The clip cage conundrum: Assessing the interplay of confinement method and aphid genotype in fitness studies

Abstract

Behavior and fitness are important ecological traits frequently measured in insect bioassays. A common method to measure them in soft-bodied herbivorous insects involves confining individuals to plant leaves using clip cages. Although studies have previously highlighted the negative effects of clip cages on leaf physiology, little is known about the impact that using this confinement method has on insect fitness. The responses of different aphid genotypes/clones to different containment methods have not previously been investigated. Here we measured key fitness traits (intrinsic rate of natural increase, mean relative growth rate, time to reach reproductive adulthood and population doubling time) in the potato aphid,Macrosiphum euphorbiae Thomas (Hemiptera: Aphididae), when confined to plants using two methods: (1) clip cages to confine aphids to individual strawberry leaves and (2) a mesh bag to confine aphids to whole strawberry plants. Our study identified a strong negative impact on all the measured aphid fitness traits when using clip cages instead of mesh bags. We also identified genotype-specific differences in response to confinement method, where clip cage confinement differentially affected the fitness of a given aphid genotype compared to the same genotype on whole plants. These results suggest that clip cage use should be carefully considered when experiments seek to quantify insect fitness and that whole plants should be used wherever possible. Given the prevalence of clip cage use in insect bioassays, our results highlight the need for caution when interpreting the existing literature as confinement method significantly impacts aphid fitness depending on their genotype.

Keywords

aphid clonal variation / aphid fitness / clip cages / confinement method / feeding site / insect bioassays

References

[1] Beetge, L. and Krüger, K. (2019) Drought and heat waves associated with climate change affect performance of the potato aphid Macrosiphum euphorbiae. Scientific Reports,9,3645.

[2] Beukeboom, L.W. (2018) Size matters in insects-an introduction. Entomologia Experimentalis Et Applicata,166,2-3.

[3] Birkle, L.M. and Douglas, A.E. (1999) Low genetic diversity among pea aphid (Acyrthosiphon pisum) biotypes of different plant affiliation. Heredity,82,605-612.

[4] Boulain, H.,Legeai, F.,Jaquiéry, J.,Guy, E.,Morlière, S.,Simon, J.C. et al. (2019) Differential expression of candidate salivary effector genes in pea aphid biotypes with distinct host plant specificity. Frontiers in Plant Science,10,1301.

[5] Braendle, C. and Weisser, W.W. (2001) Variation in escape behavior of red and green clones of the pea aphid. Journal of Insect Behavior,14,497-509.

[6] Caillaud, C.M. and Niemeyer, H.M. (1996) Possible involvement of the phloem sealing system in the acceptance of a plant as host by an aphid. Experientia,52,927-931.

[7] Cao, H.H.,Zhang, Z.F.,Wang, X.F. and Liu, T.X. (2018) Nutrition versus defense: why Myzus persicae (green peach aphid) prefers and performs better on young leaves of cabbage. PLoS ONE,13,e0196219.

[8] Castel, S.J. and Berger, P.H. (1993) Rates of growth and increase of Myzus persicae on virus-infected potatoes according to type of virus-vector relationship. Entomologia Experimentalis et Applicata,69,51-60.

[9] Cayetano, L.,Rothacher, L.,Simon, J.C. and Vorburger, C. (2015) Cheaper is not always worse: strongly protective isolates of a defensive symbiont are less costly to the aphid host. Proceedings of the Royal Society B: Biological Sciences,282,20142333.

[10] Clarke, H.V. (2013) Genotypic and endosymbiont-mediated variation in parasitoid susceptibility and other fitness traits of the potato aphid, Macrosiphum euphorbiae. PhD dissertation. University of Dundee.

[11] Clarke, H.V. Cullen, D.,Hubbard, S.F. and Karley, A.J. (2017) Susceptibility of Macrosiphum euphorbiae to the parasitoid Aphidius ervi: larval development depends on host aphid genotype. Entomologia Experimentalis et Applicata,162,148-158.

[12] Coutand, C. (2020) The effect of mechanical stress on plant susceptibility to pests: a mini opinion review. Plants,9,632.

[13] Crafts-Brandner, S.J. and Chu, C.C. (1999) Insect clip cages rapidly alter photosynthetic traits of leaves. Crop Science,39,1896-1899.

