Biological Invasions

, Volume 17, Issue 9, pp 2779–2791 | Cite as

A novel impact of a novel weapon: allelochemicals in Alliaria petiolata disrupt the legume-rhizobia mutualism

  • Cristina Portales-Reyes
  • Tina Van Doornik
  • Elizabeth H. Schultheis
  • Tomomi Suwa
Original Paper


Some introduced species become invasive by releasing novel allelochemicals into the soil, directly harming nearby plants and soil microbes. Alliaria petiolata (garlic mustard) is an invasive plant, well known to excrete a suite of phytotoxic and anti-microbial allelochemicals, including allyl isothiocyanate (AITC) and benzyl isothiocyanate (BITC). While the effects of these chemicals on plant-mycorrhizae mutualisms are well documented, the effects on other plant-soil microbe interactions, such as the legume-rhizobia mutualism, have not yet been tested. Here, we performed laboratory and greenhouse experiments with both synthetic chemicals and leaf extracts to investigate the effects of allelochemicals in A. petiolata on a native leguminous plant, Amphicarpaea bracteata, and its rhizobia mutualists. We found that BITC reduced rhizobia growth rate in the lab, but had no effect on nodulation in the greenhouse when rhizobia were grown in the presence of plants. AITC did not directly harm either plants or rhizobia, though plants and rhizobia grown in the presence of AITC showed reduced nodulation, indicating that it disrupted the formation of the mutualism itself. We found no effects of A. petiolata allelochemical leaf extracts on plant performance or nodulation. Our data suggest that AITC causes mutualism disruption in this system by preventing the formation of nodules, which reduces plant growth and could threaten the long-term performance of rhizobia. Our study shows that the allelochemicals in A. petiolata disrupt the legume-rhizobia resource mutualism, adding another impact of these novel weapons in addition to their well-documented role in disrupting plant-mycorrhizae symbioses.


Garlic mustard Amphicarpaea bracteata Allelopathy Plant invasion Mutualism disruption 



We thank J.A. Lau, J.P. Martina, R. Tinghitella, S. Murphy, A. Sher, J. Morris, and colleagues at the Kellogg Biological Station (KBS) and University of Denver who provided comments that substantially improved this manuscript. We would also like to thank the three anonymous reviewers who provided useful feedback on earlier versions of this paper. For the use of their field sites, we thank the Kalamazoo Nature Center, Pierce Cedar Creek Institute, Southwest Michigan Land Conservancy, Michigan Nature Association, and KBS. This work was funded by an NSF-BEACON REU fellowship awarded to C. Portales-Reyes, KBS Undergraduate Research Apprenticeships awarded to C. Portales-Reyes and T. Van Doornik, and by a George H. Lauff Research Award awarded to E.H. Schultheis and T. Suwa. This is KBS publication #1793.

Supplementary material

10530_2015_913_MOESM1_ESM.docx (18.1 mb)
Supplementary material 1 (DOCX 18486 kb)


