, Volume 226, Issue 2, pp 395–404 | Cite as

Investigating the mechanisms of glyphosate resistance in Lolium multiflorum

  • Alejandro Perez-JonesEmail author
  • Kee-Woong Park
  • Nick Polge
  • Jed Colquhoun
  • Carol A. Mallory-Smith
Original Article


Evolved resistance to the herbicide glyphosate has been reported in eleven weed species, including Lolium multiflorum. Two glyphosate-resistant L. multiflorum populations were collected, one from Chile (SF) and one from Oregon, USA (OR), and the mechanisms conferring glyphosate resistance were studied. Based on a Petri dish dose–response bioassay, the OR and the SF populations were two and fivefold more resistant to glyphosate when compared to the susceptible (S) population, respectively; however, based on a whole-plant dose–response bioassay, both OR and SF populations were fivefold more resistant to glyphosate than the S population, implying that different resistance mechanisms might be involved. The S population accumulated two and three times more shikimic acid in leaf tissue 96 h after glyphosate application than the resistant OR and SF populations, respectively. There were no differences between the S and the glyphosate-resistant OR and SF populations in 14C-glyphosate leaf uptake; however, the patterns of 14C-glyphosate translocation were significantly different. In the OR population, a greater percentage of 14C-glyphosate absorbed by the plant moved distal to the treated section and accumulated in the tip of the treated leaf. In contrast, in the S and in the SF populations, a greater percentage of 14C-glyphosate moved to non-treated leaves and the stem. cDNA sequence analysis of the EPSP synthase gene indicated that the glyphosate-resistant SF population has a proline 106 to serine amino acid substitution. Here, we report that glyphosate resistance in L. multiflorum is conferred by two different mechanisms, limited translocation (nontarget site-based) and mutation of the EPSP synthase gene (target site-based).


Glyphosate resistance Lolium multiflorum Translocation EPSP synthase Shikimic acid 






The 14C-glyphosate leaf uptake and translocation experiments were carried out at Syngenta Vero Beach Research Center. We thank Syngenta Crop Protection for their economic and technical support.