[14] Dixon, A.F.G. and Wratten, S.D. (1971) Laboratory studies on aggregation, size and fecundity in the black bean aphid Aphis fabae Scop. Bulletin of Entomological Research,61,97-111.

[15] Douglas, A.E. (2008) The nutritional quality of phloem sap utilized by natural aphid populations. Ecological Entomology,18,31-38.

[16] Elzinga, D.A.,De Vos, M. and Jander, G. (2014) Suppression of plant defenses by a Myzus persicae (green peach aphid) salivary effector protein. Molecular Plant-Microbe Interactions,27,747-756.

[17] Gish, M. (2021) Aphids detect approaching predators using plant-borne vibrations and visual cues. Journal of Pest Science,94,1209-1219.

[18] Haas, J.,Lozano, E.R. and Poppy, G.M. (2018) A simple, light clip-cage for experiments with aphids. Agricultural and Forest Entomology,20,589-592.

[19] Harrison, K.V. and Preisser, E.L. (2016) Dropping behavior in the pea aphid (Hemiptera: Aphididae): how does environmental context affect antipredator responses? Journal of Insect Science,16,89.

[20] Hu, X.S.,Zhang, Z.F.,Zhu, T.Y.,Song, Y.,Wu, L.J.,Liu, X.F. et al. (2018) Maternal effects of the English grain aphids feeding on the wheat varieties with different resistance traits. Scientific Reports,8,7344.

[21] Humphreys, R.K.,Ruxton, G.D. and Karley, A.J. (2021a) Drop when the stakes are high: adaptive, flexible use of dropping behaviour by aphids. Behaviour,158,603-623.

[22] Humphreys, R.K.,Ruxton, G.D. and Karley, A.J. (2021b) Post-dropping behavior of potato aphids (Macrosiphum euphorbiae). Journal of Insect Behavior,34,223-239.

[23] Jonckheere, W.,Dermauw, W.,Zhurov, V.,Wybouw, N.,Van Den Bulcke, J.,Villarroel, C.A. et al. (2016) The salivary protein repertoire of the polyphagous spider mite Tetranychus urticae: a quest for effectors. Molecular and Cellular Proteomics,15,3594-3613.

[24] Kaech, H.,Jud, S. and Vorburger, C. (2022) Similar cost of Hamiltonella defensa in experimental and natural aphid-endosymbiont associations. Ecology and Evolution,12,e8551.

[25] Karley, A.J.,Emslie-Smith, M. and Bennett, A.E. (2017) Potato aphid Macrosiphum euphorbiae performance is determined by aphid genotype and not mycorrhizal fungi or water availability. Insect Science,24,1015-1024.

[26] Kou, X.,Bai, S.,Luo, Y.,Yu, J.,Guo, H.,Wang, C. et al. (2022) Construction of a modified clip cage and its effects on the life-history parameters of Sitobion avenae (Fabricius) and defense responses of Triticum aestivum. Insects,13,777.

[27] Lamb, R.J.,MacKay, P.A. and Migui, S.M. (2009) Measuring the performance of aphids: fecundity versus biomass. Canadian Entomologist,141,401-405.

[28] Liu, D.,Dai, P.,Li, S.,Ahmed, S.S.,Shang, Z. and Shi, X. (2018) Life-history responses of insects to water-deficit stress: a case study with the aphid Sitobion avenae. BMC Ecology,18,17.

[29] Liu, S.,Liu, X.,Zhang, T.,Bai, S.,He, K.,Zhang, Y. et al. (2023) Secondary symbionts affect aphid fitness and the titer of primary symbiont. Frontiers in Plant Science,14,1096750.

[30] Markovic, D.,Glinwood, R.,Olsson, U. and Ninkovic, V. (2014) Plant response to touch affects the behaviour of aphids and ladybirds. Arthropod-Plant Interactions,8,171-181.

[31] Mehrparvar, M.,Rajaei, A.,Rokni, M.,Balog, A. and Loxdale, H.D. (2019) Bottom-up effects in a tritrophic plant-aphid-parasitoid system: why being the perfect host can have its disadvantages. Bulletin of Entomological Research,109,831-839.