  1. Alford ÉR, Vivanco JM, Paschke MW (2009) The effects of flavonoid allelochemicals from knapweeds on legume-rhizobia candidates for restoration. Restor Ecol 17(4):506–514CrossRefGoogle Scholar
  2. Anderson RC, Dhillion SS, Kelley TM (1996) Aspects of the ecology of an invasive plant, garlic mustard (Alliaria petiolata), in central Illinois. Restoration Ecol 4:181–191Google Scholar
  3. Bergersen FJ (1982) Root nodules of legumes: structure and functions. Research Studies Press, Wiley, New YorkGoogle Scholar
  4. Blair AC, Hanson BD, Brunk GR, Marrs RA, Westra P, Nissen SJ, Hufbauer RA (2005) New techniques and findings in the study of a candidate allelochemical implicated in invasion success. Ecol Lett 8:1039–1047CrossRefGoogle Scholar
  5. Blair AC, Nissen SC, Brunk GR, Hufbauer RA (2006) A lack of evidence for an ecological role of the putative allelochemical (±)-catechin in spotted knapweed invasion success. J Chem Ecol 32:2327–2331PubMedCrossRefGoogle Scholar
  6. Blažević I, Mastelić J (2008) Free and bound volatiles of garlic mustard (Alliaria petiolata). Croat Chem Acta 81:607–613Google Scholar
  7. Blossey B, Nuzzo V, Hinz HL, Gerber E (2001) Developing biological control of Alliaria petiolata (M. Bieb.) Cavara and Grande (garlic mustard). Nat Areas J 21(4):357–367Google Scholar
  8. Borek V, Morra MJ, Brown PJ, McCaffery JP (1995) Transformation of the glucosinolate-derived allelochemicals allyl isothiocyanate and allylnitrile in soil. J Agric Food Chem 43:1935–1940CrossRefGoogle Scholar
  9. Bottomley PJ, Myrold DD (2007) Biological N inputs. In: Paul EA (ed) Soil microbiology, ecology, and biochemistry, 3rd edn. Academic Press, MA, USAGoogle Scholar
  10. Brown PD, Morra MJ (1997) Control of soil-borne plant pests using glucosinolate-containing plants. Adv Agron 61:167–231CrossRefGoogle Scholar
  11. Brown PD, Morra MJ, McCaffrey JP, Auld DL, Williams L III (1991) Allelochemicals produced during glucosinolate degradation in soil. J Chem Ecol 17(10):2021–2034PubMedCrossRefGoogle Scholar
  12. Bullock DG, Anderson DS (1998) Evaluation of the Minolta SPAD-502 chlorophyll meter for nitrogen management in corn. J Plant Nutr 21(4):741–755CrossRefGoogle Scholar
  13. Burke DJ (2008) Effects of Alliaria petiolata (garlic mustard; Brassicaceae) on mycorrhizal colonization and community structure in three herbaceous plants in a mixed deciduous forest. Am J Bot 95(11):1416–1425PubMedCrossRefGoogle Scholar
  14. Callaway RM, Aschehoug ET (2000) Invasive plants versus their new and old neighbors: a mechanism for exotic invasion. Science 290(5491):521–523PubMedCrossRefGoogle Scholar
  15. Callaway RM, Ridenour WM (2004) Novel weapons: invasive success and the evolution of increased competitive ability. Front Ecol Environ 2(8):436–443CrossRefGoogle Scholar
  16. Callaway RM, Cipollini D, Barto EK, Thelen GC, Hallett SG, Prati D, Stinson KA, Klironomos J (2008) Novel weapons: invasive plant suppresses fungal mutualists in America but not in its native Europe. Ecology 89(4):1043–1055PubMedCrossRefGoogle Scholar
  17. Cantor A, Hale A, Aaron J, Traw MB, Kalisz S (2011) Low allelochemical concentrations detected in garlic-mustard invaded forest soils inhibit fungal growth and AMF spore germination. Biol. Inv. 13(12):3015–3025CrossRefGoogle Scholar
  18. Cappuccino N, Arnason JT (2006) Novel chemistry of invasive exotic plants. Biol Lett 2:189–193PubMedCentralPubMedCrossRefGoogle Scholar
  19. Cipollini D, Gruner B (2007) Cyanide in the chemical arsenal of garlic mustard, Alliaria petiolata. J Chem Ecol 33:85–94PubMedCrossRefGoogle Scholar
