the substratum by the developing stalk, the pre-spore cells are irregular in shape and are similar in appearance to cells in aggregates at earlier stages of development. As the stalk lengthens, the pre-spore cells gradually separate from one another and become rounded and elongate, but mature spores are not visible until the fruiting body reaches its maximum height. It is concluded that, contrary to previous reports, spore maturation is a slow process and is not completed until the sorus becomes pigmented. The mature stalk is surrounded by a smooth cellulose sheath but this does not envelop the cells of the basal disc, which remain discrete. The fruiting body is enclosed in a slime sheath and this may be important in holding together the mass of spores.

Isolation of telomere DNA from Neurospora crassa

1987 ◽  
Vol 7 (9) ◽  
pp. 3168-3177
Author(s):  
M G Schechtman
Keyword(s):  
Linkage Group ◽  
Neurospora Crassa ◽  
Genetic Background ◽  
Type Strain ◽  
Wild Type Strain ◽  
The Other ◽  
Wild Type ◽  
Oak Ridge ◽  
Entire Chromosome ◽  
Different Genetic Background

The most distal known gene on Neurospora crassa linkage group VR, his-6, was cloned. A genomic walk resulted in isolation of the telomere at VR. It was obtained from a library in which the endmost nucleotides of the chromosome had not been removed by nuclease treatment before being cloned, and mapping indicates that the entire chromosome end has probably been cloned. Sequences homologous to the terminal 2.5 kilobases of DNA from VR from these Oak Ridge N. crassa strains are found at other sites in the genome. To characterize these sites, I crossed an Oak Ridge-derived his-6 strain with a wild-type strain of different genetic background (Mauriceville) and characterized the hybridization patterns seen in the progeny. It appears that the sequences homologous to the VR terminus are found at genetically different sites in the two parental strains, and no hybridization to the VR telomere from Mauriceville was detected. The other genomic copies identified in the Oak Ridge parent were not telomeres. I suggest that any repeating sequence blocks found immediately adjacent to the VR terminus in Oak Ridge strains must be small and that the repeating element identified in that background may be an N. crassa transposable element integrated near the the chromosome end at VR.

motA1552, a mutation of Dictyostelium discoideum having pleiotropic effects on motility and discoidin I regulation.

Genetics ◽  
1988 ◽  
Vol 118 (3) ◽  
pp. 425-436
Author(s):  
S C Kayman ◽  
R Birchman ◽  
M Clarke
Keyword(s):  
Linkage Group ◽  
Dictyostelium Discoideum ◽  
Growth Temperature ◽  
Segregation Analysis ◽  
Pleiotropic Effects ◽  
Temperature Sensitive ◽  
Temperature Induction ◽  
Group Ii ◽  
Unstable Mutations ◽  
Axenic Growth

Abstract The Dictyostelium discoideum mutant MC2 exhibits temperature-sensitive growth, temperature-sensitive motility, and temperature induction of discoidin I synthesis. These three phenotypes of MC2 were not separated in the genetic experiments reported here. They were therefore assigned to the mutation motA1552, which was mapped to linkage group II by segregation analysis and by analysis of mitotic recombinant diploids. In one motA1552 strain, loss of motility preceded accumulation of discoidin I by 3 hr, indicating that discoidin I is not involved in generation of the motility defect. Expression of motA1552 phenotypes varied both among strains carrying the mutation, and among different clones of a particular strain. MC2 and its derivatives displayed elevated levels of recombination between whiA and acrA on linkage group II, and yielded highly unstable mutations at the acrA locus. Accumulation of large amounts of discoidin I during axenic growth of strain AX3 was found to depend on the presence of a second linkage group II mutation, daxA1551. This mutation was already present in the strain mutagenized to isolate motA1552, complicating explication of motA1552 action.

In vitro stalk cell differentiation in wild-type and ‘slugger’ mutants of Dictyostelium discoideum

Development ◽  
1993 ◽  
Vol 118 (2) ◽  
pp. 523-526 ◽  
Author(s):  
K. Inouye ◽  
J. Gross
Keyword(s):  
Dictyostelium Discoideum ◽  
Fruiting Body ◽  
Cell Maturation ◽  
Stalk Cell ◽  
Wild Type ◽  
Fruiting Body Formation ◽  
Body Formation ◽  
Close Relationship ◽  
Long Time

In ‘slugger’ mutants of Dictyostelium discoideum, aggregates of cells remain for an abnormally long time in the migratory phase under conditions where wild-type aggregates form fruiting bodies. In the present work, we have examined the relationship between the defect in fruiting body formation in these mutants and their ability to form mature stalk cells. We dissociated anterior cells from slugs of the mutants and their parents and tested their ability to form stalk cells when incubated at low density in the presence of (1) the stalk cell morphogen Differentiation Inducing Factor-1 (DIF-1) together with cyclic AMP, or (2) 8-Br-cAMP, which is believed to penetrate cell membrane and activate cAMP- dependent protein kinase (PKA). Most of the mutants were markedly defective in forming stalk cells in response to DIF-1 plus cAMP, confirming a close relationship between fruiting body formation and stalk cell maturation. On the other hand, many of these same mutants formed stalk cells efficiently in response to 8-Br-cAMP. This supports evidence for an essential role of PKA in stalk cell maturation and fruiting body formation. It also indicates that many of the mutants owe their slugger phenotype to defects in functions required for optimal adenylyl cyclase activity.