[32] Moore, J.P.,Taylor, J.E.,Paul, N.D. and Whittaker, B. (2003) The use of clip cages to restrain insects reduces leaf expansion systemically in Rumex obtusifolius. Ecological Entomology,28,239-242.

[33] Mowry, T.M. (1993) A method for confining small insects to plant surfaces. Journal of Agricultural Entomology,10,181-184.

[34] Muratori, F.B.,Rouyar, A. and Hance, T. (2014) Clonal variation in aggregation and defensive behavior in pea aphids. Behavioral Ecology,25,901-908.

[35] Nalam, V.J.,Han, J.,Pitt, W.J.,Acharya, S.R. and Nachappa, P. (2021) Location, location, location: feeding site affects aphid performance by altering access and quality of nutrients. PLoS ONE,16,e0245380.

[36] Nelson, E.H. (2007) Predator avoidance behavior in the pea aphid: costs, frequency, and population consequences. Oecologia,151,22-32.

[37] Pompon, J.,Li, X.Q. and Pelletier, Y. (2011) Resistance level to an aphid potato pest varies between genotypes from the same Solanum accession. Journal of Economic Entomology,104,1075-1079.

[38] R. Core Team. (2021) R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing,Vienna, Austria. https://www.R-project.org/

[39] Routray, S.,Hariprasad, K.V.,Manjula, K.,A. Kumar and Latha, P. (2020) PerfORmance of aphid,Aphis craccivora on different host plants. Indian Journal of Plant Protection,46,2-4.

[40] Saguez, J.,Giordanengo, P. and Vincent, C. (2013) Aphids as major potato pests. In Insect Pests of Potato (eds. P. Giordanengo & C. Vincent), pp. 31-63. Elsevier Inc, San Diego,CA.

[41] Shrestha, G.,Skovgård, H.,Reddy, G.V.P.,Steenberg, T. and Enkegaard, A. (2017) Role of the aphid species and their feeding locations in parasitization behavior of Aphelinus abdominalis, a parasitoid of the lettuce aphid Nasonovia ribisnigri. PLoS ONE,12,e0184080.

[42] Simon, J.C.,Boutin, S.,Tsuchida, T.,Koga, R.,Le Gallic, J.F.,Frantz, A. et al. (2011) Facultative symbiont infections affect aphid reproduction. PLoS ONE,6,e21831.

[43] Srinivasan, R. (2011) Specialized host utilization of Macrosiphum euphorbiae on a nonnative weed host,Solanum sarrachoides, and competition with Myzus persicae. Environmental Entomology,40,350-356.

[44] Srisakrapikoop, U.,Pirie, T.J. and Fellowes, M.D.E. (2021) Aphids show interspecific and intraspecific variation in life history responses to host plant infection by the fungal pathogen Botrytis cinerea. Entomological Science,24,228-234.

[45] Stamp, N.E. and Bowers, M.D. (1994) Effects of cages, plant age and mechanical clipping on plantain chemistry. Oecologia,99,66-71.

[46] Taravati, S. and Mannion, C. (2016) Effect of aggregation and cage setting on some life-history parameters of Aleurodicus Rugioperculatus (Hemiptera: Aleyrodidae). Journal of Economic Entomology,109,249-254.

[47] Taylor, M. (2013) Bayer Expert Guide Aphids. Bayer CropScience Ltd.

[48] van Emden, H.F. and Harrington, R. (2017) Aphids as Crop Pests. Wallingford, Oxfordshire, UK: CAB International.

[49] Vilcinskas, A. (2016) Biology and Ecology of Aphids. CRC Press.

[50] War, A.R.,Taggar, G.K.,Hussain, B.,Taggar, M.S.,Nair, R.M. and Sharma, H.C. (2018) Plant defence against herbivory and insect adaptations. AoB Plants,10,ply037.

[51] Whitehead, M.A. (2019) Genomics for optimal aphid biocontrol. PhD dissertation. University of Liverpool.

[52] Zvereva, E.L. and Kozlov, M.V. (2016) The costs and effectiveness of chemical defenses in herbivorous insects: a meta-analysis. Ecological Monographs,86,107-124.

RIGHTS & PERMISSIONS

2024 The Authors. Insect Science published by John Wiley & Sons Australia, Ltd on behalf of Institute of Zoology, Chinese Academy of Sciences.