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Long-term studies on the evolution of resistance of Myzus persicae (Hemiptera: Aphididae) to insecticides in Greece

Published online by Cambridge University Press:  16 June 2020

John T. Margaritopoulos*
Affiliation:
Department of Plant Protection, Institute of Industrial and Fodder Crops, Hellenic Agricultural Organization–DEMETER, Volos, Greece
A.N. Kati
Affiliation:
Plant Pathology Laboratory, School of Agriculture, Aristotle University of Thessaloniki, Thessaloniki, Greece
C.Ch. Voudouris
Affiliation:
Department of Plant Protection, Institute of Industrial and Fodder Crops, Hellenic Agricultural Organization–DEMETER, Volos, Greece
P.J. Skouras
Affiliation:
Laboratory of Agricultural Entomology and Zoology, Department of Agricultural Technologies, University of Peloponnese, Antikalamos, Greece
J.A. Tsitsipis
Affiliation:
University of Thessaly, Volos, Greece
*
Author for correspondence: John T. Margaritopoulos, Email: johnmargaritopoulos@gmail.com

Abstract

The aphid Myzus persicae s.l. (Hemiptera: Aphididae) is an important pest of many crops worldwide with a complex life cycle, intensely controlled by chemical pesticides, and has developed resistance to almost all used insecticides. In Greece, the aphid exhibits high genetic variation and adaptability and it is a classic example of evolution in the making. We have been studying M. persicae for over 20 years, on different host plants and varying geographical areas, analyzing its bio-ecology and the ability to develop resistance to insecticides. In this review, we present new and historical data on the effectiveness of insecticides from seven chemical groups used to control the aphid in Greece and the incidence of seven resistance mechanisms, including the new fast-spreading R81T point mutation of the postsynaptic nicotinic acetylcholine receptor. Thousands of samples were tested by biological, biochemical and molecular assays. The aphid populations were found to have developed and maintain resistance at medium to high levels to organophosphates, carbamates, pyrethroids and neonicotinoids for decades. In the latter group, a marked increase is recorded during an ~10-year period. The data analyzed and the extensive bibliography, advocate the difficulty to control the aphid making the design and application of IPM/IRM programs a challenge. We discuss principles and recommendations for the management of resistance, including the use of compounds such as flonicamid, spirotetramat, flupyradifurone and sulfoxaflor. We emphasize that resistance is a dynamic phenomenon, changing in time and space, requiring, therefore, continuous monitoring.

Type
Review Article
Copyright
Copyright © The Author(s) 2020. Published by Cambridge University Press

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Footnotes

*

Present address: Mainalou 4, 15235 Vrilissia Athens, Greece

References

Anstead, JA, Williamson, MS, Eleftherianos, I and Denholm, I (2004) High-throughput detection of knockdown resistance in Myzus Persicae using allelic discriminating quantitative PCR. Insect Biochemistry and Molecular Biology 34, 871877.CrossRefGoogle ScholarPubMed
Anstead, JA, Mallet, J and Denholm, I (2007) Temporal and spatial incidence of alleles conferring knockdown resistance to pyrethroids in the peach-potato aphid, Myzus Persicae (Hemiptera: Aphididae), and their association with other insecticide resistance mechanisms. Bulletin of entomological research 97, 243252.CrossRefGoogle Scholar
Anthon, EW (1955) Evidence for green peach aphid resistance to organo-phosphorous insecticides. Journal of Economic Entomology 48, 5657.CrossRefGoogle Scholar
