Silencing Cytochrome P51 Gene With DSRNA to Combat Peach Fungal Pathogens

School Name

South Carolina Governor's School for Science & Mathematics

Grade Level

12th Grade

Presentation Topic

Microbiology

Presentation Type

Mentored

Oral Presentation Award

2nd Place

Abstract

California, South Carolina, and Georgia are large peach producers worldwide. They face adversity with fungal peach pathogens Armilaria mellea and Botrytis cinerea. These fungi cause millions of dollars in crop loss annually. A. mellea causes root rot. B. cinerea attacks developed fruit and reduces shelf-life. The current prevention method is chemical fungicides which are expensive, harmful to the environment, and ineffective. In hopes of creating a better fungicide, we focused on the Cytochrome p51 gene in A. mellea that controls cell wall synthesis. If the gene were silenced, it would prevent the fungus from being able to grow. Genomic DNA containing Cyp51 was replicated by PCR and used to create double stranded RNA (dsRNA). Previous studies indicate dsRNA can be absorbed topically into the fungal cells creating RNA interference and reducing gene expression. The dsRNA was applied to PDA media and plated with live fungal cells. After 20 days of cultivation, the radial growth was measured and compared to the radial growth of the control, which was plated with water. dsRNA treatment did slightly reduce the radial growth of the fungus and silence the Cyp51 gene. This experiment will be replicated with increased sample size and a new variety of concentrations. It will also be repeated with B. cinerea and target it’s DCL genes, which control methylation in a cell. The use of dsRNA as a fungicide in the future may provide more efficient, eco-friendly, and fungus specific methods of fungal control.

Location

Founders Hall 213 B

Start Date

3-30-2019 11:30 AM

Presentation Format

Oral Only

Group Project

No

COinS
 
Mar 30th, 11:30 AM

Silencing Cytochrome P51 Gene With DSRNA to Combat Peach Fungal Pathogens

Founders Hall 213 B

California, South Carolina, and Georgia are large peach producers worldwide. They face adversity with fungal peach pathogens Armilaria mellea and Botrytis cinerea. These fungi cause millions of dollars in crop loss annually. A. mellea causes root rot. B. cinerea attacks developed fruit and reduces shelf-life. The current prevention method is chemical fungicides which are expensive, harmful to the environment, and ineffective. In hopes of creating a better fungicide, we focused on the Cytochrome p51 gene in A. mellea that controls cell wall synthesis. If the gene were silenced, it would prevent the fungus from being able to grow. Genomic DNA containing Cyp51 was replicated by PCR and used to create double stranded RNA (dsRNA). Previous studies indicate dsRNA can be absorbed topically into the fungal cells creating RNA interference and reducing gene expression. The dsRNA was applied to PDA media and plated with live fungal cells. After 20 days of cultivation, the radial growth was measured and compared to the radial growth of the control, which was plated with water. dsRNA treatment did slightly reduce the radial growth of the fungus and silence the Cyp51 gene. This experiment will be replicated with increased sample size and a new variety of concentrations. It will also be repeated with B. cinerea and target it’s DCL genes, which control methylation in a cell. The use of dsRNA as a fungicide in the future may provide more efficient, eco-friendly, and fungus specific methods of fungal control.