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Expressed sequence tags reveal genetic diversity and putative virulence factors of the pathogenic oomycete Pythium
Theerapong Krajaejun1, Rommanee Khositnithikul, Tassanee Lerksuthirat
1Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand. mr_en@hotmail.com
Abstract:
Oomycetes are unique eukaryotic microorganisms that share a mycelial morphology with fungi. Many oomycetes are pathogenic to plants, and a more limited number are pathogenic to animals. Pythium insidiosum is the only oomycete that is capable of infecting both humans and animals, and causes a life-threatening infectious disease, called "pythiosis". In the majority of pythiosis patients life-long handicaps result from the inevitable radical excision of infected organs, and many die from advanced infection. Better understanding P. insidiosum pathogenesis at molecular levels could lead to new forms of treatment. Genetic and genomic information is lacking for P. insidiosum, so we have undertaken an expressed sequence tag (EST) study, and report on the first dataset of 486 ESTs, assembled into 217 unigenes. Of these, 144 had significant sequence similarity with known genes, including 47 with ribosomal protein homology. Potential virulence factors included genes involved in antioxidation, thermal adaptation, immunomodulation, and iron and sterol binding. Effectors resembling pathogenicity factors of plant-pathogenic oomycetes were also discovered, such as, a CBEL-like protein (possible involvement in host cell adhesion and hemagglutination), a putative RXLR effector (possibly involved in host cell modulation) and elicitin-like (ELL) proteins. Phylogenetic analysis mapped P. insidiosum ELLs to several novel clades of oomycete elicitins (ELIs), and homology modeling predicted that P. insidiosum ELLs should bind sterols. Most of the P. insidiosum ESTs showed homology to sequences in the genome or EST databases of other oomycetes, but one putative gene, with unknown function, was found to be unique to P. insidiosum. The EST dataset reported here represents the first steps in identifying genes of P. insidiosum and beginning transcriptome analysis. This genetic information will facilitate understanding of pathogenic mechanisms of this devastating pathogen.
Insights
This study identifies novel genes in Pythium insidiosum, a pathogen causing life-threatening pythiosis in humans and animals. The findings provide crucial genetic insights for developing new treatments against this devastating oomycete infection.
Area of Science:
- Microbiology
- Molecular Biology
- Genomics
Background:
- Oomycetes are eukaryotic microorganisms with fungal-like morphology, some causing plant and animal diseases.
- Pythium insidiosum is the sole oomycete infecting both humans and animals, causing severe, often fatal, pythiosis.
- Limited genetic and genomic data for P. insidiosum hinders understanding of its pathogenesis and treatment development.
Purpose of the Study:
- To initiate the genetic and genomic characterization of Pythium insidiosum.
- To identify potential virulence factors and unique genes within P. insidiosum.
- To lay the groundwork for transcriptome analysis and understanding pathogenic mechanisms.
Main Methods:
- Expressed Sequence Tag (EST) sequencing was performed on P. insidiosum.
- ESTs were assembled into unigenes, and sequence similarity searches were conducted against known gene databases.
- Phylogenetic analysis and homology modeling were used to characterize specific effector proteins.
Main Results:
- A dataset of 486 ESTs, assembled into 217 unigenes, was generated.
- 144 unigenes showed similarity to known genes, including 47 ribosomal protein homologs.
- Potential virulence factors identified include genes for antioxidation, thermal adaptation, immunomodulation, and sterol binding, alongside oomycete effectors like CBEL-like, RXLR, and elicitin-like (ELL) proteins.
- Phylogenetic analysis revealed novel clades of P. insidiosum ELLs, predicted to bind sterols.
- One unique putative gene, not found in other oomycetes, was identified.
Conclusions:
- The generated EST dataset is the first genetic resource for P. insidiosum, enabling further transcriptome analysis.
- The identified genes and potential virulence factors offer insights into P. insidiosum pathogenesis.
- This genetic information is crucial for advancing the understanding and treatment of pythiosis.
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