Generation of protoplasts provides a powerful experimental research tool for biological and pathogenicity studies of Pythium insidiosum
Issued Date
2023-11-01
Resource Type
ISSN
11565233
eISSN
17730449
Scopus ID
2-s2.0-85170565275
Pubmed ID
37678114
Journal Title
Journal of Medical Mycology
Volume
33
Issue
4
Rights Holder(s)
SCOPUS
Bibliographic Citation
Journal of Medical Mycology Vol.33 No.4 (2023)
Suggested Citation
Sae-Chew P., Rujirawat T., Lohnoo T., Yingyong W., Kumsang Y., Payattikul P., Yurayart N., Yurayart C., Krajaejun T. Generation of protoplasts provides a powerful experimental research tool for biological and pathogenicity studies of Pythium insidiosum. Journal of Medical Mycology Vol.33 No.4 (2023). doi:10.1016/j.mycmed.2023.101430 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/90068
Title
Generation of protoplasts provides a powerful experimental research tool for biological and pathogenicity studies of Pythium insidiosum
Author's Affiliation
Other Contributor(s)
Abstract
Introduction: Pythiosis is a high-mortality infectious condition in humans and animals. The etiologic agent is Pythium insidiosum. Patients present with an ocular, vascular, cutaneous/subcutaneous, or gastrointestinal infection. Antifungal medication often fails to fight against P. insidiosum. The effective treatment is limited to radical surgery, resulting in organ loss. Fatal outcomes are observed in advanced cases. Pythiosis needs to be studied to discover novel methods for disease control. Genome data of P. insidiosum is publicly available. However, information on P. insidiosum biology and pathogenicity is still limited due to the lack of a cost-effective animal model and molecular tools. Materials and methods: We aimed to develop a high-efficiency protocol for generating P. insidiosum protoplast, and used it to set up an animal model, in vitro drug susceptibility assay, and DNA transformation for this pathogen. Results: P. insidiosum protoplast was successfully generated to establish a feasible pythiosis model in embryonic chicken eggs and an efficient in vitro drug susceptibility assay. DNA transformation is a critical method for gene manipulation necessary for functional genetic studies in pathogens. Attempts to establish a DNA transformation method for P. insidiosum using protoplast were partly successful. Significant work needs to be done for genetically engineering a more robust selection marker to generate stable transformants at increased efficiency. Conclusion: This study is the first to report an efficient P. insidiosum protoplast production for clinical and research applications. Such advances are crucial to speeding up the pathogen's biology and pathogenicity exploration.