Plant Ecology

, Volume 220, Issue 1, pp 29–39 | Cite as

Root-associated fungi increase male fitness, while high simulated herbivory decreases indirect defenses in Croton lachnostachyus plants

  • Mariana Pereyra
  • Gabriel GrilliEmail author
  • Leonardo Galetto


Plants interact with a diverse array of organisms below and above ground; some interactions with ants allow plants to be protected against herbivorous insects, influencing their growth or reproduction. In addition, indirect plant defenses—such as those mediated by extrafloral nectaries (EFNs)—could be affected by plant root symbionts. However, it is not clear how the suppression of root symbionts might affect extrafloral nectar (EFN) production and plant reproductive output. We made an experimental study with a shrub species with EFNs. Firstly, we tested if root-associated fungi (i.e., comparing plants with and without fungicide) increased the production of pollen (male function) and EFN (volume, nectar concentration, and total sugar content) in Croton lachnostachyus. Subsequently, we implemented a second experiment on the same plants, adding different levels of simulated herbivory (none, low, high) to assess the combined effects of root-associated fungi and herbivory. While we found high levels of mycorrhizal colonization, we found no signs of pathogenic fungi and negligible values of dark septate endophytes in roots so we attributed our results mostly to arbuscular mycorrhizal fungi (AMF). The first experiment showed that plants without the fungicide treatment increased pollen production and secreted a lower mean volume of EFN with higher concentration of dissolved soluble solids. In the second experiment, high levels of simulated herbivory showed a diminution on EFN variables; also, we detected a lower shoot dry mass on plants with low levels of herbivory and no interactions with AMF. Our results suggest complex ecological responses related to the root-associated fungal community and simulated herbivory.


Extrafloral nectar (EFN) Pollen Root-associated fungi Shoot dry mass Simulated herbivory Arbuscular mycorrhizal fungi (AMF) 



We thank two anonymous reviewers and Marina Omacini for useful comments and suggestions on a previous version of this manuscript. Also, we thank Romina Fernández for improving the English. This work was funded by FONCYT, the National University of Cordoba (SECyT—UNC), and CONICET. MP is a postdoctoral fellow in FONCYT. GG and LG are researchers from CONICET. Also, LG and MP are professors at the Universidad Nacional de Córdoba.