  20. Cipollini K, Titus K, Wagner C (2012a) Allelopathic effects of invasive species (Alliaria petiolata, Lonicera maackii, Ranunculus ficaria) in the Midwestern United States. Allelopathy J 29(1):63–76Google Scholar
  21. Cipollini D, Rigsby CM, Barto EK (2012b) Microbes as targets and mediators of allelopathy in plants. J Chem Ecol 38:714–727PubMedCrossRefGoogle Scholar
  22. Cleveland CC, Townsend AR, Schimel DS, Fisher H, Howarth RW, Hedin LO, Perakis SS, Latty EF, Fischer JCV, Elseroad A, Wasson MF (1999) Global patterns of terrestrial biological nitrogen (N 2) fixation in natural ecosystems. Global Biogeochem Cycles 13:623–645CrossRefGoogle Scholar
  23. Denison RF, Kiers TE (2004) Why are most rhizobia beneficial to their plant hosts, rather than parasitic? Microbes Infect 6(13):1235–1239PubMedCrossRefGoogle Scholar
  24. Fahey JW, Zalcmann AT, Talalay P (2001) The chemical diversity and distribution of glucosinolates and isothiocyanates among plants. Phytochemistry 56:5–51PubMedCrossRefGoogle Scholar
  25. Gáborčík N (2003) Relationship between contents of chlorophyll (a + b) (SPAD values) and nitrogen of some temperate grasses. Photosynthetica 41:285–287. doi: 10.1023/B:PHOT.0000011963.43628.15 CrossRefGoogle Scholar
  26. Gimsing AL, Kirkegaard JA (2006) Glucosinolate and isothiocyanate concentration in soil following incorporation of Brassica biofumigants. Soil Biol Biochem 38:2255–2264CrossRefGoogle Scholar
  27. Gimsing AL, Kirkegaard JA (2009) Glucosinolates and biofumigation: fate of glucosinolates and their hydrolysis products in soil. Phytochem Rev 8:299–310CrossRefGoogle Scholar
  28. Grieve M (1959) A modern herbal, vol 2. Hafner, New YorkGoogle Scholar
  29. Hale AN, Kalisz S (2012) Perspectives on allelopathic disruption of plant mutualisms: a framework for individual- and population-level fitness consequences. Plant Ecol 213(8):1991–2006CrossRefGoogle Scholar
  30. Hale AN, Tonsor SJ, Kalisz S (2011) Testing the mutualism disruption hypothesis: physiological mechanisms for invasion of intact perennial plant communities. Ecosphere 2(10):art110Google Scholar
  31. Heath KD, Tiffin P (2007) Context dependence in the coevolution of plant and rhizobial mutualists. Proc R Soc B 274(1620):1905–1912PubMedCentralPubMedCrossRefGoogle Scholar
  32. Kiers TE, Rousseau RA, West SA, Denison RF (2003) Host sanctions and the legume-rhizobium mutualism. Nature 425:78–81PubMedCrossRefGoogle Scholar
  33. Klironomos JN (2002) Feedback with soil biota contributes to plant rarity and invasiveness in communities. Nature 417:67–70PubMedCrossRefGoogle Scholar
  34. Lankau RA (2011) Intraspecific variation in allelochemistry determines an invasive species’ impact on soil microbial communities. Oecologia 165:453–463PubMedCrossRefGoogle Scholar
  35. Lankau RA (2012) Coevolution between invasive and native plants driven by chemical competition and soil biota. Proc Natl Acad Sci 109:11240–11245PubMedCentralPubMedCrossRefGoogle Scholar
  36. Lankau RA, Nuzzo V, Spyreas G, Davis AS (2009) Evolutionary limits ameliorate the negative impact of an invasive plant. Proc Natl Acad Sci USA 106:15362–15367Google Scholar
  37. Larsen PO (1981) Glucosinolates. In: Conn EE (ed) The biochemistry of plants, secondary plant products, vol 7. Academic Press, New York, pp 501–525Google Scholar
  38. Larsen LM, Olsen O, Plöger A, Søreson H (1983) Sinapine-O-ß-D-gluco-pyranoside in seeds of Alliaria officinalis. Phytochemistry 22:219–222CrossRefGoogle Scholar
  39. Lennon JT, Khatana SAM, Marston MF, Martiny JBH (2007) Is there a cost of virus resistance in marine cyanobacteria? ISME J 1:300–312PubMedGoogle Scholar
