Arthropod-Plant Interactions

, Volume 12, Issue 4, pp 567–574 | Cite as

A PCR-based analysis of plant DNA reveals the feeding preferences of Apolygus lucorum (Heteroptera: Miridae)

  • Qian Wang
  • Weifang Bao
  • Fan Yang
  • Yizhong Yang
  • Yanhui Lu
Original Paper


The mirid bug Apolygus lucorum (Meyer-Dür) (Heteroptera: Miridae) is a severe pest of cotton and other crops in China. The feeding preferences of this pest are unclear due to its frequent movement among different host plants and the inconspicuous signs of its feeding. Here, we present results of a field trial that used direct observation of bug densities and a PCR-based molecular detection assay to detect plant DNA in bugs to explore relationships between A. lucorum population abundance and its feeding preference between two host plants, Humulus scandens (Loureiro) Merrill and Medicago sativa L. The field-plot samples showed that A. lucorum adults generally prefer flowering host plants. Its density was significantly higher on flowering H. scandens than on seedlings of M. sativa, and a similarly higher bug density was observed on flowering M. sativa than on seedlings of H. scandens. In the laboratory, we designed two pairs of species-specific primers targeting the trnL-F region for H. scandens and M. sativa, respectively. The detectability of plant DNA generally decreased with time post-feeding, and the half-life of plant DNA detection (DS50) in the gut was estimated as 6.26 h for H. scandens and 3.79 h for M. sativa with significant differences between each other. In mirid bugs exposed to seedlings of H. scandens and flowering M. sativa, the detection rate of M. sativa DNA was significantly higher than that of H. scandens. Meanwhile, in mirid bugs exposed to seedlings of M. sativa and flowering H. scandens, a significantly higher detection rate of H. scandens DNA was found. We developed a useful tool to detect the remaining plant food species specifically from the gut of A. lucorum in the current study. We provided direct evidence of its feeding preference between H. scandens and M. sativa at different growth stages, which strongly supported a positive correlation between population abundance and feeding preference of A. lucorum on different plants under field conditions. The findings provide new insights into the understanding of A. lucorum’s feeding preference, and are helpful for developing the strategies to control this pest.


Herbivory Population abundance Host plant preference Plant DNA Species-specific primers 



We greatly thank Dr. Corinna Wallinger for helpful comments and suggestions on the manuscript revision. This research was supported by the National Natural Science Funds of China (Nos. 31321004, 31222046), the National Key Research and Development Program of China (2017YFD0201900), and China Agriculture Research System (CARS-15-19).

Supplementary material

11829_2018_9604_MOESM1_ESM.docx (488 kb)
Supplementary material 1 (DOCX 488 KB)