Bass, C, Puinean, AM, Andrews, M, Cutler, P, Daniels, M, Elias, J, Paul, VL, Crossthwaite, AJ, Denholm, I, Field, LM, Foster, SP, Lind, R, Williamson, MS and Slater, R (2011) Mutation of a nicotinic acetylcholine receptor β subunit is associated with resistance to neonicotinoid insecticides in the aphid Myzus persicae. BMC Neuroscience 12, 51.CrossRefGoogle ScholarPubMed
Bass, C, Puinean, AM, Zimmer, CT, Denholm, I, Field, LM, Foster, SP, Gutbrod, O, Nauen, R, Slater, R and Williamson, MS (2014) The evolution of insecticide resistance in the peach potato aphid, Myzus persicae. Insect Biochemistry and Molecular Biology 51, 4151.CrossRefGoogle ScholarPubMed
Bass, C, Denholm, I, Williamson, MS and Nauen, R (2015) The global status of insect resistance to neonicotinoid insecticides. Pesticide Biochemistry and Physiology 121, 7887.CrossRefGoogle ScholarPubMed
Blackman, RL and Eastop, VF (2017) Taxonomic issues. In van Emden, HF and Harrington, R (eds), Aphids as Crop Pests. Wallingford, Oxfordshire, UK: CAB International, pp. 136.Google Scholar
Blackman, RL, Spence, JM, Field, LM and Devonshire, AL (1999) Variation in the chromosomal distribution of amplified esterase (FE4) genes in Greek field populations of Myzus Persicae (Sulzer). Heredity 82, 180186.CrossRefGoogle Scholar
Blackman, RL, Malarky, G, Margaritopoulos, JT and Tsitsipis, JA (2007) Distribution of common genotypes of Myzus Persicae (Hemiptera: Aphididae) in Greece, in relation to life cycle and host plant. Bulletin of entomological research 97, 253263.CrossRefGoogle ScholarPubMed
Brück, E, Elbert, A, Fischer, R, Krueger, S, Kühnhold, J, Klueken, AM, Nauen, R, Niebes, J-F, Reckmann, U, Schnorbach, H-J, Steffens, R and van Waetermeulen, X (2009) Movento®, an innovative ambimobile insecticide for sucking insect pest control in agriculture: biological profile and field performance. Crop Protection 28, 838844.CrossRefGoogle Scholar
Charaabi, K, Boukhris-Bouhachem, S, Makni, M and Denholm, I (2018) Occurrence of target-site resistance to neonicotinoids in the aphid Myzus Persicae in Tunisia, and its status on different host plants. Pest Management Science 74, 12971301.CrossRefGoogle ScholarPubMed
Chen, X, Li, F, Chen, A, Ma, K, Liang, P, Liu, Y, Song, D and Gao, X (2017) Both point mutations and low expression levels of the nicotinic acetylcholine receptor β1 subunit are associated with imidacloprid resistance in an Aphis gossypii (Glover) population from a Bt cotton field in China. Pesticide Biochemistry and Physiology 141, 18.CrossRefGoogle Scholar
Cox, D, Denholm, I and Devonshire, A (2004) Monitoring of insecticide resistance in Myzus persicae from Greece. In Simon, J-C, Dedryver, CA, Rispe, C and Hullé, M (eds), Aphids in A new Millennium. Paris: INRA Editions, pp. 275280.Google Scholar
Cutler, P, Slater, R, Edmunds, AJF, Maienfisch, P, Hall, RG, Earley, FGP, Pitterna, T, Pal, S, Paul, V-L, Goodchild, J, Blacker, M, Hagmann, L and Crossthwaite, AJ (2013) Investigating the mode of action of sulfoxaflor: a fourth-generation neonicotinoid. Pest Management Science 69, 607619.CrossRefGoogle ScholarPubMed
Davies, TGE, Field, LM, Usherwood, PNR and Williamson, MS (2007) DDT, pyrethrins, pyrethroids and insect sodium channels. IUBMB Life 59, 151162.CrossRefGoogle ScholarPubMed
de Little, SC, Edwards, O, van Rooyen, AR, Weeks, A and Umina, PA (2017) Discovery of metabolic resistance to neonicotinoids in green peach aphids (Myzus Persicae) in Australia. Pest Management Science 73, 16111617.CrossRefGoogle ScholarPubMed
