Exploring the Effectiveness and Durability of Trans-Kingdom Silencing of Fungal Genes in the Vascular Pathogen <i>Verticillium dahliae</i>
Host-induced gene silencing (HIGS) based on trans-kingdom RNA interference (RNAi) has been successfully exploited to engineer host resistance to pests and pathogens, including fungi and oomycetes. However, revealing the mechanisms underlying trans-kingdom RNAi between hosts and pathogens lags behind...
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MDPI AG
2022-03-01
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| Series: | International Journal of Molecular Sciences |
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| author | Tao Zhang Jian-Hua Zhao Yuan-Yuan Fang Hui-Shan Guo Yun Jin |
| author_facet | Tao Zhang Jian-Hua Zhao Yuan-Yuan Fang Hui-Shan Guo Yun Jin |
| author_sort | Tao Zhang |
| collection | DOAJ |
| description | Host-induced gene silencing (HIGS) based on trans-kingdom RNA interference (RNAi) has been successfully exploited to engineer host resistance to pests and pathogens, including fungi and oomycetes. However, revealing the mechanisms underlying trans-kingdom RNAi between hosts and pathogens lags behind applications. The effectiveness and durability of trans-kingdom silencing of pathogenic genes are uncharacterized. In this study, using our transgenic <i>35S-VdH1i</i> cotton plants in which dsVdH1-derived small RNAs (siVdH1) accumulated, small RNA sequencing analysis revealed that siVdH1s exclusively occur within the double-stranded (ds)VdH1 region, and no transitive siRNAs were produced beyond this region in recovered hyphae of <i>Verticillium dahliae</i> (<i>V. dahliae</i>). Accordingly, we found that <i>VdH1</i> silencing was reduced over time in recovered hyphae cultured in vitro, inferring that once the fungus got rid of the <i>35S-VdH1i</i> cotton plants would gradually regain their pathogenicity. To explore whether continually exporting dsRNAs/siRNAs from transgenic plants into recipient fungal cells guaranteed the effectiveness and stability of HIGS, we created GFP/RFP double-labeled <i>V. dahliae</i> and transgenic <i>Arabidopsis</i> expressing dsGFP (<i>35S</i>-<i>GFPi</i> plants). Confocal images visually demonstrate the efficient silencing of <i>GFP</i> in <i>V. dahliae</i> that colonized host vascular tissues. Taken together, our results demonstrate that HIGS effectively triggers long-lasting trans-kingdom RNAi during plant vasculature <i>V. dahliae</i> interactions, despite no amplification or transitivity of RNAi being noted in this soil-borne fungal pathogen. |
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| language | English |
| last_indexed | 2024-03-09T20:36:47Z |
| publishDate | 2022-03-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-4508f5bd3ab1453683133e8a12d1fbab2023-11-23T23:08:58ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-03-01235274210.3390/ijms23052742Exploring the Effectiveness and Durability of Trans-Kingdom Silencing of Fungal Genes in the Vascular Pathogen <i>Verticillium dahliae</i>Tao Zhang0Jian-Hua Zhao1Yuan-Yuan Fang2Hui-Shan Guo3Yun Jin4State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, CAS Center for Excellence in Biotic Interactions, University of the Chinese Academy of Sciences, Beijing 100049, ChinaState Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, CAS Center for Excellence in Biotic Interactions, University of the Chinese Academy of Sciences, Beijing 100049, ChinaState Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, CAS Center for Excellence in Biotic Interactions, University of the Chinese Academy of Sciences, Beijing 100049, ChinaState Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, CAS Center for Excellence in Biotic Interactions, University of the Chinese Academy of Sciences, Beijing 100049, ChinaState Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, CAS Center for Excellence in Biotic Interactions, University of the Chinese Academy of Sciences, Beijing 100049, ChinaHost-induced gene silencing (HIGS) based on trans-kingdom RNA interference (RNAi) has been successfully exploited to engineer host resistance to pests and pathogens, including fungi and oomycetes. However, revealing the mechanisms underlying trans-kingdom RNAi between hosts and pathogens lags behind applications. The effectiveness and durability of trans-kingdom silencing of pathogenic genes are uncharacterized. In this study, using our transgenic <i>35S-VdH1i</i> cotton plants in which dsVdH1-derived small RNAs (siVdH1) accumulated, small RNA sequencing analysis revealed that siVdH1s exclusively occur within the double-stranded (ds)VdH1 region, and no transitive siRNAs were produced beyond this region in recovered hyphae of <i>Verticillium dahliae</i> (<i>V. dahliae</i>). Accordingly, we found that <i>VdH1</i> silencing was reduced over time in recovered hyphae cultured in vitro, inferring that once the fungus got rid of the <i>35S-VdH1i</i> cotton plants would gradually regain their pathogenicity. To explore whether continually exporting dsRNAs/siRNAs from transgenic plants into recipient fungal cells guaranteed the effectiveness and stability of HIGS, we created GFP/RFP double-labeled <i>V. dahliae</i> and transgenic <i>Arabidopsis</i> expressing dsGFP (<i>35S</i>-<i>GFPi</i> plants). Confocal images visually demonstrate the efficient silencing of <i>GFP</i> in <i>V. dahliae</i> that colonized host vascular tissues. Taken together, our results demonstrate that HIGS effectively triggers long-lasting trans-kingdom RNAi during plant vasculature <i>V. dahliae</i> interactions, despite no amplification or transitivity of RNAi being noted in this soil-borne fungal pathogen.https://www.mdpi.com/1422-0067/23/5/2742host-induced gene silencingtrans-kingdom RNA interference<i>Verticillium dahliae</i> |
| spellingShingle | Tao Zhang Jian-Hua Zhao Yuan-Yuan Fang Hui-Shan Guo Yun Jin Exploring the Effectiveness and Durability of Trans-Kingdom Silencing of Fungal Genes in the Vascular Pathogen <i>Verticillium dahliae</i> International Journal of Molecular Sciences host-induced gene silencing trans-kingdom RNA interference <i>Verticillium dahliae</i> |
| title | Exploring the Effectiveness and Durability of Trans-Kingdom Silencing of Fungal Genes in the Vascular Pathogen <i>Verticillium dahliae</i> |
| title_full | Exploring the Effectiveness and Durability of Trans-Kingdom Silencing of Fungal Genes in the Vascular Pathogen <i>Verticillium dahliae</i> |
| title_fullStr | Exploring the Effectiveness and Durability of Trans-Kingdom Silencing of Fungal Genes in the Vascular Pathogen <i>Verticillium dahliae</i> |
| title_full_unstemmed | Exploring the Effectiveness and Durability of Trans-Kingdom Silencing of Fungal Genes in the Vascular Pathogen <i>Verticillium dahliae</i> |
| title_short | Exploring the Effectiveness and Durability of Trans-Kingdom Silencing of Fungal Genes in the Vascular Pathogen <i>Verticillium dahliae</i> |
| title_sort | exploring the effectiveness and durability of trans kingdom silencing of fungal genes in the vascular pathogen i verticillium dahliae i |
| topic | host-induced gene silencing trans-kingdom RNA interference <i>Verticillium dahliae</i> |
| url | https://www.mdpi.com/1422-0067/23/5/2742 |
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