  1. Bernays E, Graham M (1988) On the evolution of host specificity in phytophagous arthropods. Ecology 69:886–892CrossRefGoogle Scholar
  2. Bernays EA, Bright KL, Gonzalez N, Angel J (1994) Dietary mixing in a generalist herbivore: tests of two hypotheses. Ecology 75:1997–2006CrossRefGoogle Scholar
  3. Borsch T, Hilu KW, Quandt D, Wilde V, Neinhuis C, Barthlott W (2003) Noncoding plastid trnT-trnF sequences reveal a well resolved phylogeny of basal angiosperms. J Evolution Biol 16(4):558–576CrossRefGoogle Scholar
  4. Chu HF, Meng HL (1958) Studies on three species of cotton plant bugs, Adelphocoris taeniophorus Reuter, A. lineolatus (Goeze), and Lygus lucorum Meyer-Dür (Hemiptera: Miridae). Acta Entomol Sin 8:97–118Google Scholar
  5. Dong JW, Pan HS, Lu YH, Yang YZ (2013) Nymphal performance correlated with adult preference for flowering host plants in a polyphagous mirid bug, Apolygus lucorum. (Heteroptera: Miridae). Arthropod 7:83–91CrossRefGoogle Scholar
  6. Geng HH, Pan HS, Lu YH, Yang YZ (2012) Nymphal and adult performance of Apolygus lucorum (Hemiptera: Miridae) on a preferred host plant, mungbean Vigna radiata. Appl Entomol Zool 47:191–197CrossRefGoogle Scholar
  7. Greenstone MH, Rowley DL, Weber DC, Payton ME, Hawthorne DJ (2007) Feeding mode and prey detectability half-lives in molecular gut-content analysis: an example with two predators of the Colorado potato beetle. B Entomol Res 97:201–209CrossRefGoogle Scholar
  8. Greenstone MH, Szendrei Z, Payton ME, Rowley DC, Coudron TC, Weber DC (2010) Choosing natural enemies for conservation biological control: use of the prey detectability half-life to rank key predators of Colorado potato beetle. Entomol Exp Appl 136:97–107CrossRefGoogle Scholar
  9. Greenstone MH, Payton ME, Weber DC, Simmons AM (2014) The detectability half-life in arthropod predator-prey research: what it is, why we need it, how to measure it, and how to use it. Mol Ecol 23:3799–3813CrossRefPubMedGoogle Scholar
  10. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98Google Scholar
  11. Jiang YY, Lu YH, Zeng J (2015) Forecast and management of mirid bugs in multiple agroecosystem of China. China Agriculture Press, BeijingGoogle Scholar
  12. Jurado-Rivera JA, Vogler AP, Reid CA, Petitpierre E, Gomez-Zurita J (2009) DNA barcoding insect-host plant associations. Proc Biol Sci Royal Soc 276:639–648CrossRefGoogle Scholar
  13. Kennedy GG, Storer NP (2000) Life systems of polyphagous arthropod pests in temporally unstable cropping systems. Annu Rev Entomol 45:467–493CrossRefPubMedGoogle Scholar
  14. King RA, Read DS, Traugott M, Symondson WOC (2008) Molecular analysis of predation: a review of best practice for DNA-based approaches. Mol Ecol 17:947–963CrossRefPubMedGoogle Scholar
  15. Lu YH, Qiu F, Feng HQ, Li HB, Yang ZC, Wyckhuys KAG, Wu KM (2008) Species composition and seasonal abundance of pestiferous plant bugs (Hemiptera: Miridae) on Bt cotton in China. Crop Prot 27:465–472CrossRefGoogle Scholar
  16. Lu YH, Wu KM, Wyckhuys KAG, Guo YY (2009) Potential of mungbean, Vigna radiatus as a trap crop for managing Apolygus lucorum (Hemiptera: Miridae) on Bt cotton. Crop Prot 28:77–81CrossRefGoogle Scholar
  17. Lu YH, Wu KM, Wyckhuys KAG, Guo YY (2010a) Overwintering hosts of Apolygus lucorum (Hemiptera: Miridae) in northern China. Crop Prot 29:1026–1033CrossRefGoogle Scholar
  18. Lu YH, Wu KM, Jiang YY, Xia B, Li P, Feng HQ, Wyckhuys KAG, Guo YY (2010b) Mirid bug outbreaks in multiple crops correlated with wide-scale adoption of Bt cotton in China. Science 328:1151–1154CrossRefPubMedGoogle Scholar
  19. Lu Y, Jiao Z, Wu K (2012) Early season host plants of Apolygus lucorum (Heteroptera: Miridae) in northern China. J Econ Entomol 105:1603–1611CrossRefPubMedGoogle Scholar
  20. Matheson C, Muller G, Junnila A, Vernon K, Hausmann A, Miller M, Greenblatt C, Schlein Y (2008) A PCR method for detection of plant meals from the guts of insects. Org Divers Evol 7:294–303CrossRefGoogle Scholar
  21. Mitter C, Farrell B, Futuyma DJ (1991) Phylogenetic studies of insect-plant interactions: Insights into the genesis of diversity. Trends Ecol Evol 6:290–293CrossRefPubMedGoogle Scholar
  22. Navarro SP, Jurado-Rivera JA, Gómez-Zurita J, Lyal CHC, Vogler AP (2010) DNA profiling of host-herbivore interactions in tropical forests. Ecol Entomol 35(s1):18–32CrossRefGoogle Scholar
  23. Novotny V, Drozd P, Miller SE, Kulfan M, Janda M, Basset Y, Weiblen GD (2006) Why are there so many species of herbivorous insects in tropical rainforests? Science 313:1115–1118CrossRefPubMedGoogle Scholar
  24. Pan HS, Lu YH, Wyckhuys KAG, Wu KM (2013) Preference of a polyphagous mirid bug, Apolygus lucorum (Meyer-Dür) for flowering host plants. PLoS ONE 8:e68980CrossRefPubMedPubMedCentralGoogle Scholar
