Arthropod-Plant Interactions

, Volume 11, Issue 6, pp 901–909 | Cite as

1H NMR analysis of Citrus macrophylla subjected to Asian citrus psyllid (Diaphorina citri Kuwayama) feeding

  • Elizabeth Chin
  • Kris Godfrey
  • MaryLou Polek
  • Carolyn Slupsky
Original Paper


The Asian citrus psyllid (ACP) is a phloem-feeding insect that can host and transmit the bacterium Candidatus Liberibacter asiaticus (CLas), which is the putative causative agent of the economically important citrus disease, Huanglongbing (HLB). ACP are widespread in Florida, and are spreading in California; they are the primary mode of CLas transmission in citrus groves. To understand the effects of ACP feeding, different numbers of ACP [0 ACP (control), 5 ACP (low), 15–20 ACP (medium), and 25–30 ACP (high)] were allowed to feed on Citrus macrophylla greenhouse plants. After 7 days of feeding, leaves were collected and analyzed using 1H NMR. Metabolite concentrations from leaves of trees with ACP feeding had higher variability than control trees. Many metabolites were higher in concentration in the low ACP feeding group relative to control; however, leaves from trees with high ACP feeding had lower concentrations of many metabolites relative to control, including many amino acids such as phenylalanine, arginine, isoleucine, valine, threonine, and leucine. These results suggest ACP density-dependent changes in primary metabolism that can be measured by 1H NMR. The implications in plant defense are discussed.


Huanglongbing (HLB) Asian citrus psyllid (ACP) Candidatus Liberibacter asiaticus (CLas) 



The authors gratefully acknowledge support for this project from the Citrus Research Board. This project was made possible in part by support from the USDA National Institute of Food and Agriculture Hatch Project 1005945. We thank Steve Stearns for assistance with sample preparation, Dr. Cythia LeVesque for performing qPCR on the ACP colonies, Drs. Michelle Cilia, John Ramsey, and Greg McCollum for their suggestions and critical reading of the manuscript.

Supplementary material

11829_2017_9546_MOESM1_ESM.pdf (488 kb)
Supplementary material 1 (PDF 488 kb)


