Crowdsourced Identification of Potential Target Genes for CTV Induced Gene Silencing for Controlling the Citrus Greening Vector Diaphorina citri
| Autor: | Dov Borovsky, William O. Dawson, Charles A. Powell, Siddarame Gowda, Ritesh G. Jain, Robert G. Shatters, John E. Ramos |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Genetics
lcsh:QP1-981 biology Candidatus Liberibacter Physiology Diaphorina citri Transgene Rieske down regulation Citrus tristeza virus food and beverages biology.organism_classification lcsh:Physiology RNA silencing RNA interference Citrus tristeza virus engineering Physiology (medical) Gene silencing gene silencing with dsRNA Gene Original Research |
| Zdroj: | Frontiers in Physiology Frontiers in Physiology, Vol 12 (2021) |
| ISSN: | 1664-042X |
| Popis: | Citrus Greening or Huanglongbing (HLB) is a disease of citrus, causing high reduction in citrus production and is transmitted by the Asian citrus psyllidDiaphorina citriKuwayama vectoring a phloem-limited bacteriumCandidatus Liberibactersp. We report research results using crowdsourcing challenge strategy identifying potential gene targets inD. citrito control the insect using RNA interference (RNAi). From 63 submitted sequences, 43 were selected and tested by feeding them toD. citriusing artificial diet assays. After feeding on artificial diet, the three most effective dsRNAs causing 30% mortality above control silenced genes expressing iron-sulfur cluster subunit of the mitochondrial electron transport chain complex (Rieske), heme iron-binding terminal oxidase enzyme (Cytochrome P450) and tetrahydrobiopterin (BH4) pathway enzyme (Pterin 4α-Carbinolamine Dehydratase). These sequences were cloned into a citrus phloem-limited virus (Citrus tristeza virus, CTV T36) expressing dsRNA against these target genes in citrus. The use of a viral mediated “para-transgenic” citrus plant system caused higher mortality to adultD. citrithan what was observed using artificial diet, reaching 100% when detached citrus leaves with the engineered CTV expressing dsRNA were fed to adultD. citri. Using this approach, a virus-induced gene silencing (VIGS) can be used to test future transgenic cultivars before genetically engineering citrus. RNA Seq analysis after feedingD. citriCTV-RIE on infected leaves identified transcriptionally modified genes located upstream and downstream of the targeted RIE gene. These genes were annotated showing that many are associated with the primary function of the Rieske gene that was targeted by VIGS. |
| Databáze: | OpenAIRE |
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