Gene Mining for Conserved, Non-Annotated Proteins of <i>Podosphaera xanthii</i> Identifies Novel Target Candidates for Controlling Powdery Mildews by Spray-Induced Gene Silencing
The powdery mildew fungus <i>Podosphaera xanthii</i> is one of the most important limiting factors for cucurbit production worldwide. Despite the significant efforts made by breeding and chemical companies, effective control of this pathogen remains elusive to growers. In this work, we e...
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MDPI AG
2021-09-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/7/9/735 |
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| author | Laura Ruiz-Jiménez Álvaro Polonio Alejandra Vielba-Fernández Alejandro Pérez-García Dolores Fernández-Ortuño |
| author_facet | Laura Ruiz-Jiménez Álvaro Polonio Alejandra Vielba-Fernández Alejandro Pérez-García Dolores Fernández-Ortuño |
| author_sort | Laura Ruiz-Jiménez |
| collection | DOAJ |
| description | The powdery mildew fungus <i>Podosphaera xanthii</i> is one of the most important limiting factors for cucurbit production worldwide. Despite the significant efforts made by breeding and chemical companies, effective control of this pathogen remains elusive to growers. In this work, we examined the suitability of RNAi technology called spray-induced gene silencing (SIGS) for controlling cucurbit powdery mildew. Using leaf disc and cotyledon infiltration assays, we tested the efficacy of dsRNA applications to induce gene silencing in <i>P. xanthii</i>. Furthermore, to identify new target candidate genes, we analyzed sixty conserved and non-annotated proteins (CNAPs) deduced from the <i>P. xanthii</i> transcriptome in silico. Six proteins presumably involved in essential functions, specifically respiration (CNAP8878, CNAP9066, CNAP10905 and CNAP30520), glycosylation (CNAP1048) and efflux transport (CNAP948), were identified. Functional analysis of these CNAP coding genes by dsRNA-induced gene silencing resulted in strong silencing phenotypes with large reductions in fungal growth and disease symptoms. Due to their important contributions to fungal development, the <i>CNAP1048</i>, <i>CNAP10905</i> and <i>CNAP30520</i> genes were selected as targets to conduct SIGS assays under plant growth chamber conditions. The spray application of these dsRNAs induced high levels of disease control, supporting that SIGS could be a sustainable approach to combat powdery mildew diseases. |
| first_indexed | 2024-03-10T07:32:06Z |
| format | Article |
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| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-10T07:32:06Z |
| publishDate | 2021-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-972a97076f674c9ba0947bbb470963032023-11-22T13:48:10ZengMDPI AGJournal of Fungi2309-608X2021-09-017973510.3390/jof7090735Gene Mining for Conserved, Non-Annotated Proteins of <i>Podosphaera xanthii</i> Identifies Novel Target Candidates for Controlling Powdery Mildews by Spray-Induced Gene SilencingLaura Ruiz-Jiménez0Álvaro Polonio1Alejandra Vielba-Fernández2Alejandro Pérez-García3Dolores Fernández-Ortuño4Departamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, 29071 Málaga, SpainDepartamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, 29071 Málaga, SpainDepartamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, 29071 Málaga, SpainDepartamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, 29071 Málaga, SpainDepartamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, 29071 Málaga, SpainThe powdery mildew fungus <i>Podosphaera xanthii</i> is one of the most important limiting factors for cucurbit production worldwide. Despite the significant efforts made by breeding and chemical companies, effective control of this pathogen remains elusive to growers. In this work, we examined the suitability of RNAi technology called spray-induced gene silencing (SIGS) for controlling cucurbit powdery mildew. Using leaf disc and cotyledon infiltration assays, we tested the efficacy of dsRNA applications to induce gene silencing in <i>P. xanthii</i>. Furthermore, to identify new target candidate genes, we analyzed sixty conserved and non-annotated proteins (CNAPs) deduced from the <i>P. xanthii</i> transcriptome in silico. Six proteins presumably involved in essential functions, specifically respiration (CNAP8878, CNAP9066, CNAP10905 and CNAP30520), glycosylation (CNAP1048) and efflux transport (CNAP948), were identified. Functional analysis of these CNAP coding genes by dsRNA-induced gene silencing resulted in strong silencing phenotypes with large reductions in fungal growth and disease symptoms. Due to their important contributions to fungal development, the <i>CNAP1048</i>, <i>CNAP10905</i> and <i>CNAP30520</i> genes were selected as targets to conduct SIGS assays under plant growth chamber conditions. The spray application of these dsRNAs induced high levels of disease control, supporting that SIGS could be a sustainable approach to combat powdery mildew diseases.https://www.mdpi.com/2309-608X/7/9/735cucurbitsdsRNA<i>Podosphaera xanthii</i>powdery mildewsRNAispray-induced gene silencing (SIGS) |
| spellingShingle | Laura Ruiz-Jiménez Álvaro Polonio Alejandra Vielba-Fernández Alejandro Pérez-García Dolores Fernández-Ortuño Gene Mining for Conserved, Non-Annotated Proteins of <i>Podosphaera xanthii</i> Identifies Novel Target Candidates for Controlling Powdery Mildews by Spray-Induced Gene Silencing Journal of Fungi cucurbits dsRNA <i>Podosphaera xanthii</i> powdery mildews RNAi spray-induced gene silencing (SIGS) |
| title | Gene Mining for Conserved, Non-Annotated Proteins of <i>Podosphaera xanthii</i> Identifies Novel Target Candidates for Controlling Powdery Mildews by Spray-Induced Gene Silencing |
| title_full | Gene Mining for Conserved, Non-Annotated Proteins of <i>Podosphaera xanthii</i> Identifies Novel Target Candidates for Controlling Powdery Mildews by Spray-Induced Gene Silencing |
| title_fullStr | Gene Mining for Conserved, Non-Annotated Proteins of <i>Podosphaera xanthii</i> Identifies Novel Target Candidates for Controlling Powdery Mildews by Spray-Induced Gene Silencing |
| title_full_unstemmed | Gene Mining for Conserved, Non-Annotated Proteins of <i>Podosphaera xanthii</i> Identifies Novel Target Candidates for Controlling Powdery Mildews by Spray-Induced Gene Silencing |
| title_short | Gene Mining for Conserved, Non-Annotated Proteins of <i>Podosphaera xanthii</i> Identifies Novel Target Candidates for Controlling Powdery Mildews by Spray-Induced Gene Silencing |
| title_sort | gene mining for conserved non annotated proteins of i podosphaera xanthii i identifies novel target candidates for controlling powdery mildews by spray induced gene silencing |
| topic | cucurbits dsRNA <i>Podosphaera xanthii</i> powdery mildews RNAi spray-induced gene silencing (SIGS) |
| url | https://www.mdpi.com/2309-608X/7/9/735 |
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