  1. Baldwin IT, Halitschke R, Kessler A, Schittko U (2001) Merging molecular and ecological approaches in plant–insect interactions. Curr Opin Plant Biol 4:351–358CrossRefPubMedGoogle Scholar
  2. Bernards MA, Båstrup-Spohr L (2008) Phenylpropanoid metabolism induced by wounding and insect herbivory. In: Schaller A (ed) Induced plant resistance to herbivory. Springer, Dordrecht, pp 189–211CrossRefGoogle Scholar
  3. Chin EL, Mishchuk DO, Breksa AP, Slupsky CM (2014) Metabolite signature of Candidatus Liberibacter asiaticus infection in two citrus varieties. J Agric Food Chem 62:6585–6591CrossRefPubMedGoogle Scholar
  4. Dadd R, Krieger D (1968) Dietary amino acid requirements of the aphid, Myzus persicae. J Insect Physiol 14:741–764CrossRefGoogle Scholar
  5. Douglas A (2006) Phloem-sap feeding by animals: problems and solutions. J Exp Bot 57:747–754CrossRefPubMedGoogle Scholar
  6. Fan J, Chen C, Brlansky R, Gmitter F Jr, Li ZG (2010) Changes in carbohydrate metabolism in Citrus sinensis infected with ‘Candidatus Liberibacter asiaticus’. Plant Pathol 59:1037–1043CrossRefGoogle Scholar
  7. Giordanengo P et al (2010) Compatible plant–aphid interactions: how aphids manipulate plant responses. C R Biol 333:516–523CrossRefPubMedGoogle Scholar
  8. Gómez S, Ferrieri RA, Schueller M, Orians CM (2010) Methyl jasmonate elicits rapid changes in carbon and nitrogen dynamics in tomato. New Phytol 188:835–844CrossRefPubMedGoogle Scholar
  9. Grafton-Cardwell EE, Godfrey KE, Rogers ME, Childers CC, Stansly PA (2005) Asian citrus psyllid. UCANR Publications. Accessed 27 July 2016
  10. Grafton-Cardwell EE, Stelinski LL, Stansly PA (2013) Biology and management of Asian citrus psyllid, vector of the huanglongbing pathogens. Annu Rev Entomol 58:413–432CrossRefPubMedGoogle Scholar
  11. Halbert SE, Manjunath KL (2004) Asian citrus psyllids (Sternorrhyncha: Psyllidae) and greening disease of citrus: a literature review and assessment of risk in Florida. Fla Entomol 87:330–353CrossRefGoogle Scholar
  12. Heidel A, Baldwin I (2004) Microarray analysis of salicylic acid- and jasmonic acid-signalling in responses of Nicotiana attenuata to attack by insects from multiple feeding guilds. Plant, Cell Environ 27:1362–1373CrossRefGoogle Scholar
  13. Heil M (2002) Ecological costs of induced resistance. Curr Opin Plant Biol 5:345–350CrossRefPubMedGoogle Scholar
  14. Howe GA, Jander G (2008) Plant immunity to insect herbivores. Annu Rev Plant Biol 59:41–66CrossRefPubMedGoogle Scholar
  15. Kant M et al (2015) Mechanisms and ecological consequences of plant defence induction and suppression in herbivore communities. Ann Bot London 115:1015–1051CrossRefGoogle Scholar
  16. Kessler A, Baldwin IT (2002) Plant responses to insect herbivory: the emerging molecular analysis. Annu Rev Plant Biol 53:299–328CrossRefPubMedGoogle Scholar
  17. Kim JS, Sagaram US, Burns JK, Li JL, Wang N (2009) Response of sweet orange (Citrus sinensis) to ‘Candidatus Liberibacter asiaticus’ infection: microscopy and microarray analyses. Phytopathology 99:50–57CrossRefPubMedGoogle Scholar
  18. Korth KL, Dixon RA (1997) Evidence for chewing insect-specific molecular events distinct from a general wound response in leaves. Plant Physiol 115:1299–1305CrossRefPubMedPubMedCentralGoogle Scholar
  19. Lee JA, Halbert SE, Dawson WO, Robertson CJ, Keesling JE, Singer BH (2015) Asymptomatic spread of huanglongbing and implications for disease control. Proc Natl Acad Sci USA 112:7605–7610CrossRefPubMedPubMedCentralGoogle Scholar
  20. Leiss KA, Maltese F, Choi YH, Verpoorte R, Klinkhamer PG (2009) Identification of chlorogenic acid as a resistance factor for thrips in chrysanthemum. Plant Physiol 150:1567–1575CrossRefPubMedPubMedCentralGoogle Scholar