  1. Agrawal AA (2011) Current trends in the evolutionary ecology of plant defense. Funct Ecol 25:420–432. CrossRefGoogle Scholar
  2. Aslan CE, Zavaleta ES, Tershy B, Croll D (2013) Mutualism disruption threatens global plant biodiversity: a systematic review. PLoS ONE 8:e66993. CrossRefGoogle Scholar
  3. Ballhorn DJ, Kay J, Kautz S (2014) Quantitative effects of leaf area removal on indirect defense of Lima bean (Phaseolus lunatus) in nature. J Chem Ecol 40:294–296. CrossRefGoogle Scholar
  4. Barber NA, Soper Gorden NL (2014) How do belowground organisms influence plant–pollinator interactions? J Plant Ecol 8:1–11. CrossRefGoogle Scholar
  5. Bardgett RD, van der Putten WH (2014) Belowground biodiversity and ecosystem functioning. Nature 515:505–511. CrossRefGoogle Scholar
  6. Barto K, Rillig MC (2010) Does herbivory really suppress mycorrhiza? A meta-analysis. J Ecol 98:745–753. CrossRefGoogle Scholar
  7. Bennett AE, Bever JD (2007) Mycorrhizal species differentially alter plant growth and response to herbivory. Ecology 88:210–218.;2 CrossRefGoogle Scholar
  8. Bezemer TM, van Dam NM (2005) Linking aboveground and belowground interactions via induced plant defenses. Trends Ecol Evol 20:617–624. CrossRefGoogle Scholar
  9. Brundrett M, Bougher N, Dell B, Grove T (1996) Working with mycorrhizas in forestry and agriculture. AClAR Mg S 32:374Google Scholar
  10. Carrillo J, Wang Y, Ding J, Klootwyk K, Siemann E (2012) Decreased indirect defense in the invasive tree, Triadica sebifera. Plant Ecol 13:945–954. CrossRefGoogle Scholar
  11. Casas C, Torretta JP, Exeler N, Omacini M (2016) What happens next? Legacy effects induced by grazing and grass-endophyte symbiosis on thistle plants and their floral visitors. Plant Soil 405:211–229. CrossRefGoogle Scholar
  12. Croizat L (1941) Preliminaries for the study of Argentine and Uruguayan species of Croton. Darwiniana 5:417–462Google Scholar
  13. Delph LF, Johannsson MH, Stephenson AG (1997) How environmental factors affect pollen performance: ecological and evolutionary perspectives. Ecology 78:1632–1639.;2 CrossRefGoogle Scholar
  14. Fracchia S, Aranda A, Gopar A, Silvani V, Fernandez L, Godeas A (2009) Mycorrhizal status of plant species in the Chaco Serrano Woodland from central Argentina. Mycorrhiza 19:205–214. CrossRefGoogle Scholar
  15. Freitas L, Galetto L, Bernardello G, Paoli AA (2000) Ant exclusion and reproduction of Croton sarcopetalus (Euphorbiaceae). Flora (Jena) 195:398–402. CrossRefGoogle Scholar
  16. Freitas L, Bernardello G, Galetto L, Paoli AA (2001) Nectaries and reproductive biology of Croton sarcopetalus (Euphorbiaceae). Bot J Linn Soc 136:267–277. CrossRefGoogle Scholar
  17. Galetto L, Bernardello G (2005) Rewards in flowers: Nectar. In: Dafni A, Kevan PG, Husband BC (eds) Practical pollination biology. Enviroquest Ltd, Canada, pp 261–313Google Scholar
  18. Gange AC, Smith AK (2005) Arbuscular mycorrhizal fungi influence visitation rates of pollinating insects. Ecol Entomol 30:600–606. CrossRefGoogle Scholar
  19. Godschalx AL, Schädler M, Trisel JA, Balkan MA, Ballhorn DJ (2015) Ants are less attracted to the extrafloral nectar of plants with symbiotic, nitrogen-fixing rhizobia. Ecology 96:348–354. CrossRefGoogle Scholar
  20. Grace C, Stribley DP (1991) A safer procedure for routine staining of vesicular-arbuscular mycorrhizal fungi. Mycol Res 95:1160–1162. CrossRefGoogle Scholar
  21. Grilli G, Urcelay C, Longo MS, Galetto L (2014) Mycorrhizal fungi affect plant growth: experimental evidence comparing native and invasive hosts in the context of forest fragmentation. Plant Ecol 215:1513–1525. CrossRefGoogle Scholar
  22. Grilli G, Urcelay C, Galetto L, Davison J, Vasar M, Saks Ü, Jarius T, Öpik M (2015) The composition of arbuscular mycorrhizal fungal communities in the roots of a ruderal forb is not related to the forest fragmentation process. Environ Microbiol 17:2709–2720. CrossRefGoogle Scholar
  23. Harvell CD (1990) The ecology and evolution of inducible defenses. Q Rev Biol 65:323–340CrossRefGoogle Scholar
  24. Johnson NC, Graham JH (2013) The continuum concept remains a useful framework for studying mycorrhizal functioning. Plant Soil 363:411–419. CrossRefGoogle Scholar
  25. Jones IM, Koptur S (2015) Dynamic extrafloral nectar production: the timing of leaf damage affects the defensive response in Senna mexicana var. chapmanii (Fabaceae). Am J Bot 102:58–66. CrossRefGoogle Scholar