Devonshire, AL, Devine, GJ and Moores, GD (1992) Comparison of microplate esterase assays and immunoassay for identifying insecticide resistant variants of Myzus Persicae (Homoptera: Aphididae). Bulletin of Entomological Research 82, 459463.CrossRefGoogle Scholar
Devonshire, AL, Field, LM, Foster, SP, Moores, GD, Williamson, MS and Blackman, RL (1998) The evolution of insecticide resistance in the peach-potato aphid, Myzus persicae. Philosophical Transactions of the Royal Society B: Biological Sciences 353, 16771684.CrossRefGoogle Scholar
Elbert, A, Nauen, R and Salmon, E (2008) Resistance management guidelines for the new ketoenol insecticide Movento®. Bayer Crop Science Journal 61, 403416.Google Scholar
Eleftherianos, I, Foster, SP, Williamson, MS and Denholm, I (2008) Characterization of the M918T sodium channel gene mutation associated with strong resistance to pyrethroid insecticides in the peach-potato aphid, Myzus Persicae (Sulzer). Bulletin of Entomological Research 98, 183191.CrossRefGoogle Scholar
Fenton, B, Kasprowicz, L, Malloch, G and Pickup, J (2010 a) Reproductive performance of asexual clones of the peach-potato aphid, (Myzus Persicae, Homoptera: Aphididae), colonising Scotland in relation to host plant and field ecology. Bulletin of Entomological Research 100, 451460.CrossRefGoogle Scholar
Fenton, B, Margaritopoulos, JT, Malloch, GL and Foster, SP (2010 b) Micro-evolutionary change in relation to insecticide resistance in the peach–potato aphid, Myzus persicae. Ecological Entomology 35, 131146.CrossRefGoogle Scholar
Field, LM and Foster, SP (2002) Amplified esterase genes and their relationship with other insecticide resistance mechanisms in English field populations of the aphid, Myzus Persicae (Sulzer). Pest Management Science 58, 889894.CrossRefGoogle Scholar
Fontaine, S, Caddoux, L, Brazier, C, Bertho, C, Bertolla, P, Micoud, A and Roy, L (2011) Uncommon associations in target resistance among French populations of Myzus Persicae from oilseed rape crops. Pest Management Science 67, 881885.CrossRefGoogle ScholarPubMed
Foster, SP, Denholm, I and Devonshire, AL (2000) The ups and downs of insecticide resistance in peach-potato aphids (Myzus Persicae) in the UK. Crop Protection 19, 873879.CrossRefGoogle Scholar
Foster, SP, Denholm, I and Thompson, R (2002 a) Bioassay and field-simulator studies of the efficacy of pymetrozine against peach-potato aphids, Myzus Persicae (Hemiptera: Aphididae), possessing different mechanisms of insecticide resistance. Pest Management Science 58, 805810.CrossRefGoogle Scholar
Foster, SP, Harrington, R, Dewar, AM, Denholm, I and Devonshire, AL (2002 b) Temporal and spatial dynamics of insecticide resistance in Myzus Persicae (Hemiptera: Aphididae). Pest Management Science 58, 895907.CrossRefGoogle Scholar
Foster, SP, Devine, G and Devonshire, AL (2017) Insecticide resistance. In van Emden, HF and Harrington, R (eds), Aphids as Crop Pests. Wallingford, Oxfordshire, UK: CAB International, pp. 426446.CrossRefGoogle Scholar
Hance, T, Kohandani-Tafresh, F and Munaut, F (2017) Biological control. In van Emden, HF and Harrington, R (eds), Aphids as Crop Pests. Wallingford, Oxfordshire, UK: CAB International, pp. 448493.CrossRefGoogle Scholar
Harrewijn, P and Piron, P (1994) Pymetrozine, a novel agent for reducing virus transmission by Myzus persicae. in Proceedings Brighton Crop Protection Conference - Pests and Diseases. BCPC, Farnham, Surrey, UK, pp. 923928.Google Scholar
Hirata, K, Kiyota, R, Matsuura, A, Toda, S, Yamamoto, A and Iwasa, T (2015) Association between the R81T mutation in the nicotinic acetylcholine receptor β1 subunit of Aphis gossypii and the differential resistance to acetamiprid and imidacloprid. Journal of Pesticide Science 40, 2531.CrossRefGoogle Scholar
