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Mating-type genes from asexual phytopathogenic ascomycetes Fusarium oxysporum and Alternaria alternata
1Microbial Toxicology Laboratory, RIKEN, Wako, Saitama, Japan. arie@cc.tuat.ac.jp
Abstract:
Mating-type (MAT) loci were cloned from two asexual (mitosporic) phytopathogenic ascomycetes, Fusarium oxysporum (a pyrenomycete) and Alternaria alternata (a loculoascomycete), by a polymerase chain reaction (PCR)-based strategy. The conserved high mobility group (HMG) box domain found in the MAT1-2-1 protein was used as a starting point for cloning and sequencing the entire MAT1-2 idiomorph plus flanking regions. Primer pairs designed to both flanking regions were used to amplify the opposite MAT1-1 idiomorph. The MAT1-1 and MAT1-2 idiomorphs were approximately 4.6 and 3.8 kb in F. oxysporum and approximately 1.9 and 2.2 kb in A. alternata, respectively. In both species, the MAT1-1 idiomorph contains at least one gene that encodes a protein with a putative alpha box domain and the MAT1-2 idiomorph contains one gene that encodes a protein with a putative HMG box domain. MAT-specific primers were used to assess the mating type of F. oxysporum and A. alternata field isolates by PCR. MAT genes from A. alternata were expressed. The A. alternata genes were confirmed to be functional in a close sexual relative, Cochliobolus heterostrophus, by heterologous expression.
Insights
Researchers cloned mating-type (MAT) loci from two fungal pathogens, Fusarium oxysporum and Alternaria alternata, using PCR. They identified key genes and confirmed their function, advancing fungal mating-type research.
Area of Science:
- Mycology
- Molecular Biology
- Genetics
Background:
- Asexual (mitosporic) fungi pose significant challenges in agriculture and medicine.
- Understanding mating-type (MAT) loci is crucial for fungal reproduction and genetic studies.
Purpose of the Study:
- To clone and characterize the mating-type (MAT) loci from two phytopathogenic fungi: Fusarium oxysporum and Alternaria alternata.
- To investigate the structure and potential function of MAT genes in these asexual fungi.
Main Methods:
- Polymerase chain reaction (PCR)-based cloning strategy utilizing conserved domains.
- Sequencing of MAT idiomorphs and flanking regions.
- Heterologous expression in Cochliobolus heterostrophus to confirm gene function.
Main Results:
- Successfully cloned and sequenced MAT1-1 and MAT1-2 idiomorphs from F. oxysporum and A. alternata.
- Identified genes encoding putative alpha box and HMG box proteins within the MAT loci.
- Confirmed expression and functionality of A. alternata MAT genes through heterologous expression.
Conclusions:
- The study successfully identified and characterized MAT loci in two important fungal pathogens.
- The cloned MAT genes are conserved and functional, providing tools for further research into fungal mating and evolution.
- This work contributes to understanding the genetic basis of mating in agriculturally significant fungi.