  21. Li W, Hartung JS, Levy L (2006) Quantitative real-time PCR for detection and identification of Candidatus Liberibacter species associated with citrus huanglongbing. J Microbiol Meth 66:104–115CrossRefGoogle Scholar
  22. Liu D, Johnson L, Trumble JT (2006) Differential responses to feeding by the tomato/potato psyllid between two tomato cultivars and their implications in establishment of injury levels and potential of damaged plant recovery. Insect Sci 13:195–204CrossRefGoogle Scholar
  23. Malik NS, Perez JL, Kunta M, Patt JM, Mangan RL (2014) Changes in free amino acids and polyamine levels in Satsuma leaves in response to Asian citrus psyllid infestation and water stress. Insect Sci 21:707–716CrossRefPubMedGoogle Scholar
  24. Manjunath KL, Halbert SE, Ramadugu C, Webb S, Lee RF (2008) Detection of ‘Candidatus Liberibacter asiaticus’ in Diaphorina citri and its importance in the management of citrus huanglongbing in Florida. Phytopathology 98:387–396CrossRefPubMedGoogle Scholar
  25. Mattson WJ (1980) Herbivory in relation to plant nitrogen content. Annu Rev Ecol Syst 11:119–161CrossRefGoogle Scholar
  26. Morath S, Pratt P, Silvers C, Center T (2006) Herbivory by Boreioglycaspis melaleucae (Hemiptera: Psyllidae) accelerates foliar senescence and abscission in the invasive tree Melaleuca quinquenervia. Environ Entomol 35:1372–1378CrossRefGoogle Scholar
  27. Mozoruk J, Hunnicutt LE, Cave RD, Hunter WB, Bausher MG (2006) Profiling transcriptional changes in Citrus sinensis (L.) Osbeck challenged by herbivory from the xylem-feeding leafhopper Homalodisca coagulata (Say) by cDNA macroarray analysis. Plant Sci 170:1068–1080CrossRefGoogle Scholar
  28. Newingham BA, Callaway RM, Bassirirad H (2007) Allocating nitrogen away from a herbivore: a novel compensatory response to root herbivory. Oecologia 153:913–920CrossRefPubMedGoogle Scholar
  29. Orians CM, Thorn A, Gómez S (2011) Herbivore-induced resource sequestration in plants: why bother? Oecologia 167:1–9CrossRefPubMedGoogle Scholar
  30. Pelz-Stelinski KS, Brlansky RH, Ebert TA, Rogers ME (2010) Transmission parameters for Candidatus Liberibacter asiaticus by Asian citrus psyllid (Hemiptera: Psyllidae). J Econ Entomol 103:1531–1541CrossRefPubMedGoogle Scholar
  31. Polek M, Vidalakis G, Godfrey K (2007) Citrus bacterial canker disease and Huanglongbing (citrus greening). UCANR Publications. Accessed 26 July 2016
  32. Ray P, Hill MP (2016) More is not necessarily better: the interaction between insect population density and culture age of fungus on the control of invasive weed water hyacinth. Hydrobiologia 766:189–200CrossRefGoogle Scholar
  33. Sandström J, Telang A, Moran N (2000) Nutritional enhancement of host plants by aphids—a comparison of three aphid species on grasses. J Insect Physiol 46:33–40CrossRefPubMedGoogle Scholar
  34. Sardans J, Gargallo-Garriga A, Pérez-Trujillo M, Parella T, Seco R, Filella I, Peñuelas J (2014) Metabolic responses of Quercus ilex seedlings to wounding analysed with nuclear magnetic resonance profiling. Plant Biol 16:395–403CrossRefPubMedGoogle Scholar
  35. Schultz JC, Appel HM, Ferrieri AP, Arnold TM (2013) Flexible resource allocation during plant defense responses. Front Plant Sci 4:1–11CrossRefGoogle Scholar
  36. Schweiger R, Heise AM, Persicke M, Müller C (2014) Interactions between the jasmonic and salicylic acid pathway modulate the plant metabolome and affect herbivores of different feeding types. Plant Cell Environ 37:1574–1585CrossRefPubMedGoogle Scholar
  37. Seigler DS (1998) Phenylpropanoids. Plant secondary metabolism. Springer, Boston, pp 106–129CrossRefGoogle Scholar
  38. Sétamou M, Flores D, French JV, Hall DG (2008) Dispersion patterns and sampling plans for Diaphorina citri (Hemiptera: Psyllidae) in citrus. J Econ Entomol 101:1478–1487CrossRefPubMedGoogle Scholar