  26. Klironomos JN (2003) Variation in plant response to native and exotic arbuscular mycorrhizal fungi. Ecology 84:2292–2301. CrossRefGoogle Scholar
  27. Koorem K, Saks Ü, Sõber V, Uibopuu A, Öpik M, Zobel M, Moora M (2012) Effects of arbuscular mycorrhiza on community composition and seedling recruitment in temperate forest understory. Basic Appl Ecol 13:663–672. CrossRefGoogle Scholar
  28. Koptur S (1989) Is extrafloral nectar production an inducible defense. In: Bloc JH, Linhart YB (eds) The evolutionary ecology of plants. Westview Press, Boulder, pp 323–339Google Scholar
  29. Koptur S (1992) Extrafloral nectary-mediated interactions between insects and plants. In: Bernays EA (ed) Insect-plant interactions. CRC Press, Boca Raton, pp 81–129Google Scholar
  30. Koricheva J, Gange AC, Jones T (2009) Effects of mycorrhizal fungi on insect herbivores: a meta-analysis. Ecology 90:2088–2097. CrossRefGoogle Scholar
  31. Kula AA, Hartnett DC, Wilson GW (2005) Effects of mycorrhizal symbiosis on tallgrass prairie plant–herbivore interactions. Ecol Lett 8:61–69. CrossRefGoogle Scholar
  32. Laird RA, Addicott JF (2007) Arbuscular mycorrhizal fungi reduce the construction of extrafloral nectaries in Vicia faba. Oecologia 152:541–551. CrossRefGoogle Scholar
  33. Laird RA, Addicott JF (2009) Testing for mycorrhizal fungi-plant-ant indirect effects. J Plant Interact 4:7–14. CrossRefGoogle Scholar
  34. Leitner M, Kaiser R, Hause B, Boland W, Mithöfer A (2010) Does mycorrhization influence herbivore-induced volatile emission in Medicago truncatula? Mycorrhiza 20:89–101. CrossRefGoogle Scholar
  35. Li T, Holopainen JK, Kokko H, Tervahauta AI, Blande JD (2012) Herbivore induced aspen volatiles temporally regulate two different indirect defenses in neighbouring plants. Funct Ecol 26:1176–1185. CrossRefGoogle Scholar
  36. Massad TJ, Fincher RM, Smilanich AM, Dyer L (2011) A quantitative evaluation of major plant defense hypotheses, nature versus nurture, and chemistry versus ants. Arthropod-Plant Interact 5:125–139. CrossRefGoogle Scholar
  37. McGonigle TP, Miller MH, Evans DG, Fairchild GL, Swan JA (1990) A new method which gives an objective measure of colonization of roots by vesicular-arbuscular mycorrhizal fungi. New Phytol 115:495–501. CrossRefGoogle Scholar
  38. Mondor EB, Addicott JF (2003) Conspicuous extrafloral nectaries are inducible in Vicia faba. Ecol Lett 6:495–497. CrossRefGoogle Scholar
  39. Mondor EB, Tremblay MN, Messing RH (2006) Extrafloral nectary phenotypic plasticity is damage-and resource-dependent in Vicia faba. Biol Lett 2:583–585. CrossRefGoogle Scholar
  40. Montesinos-Navarro A, Valiente-Banuet A, Verdú M (2018) Mycorrhizal symbiosis increases the benefits of plant facilitative interactions. Ecography. Google Scholar
  41. Ness J (2003) Catalpa bignonioides alters extrafloral nectar production after herbivory and attracts ant bodyguards. Oecologia 134:210–218. CrossRefGoogle Scholar
  42. Oliveira PS, Brandão CRF (1991) The ant community associated with extrafloral nectaries in the Brazilian cerrados. In: Huxley CR, Cutler DF (eds) Ant-plant interactions. Oxford University Press, Oxford, pp 198–212Google Scholar
  43. Paul ND, Ayres PG, Wyness LE (1989) On the use of fungicides for experimentation in natural vegetation. Funct Ecol 3:59–769. CrossRefGoogle Scholar
  44. Pereyra M, Pol RG, Galetto L (2015) Does edge effect and patch size affect the interaction between ants and Croton lachnostachyus in fragmented landscapes of Chaco forest? Arthropod-Plant Interact 9:175–186. CrossRefGoogle Scholar
  45. Philip LJ, Posluszny U, Klironomos JN (2001) The influence of mycorrhizal colonization on the vegetative growth and sexual reproductive potential of Lythrum salicaria L. Can J Bot 79:381–388. Google Scholar
  46. Poulton JL, Bryla D, Koide RT, Stephenson AG (2002) Mycorrhizal infection and high soil phosphorus improve vegetative growth and the female and male functions in tomato. New Phytol 154:255–264. CrossRefGoogle Scholar
  47. Poveda K, Steffan-Dewenter I, Scheu S, Tscharntke T (2003) Effects of below-and above-ground herbivores on plant growth, flower visitation and seed set. Oecologia 135:601–605. CrossRefGoogle Scholar
  48. Pulice CE, Packer AA (2008) Simulated herbivory induces extrafloral nectary production in Prunus avium. Funct Ecol 22:801–807. CrossRefGoogle Scholar
  49. R Core Team (2017) R: a language and environment for statistical computing. R Foundation for Statistical Computing, ViennaGoogle Scholar