PARASEXUAL GENETIC ANALYSIS OF CELL PROPORTIONING MUTANTS OF DICTYOSTELIUM DISCOIDEUM

Genetics ◽  
1981 ◽  
Vol 99 (2) ◽  
pp. 183-196
Author(s):  
James H Morrissey ◽  
William F Loomis
Keyword(s):  
Cell Differentiation ◽  
Linkage Group ◽  
Genetic Analysis ◽  
Dictyostelium Discoideum ◽  
Double Mutant ◽  
Complementation Group ◽  
Stalk Cell ◽  
Recessive Mutation ◽  
Group Vi ◽  
Recessive Mutations

ABSTRACT Eight independently isolated mutants of Dictyostelium discoideum that differentiate exclusively into stalk cells make up one complementation group and carry single recessive mutations at the stalky locus, stkA, located on linkage group II. KY19, a previously described strain that differentiates into spores, but not stalk cells, was found to possess a recessive mutation defining the stalkless locus, stlA, located on linkage group VI. An analysis of the properties of these mutants, together with the phenotype of a haploid double mutant carrying stkA and stlA indicates that stlA results in poorly organized stalk tubes and incomplete stalk cell differentiation, while stkA causes all of the cells to differentiate into stalk cells, even when not enclosed in the stalk tube. The significance of these results is discussed in relation to current theories of pattern formation in D. discoideum.

scholarly journals Imaging Promoter Assay of Adenylyl Cyclase A Gene in Dictyostelium discoideum during Fruiting Body Formation by Dual-Color Bioluminescence Microscopy

2021 ◽  
Author(s):  
Taro Hayashi ◽  
Katsunori Ogoh ◽  
Hirobumi Suzuki
Keyword(s):  
Adenylyl Cyclase ◽  
Dictyostelium Discoideum ◽  
Fruiting Body ◽  
Cell Aggregation ◽  
Adenosine Monophosphate ◽  
Fruiting Body Formation ◽  
Promoter Assay ◽  
Body Formation ◽  
Secondary Messenger ◽  
Dual Color

Cyclic adenosine monophosphate (cAMP), which is derived from adenosine triphosphate through adenylyl cyclase A (acaA), acts as an intracellular secondary messenger and an extracellular chemotactic substance in important biological processes. In the social amoebae Dictyostelium discoideum, cAMP mediates cell aggregation, development, and differentiation to spore and stalk cells during fruiting body formation. The acaA gene is transcribed under the control of three different alternative promoters. This study aimed to develop a promoter assay for acaA in D. discoideum using bioluminescence microscopy. Here, we inserted green- and red-emitting luciferase genes into downstream of promoter regions 1 and 3, respectively. Promoter activities were visualized by bioluminescence microscopy. We confirmed the differential expression of acaA under the control of promoters 1 and 3 at the different stages of D. discoideum development. We also demonstrated the application of dual-color bioluminescence imaging in the development of an imaging promoter assay.

scholarly journals THE USE OF RESTRICTION FRAGMENT LENGTH POLYMORPHISMS AND DNA DUPLICATIONS TO STUDY THE ORGANIZATION OF THE ACTIN MULTIGENE FAMILY IN DICTYOSTELIUM DISCOIDEUM

Genetics ◽  
1986 ◽  
Vol 112 (1) ◽  
pp. 27-42
Author(s):  
Dennis L Welker ◽  
K Peter Hirth ◽  
Patricia Romans ◽  
Angelika Noegel ◽  
Richard A Firtel ◽  
...  
Keyword(s):  
Linkage Group ◽  
Dictyostelium Discoideum ◽  
Restriction Fragment ◽  
Fragment Length ◽  
Gene Copy ◽  
Actin Genes ◽  
Group I ◽  
Group Ii ◽  
Cloned Gene ◽  
Restriction Fragment Length

ABSTRACT The techniques of restriction fragment length polymorphism analysis and examination of gene copy number in duplication-bearing Dictyostelium discoideum strains have been used to map four actin genes of the wild-type strain NC4 to specific linkage groups. In part, this was accomplished by identification of restriction fragments corresponding to particular cloned actin genes using genespecific probes from unique sequence 5' and 3' untranslated regions. Cloned gene Actin 8 (designation act-8) maps to linkage group I; Actins 12 (act-12) and M6 (actM6) to linkage group II. An uncloned gene (act-100) also maps to linkage group II in the same region as actM6, as defined by a chromosomal duplication. From analysis of other wild isolates of D. discoideum, it was determined that in these isolates at least two actin genes map to linkage group I and at least four map to linkage group II. These results demonstrate the utility of molecular techniques in genetic analysis of Dictyostelium, particularly for developmentally regulated genes that have been cloned but that have no identified mutant phenotypes.

scholarly journals GENETIC ANALYSIS OF THE GENE FOR N-ACETYLGLUCOSAMINIDASE IN DICTYOSTELIUM DISCOIDEUM

Genetics ◽  
1978 ◽  
Vol 88 (2) ◽  
pp. 277-284 ◽  
Author(s):  
William F Loomis
Keyword(s):  
Linkage Group ◽  
Genetic Analysis ◽  
Dictyostelium Discoideum ◽  
Structural Gene ◽  
Regulatory Protein ◽  
Group Iv ◽  
Single Locus ◽  
Stages Of Development ◽  
Separate Locus

ABSTRACT Three independent mutations affecting N-acetylglucosaminidase in Dictyostelium discoideum were mapped by the parasexual system and found to lie on linkage group IV. These mutations as well as two others were found to be recessive and noncomplementing in heterozygous diploids. Thus they all appear to affect the nagA locus. Since two of the mutations give rise to thermolabile enzyme, this defines the structural gene for N-acetylglucosaminidase. The enzyme is a homodimer of a 68,000 dalton subunit and thus would be expected to be determined by a single locus. The expression of this gene is regulated by the stages of development; however, it should be mentioned that none of the mutations fell in a separate locus that might determine a specific positive regulatory protein.

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