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Ilan Sela
Hadassah Medical Center
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Nitzan Paldi
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SOUTHWESTERN ENTOMOLOGIST
SCIENTIFIC NOTE
MAR. 2012
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Fig. 1. Detection of dsRNA molecules in Citrus trees by Real time RT-PCR. Meltcurve analyses are shown of replicated samples using dsRNA-specific primers on
total RNA isolated at 35 and 57 days post-treatment, dsRNA (81C). Melt Curve
temperatures listed at top of curves are mean values of three repeated samples,
from two replicates. Control samples for citrus dehydrin occurred at (84.8C).
RNAi effects from species specific dsRNAs, made to arginine kinase, under
laboratory conditions, reduced survival of two psyllids species: Asian citrus psyllid,
Diaphorina citri Kuwayama, and potato psyllid, Bactericera cockerelli (Sulc)
(Hemiptera: Psyllidae), and the glassy-winged sharpshooter, Homalodisca
vitripennis (Germar) (Hemiptera: Cicadellidae). The RNAi-increased mortality,
worked in feeding assays of dsRNAs in sucrose solutions, cut plant tissue
absorption, and rooted plantlets. Treatments of Citrus trees and grapevines
demonstrated uptake into whole plant systems by root drench and injections.
Introduced dsRNA was detected in Citrus trees, 6-year-old Mexican limes,
approximately 2.5 m tall, 7 weeks post treatment, after a single treatment (2 g
dsRNA in 15 liters of water). Previous work supporting the efficacy of RNAi
technology also showed silencing endogenous insect genes both by injection and
feeding, to a wide range of insects; honey bees, Apis mellifera L.; beetles;
whiteflies; mosquitoes; flies; and others (Price and Gatehouse 2008, Tomoyasu et
al. 2008, Maori et al. 2009, Whyard et al. 2009, Hunter et al. 2010c). The evidence
supports development of RNAi approaches for area-wide, host-delivered, population
suppression strategies of insect pests. RNAi-based pest suppression, HiSPeC,
may be suitable to reduce insects and transmitted disease pathogens (i.e.,
Huanglongbing in citrus, Pierces disease of grapevine, and zebra chip of potato,
Solanum tuberosum L.). In conclusion, this is the first report on the delivery and
stability of dsRNA treatments in field conditions, i.e., Citrus trees and grapevine,
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over a 3-month period. This discovery, while not yet fully understood, is a
breakthrough that expands current understanding of the use and applications of
RNAi management of insect pests and plant diseases.
Acknowledgment
The authors thank Maria Gonzalez, Biological Science Technician, USDA,
ARS, U.S. Horticultural Research Lab, Fort Pierce, FL 34945.
References Cited
Bells, X. 2010. Beyond Drosophila: RNAi in vivo and functional genomics in
insects. Annu. Rev. Entomol. 55: 111-128.
Castanotto, D., and J. J. Rossi. 2009. The promises and pitfalls of RNAinterference-based therapeutics. Nature 457: 426-433.
Hunter, W. B., B. R. Bextine, R. G. Shatters, Jr., and D. G. Hall. 2010a. Emerging
psyllid genomes increasing RNAinterference targets for insect management.
DSP1. 93rd Annual Meeting Florida Entomol. Soc., 25-28 July.
http://flaentsoc.org/2010annmeetabstracts.pdf.
Hunter, W. B., E. Glick, B. R. Bextine, and N. Paldi. 2010b. RNAinterference to
reducing insect pests: plant infusions. DSP2. 93rd Annual meeting Florida
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Hunter, W., J. Ellis, D. vanEngelsdorp, J. Hayes, D. Westervelt, E. Glick, M.
Williams, I. Sela, E. Maori, J. Pettis, D. Cox-Foster, and N. Paldi. 2010c.
Large-Scale Field Application of RNAi Technology Reducing Israeli Acute
Paralysis Virus Disease in Honey Bees (Apis mellifera, Hymenoptera: Apidae).
PLoS Pathogens 6(12): e1001160.
Huvenne, H., and G. Smagghe. 2010. Mechanisms of dsRNA uptake in insects
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Maori E., N. Paldi, S. Shafir, H. Kalev, E. Tsur, E. Glick, and I. Sela. 2009. IAPV, a
bee-affecting virus associated with colony collapse disorder can be silenced by
dsRNA ingestion. Ins. Mol. Biol. 18: 55-60.
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insects. Trends in Biotechnol. 26: 393-400.
Saleh, M. C., R. P. van Rij, A. Hekele, A. Gillis, E. Foley, P. H. OFarrell, and R.
Andino. 2006. The endocytic pathway mediates cell entry of dsRNA to induce
RNAi silencing. Nature Cell Biology 8: 793-802.
Siomi, H., and M. C. Siomi. 2009. On the road to reading the RNA-interference
code. Nature 457: 396-404.
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Bucher. 2008. Exploring systemic RNA interference in insects: a genomewide survey for RNAi genes in Tribolium. Genome Biology 9, R10.
Whyard, S., A. D. Singh, and S. Wong. 2009. Ingested double-stranded RNAs can
act as species-specific insecticides Ins. Biochem. Mol. Biol. 39: 824-832.
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