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Spheroplast fusion and heterokaryon formation in Mucor racemosus
Abstract:
Heterokaryons of Mucor racemosus were produced by fusion of spheroplasts from two auxotrophic strains of the fungus. Germinated sporangiospores were converted to spheroplasts by using commercial chitinase and Myxobacter AL-1 chitosanase. Spheroplasts from the auxotrophic strains were mixed in a buffered Ca(NO3)2 solution and fusion occurred. After cell wall regeneration, prototrophs were isolated. The frequency of heterokaryon formation was 1.45 X 10(-4). Prototrophic isolates segregated parental nuclei at a high frequency, indicating that heterokaryons had formed.
Insights
Researchers fused auxotrophic strains of Mucor racemosus to create heterokaryons. This fungal fusion technique successfully produced prototrophic colonies, demonstrating successful genetic exchange.
Area of Science:
- Mycology
- Fungal Genetics
- Cell Biology
Background:
- Auxotrophic strains of Mucor racemosus were utilized.
- Understanding fungal heterokaryon formation is crucial for genetic studies.
Purpose of the Study:
- To produce heterokaryons of Mucor racemosus through protoplast fusion.
- To investigate the frequency and stability of heterokaryon formation.
Main Methods:
- Germinated sporangiospores were converted to spheroplasts using chitinase and chitosanase.
- Spheroplasts from auxotrophic strains were induced to fuse in a buffered Ca(NO3)2 solution.
- Prototrophs were isolated after cell wall regeneration.
Main Results:
- Heterokaryons of Mucor racemosus were successfully produced.
- The frequency of heterokaryon formation was determined to be 1.45 X 10(-4).
- Prototrophic isolates exhibited high-frequency segregation of parental nuclei, confirming heterokaryon formation.
Conclusions:
- Protoplast fusion is an effective method for generating heterokaryons in Mucor racemosus.
- The observed segregation patterns indicate the presence and genetic instability of the formed heterokaryons.