Direct Nanopore Sequencing of Novel RNA Viruses within Yeasts

Additional Funding Sources

The project described was supported by a student grant from the UI Office of Undergraduate Research.

Presentation Date

7-2019

Abstract

Fungal diseases are a constant threat to agriculture and can lead to large annual crop losses of up to 35%. Mycoviruses from the family Partitiviridae have been shown to confer a hypovirulent phenotype to their fungal hosts, which can lead to a lower infection rate and reduced crop loss. After the Rowley lab’s recent discovery of a novel group of double-stranded RNA (dsRNA) partitiviruses in Saccharomyces cerevisiae, we have begun to survey and characterize their diversity and prevalence in the Saccharomyces genus. We have developed a new protocol to extract dsRNAs from yeasts to determine the presence of partitiviruses in more than 300 strains of yeast. Currently, we have identified dsRNAs consistent with the presence of partitiviruses within ~14% of the surveyed strains. 11% of strains contain dsRNAs derived from partitivirus, while 3% are co-infected with multiple viruses from different families. 90.4% of all partitiviruses were found within yeast strains isolated from coffee and cacao beans. Many of these viruses consist of two dsRNA strands, ranging between 1.3kb and 1.6kb. Currently, we are developing a long-read sequencing method to rapidly determine the sequence cDNAs derived from partitivirus dsRNA using Oxford’s Nanopore technology. Infected strains have the potential as a new model to study the mechanisms and functions of partitiviruses. This will lead to their more effective usage as a hypovirulent-inducing mycovirus to treat fungal diseases.

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Direct Nanopore Sequencing of Novel RNA Viruses within Yeasts

Fungal diseases are a constant threat to agriculture and can lead to large annual crop losses of up to 35%. Mycoviruses from the family Partitiviridae have been shown to confer a hypovirulent phenotype to their fungal hosts, which can lead to a lower infection rate and reduced crop loss. After the Rowley lab’s recent discovery of a novel group of double-stranded RNA (dsRNA) partitiviruses in Saccharomyces cerevisiae, we have begun to survey and characterize their diversity and prevalence in the Saccharomyces genus. We have developed a new protocol to extract dsRNAs from yeasts to determine the presence of partitiviruses in more than 300 strains of yeast. Currently, we have identified dsRNAs consistent with the presence of partitiviruses within ~14% of the surveyed strains. 11% of strains contain dsRNAs derived from partitivirus, while 3% are co-infected with multiple viruses from different families. 90.4% of all partitiviruses were found within yeast strains isolated from coffee and cacao beans. Many of these viruses consist of two dsRNA strands, ranging between 1.3kb and 1.6kb. Currently, we are developing a long-read sequencing method to rapidly determine the sequence cDNAs derived from partitivirus dsRNA using Oxford’s Nanopore technology. Infected strains have the potential as a new model to study the mechanisms and functions of partitiviruses. This will lead to their more effective usage as a hypovirulent-inducing mycovirus to treat fungal diseases.