  39. Sétamou M, Alabi OJ, Kunta M, Jifon JL, da Graça JV (2016) Enhanced acquisition rates of ‘Candidatus Liberibacter asiaticus’ by the Asian citrus psyllid (Hemiptera: Liviidae) in the presence of vegetative flush growth in citrus. J Econ Entomol 109:1973–1978CrossRefPubMedGoogle Scholar
  40. Simmonds MS (2003) Flavonoid–insect interactions: recent advances in our knowledge. Phytochemistry 64:21–30CrossRefPubMedGoogle Scholar
  41. Singerman A, Useche P (2016) Impact of citrus greening on citrus operations in Florida. University of Florida, IFAS Citrus Extension. Accessed 24 Jan 2017
  42. Steinbauer M, Burns A, Hall A, Riegler M, Taylor G (2014) Nutritional enhancement of leaves by a psyllid through senescence-like processes: insect manipulation or plant defence? Oecologia 176:1061–1074CrossRefPubMedPubMedCentralGoogle Scholar
  43. Steinbrenner AD, Gómez S, Osorio S, Fernie AR, Orians CM (2011) Herbivore-induced changes in tomato (Solanum lycopersicum) primary metabolism: a whole plant perspective. J Chem Ecol 37:1294–1303CrossRefPubMedGoogle Scholar
  44. Tomlin E, Sears M (1992) Effects of Colorado potato beetle and potato leafhopper on amino acid profile of potato foliage. J Chem Ecol 18:481–488CrossRefPubMedGoogle Scholar
  45. USDA, APHIS, PPQ (2014) National quarantine boundaries for Asian citrus psyllid and citrus greening. Accessed 01 Aug 2017
  46. Walling LL (2000) The myriad plant responses to herbivores. J Plant Growth Regul 19:195–216PubMedGoogle Scholar
  47. Walling LL (2008) Avoiding effective defenses: strategies employed by phloem-feeding insects. Plant Physiol 146:859–866CrossRefPubMedPubMedCentralGoogle Scholar
  48. Weljie AM, Newton J, Mercier P, Carlson E, Slupsky CM (2006) Targeted profiling: quantitative analysis of 1H NMR metabolomics data. Anal Chem 78:4430–4442CrossRefPubMedGoogle Scholar
  49. Widarto HT, Van Der Meijden E, Lefeber AW, Erkelens C, Kim HK, Choi YH, Verpoorte R (2006) Metabolomic differentiation of Brassica rapa following herbivory by different insect instars using two-dimensional nuclear magnetic resonance spectroscopy. J Chem Ecol 32:2417–2428CrossRefPubMedGoogle Scholar
  50. Will T, Furch AC, Zimmermann MR (2013) How phloem-feeding insects face the challenge of phloem-located defenses. Front Plant Sci 4:1–12CrossRefGoogle Scholar
  51. Wu J, Baldwin IT (2010) New insights into plant responses to the attack from insect herbivores. Annu Rev Genet 44:1–24CrossRefPubMedGoogle Scholar
  52. Xia J, Wishart DS (2016) Using MetaboAnalyst 3.0 for comprehensive metabolomics data analysis. Curr Protoc Bioinform 55:14.10.1–14.10.91CrossRefGoogle Scholar
  53. Xu C, Xia Y, Li K, Ke C (1988) Further study of the transmission of citrus huanglongbing by a psyllid, Diaphorina citri Kuwayama. In: LW Timmer, SM Garnsey, and L. Navarro (eds) Proceedings of the 10th conference of the international organization of citrus virologists. University of California, Riverside, pp 243–248Google Scholar
  54. Zhang X, Breksa AP, Mishchuk DO, Fake CE, O’Mahony MA, Slupsky CM (2012) Fertilisation and pesticides affect mandarin orange nutrient composition. Food Chem 134:1020–1024CrossRefPubMedGoogle Scholar
  55. Zhu-Salzman K, Salzman RA, Ahn J-E, Koiwa H (2004) Transcriptional regulation of sorghum defense determinants against a phloem-feeding aphid. Plant Physiol 134:420–431CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  • Elizabeth Chin
    • 1
  • Kris Godfrey
    • 2
  • MaryLou Polek
    • 3
  • Carolyn Slupsky
    • 1
  1. 1.Department of Food Science and TechnologyUniversity of California, DavisDavisUSA
  2. 2.Contained Research FacilityUniversity of California, DavisDavisUSA
  3. 3.National Clonal Germplasm Repository for Citrus & DatesRiversideUSA

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