Knockdown of <i>Bmp1</i> and <i>Pls1</i> Virulence Genes by Exogenous Application of RNAi-Inducing dsRNA in <i>Botrytis cinerea</i>
<i>Botrytis cinerea</i> is a pathogen of wide agronomic and scientific importance partly due to its tendency to develop fungicide resistance. Recently, there has been great interest in the use of RNA interference as a control strategy against <i>B. cinerea</i>. In order to re...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-03-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/5/4869 |
| Summary: | <i>Botrytis cinerea</i> is a pathogen of wide agronomic and scientific importance partly due to its tendency to develop fungicide resistance. Recently, there has been great interest in the use of RNA interference as a control strategy against <i>B. cinerea</i>. In order to reduce the possible effects on non-target species, the sequence-dependent nature of RNAi can be used as an advantage to customize the design of dsRNA molecules. We selected two genes related to virulence: <i>BcBmp1</i> (a MAP kinase essential for fungal pathogenesis) and <i>BcPls1</i> (a tetraspanin related to appressorium penetration). After performing a prediction analysis of small interfering RNAs, dsRNAs of 344 (<i>BcBmp1</i>) and 413 (<i>BcPls1</i>) nucleotides were synthesized in vitro. We tested the effect of topical applications of dsRNAs, both in vitro by a fungal growth assay in microtiter plates and in vivo on artificially inoculated detached lettuce leaves. In both cases, topical applications of dsRNA led to gene knockdown with a delay in conidial germination for <i>BcBmp1</i>, an evident growth retardation for <i>BcPls1</i>, and a strong reduction in necrotic lesions on lettuce leaves for both genes. Furthermore, a strongly reduced expression of the <i>BcBmp1</i> and <i>BcPls1</i> genes was observed in both in vitro and in vivo experiments, suggesting that these genes could be promising targets for the development of RNAi-based fungicides against <i>B. cinerea</i>. |
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| ISSN: | 1661-6596 1422-0067 |