  50. Radhika V, Kost C, Bartram S, Heil M, Boland W (2008) Testing the optimal defense hypothesis for two indirect defenses: extrafloral nectar and volatile organic compounds. Planta 228:449–457. CrossRefGoogle Scholar
  51. Rasmann S, Bennett A, Biere A, Karley A, Guerrieri E (2017) Root symbionts: powerful drivers of plant above and belowground indirect defenses. Insect Sci 2:4. Google Scholar
  52. Rico-Gray V, Oliveira PS (2007) The ecology and evolution of ant-plant interactions. University of Chicago Press, ChicagoCrossRefGoogle Scholar
  53. Rutter MT, Rausher MD (2004) Natural selection on extrafloral nectar production in Chamaecrista fasciculata: the costs and benefits of a mutualism trait. Evolution 58:2657–2668. CrossRefGoogle Scholar
  54. Schädler M, Ballhorn DJ (2017) Beneficial soil microbiota as mediators of the plant defensive phenotype and aboveground plant-herbivore interactions. Prog Bot 78:305–343. Google Scholar
  55. Schat M, Blossey B (2005) Influence of natural and simulated leaf beetle herbivory on biomass allocation and plant architecture of purple loosestrife (Lythrum salicaria L.). Environ Entomol 34:906–914. CrossRefGoogle Scholar
  56. Smith SE, Read DJ (2008) Mycorrhizal Symbiosis, 3rd edn. Academic, San DiegoGoogle Scholar
  57. Smith LL, Lanza J, Smith GC (1990) Amino acid concentrations in extrafloral nectar of Impatiens sultani increase after simulated herbivory. Ecology 71:107–115. CrossRefGoogle Scholar
  58. Smith SE, Smith FA, Jakobsen I (2003) Mycorrhizal fungi can dominate phosphate supply to plants irrespective of growth responses. Plant Physiol 133:16–20. CrossRefGoogle Scholar
  59. Smith SE, Smith FA, Jakobsen I (2004) Functional diversity in arbuscular mycorrhizal (AM) symbioses: the contribution of the mycorrhizal P uptake pathway is not correlated with mycorrhizal responses in growth or total P uptake. New Phytol 162:511–524. CrossRefGoogle Scholar
  60. Strauss SY, Agrawal AA (1999) The ecology and evolution of plant tolerance to herbivory. Trends Ecol Evol 14:179–185. CrossRefGoogle Scholar
  61. Varga S (2010) Effects of arbuscular mycorrhizas on reproductive traits in sexually dimorphic plants: a review. Span J Agric Res 8:11–24. CrossRefGoogle Scholar
  62. Varga S, Kytöviita MM (2010) Gender dimorphism and mycorrhizal symbiosis affect floral visitors and reproductive output in Geranium sylvaticum. Funct Ecol 24:750–758. CrossRefGoogle Scholar
  63. Wäckers FL, Zuber D, Wunderlin R, Keller F (2001) The effect of herbivory on temporal and spatial dynamics of foliar nectar production in cotton and castor. Ann Bot 87:365–370. CrossRefGoogle Scholar
  64. Wardle DA (2002) Communities and ecosystems: linking the aboveground and belowground components. Princeton University Press, New JerseyGoogle Scholar
  65. Weber MG, Keeler KH (2013) The phylogenetic distribution of extrafloral nectaries in plants. Ann Bot 111:1251–1261. CrossRefGoogle Scholar
  66. Yamawaki K, Matsumura A, Hattori R, Tarui A, Hossain MA, Ohashi Y, Daimon H (2013) Effect of inoculation with arbuscular mycorrhizal fungi on growth, nutrient uptake and curcumin production of turmeric (Curcuma longa L.). Agric Sci 4:66. Google Scholar
  67. Yamawo A, Katayama N, Suzuki N, Hada Y (2012) Plasticity in the expression of direct and indirect defense traits of young plants of Mallotus japonicus in relation to soil nutritional conditions. Plant Ecol 213:127–132. CrossRefGoogle Scholar
  68. Zangerl AR, Hamilton JG, Miller TJ, Crofts AR, Oxborough K, Berenbaum MR, De Lucia EH (2002) Impact of folivory on photosynthesis is greater than the sum of its holes. Proc Natl Acad Sci USA 99:1088–1091. CrossRefGoogle Scholar
  69. Zhang W, Chen XX, Liu YM, Liu DY, Chen XP, Zou CQ (2017) Zinc uptake by roots and accumulation in maize plants as affected by phosphorus application and arbuscular mycorrhizal colonization. Plant Soil 413:59–71. CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Mariana Pereyra
    • 1
  • Gabriel Grilli
    • 1
    Email author
  • Leonardo Galetto
    • 1
    • 2
  1. 1.Instituto Multidisciplinario de Biología Vegetal, FCEFyN, (CONICET-Universidad Nacional de Córdoba)CórdobaArgentina
  2. 2.Departamento de Diversidad Biológica y Ecología, Facultad de Ciencias ExactasFísicas y Naturales Universidad Nacional de CórdobaCórdobaArgentina

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