  1. Arnaud L, Nurit F, Ravanel P, Tissut M (1994) Distribution of glyphosate and of its target enzyme inside wheat plants. Pestic Sci 40:217–223CrossRefGoogle Scholar
  2. Baerson SR, Rodriguez D, Tran M, Feng Y, Biest NA, Dill GM (2002a) Glyphosate-resistant goosegrass. Identification of a mutation in the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase. Plant Physiol 129:1265–1275PubMedCrossRefGoogle Scholar
  3. Baerson SR, Rodriguez DJ, Biest NA, Tran M, You J, Kreuger RW, Dill GM, Pratley JE, Gruys KJ (2002b) Investigating the mechanism of glyphosate resistance in rigid ryegrass (Lolium rigidum). Weed Sci 50:721–730CrossRefGoogle Scholar
  4. Baylis A (2000) Why glyphosate is a global herbicide: strengths, weaknesses and prospects. Pest Manag Sci 56:299–308CrossRefGoogle Scholar
  5. Bradshaw LD, Padgette SR, Kimball SL, Wells BH (1997) Perspectives on glyphosate resistance. Weed Technol 11:189–198Google Scholar
  6. Caseley JC, Coupland D (1985) Environmental and plant factors affecting glyphosate uptake, movement and activity. In: Grossbard E, Atkinson D (eds) The herbicide glyphosate. Butterworths, London, pp 92–123Google Scholar
  7. Cromartie TH, Polge ND (2000) An improved assay for shikimic acid and its use as a monitor for the activity of sulfosate. Proc Weed Sci Soc Am 40:291Google Scholar
  8. Culpepper AS, Grey TL, Vencill WK, Kichler JM, Webster TM, Brown SM, York AC, Davis JW, Hanna WW (2006) Glyphosate-resistant palmer amaranth (Amaranthus palmeri) confirmed in Georgia. Weed Sci 54:620–626CrossRefGoogle Scholar
  9. Dinelli G, Marotti I, Bonetti A, Minelli M, Catizone P, Barnes J (2006) Physiological and molecular insight on the mechanisms of resistance to glyphosate in Conyza canadensis (L.) Cronq. biotypes. Pest Biochem Physiol 86:30–41CrossRefGoogle Scholar
  10. Duke SO, Baerson SR, Rimando AM (2003) Glyphosate. Encyclopedia of agrochemicals. Wiley, New York. Article on line.
  11. Feng PCC, Pratley JE, Bohn JA (1999) Resistance to glyphosate in Lolium rigidum. II. Uptake, translocation, and metabolism. Weed Sci 47:412–415Google Scholar
  12. Feng PCC, Tran M, Chiu T, Sammons RD, Heck GR, Cajacob CA (2004) Investigations into glyphosate-resistant horseweed (Conyza canadensis): retention, uptake, translocation, and metabolism. Weed Sci 52:498–505CrossRefGoogle Scholar
  13. Franz JE, Mao MK, Sikorski JA (1997) Glyphosate: a unique global herbicide. ACS Monograph 189. American Chemical Society, Washington USAGoogle Scholar
  14. Geiger DR, Fuchs MA (2002) Inhibitors of aromatic amino acid biosynthesis (glyphosate). In: Böger P, Wakabayashi K, Hirai K (eds) Herbicide classes in development. Springer, Heidelberg, pp 59–85Google Scholar
  15. Heap IM (2006) International survey of herbicide-resistant weeds. Available at (accessed October, 2006)
  16. Hall GJ, Hart CA, Jones CA (2000) Plants as sources of cations antagonistic to glyphosate. Pest Manag Sci 56:351–358CrossRefGoogle Scholar
  17. Hess FD (1985) Herbicide absorption and translocation and their relationship to plant tolerances and susceptibility. In: Duke SO (ed) Weed physiology, vol II. Herbicide physiology. CRC Press, Boca Raton, pp 191–214Google Scholar
  18. Kishore GM, Shah DM (1988) Amino acid biosynthesis inhibitors as herbicides. Annu Rev Biochem 57:627–663PubMedCrossRefGoogle Scholar
  19. Koger CH, Poston DH, Hayes RM, Montgomery RF (2004) Glyphosate-resistant horseweed in Mississippi. Weed Technol 18:820–825CrossRefGoogle Scholar
  20. Koger CH, Reddy KN (2005) Role of absorption and translocation in the mechanism of glyphosate resistance in horseweed (Conyza canadensis). Weed Sci 53:84–89CrossRefGoogle Scholar
