Crowdsourced Identification of Potential Target Genes for CTV Induced Gene Silencing for Controlling the Citrus Greening Vector Diaphorina citri .

Autor: Ramos JE; U.S. Horticultural Research Laboratory (USDA-ARS), Fort Pierce, FL, United States., Jain RG; Indian River Research and Education Center, UF/IFAS, Fort Pierce, FL, United States., Powell CA; Indian River Research and Education Center, UF/IFAS, Fort Pierce, FL, United States., Dawson WO; Citrus Research and Education Center, UF/IFAS, Lake Alfred, FL, United States., Gowda S; Citrus Research and Education Center, UF/IFAS, Lake Alfred, FL, United States., Borovsky D; Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, CO, United States., Shatters RG Jr; U.S. Horticultural Research Laboratory (USDA-ARS), Fort Pierce, FL, United States.
Jazyk: angličtina
Zdroj: Frontiers in physiology [Front Physiol] 2021 Apr 09; Vol. 12, pp. 571826. Date of Electronic Publication: 2021 Apr 09 (Print Publication: 2021).
DOI: 10.3389/fphys.2021.571826
Abstrakt: Citrus Greening or Huanglongbing (HLB) is a disease of citrus, causing high reduction in citrus production and is transmitted by the Asian citrus psyllid Diaphorina citri Kuwayama vectoring a phloem-limited bacterium Candidatus Liberibacter sp. We report research results using crowdsourcing challenge strategy identifying potential gene targets in D. citri to control the insect using RNA interference (RNAi). From 63 submitted sequences, 43 were selected and tested by feeding them to D. citri using 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 (BH 4 ) 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 adult D. citri than what was observed using artificial diet, reaching 100% when detached citrus leaves with the engineered CTV expressing dsRNA were fed to adult D. 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 feeding D. citri CTV-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.
Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
(Copyright © 2021 Ramos, Jain, Powell, Dawson, Gowda, Borovsky and Shatters.)
Databáze: MEDLINE
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