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Growth of large plasmodia of the myxomycete Physarum polycephalum
Abstract:
A method has been developed for growing Physarum polycephalum plasmodia that are 8 to 10 times larger than those obtained in the petri dish cultures used by Nygaard, Guttes, and Rusch. In the large-scale procedure, plasmodia were grown in metal trays on a membrane supported by filter paper on stainless-steel screen. Plasmodia were started from a ring of inoculum to allow inward and outward migration and were incubated on a rocker so that nutrient medium would flow back and forth, wetting the undersurface of the plasmodium. Rocker and petri dish cultures had similar growth characteristics: (i) the interphase time between mitoses I and II and between II and III was about 8 hr; (ii) ribonucleic acid and protein increased essentially logarithmically throughout the cell cycle; and (iii) deoxyribonucleic acid increased only during early interphase and it doubled in approximately 3 hr after each mitosis. Rocker cultures were not as nearly synchronous as petri dish cultures and had a range in metaphase time (at mitosis III) within individual plasmodia of 15 to 45 min, as compared with 5 to 10 min in petri dish cultures.
Insights
Researchers developed a novel method to grow larger Physarum polycephalum plasmodia. This technique supports large-scale cultures with similar cell cycle characteristics to smaller petri dish experiments.
Area of Science:
- Cell Biology
- Mycology
Background:
- Physarum polycephalum is a model organism for studying cell growth and division.
- Previous methods limited the size of P. polycephalum plasmodia cultures.
Purpose of the Study:
- To develop a method for growing significantly larger Physarum polycephalum plasmodia.
- To compare the growth characteristics of large-scale cultures with traditional petri dish methods.
Main Methods:
- Plasmodia were cultured in metal trays on a membrane supported by filter paper and stainless-steel screen.
- Inoculum was applied as a ring to allow migration, and cultures were incubated on a rocker for nutrient flow.
- Growth characteristics including cell cycle timing, RNA, protein, and DNA synthesis were monitored.
Main Results:
- The new method produced plasmodia 8 to 10 times larger than petri dish cultures.
- Both rocker and petri dish cultures exhibited similar cell cycle timings and macromolecule synthesis patterns.
- Rocker cultures showed slightly less synchrony in metaphase compared to petri dish cultures.
Conclusions:
- The developed method enables large-scale cultivation of Physarum polycephalum plasmodia.
- This technique maintains key cellular growth characteristics observed in smaller cultures.
- The findings facilitate further research into P. polycephalum biology at a larger scale.