  21. Lee LJ, Ngim J (2000) A first report of glyphosate-resistant goosegrass (Eleusine indica (L) Gaertn) in Malaysia. Pest Manag Sci 56:336–339CrossRefGoogle Scholar
  22. Lorraine-Colwill DF, Powles SB, Hawkes TR, Hollinshead PH, Warner SAJ, Preston C (2003) Investigations into the mechanism of glyphosate resistance in Lolium rigidum. Pestic Biochem Physiol 74:62–72CrossRefGoogle Scholar
  23. Main CL, Mueller TC, Hayes RM, Wilkerson JB (2004) Response of selected horseweed (Conyza canadensis (L.) Cronq.) populations to glyphosate. J Agric Food Chem 52:879–883PubMedCrossRefGoogle Scholar
  24. McWhorter CG, Jordan TN, Will GD (1980) Translocation of 14C-glyphosate in soybean (Glycine max) and johnsongrass (Sorghum halepense). Weed Sci 28:113–118Google Scholar
  25. Mueller TC, Massey JH, Hayes RM, Main CL, Stewart N Jr (2003) Shikimate accumulates in both glyphosate-sensitive and glyphosate-resistant horseweed (Conyza canadensis L. Cronq.) J Agric Food Chem 51:680–684PubMedCrossRefGoogle Scholar
  26. Neve P, Sadler J, Powles SB (2004) Multiple herbicide resistance in a glyphosate-resistant rigid ryegrass (Lolium rigidum) population. Weed Sci 52:920–928CrossRefGoogle Scholar
  27. Ng CH, Wickneswari R, Salmijah S, Teng YT, Ismail BS (2003) Gene polymorphisms in glyphosate-resistant and -susceptible biotypes of Eleusine indica from Malaysia. Weed Res 43:108–115CrossRefGoogle Scholar
  28. Padgette SR, Re DB, Gasser CS, Eichholtz DA, Frazier RB, Hironaka CM, Levine EB, Shah DM, Fraley RT, Kishore GM (1991) Site-directed mutagenesis of a conserved region of the 5-enolpyruvylshikimate-3-phosphate synthase active site. J Biol Chem 266:22364–22369PubMedGoogle Scholar
  29. Padgette SR, Re DB, Barry GF, Eichholtz DE, Delannay X, Fuchs RL, Kishore GM, Fraley RT (1996) New weed control opportunities: development of soybeans with a Roundup ReadyTM gene. In: Duke SO (ed) Herbicide-resistant crops: agricultural, environmental, economic, regulatory, and technical aspects. CRC Press, Inc. Boca Raton, pp 53–84Google Scholar
  30. Perez A, Kogan M (2003) Glyphosate-resistant Lolium multiflorum in Chilean orchards. Weed Res 43:12–19CrossRefGoogle Scholar
  31. Perez-Jones A, Park KW, Colquhoun J, Mallory-Smith C, Shaner D (2005) Identification of glyphosate resistant Italian ryegrass (Lolium multiflorum) in Oregon. Weed Sci 53:775–779CrossRefGoogle Scholar
  32. Pline WA, Wilcut JW, Duke SO, Edmisten KL, Wells R (2002) Tolerance and accumulation of shikimic acid in response to glyphosate applications in glyphosate-resistant and nonglyphosate-resistant cotton (Gossypium hirsutum L.) J Agric Food Chem 50:506–512PubMedCrossRefGoogle Scholar
  33. Powles SB, Lorraine-Colwill DF, Dellow JJ, Preston C (1998) Evolved resistance to glyphosate in rigid ryegrass (Lolium rigidum) in Australia. Weed Sci 46:604–607Google Scholar
  34. Pratley J, Baines P, Eberbach P, Incerti M, Broster J (1996) Glyphosate resistance in annual ryegrass. In: Virgona J, Michalk D (eds) Proceedings of the 11th annual conference of the Grassland Society of New South Wales. Wagga Wagga, Australia, p 126Google Scholar
  35. Pratley J, Urwin N, Stanton R, Baines P, Broster J, Cullis K, Schafer D, Bohn J, Krueger R (1999) Resistance to glyphosate in Lolium rigidum. I. Bioevaluation. Weed Sci 47:405–411Google Scholar
  36. Seefeldt S, Jensen JE, Fuerst EP (1995) Log–logistic analysis of herbicide dose–response relationships. Weed Technol 9:218–225Google Scholar
  37. Sellers BA, Pollard JM, Smeda RJ (2005) Two common ragweed (Ambrosia artemisiifolia) biotypes differ in biology and response to glyphosate. Proc Weed Sci Soc 45:156Google Scholar
  38. Shaner DL (2000) The impact of glyphosate-tolerant crops on the use of other herbicides and on resistance management. Pest Manag Sci 56:320–326CrossRefGoogle Scholar
