RNA interference: potential uses on functional genomics and genetic control of Hypothenemus hampei (Coleoptera: Scolytinae)
Main Article Content
Eukaryotic organisms, including insects, have a mechanism for specific gene silencing by short sequences of double-stranded RNA (dsRNA), such a process is known as RNA interference (RNAi). The objective of this research was to follow up this technique to screen candidate genes for the genetic control of the coffee berry borer Hypothenemus hampei. A methodology of feeding by oral drops containing different dsRNAs and dsRNAs´ dosages that were placed in the preoral cavity of first instar larvae was used. Then, the larvae were reared on an artificial diet supplemented with the respective dsRNA. Previously, we verified the effectiveness of the dsRNA feeding using two control strategies: a) intake of a fluorescent compound (CalcofluorTM) and b) intake of a commercial mixture of protease inhibitors. The genes coding for mannanase, xylanase, cytochrome P450 mono-oxygenase (two genes), ATPase D, α-tubulin and actin, expressed in the coffee berry borer, were selected for silencing. We found that dsRNA of cytochromes P450 produced the greatest effect, causing larval mortality of 64% and 52% respectively, followed by dsRNA of mannanase and xylanase, with a mortality rate of 37% and 33%, respectively, for the highest concentration tested.
ASHIHARA, H.; SANO, H.; CROZIER, A. 2008. Caffeine and related purine alkaloids: Biosynthesis, catabolism, function and genetic engineering. Phytochemstry 69: 841-856.
BAKER, P.S. 1999. La broca del café en Colombia. Informe final del proyecto MIP para el café DFID-Cenicafé-CABI Bioscience (CNTR 93/1536 A). Chinchiná, Colombia, DFID. 154 p.
BAUM, J.; BOGAERT, T.; CLINTON, W.; HECK, G.R.; FELDMANN, P.; ILAGAN, O.; JOHNSON, S.; PLAETINCK, G.; MUNYIKWA, T.; PLEAU, M.; VAUGHN1, T.Y.; ROBERTS, J. 2007. Control of coleopteran insect pests through RNA interference. Nature Biotechnology 25: 1322-1326.
BAUTISTA, M.A.M.; MIYATA, T.; MIURA, K.; TANAKA, T. 2009. RNA interference-mediated knockdown of a cytochrome P450, CYP6BG1, from the diamondback moth, Plutella xylostella, reduces larval resistance to permethrin. Insect Biochemistry Molecular Biology 39:38-46.
BUSTILLO P, A. 2008. Aspectos sobre la broca del café, Hypothenemus hampei, en Colombia. pp. 388-418. En: Bustillo P, A.E. (Ed.). Los insectos y su manejo en la caficultora colombiana. Federación Nacional de Cafeteros. Chinchiná. Colombia. 466 p.
BUSTILLO, A.E.; CÁRDENAS, R.; VILLALBA, D.A.; BENAVIDES, P.; OROZCO, J.; POSADA, F.J. 1998. Manejo Integrado de la Broca, Hypothenemus hampei (Ferrari), en Colombia. Cenicafe, Chinchiná, Colombia. 134 p.
CADENA, G.G. 1991. Sostenibilidad de la producción cafetera, el control de plagas y enfermedades. Ensayos Sobre Economía Cafetera 4 (6): 19-26.
CAMAYO, G.; ARCILA, J. 1997. Desarrollo floral del cafeto en condiciones de la zona cafetera colombiana (Chinchiná Caldas). Cenicafé , Avances técnicos, No. 245. Chinchiná, Colombia. 8 p.
CHAVES, B.; RILEY, J. 2001. Determination of factors influencing integrated pest management adoption in coffee berry borer in Colombian farms. Agriculture, Ecosystems and Environment 87: 159-177.
CHEN, M-S. 2008. Inducible direct plant defense against insect herbivores: A review. Insect science 15: 101-114.