Jansen, JP, Defrance, T and Warnier, AM (2011) Side effects of flonicamide and pymetrozine on five aphid natural enemy species. BioControl 56, 759770.CrossRefGoogle Scholar
Karatolos, N, Williamson, MS, Denholm, I, Gorman, K, ffrench-Constant, R and Nauen, R (2012) Resistance to spiromesifen in Trialeurodes Vaporariorum is associated with a single amino acid replacement in its target enzyme acetyl-coenzyme A carboxylase: Spiromesifen resistance in the greenhouse whitefly. Insect Molecular Biology 21, 327334.CrossRefGoogle Scholar
Kati, AN, Mandrioli, M, Skouras, PJ, Malloch, GL, Voudouris, CC, Venturelli, M, Manicardi, GC, Tsitsipis, JA, Fenton, B and Margaritopoulos, JT (2014) Recent changes in the distribution of carboxylesterase genes and associated chromosomal rearrangements in Greek populations of the tobacco aphid Myzus persicae nicotianae. Biological Journal of the Linnean Society 113, 455470.CrossRefGoogle Scholar
Kaufmann, L, Schürmann, F, Yiallouros, M, Harrewijn, P and Kayser, H (2004) The serotonergic system is involved in feeding inhibition by pymetrozine. Comparative studies on a locust (Locusta Migratoria) and an aphid (Myzus Persicae). Comparative Biochemistry and Physiology. Toxicology & Pharmacology: CBP 138, 469483.CrossRefGoogle Scholar
Kramer, T and Nauen, R (2011) Monitoring of spirodiclofen susceptibility in field populations of European red mites, Panonychus ulmi (Koch) (Acari: Tetranychidae), and the cross-resistance pattern of a laboratory-selected strain. Pest Management Science 67, 12851293.CrossRefGoogle Scholar
Margaritopoulos, JT, Tsitsipis, JA, Zintzaras, E and Blackman, RL (2000) Host-correlated morphological variation of Myzus Persicae (Hemiptera: Aphididae) populations in Greece. Bulletin of Entomological Research 90, 233244.CrossRefGoogle Scholar
Margaritopoulos, JT, Blackman, RL, Tsitsipis, JA and Sannino, L (2003) Co-existence of different host-adapted forms of the Myzus Persicae group (Hemiptera: Aphididae) in southern Italy. Bulletin of Entomological Research 93, 131135.CrossRefGoogle Scholar
Margaritopoulos, JT, Malarky, G, Tsitsipis, JA and Blackman, RL (2007 a) Microsatellite DNA and behavioural studies provide evidence of host-mediated speciation in Myzus Persicae (Hemiptera: Aphididae). Biological Journal of the Linnean Society 91, 687702.CrossRefGoogle Scholar
Margaritopoulos, JT, Skouras, PJ, Nikolaidou, P, Manolikaki, J, Maritsa, K, Tsamandani, K, Kanavaki, OM, Bacandritsos, N, Zarpas, KD and Tsitsipis, JA (2007 b) Insecticide resistance status of Myzus Persicae (Hemiptera: Aphididae) populations from peach and tobacco in mainland Greece. Pest Management Science 63, 821829.CrossRefGoogle ScholarPubMed
Margaritopoulos, JT, Kasprowicz, L, Malloch, GL and Fenton, B (2009) Tracking the global dispersal of a cosmopolitan insect pest, the peach potato aphid. BMC Ecology 9, 13.CrossRefGoogle ScholarPubMed
Margaritopoulos, JT, Tsamandani, K, Kanavaki, OM, Katis, NI and Tsitsipis, JA (2010) Efficacy of pymetrozine against Myzus persicae and in reducing potato virus Y transmission on tobacco plants. Journal of Applied Entomology 134, 323332.CrossRefGoogle Scholar
Martinez-Torres, D, Foster, SP, Field, LM, Devonshire, AL and Williamson, MS (1999) A sodium channel point mutation is associated with resistance to DDT and pyrethroid insecticides in the peach-potato aphid, Myzus Persicae (Sulzer) (Hemiptera: Aphididae). Insect Molecular Biology 8, 339346.CrossRefGoogle Scholar
Meng, J, Chen, X and Zhang, C (2019) Transcriptome-based identification and characterization of genes responding to imidacloprid in Myzus persicae. Scientific Reports 9, 18.CrossRefGoogle ScholarPubMed
