Double-Stranded RNA Uptake through Topical Application, Mediates Silencing of Five CYP4 Genes and Suppresses Insecticide Resistance in Diaphorina citri

Killiny, Nabil; Hajeri, Subhas; Tiwari, Siddharth; Gowda, Siddarame; Stelinski, Lukasz L.

doi:10.1371/journal.pone.0110536

Cited by 141 publications

(166 citation statements)

References 41 publications

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“…Similar results from RNAi studies in ACP have also been reported1730313245. However, the results presented here also indicate that, apparently, ACP is very sensitive to RNAi, as a small dose of dsRNA (100 ng) administered through a citrus flush (new growth tissue between 0.25 g to 0.35 g) could induce significant RNAi responses (target gene suppression and death).…”

Section: Discussionsupporting

confidence: 89%

Asian Citrus Psyllid RNAi Pathway – RNAi evidence

Taning

Andrade

Hunter

et al. 2016

Sci Rep

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Diaphorina citri, known as the Asian citrus psyllid, is an important pest of citrus because it transmits a phloem-limited bacteria strongly implicated in huanglongbing (citrus greening disease). Emerging biotechnologies, such as RNA interference, could provide a new sustainable and environmentally friendly strategy for the management of this pest. In this study, genome and functional analysis were performed to verify whether the RNAi core genes are present in the Asian psyllid genome and if the RNAi machinery could be exploited to develop a management strategy for this pest. Analyses of RNAi-related genes in the Asian citrus psyllid genome showed an absence of sequences encoding R2D2, a dsRNA-binding protein that functions as a cofactor of Dicer-2 in Drosophila. Nevertheless, bioassays using an in Planta System showed that the Asian citrus psyllid was very sensitive to ingested dsRNA, demonstrating a strong RNAi response. A small dose of dsRNA administered through a citrus flush was enough to trigger the RNAi mechanism, causing significant suppression of the targeted transcript, and increased psyllid mortality. This study provides evidence of a functional RNAi machinery, which could be further exploited to develop RNAi based management strategies for the control of the Asian citrus psyllid.

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Section: Discussionsupporting

confidence: 89%

Asian Citrus Psyllid RNAi Pathway – RNAi evidence

Taning

Andrade

Hunter

et al. 2016

Sci Rep

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“…The RISC complex uses the remaining guide strand to bind to complementary viral mRNAs, cleave them, and inhibit viral replication. Silencing insect genes using RNAi is a promising new tool for D. citri management (El-Shesheny et al, 2013; Killiny et al, 2014). The primary components of the RNAi system, Argonaut, Dicer, and SID-1, were identified in D. citri (Table 5).…”

Section: Resultsmentioning

confidence: 99%

Annotation of the Asian Citrus Psyllid Genome Reveals a Reduced Innate Immune System

2016

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Citrus production worldwide is currently facing significant losses due to citrus greening disease, also known as Huanglongbing. The citrus greening bacteria, Candidatus Liberibacter asiaticus (CLas), is a persistent propagative pathogen transmitted by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). Hemipterans characterized to date lack a number of insect immune genes, including those associated with the Imd pathway targeting Gram-negative bacteria. The D. citri draft genome was used to characterize the immune defense genes present in D. citri. Predicted mRNAs identified by screening the published D. citri annotated draft genome were manually searched using a custom database of immune genes from previously annotated insect genomes. Toll and JAK/STAT pathways, general defense genes Dual oxidase, Nitric oxide synthase, prophenoloxidase, and cellular immune defense genes were present in D. citri. In contrast, D. citri lacked genes for the Imd pathway, most antimicrobial peptides, 1,3-β-glucan recognition proteins (GNBPs), and complete peptidoglycan recognition proteins. These data suggest that D. citri has a reduced immune capability similar to that observed in A. pisum, P. humanus, and R. prolixus. The absence of immune system genes from the D. citri genome may facilitate CLas infections, and is possibly compensated for by their relationship with their microbial endosymbionts.

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“…Data generated thus far suggest that cross-resistance is not likely between these 2 insecticides based on known mechanisms of resistance (Nauen et al 2013(Nauen et al , 2015. It remains to be demonstrated experimentally, but given the difference in chemical structure, it is unlikely the CYP4 P450 monooxygenases associated with imidacloprid exposure in D. citri will also be responsive to flupyradifurone (Tiwari et al 2011b;Killiny et al 2014). Interestingly, the RR50 estimates for flupyradifurone clustered with those of imidacloprid and thiamethoxam, except in 2 cases where thiamethoxam was higher and the RR50 estimate for flupyradifurone remained similar to that observed for imidacloprid (Fig.…”

Section: Flupyradifuronementioning

confidence: 99%

“…CYP genes code for a major class of enzymes (cytochrome P450 monooxygenases) frequently associated with insecticide resistance in insects (Li et al 2007). Subsequent molecular and biochemical studies using RNA interference further linked CYP4 expression and imidacloprid by demonstrating that silencing of the CYP4 genes in adult psyllids resulted in a decrease in P450 enzymatic activity with a concomitant increase in susceptibility to imidacloprid (Killiny et al 2014).…”

Section: Introductionmentioning

confidence: 99%