  39. Siehl DL (1997) Inhibitors of EPSPS synthase, glutamine synthetase and histidine synthesis. In: Roe RM, Burton JD, Kuhr RJ (eds) Herbicide activity: toxicology, biochemistry and molecular biology. IOS Press, Amsterdam, pp 37–67Google Scholar
  40. Simarmata M, Kaufmann JE, Penner D (2003) Potential basis of glyphosate resistance in California rigid ryegrass (Lolium rigidum). Weed Sci 51:678–682CrossRefGoogle Scholar
  41. Singh BK, Shaner DL (1998) Rapid determination of glyphosate injury to plants and identification of glyphosate-resistant plants. Weed Tech 12:527–530Google Scholar
  42. Schönbrunn E, Eschenburg S, Shuttleworth WA, Schloss JV, Amrhein N, Evans JNS, Kabsch W (2001) Interaction of the herbicide glyphosate with its target enzyme 5-enolpyruvylshikimate 3-phosphate synthase in atomic detail. PNAS 98:1376–1380PubMedCrossRefGoogle Scholar
  43. Sprankle P, Meggitt WF, Penner D (1975) Absorption, action, and translocation of glyphosate. Weed Sci 23:235–240Google Scholar
  44. Stalker DM, Hiatt WR, Comai L (1985) A single amino acid substitution in the enzyme 5-enolpyruvylshikimate-3-phosphate synthase confers resistance to the herbicide glyphosate. J Biol Chem 260:4724–4728PubMedGoogle Scholar
  45. Steinrücken H, Amrhein N (1980) The herbicide glyphosate is a potent inhibitor of 5-enolpyruvylshikimic acid-3-phosphate synthase. Biochem Biophys Res Commun 94:1207–1212PubMedCrossRefGoogle Scholar
  46. Sterling TM (1994) Mechanisms of herbicide absorption across plant membranes and accumulation in plant cells. Weed Sci 42:263–276Google Scholar
  47. Streibig JC (1988) Herbicide bioassay. Weed Res 28:479–484CrossRefGoogle Scholar
  48. Streibig JC, Rudemo M, Jensen JE (1993) Dose–response curves and statistical models. In: Streibig JC, Kudsk P (eds) Herbicide bioassays. CRC Press, Boca Raton, pp 29–56Google Scholar
  49. Tran M, Baerson S, Brinker R, Casagrande L, Faletti M, Feng Y, Nemeth M, Reynolds T, Rodriguez D, Shaffer D, Stalker D, Taylor N, Teng Y, Dill G (1999) Characterization of glyphosate resistant Eleusine indica biotypes from Malaysia. In: Proceedings 1 (B) of the 17th Asian-Pacific weed science society conference. The Asian-Pacific Weed Science Society, Bangkok, pp 527–536Google Scholar
  50. Urbano JM, Borrego A, Torres V, Jimenez C, Leon JM, Barnes J (2005) Glyphosate-resistant hairy fleabane (Conyza bonariensis) in Spain. Proc Weed Sci Soc 45:394Google Scholar
  51. VanGessel MJ (2001) Glyphosate-resistant horseweed from Delaware. Weed Sci 49:703–705CrossRefGoogle Scholar
  52. Wakelin AM, Preston C (2006) A target-site mutation is present in a glyphosate-resistant Lolium rigidum population. Weed Res 46:432–440CrossRefGoogle Scholar
  53. Wakelin AM, Lorraine-Colwill DF, Preston C (2004) Glyphosate resistance in four different population of Lolium rigidum is associated with reduced translocation of glyphosate to meristematic zones. Weed Res 44:453–459CrossRefGoogle Scholar
  54. Yu Q, Cairns A, Powles S (2007) Glyphosate, paraquat and ACCase multiple herbicide resistance evolved in a Lolium rigidum biotype. Planta 225:499–513PubMedCrossRefGoogle Scholar
  55. Zelaya IA, Owen MDK (2005) Differential response of Amaranthus tuberculatus (Moq ex DC) JD Sauer to glyphosate. Pest Manag Sci 61:936–950PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Alejandro Perez-Jones
    • 1
    Email author
  • Kee-Woong Park
    • 1
  • Nick Polge
    • 2
  • Jed Colquhoun
    • 3
  • Carol A. Mallory-Smith
    • 1
  1. 1.Department of Crop and Soil ScienceOregon State UniversityCorvallisUSA
  2. 2.Syngenta Crop Protection, Inc.Vero Beach Research CenterVero BeachUSA
  3. 3.Department of HorticultureUniversity of WisconsinMadisonUSA

Personalised recommendations