Moores, GD, Devine, GJ and Devonshire, AL (1994) Insecticide-insensitive acetylcholinesterase can enhance esterase-based resistance in Myzus Persicae and Myzus nicotianae. Pesticide Biochemistry and Physiology 49, 114120.CrossRefGoogle Scholar
Morita, M, Ueda, T, Yoneda, T, Koyanagi, T and Haga, T (2007) Flonicamid, a novel insecticide with a rapid inhibitory effect on aphid feeding. Pest Management Science 63, 969973.CrossRefGoogle ScholarPubMed
Morita, M, Yoneda, T and Akiyoshi, N (2014) Research and development of a novel insecticide, flonicamid. Journal of Pesticide Science 39, 179180.CrossRefGoogle Scholar
Mottet, C, Fontaine, S, Caddoux, L, Brazier, C, Mahéo, F, Simon, J-C, Micoud, A and Roy, L (2016) Assessment of the dominance level of the R81T target resistance to two neonicotinoid insecticides in Myzus Persicae (Hemiptera: Aphididae). Journal of Economic Entomology 109, 21822189.CrossRefGoogle Scholar
Nabeshima, T, Kozaki, T, Tomita, T and Kono, Y (2003) An amino acid substitution on the second acetylcholinesterase in the pirimicarb-resistant strains of the peach potato aphid, Myzus persicae. Biochemical and Biophysical Research Communications 307, 1522.CrossRefGoogle ScholarPubMed
Nauen, R, Reckmann, U, Thomzik, J and Thielert, W (2008) Biological profile of spirotetramat (Movento®) – a new two-way systemic (ambimobile) insecticide against sucking pest species. Bayer Crop Science Journal 61, 245278.Google Scholar
Nauen, R, Jeschke, P, Velten, R, Beck, ME, Ebbinghaus-Kintscher, U, Thielert, W, Wölfel, K, Haas, M, Kunz, K and Raupach, G (2015) Flupyradifurone: a brief profile of a new butenolide insecticide. Pest Management Science 71, 850862.CrossRefGoogle ScholarPubMed
Needham, PH and Sawicki, RM (1971) Diagnosis of resistance to organophosphorus insecticides in Myzus Persicae (Sulz). Nature 230, 125126.CrossRefGoogle Scholar
Pan, Y, Yang, C, Gao, X, Peng, T, Bi, R, Xi, J, Xin, X, Zhu, E, Wu, Y and Shang, Q (2015) Spirotetramat resistance adaption analysis of Aphis gossypii Glover by transcriptomic survey. Pesticide Biochemistry and Physiology 124, 7380.CrossRefGoogle ScholarPubMed
Panini, M, Dradi, D, Marani, G, Butturini, A and Mazzoni, E (2014) Detecting the presence of target-site resistance to neonicotinoids and pyrethroids in Italian populations of Myzus persicae. Pest Management Science 70, 931938.CrossRefGoogle ScholarPubMed
Peng, T, Pan, Y, Yang, C, Gao, X, Xi, J, Wu, Y, Huang, X, Zhu, E, Xin, X, Zhan, C and Shanget, Q (2016) Over-expression of CYP6A2 is associated with spirotetramat resistance and cross-resistance in the resistant strain of Aphis gossypii Glover. Pesticide Biochemistry and Physiology 126, 6469.CrossRefGoogle ScholarPubMed
Philippou, D, Field, L and Moores, G (2010) Metabolic enzyme(s) confer imidacloprid resistance in a clone of Myzus Persicae (Sulzer) (Hemiptera: Aphididae) from Greece. Pest Management Science 66, 390395.CrossRefGoogle Scholar
Puinean, AM, Foster, SP, Oliphant, L, Denholm, I, Field, LM, Millar, NS, Williamson, MS and Bass, C (2010) Amplification of a cytochrome P450 gene is associated with resistance to neonicotinoid insecticides in the aphid Myzus persicae. PLoS Genetics 6, e1000999.CrossRefGoogle ScholarPubMed
Puinean, AM, Elias, J, Slater, R, Warren, A, Field, LM, Williamson, MS and Bass, C (2013) Development of a high-throughput real-time PCR assay for the detection of the R81T mutation in the nicotinic acetylcholine receptor of neonicotinoid-resistant Myzus persicae. Pest Management Science 69, 195199.CrossRefGoogle ScholarPubMed
R Core Team (2017) R: A Language and Environment for Statistical Computing. Vienna: R Foundation for Statistical Computing.Google Scholar