  40. Lorenzo P, Pereira CS, Rodríguez-Echeverría S (2013) Differential impact on soil microbes of allelopathic compounds released by the invasive Acacia dealbata Link. Soil Biol Biochem 57:156–163CrossRefGoogle Scholar
  41. Mallik MAB, Tesfai K (1988) Allelopathic effect of common weeds on soybean growth and soybean-Bradyrhizobium symbiosis. Plant Soil 112:177–182CrossRefGoogle Scholar
  42. McCarthy BC (1997) Response of a forest understory community to experimental removal of an invasive nonindigenous plant (Alliaria petiolata Brassicaceae) assessment and management of plant invasions. Springer Series on Environmental Management, pp 117–130Google Scholar
  43. McCarthy BC, Hanson SL (1998) An assessment of the allelopathic potential of the invasive weed Alliaria petiolata (Brassicacea). Castanea 63:68–73Google Scholar
  44. Meekins JF, McCarthy BC (1999) Competitive ability of Alliaria petiolata (garlic mustard, Brassicaceae), an invasive nonindigenous forest herb. Int J Plant Sci 160:743–752CrossRefGoogle Scholar
  45. Meiners SJ, Kong C-H (2012) Introduction to the special issue on allelopathy. Plant Ecol 213(12):1857–1859CrossRefGoogle Scholar
  46. Muller RN, Borman FH (1976) Role of Erythronium americanum Ker. in energy flow and nutrient dynamics of a northern hardwood forest ecosystem. Science 193:1126–1128Google Scholar
  47. Murrell C, Gerber E, Krebs C, Parepa M, Schaffner U, Bossdorf O (2011) Invasive knotweed affects native plants through allelopathy. Am J Bot 98(1):38–43PubMedCrossRefGoogle Scholar
  48. Nuzzo VA (1993) Distribution and spread of the invasive biennial garlic mustard (Alliaria petiolata) in North America. In: McNight BN (ed) Biological pollution: the control and impact of invasive exotic species. Indiana Academy of Science, Indianapolis, pp 137–146Google Scholar
  49. Nuzzo VA (1999) Invasion pattern of the herb garlic mustard (Alliaria petiolata) in high quality forests. Biol Invasions 1:169–179CrossRefGoogle Scholar
  50. Oono R, Anderson CG, Denison RF (2011) Failure to fix nitrogen by non-reproductive symbiotic rhizobia triggers host sanctions that reduce fitness of their reproductive clonemates. Proc R Soc Lond Ser B 278:2698–2703Google Scholar
  51. Parker MA (1996) Cryptic species within Amphicarpaea bracteata (Leguminosae): evidence from isozymes, morphology, and pathogen specificity. Can J Bot 74:1640–1650CrossRefGoogle Scholar
  52. Pearse IS, Bastow JL, Tsang A (2014) Radish introduction affects soil biota and has a positive impact on the growth of a native plant. Oecologia 174(2):471–478PubMedCrossRefGoogle Scholar
  53. R Core Team (2015) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria.
  54. Reznicek AA, Voss EG, Walters BS (2011) Michigan flora online. University of Michigan. Accessed 15 Jan 2014
  55. Rice EL (1974) Allelopathy. Academic Press, New YorkGoogle Scholar
  56. Roberts KJ, Anderson RC (2001) Effect of garlic mustard [Alliaria petiolata (Beib. Cavara & Grande)] extracts on plants and arbuscular mycorrhizal (AM) fungi. Am Midl Nat 146:146–152CrossRefGoogle Scholar
  57. Rodgers VL, Stinson KA, Finzi AC (2008a) Ready or not, garlic mustard is moving in: Alliaria petiolata as a Member of Eastern North American Forests. BioScience 58(5):1–11Google Scholar
  58. Rodgers VL, Wolfe BE, Werden LK, Finzi AC (2008b) The invasive species Alliaria petiolata (garlic mustard) increases soil nutrient availability in northern hardwood-conifer forests. Oecologia 157:459–471PubMedCrossRefGoogle Scholar
  59. Rodriguez LF (2006) Can invasive species facilitate native species? Evidence of how, when, and why these impacts occur. Biol Invasions 8:927–939CrossRefGoogle Scholar
