Publicação
Designing RNA based biopesticides for the control of cork oak phytopathogens
| Resumo: | Cork oak (Quercus suber L.) is an evergreen tree species with a great ecological and economic impact along the Mediterranean basin. Although being widely distributed, Q. suber forest has been declining for the past 80 years, being further enhanced by the climate changes. An example is severe drought, which is exposing cork oaks to an extra layer of stress allowing opportunistic pathogenic attacks. An example is the charcoal diseases caused by Biscogniauxia mediterranea, and bot canker caused by Diplodia corticola. The lack of a specific treatment for both diseases and the progressive ban of chemical fungicides in the EU, calls for new and innovative alternatives. In this context, RNA interference (RNAi) based fungicides are a promising strategy. Spray-induced gene silencing (SIGS) is a state-of-the-art RNAi silencing approach that targets specific pathogen genes, following the spraying of the host plant with a solution containing double stranded RNA (dsRNA). In this work, the possibility of using such approach for controlling both cork oak pathogens was explored. For the success of this biotechnological solution, the pathogen needs to uptake dsRNA molecules and process them for silencing targeted genes. In silico results revealed that both B. mediterranea and D. corticola contained the required protein machinery to uptake and process exogenous dsRNA. This result was corroborated by testing the capacity of both fungi in uptaking a labelled sGFT-dsRNA and by using an in vitro assay with sGFP transformants for testing silencing machinery. After testing the designed dsRNAs for fungal growth impairment, differences were detected in both cork oak pathogens. B. mediterranea appears to be more recalcitrant to the use of a SIGS approach than D. corticola. This work provided evidences of the potential of SIGS in controlling cork oak pathogens, particularly D. corticola. |
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| Autores principais: | Ferreira, Pedro Miguel Bento |
| Assunto: | Cork oak RNA interference Spray-induced gene silencing (SIGS) Biscogniauxia mediterranea Diplodia corticola Sobreiro RNA de interferência |
| Ano: | 2022 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade do Minho |
| Idioma: | inglês |
| Origem: | RepositóriUM - Universidade do Minho |
| Resumo: | Cork oak (Quercus suber L.) is an evergreen tree species with a great ecological and economic impact along the Mediterranean basin. Although being widely distributed, Q. suber forest has been declining for the past 80 years, being further enhanced by the climate changes. An example is severe drought, which is exposing cork oaks to an extra layer of stress allowing opportunistic pathogenic attacks. An example is the charcoal diseases caused by Biscogniauxia mediterranea, and bot canker caused by Diplodia corticola. The lack of a specific treatment for both diseases and the progressive ban of chemical fungicides in the EU, calls for new and innovative alternatives. In this context, RNA interference (RNAi) based fungicides are a promising strategy. Spray-induced gene silencing (SIGS) is a state-of-the-art RNAi silencing approach that targets specific pathogen genes, following the spraying of the host plant with a solution containing double stranded RNA (dsRNA). In this work, the possibility of using such approach for controlling both cork oak pathogens was explored. For the success of this biotechnological solution, the pathogen needs to uptake dsRNA molecules and process them for silencing targeted genes. In silico results revealed that both B. mediterranea and D. corticola contained the required protein machinery to uptake and process exogenous dsRNA. This result was corroborated by testing the capacity of both fungi in uptaking a labelled sGFT-dsRNA and by using an in vitro assay with sGFP transformants for testing silencing machinery. After testing the designed dsRNAs for fungal growth impairment, differences were detected in both cork oak pathogens. B. mediterranea appears to be more recalcitrant to the use of a SIGS approach than D. corticola. This work provided evidences of the potential of SIGS in controlling cork oak pathogens, particularly D. corticola. |
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