Robertson, J, Russell, R, Preisler, H and Savin, N (2007) Pesticide Bioassays with Arthropods. Boca Raton, FL: CRC Press.CrossRefGoogle Scholar
Sauphanor, B and Bouvier, JC (1995) Cross resistance between benzoylureas and benzoylhydrazines in the codling moth, Cydia Pomonella L. Pesticide Science 45, 369375.CrossRefGoogle Scholar
Scott, JG and Wen, Z (2001) Cytochromes P450 of insects: the tip of the iceberg. Pest Management Science 57, 958967.CrossRefGoogle ScholarPubMed
Skouras, P (2009) Study on the bio-ecology, population genetics and insecticide resistance of the aphid Myzus persicae and its natural enemies (PhD Thessis). University of Thessaly, Volos, Greece.Google Scholar
Slater, R, Paul, VL, Andrews, M, Garbay, M and Camblin, P (2012) Identifying the presence of neonicotinoid resistant peach-potato aphid (Myzus Persicae) in the peach-growing regions of southern France and northern Spain. Pest Management Science 68, 634638.CrossRefGoogle Scholar
Sparks, TC and Nauen, R (2015) IRAC: mode of action classification and insecticide resistance management. Pesticide Biochemistry and Physiology 121, 122128.CrossRefGoogle ScholarPubMed
Unal, G and Jepson, PC (1991) The toxicity of aphicide residues to beneficial invertebrates in cereal crops. Annals of Applied Biology 118, 493502.CrossRefGoogle Scholar
Vais, H, Williamson, MS, Goodson, SJ, Devonshire, AL, Warmke, JW, Usherwood, PNR and Cohen, CJ (2000) Activation of Drosophila Sodium channels promotes modification by deltamethrin: reductions in affinity caused by knock-down resistance mutations. The Journal of General Physiology 115, 305318.CrossRefGoogle ScholarPubMed
Van Pottelberge, S, Van Leeuwen, T, Khajehali, J and Tirry, L (2009) Genetic and biochemical analysis of a laboratory-selected spirodiclofen-resistant strain of Tetranychus urticae Koch (Acari: Tetranychidae). Pest Management Science 65, 358366.CrossRefGoogle Scholar
Vorburger, C, Lancaster, M and Sunnucks, P (2003) Environmentally related patterns of reproductive modes in the aphid Myzus Persicae and the predominance of two “superclones” in Victoria, Australia. Molecular Ecology 12, 34933504.CrossRefGoogle Scholar
Voudouris, CC, Kati, AN, Sadikoglou, E, Williamson, M, Skouras, PJ, Dimotsiou, O, Georgiou, S, Fenton, B, Skavdis, G and Margaritopoulos, JT (2016) Insecticide resistance status of Myzus Persicae in Greece: long-term surveys and new diagnostics for resistance mechanisms. Pest Management Science 72, 671683.CrossRefGoogle ScholarPubMed
Voudouris, CC, Williamson, MS, Skouras, PJ, Kati, AN, Sahinoglou, AJ and Margaritopoulos, JT (2017) Evolution of imidacloprid resistance in Myzus Persicae in Greece and susceptibility data for spirotetramat. Pest Management Science 73, 18041812.CrossRefGoogle ScholarPubMed
Wang, Z-H, Gong, Y-J, Jin, G-H, Zhu, L and Wei, S-J (2016) Effects of spirotetramat on development and reproduction of Myzus Persicae (Hemiptera: Aphididae). Austral Entomology 55, 235241.CrossRefGoogle Scholar
Wang, Z-H, Gong, Y-J, Chen, J-C, Su, X-C, Cao, L-J, Hoffmann, AA and Wei, S-J (2018) Laboratory selection for resistance to sulfoxaflor and fitness costs in the green peach aphid Myzus persicae. Journal of Asia-Pacific Entomology 21, 408412.CrossRefGoogle Scholar
Zepeda-Paulo, FA, Simon, J-C, Ramírez, CC, Fuentes-Contreras, E, Margaritopoulos, JT, Wilson, ACC, Sorenson, CE, Briones, LM, Azevedo, R, Ohashi, DV, Lacroix, C, Glais, L and Figueroa, CC (2010) The invasion route for an insect pest species: the tobacco aphid in the New World. Molecular Ecology 19, 47384752.CrossRefGoogle ScholarPubMed
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