  25. Pompanon F, Deagle B, Symondson WOC, Brown DS, Jarman SN, Taberlet P (2012) Who is eating what: diet assessment using Next Generation Sequencing. Mol Ecol 21:1931–1950CrossRefPubMedGoogle Scholar
  26. Price PW, Bouton CE, Gross P, McPheron BA, Thompson JN (1987) Interactions among three trophic levels: influence of plants on interactions between insect herbivores and natural enemies. Annu Rev Ecol Syst 11(11):41–65Google Scholar
  27. Pumariño L, Alomar O, Agusti N (2011) Development of specific ITS markers for plant DNA identification within herbivorous insects. B Entomol Res 101:271–276CrossRefGoogle Scholar
  28. Remén C, Krüger M, Cassel-Lundhagen A (2010) Successful analysis of gut contents in fungal-feeding oribatid mites by combining body-surface washing and PCR. Soil Biol Biochem 42:1952–1957CrossRefGoogle Scholar
  29. Silberbauer L, Yee M, Del Socorro A, Wratten S, Gregg P, Bowie M (2004) Pollen grains as markers to track the movements of generalist predatory insects in agroecosystems. Int J Pest Manag 50:165–171CrossRefGoogle Scholar
  30. Sipos R, Szekely AJ, Palatinsky M, Révész S, Márialigeti K, Nikolausz M (2007) Effect of primer mismatch, annealing temperature and PCR cycle number on 16S rRNA gene-targetting bacterial community analysis. FEMS Microbiol Ecol 60:341–350CrossRefPubMedGoogle Scholar
  31. Spence KO, Rosenheim JA (2005) Isotopic enrichment in herbivorous insects: a comparative field-based study of variation. Oecologia 146:89–97CrossRefPubMedGoogle Scholar
  32. Staudacher K, Wallinger C, Schallhart N, Traugott M (2011) Detecting ingested plant DNA in soil-living insect larvae. Soil Biol Biochem 43:346–350CrossRefPubMedPubMedCentralGoogle Scholar
  33. Stephens AEA, Barrington AM, Bush VA, Fletcher NM, Mitchell VJ, Suckling DM (2008) Evaluation of dyes for marking painted apple moths (Teia anartoides Walker, Lep. Lymantriidae) used in a sterile insect release program. Aust J Entomol 47:131–136CrossRefGoogle Scholar
  34. Taberlet P, Gielly L, Pautou G, Bouvet J (1991) Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Mol Biol 17:1105–1109CrossRefPubMedGoogle Scholar
  35. Traugott M, Kamenova S, Ruess L, Seeber J, Plantegenest M (2013) Empirically characterising trophic networks: what emerging DNA-based methods, stable isotope and fatty acid analyses can offer. Adv Ecol Res 49:177–224CrossRefGoogle Scholar
  36. Valentini A, Pompanon F, Taberlet P (2009) DNA barcoding for ecologists. Trends Ecol Evol 24:110–117CrossRefPubMedGoogle Scholar
  37. Wallinger C, Juen A, Staudacher K, Schallhart N, Mitterrutzner E, Steiner EM, Thalinger B, Traugott M (2012) Rapid plant identification using species- and group-specific primers targeting chloroplast DNA. PLoS ONE 7:e29473CrossRefPubMedPubMedCentralGoogle Scholar
  38. Wallinger C, Staudacher K, Schallhart N, Peter E, Dresch P, Juen A, Traugott M (2013) The effect of plant identity and the level of plant decay on molecular gut content analysis in a herbivorous soil insect. Mol Ecol Res 13:75–83CrossRefGoogle Scholar
  39. Wallinger C, Staudacher K, Schallhart N, Mitterrutzner E, Steiner EM, Juen A, Traugott M (2014) How generalist herbivores exploit belowground plant diversity in temperate grasslands. Mol Ecol 23:3826–3837CrossRefPubMedGoogle Scholar
  40. Wallinger C, Sint D, Baier F, Schmid C, Mayer R, Traugott M (2015) Detection of seed DNA in regurgitates of granivorous carabid beetles. B Entomol Res 105:728–735CrossRefGoogle Scholar
  41. Wang Q, Bao WF, Zeng J, Yang YZ, Lu YH (2016) Tracking the tropic relationship between herbivorous insects and host plants by DNA-based technology. Acta Entomol Sin 59:472–480Google Scholar
  42. Wang Q, Bao WF, Yang F, Xu B, Yang YZ(2017)The specific host plant DNA detection suggests a potential migration of Apolygus lucorum from cotton to mungbean fields. PLoS ONE 12(6): e0177789Google Scholar
  43. Wanner H, Gu H, Gu Ènther D, Hein S, Dorn S (2006) Tracing spatial distribution of parasitism in fields with flowering plant strips using stable isotope marking. Biol Control 35:240–247CrossRefGoogle Scholar
  44. Wu KM, Li W, Feng HQ, Guo YY (2002) Seasonal abundance of the mirids, Lygus lucorum and Adelphocoris spp. (Hemiptera: Miridae) on Bt cotton in northern China. Crop Prot 21:997–1002CrossRefGoogle Scholar
  45. Wu KM, Lu YH, Feng HQ, Jiang YY, Zhao JZ (2008) Suppression of cotton bollworm in multiple crops in China in areas with Bt toxin-containing cotton. Science 321:1676–1678CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.College of Horticulture and Plant ProtectionYangzhou UniversityYangzhouChina
  2. 2.State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijingChina
  3. 3.College of AgricultureNortheast Agricultural UniversityHarbinChina

Personalised recommendations