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Mutations Increasing Asexual Plasmodium Formation in PHYSARUM POLYCEPHALUM
1Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139.
Abstract:
Rare plasmodia formed in clones of heterothallic amoebae were analyzed in a search for mutations affecting plasmodium formation. The results show that the proportion of mutants varies with both temperature (18 degrees , 26 degrees or 30 degrees ) and mating-type allele (mt1, mt2, mt3, mt4). At one extreme, only one of 33 plasmoida formed by mt2 amoebae at 18 degrees is mutant. At the other extreme, three of three plasmodia formed by mt1 amoebae at 30 degrees are mutant. The mutant plasmodia fall into two groups, the GAD (greater asexual differentiation) mutants and the ALC (amoebaless life cycle) mutants. The spores of GAD mutants give rise to amoebae that differentiate into plasmodia asexually at much higher frequencies than normal heterothallic amoebae. Seven of eight gad mutations analyzed genetically are linked to mt and one (gad-12) is not. The gad-12 mutation is expressed in strains with different alleles of mt. The frequency of asexual plasmodium formation is heat sensitive in some (e.g., mt3 gad-11 ), heat-insensitive in two (mt2 gad-8 and mt2 gad-9) and cold-sensitive in one (mt1 gad-12) of twelve GAD mutants analyzed phenotypically. The spores of ALC mutants give rise to plasmodia directly, thereby circumventing the amoebal phase of the life cycle. Spores from five of the seven ALC mutants give rise to occasional amoebae, as well as plasmodia. The amoebae from one of the mutants carry a mutation (alc-1) that is unlinked to mt and is responsible for the ALC phenotype in this mutant. Like gad-12, alc-1 is expressed with different mt alleles. Preliminary observations with amoebae from the other four ALC mutants suggest that two are similar to the one containing alc-1; one gives rise to revertant amoebae, and one gives rise to amoebae carrying an alc mutation and a suppressor of the mutation.
Insights
This study identified mutations affecting plasmodium formation in amoebae, revealing temperature and mating-type allele influences. Two mutant groups, GAD and ALC, exhibit altered asexual differentiation and life cycles, respectively.
Area of Science:
- Cell Biology
- Genetics
- Developmental Biology
Background:
- Plasmodium formation is a critical developmental process in heterothallic amoebae.
- Understanding mutations affecting this process is key to deciphering developmental pathways.
Purpose of the Study:
- To identify and characterize mutations that influence plasmodium formation in heterothallic amoebae.
- To investigate the effects of temperature and mating-type alleles on mutation frequency and expression.
Main Methods:
- Analysis of rare plasmodia formed in clones of heterothallic amoebae.
- Genetic analysis of identified mutant strains.
- Phenotypic analysis of mutant spores and resulting amoebae under varying temperatures.
Main Results:
- Mutation frequency varied significantly with temperature and mating-type allele.
- Two distinct mutant classes were identified: GAD (greater asexual differentiation) and ALC (amoebaless life cycle).
- GAD mutants showed increased asexual plasmodium formation, with some mutations linked to mating-type and others unlinked; temperature sensitivity varied.
- ALC mutants bypassed the amoebal phase, with some mutations unlinked to mating-type and exhibiting complex inheritance patterns.
Conclusions:
- Environmental factors like temperature and genetic background (mating-type alleles) critically influence mutation rates and phenotypes.
- Discovered mutations provide insights into the genetic regulation of asexual differentiation and life cycle progression in amoebae.
- Unlinked mutations (gad-12, alc-1) suggest complex genetic interactions and potential regulatory networks controlling plasmodium formation.
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