CLIFFORD, M. N.; IN, M. N.; CLIFFORD, K. C. 1985. Chemical and physical aspects of green coffee and coffee products. Wilson (eds), pp. 305-374.
FIRE, A.; XU,S.; MONTGOMERY, M.K.; KOSTAS, S.A.; DRIVER, S.E.; MELLO, C.C . 1998. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391(6669): 806-11.
FISCHER, M.; REIMANN, S.; TROVATO, V.; REDGWELL, R.J. 2001. Polysaccharides of green Arabica and Robusta coffee beans. Carbohydrate Research 330: 93-101.
GRIEBLER, M.; WESTERLUND, S.A.; HOFFMANN, K.H.; MEYERING-VOS, M. 2008. RNA interference with the allatoregulating neuropeptide genes from the fall armyworm Spodoptera frugiperda and its effects on the JH titer in the hemolymph. Journal of Insect Physiology 54: 997-1007.
GUERREIRO, O.; MAZZAFERA, P. 2003. Caffeine and resistance of coffee to the berry borer Hypothenemus hampei (Coleoptera: Scolytidae). Journal of Agricultural and Food Chemistry 51: 6987-6991.
GOKULAKRISHNAN, S.; CHANDRARAJ, K.; SATHYANARAYANA, N. GUMMADI. 2005 Microbial and enzymatic methods for the removal of caffeine. Enzyme and Microbial Technology 37: 225-232.
HUVENNE. H; SMAGGHE, G. 2010. Mechanisms of dsRNA uptake in insects and potential of RNAi for pest control: A review. Journal of Insect Physiology 56: 227-235.
KUMAR, M.; GUPTA, G.P.; RAJAM, M.V.2009. Silencing of acetylcholinesterase gene of Helicoverpa armigera by siRNA affects larval growth and its life cycle. Journal of Insect Physiology 55: 273-278.
MAO, Y.B.; CAI,W.J.; WANG, J.W.; HONG, G.J.; TAO, X.Y.; WANG, L.J.; HUANG, Y.P.; CHEN, X.Y. 2007. Silencing a cotton bollworm P450 monooxygenase gene by plant-mediated RNAi impairs larval tolerance of gossypol. Nature Biotechnology 25: 1307-1313.
PORTILLA, M. 1999. Desarrollo y evaluación de una dieta artificial para la cría de Hypothenemus hampei. Revista Cenicafé (Colombia) 50(1): 24-38.
REDGWELL, R.J; FISHER, M. 2006. Coffee carbohydrates. Mini review. Brazilian Journal of Plant Physiology 18(1): 165-174.
UEFUJI, H.; TATSUMI, Y.; MORIMOTO, M.; KAOTHIENNAKAYAMA, P.; OGITA, S.; SANO, H. 2005. Caffeine production in tobacco plants by simultaneous expression of three coffee N-methyltransferases and its potential as a pest repellant. Plant Molecular Biology 59: 221-227.
VANHECKE, D.; JANITZ, M. 2005. Functional genomics using high throughput RNA interference. Drug discovery Today: Targets 10: 205-212.
VILLALBA, D.A.; BUSTILLO, A.E.; CHAVEZ, B. 1995. Evaluación de insecticidas para el control de la broca del café en Colombia. Revista Cenicafé (Colombia) 46 (3): 152-163.
WOLFROM, M. L.; PATIN, D. L. 1965. Carbohydrates of the coffee bean. IV. An arabinogalactan. Journal of Organic Chemistry 30: 4060-4063.
ZHOU, X.; WHEELER, M.M.; OI, F.M.; SCHARF, M.E. 2008. RNA interference in the termite Reticulitermes flavipes through ingestion of double-stranded RNA. Insect Biochemistry and Molecular Biology 38: 805-815.
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
Authors retain the copyright on their work and are responsible for the ideas expressed in them. Once a manuscript is approved for publication, authors are asked for a publication license for the term of legal protection, for all territories that allows the use, dissemination and disclosure of the same.