  60. Sanon A, Beguiristain T, Cebron A, Berthelin J, Ndoye I, Leyval C, Sylla S, Duponnois R (2009) Changes in soil diversity and global activities following invasions of the exotic invasive plant, Amaranthus viridis L., decrease the growth of native sahelian Acacia species. FEMS Microbiol Ecol 70(1):118–131PubMedCrossRefGoogle Scholar
  61. Schnee BK, Waller DM (1986) Reproductive behavior of Amphicarpaea bracteata (Leguminosae), an amphicarpic annual. Am J Bot 73(3):376–386CrossRefGoogle Scholar
  62. Simms EL, Taylor DL (2002) Partner choice in nitrogen-fixation mutualisms of legumes and rhizobia. Integr Comp Biol 42:369–380PubMedCrossRefGoogle Scholar
  63. Somasegaran P, Hoben HJ (1994) Handbook for rhizobia: methods in legume-rhizobium technology. Springer, New York, p 450CrossRefGoogle Scholar
  64. Stinson KA, Campbell SA, Powell JR, Wolfe BE, Callaway RM, Thelen GC, Hallett SG, Prati D, Klironomos J (2006) Invasive plant suppresses the growth of native tree seedlings by disrupting belowground mutualisms. PLoS Biol 4:727–731CrossRefGoogle Scholar
  65. Stinson KA, Kaufman S, Durbin L, Lowenstein F (2007) Impacts of garlic mustard invasion on a forest understory community. Northeast Nat 14:73–88CrossRefGoogle Scholar
  66. Swiader JM, Moore A (2002) Spad-chlorophyll response to nitrogen fertilization and evaluation of nitrogen status in dryland and irrigated pumpkins. J Plant Nutr 25(5):1089–1100CrossRefGoogle Scholar
  67. van Berkum P (1990) Evidence for a third uptake hydrogenase phenotype among the soybean bradyrhizobia. Appl Environ Microbiol 5:3835–3841Google Scholar
  68. van der Heijden MGA, Bardgett RD, van Straalen NM (2008) The unseen majority: soil microbes as drivers of plant diversity and productivity in terrestrial ecosystems. Ecol Lett 11(3):296–310PubMedCrossRefGoogle Scholar
  69. Vaughn SF, Berhow MA (1999) Allelochemicals isolated from tissues of the invasive weed garlic mustard (Alliaria petiolata). J Chem Ecol 25:2495–2504CrossRefGoogle Scholar
  70. Verhoeven KJF, Biere A, Harvey JA, van der Putten WH (2009) Plant invaders and their novel natural enemies: who is naive? Ecol Lett 12(2):107–117PubMedCrossRefGoogle Scholar
  71. Weidenhamer JD, Callaway RM (2010) Direct and indirect effects of invasive plants on soil chemistry and ecosystem function. J Chem Ecol 36(1):59–69PubMedCrossRefGoogle Scholar
  72. West SA, Kiers ET, Simms EL, Denison RF (2002) Sanctions and mutualism stability: why do rhizobia fix nitrogen? Proc R Soc B 269(1492):685–694PubMedCentralPubMedCrossRefGoogle Scholar
  73. Wolfe BE, Rodgers VL, Stinson KA, Pringle A (2008) The invasive plant Alliaria petiolata (garlic mustard) inhibits ectomycorrhizal fungi in its introduced range. J Ecol 96:777–778CrossRefGoogle Scholar
  74. Yoder BJ, Pettigrew-Crosby RE (1995) Predicting nitrogen and chlorophyll content and concentrations from reflectance spectra (400–2500 nm) at leaf and canopy scales. Remote Sens Environ 53:199–211. doi: 10.1016/0034-4257(95)00135-N CrossRefGoogle Scholar
  75. Yuan Y, Wang B, Zhang S, Tang J, Tu C, Hu S, Yong JWH, Chen X (2012) Enhanced allelopathy and competitive ability of invasive plant Solidago canadensis in its introduced range. J Plant Ecol 6(3):253–263CrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Cristina Portales-Reyes
    • 1
  • Tina Van Doornik
    • 2
  • Elizabeth H. Schultheis
    • 3
  • Tomomi Suwa
    • 3
  1. 1.Department of Ecology, Evolution & BehaviorUniversity of Minnesota Twin CitiesSaint PaulUSA
  2. 2.Rosenstiel School of Marine and Atmospheric Science, Division of Marine Biology and FisheriesUniversity of MiamiCoral GablesUSA
  3. 3.W.K. Kellogg Biological Station, Department of Plant BiologyMichigan State UniversityEast LansingUSA

Personalised recommendations