Seasonal Changes in Undifilum Colonization and Swainsonine Content of Locoweeds

Abstract

Locoweeds (Astragalus and Oxytropis) are leguminous plants that are toxic due to a symbiotic association with the endophytic fungus Undifilum oxytropis. The fungus produces the alkaloid swainsonine, an α-mannosidase-inhibitor that causes serious damage to mammals when consumed. A real-time PCR technique was developed to quantify the colonization extent of Undifilum in locoweeds and to compare it to the swainsonine concentration in the plants. Amplification of the endophyte nuclear ITS region allowed reliable quantification of Undifilum DNA from field plants and in vitro cultures. Swainsonine concentration was highly correlated (ρ = 0.972, P < 0.001) with the proportion of Undifilum DNA during the first 4 weeks of in vitro culture growth. Species of Astragalus and Oxytropis were sampled seasonally in New Mexico and Colorado for two years. High swainsonine concentration in plant samples was associated with high levels of endophyte DNA, except in plant reproductive tissues.

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References

  1. Andersen, B., Smedsgaard, J., Jørring, I., Skouboe, P., and Hagsholm Pederson, L. 2006. Real-time PCR quantification of the AM-toxin gene and HPLC qualification of toxigenic metabolites from Alternaria species from apples. International J of Food Microbiology 111:105–111.

  2. Braun, K., Romero, J., Liddell, C., and Creamer, R. 2003. Production of swainsonine by fungal endophytes of locoweed. Mycol. Res. 107:980–988.

    PubMed  Article  CAS  Google Scholar 

  3. Cook, D., Gardner, D. R., Welch, K. D., Roper, J. M., Ralphs, M. H., and GREEN, B. T. 2009. Quantitative PCR method to measure the fungal endophyte in locoweeds. J. Agric. Food Chem. 57:6050–6054.

    PubMed  Article  CAS  Google Scholar 

  4. Davis, D., Schwarz, P., Hernandez, T., Mitchell, M., Warnock, B., and ELBEIN, A. D. 1984. Isolation and characterization of swainsonine from Texas locoweed (Astragalus emoryanus). Plant Physiol. 76:972–975.

    PubMed  Article  CAS  Google Scholar 

  5. FOX, W.E., ALLRED, K.W., and ROALSON, E.H. 1998. A guide to the common locoweeds and milkvetches of New Mexico. NM Agric. Exp. Sta. Circ. 557.

  6. Gachon, C. and Saindrenan, P. 2004. Real-time PCR monitoring of fungal development in Arabidopsis thaliana infected by Alternaria brassicola and Botrytis cinerea. Plant Physiol. Biochem. 42:367–371.

    PubMed  Article  CAS  Google Scholar 

  7. Gardner, D. R., Molyneux, R. J., and Ralphs, M. H. 2001. Analysis of swainsonine: Extraction methods, detection, and measurement in populations of locoweeds (Oxytropis spp.). J. Agric Food Chem 49:4573–4580.

    PubMed  Article  CAS  Google Scholar 

  8. Glynn, N. C., Ray, R., Edwards, S. G., Hare, M. C., Parry, D. W., Barnett, C. J., and Beck, J. J. 2007. Quantitative Fusarium spp. and Microdochium spp. PCR assays to evaluate seed treatment for the control of Fusarium seedling blight of wheat. J. Appl. Microbiol. 102:1645–1653.

    PubMed  Article  CAS  Google Scholar 

  9. James, L. F. and Panter, K. E. 1989. Locoweed poisoning in livestock. pp 23-28 in: Swainsonine and related glycosidase inhibitors. Edited by L. F. James, A. D. Elbein, R. J. Molyneux and C. D. Warren, Iowa State University Press, Ames.

  10. James, L. F., Gardner, D. R., Lee, S. T., Panter, K. E., Pfister, J. A., Ralphs, M. H., and Stegelmeier, B. L. 2005. Important poisonous plants on rangelands. Management strategies based on toxin level in the plant, animal susceptibility, and grazing behavior can reduce the risk of poisoning. Rangelands 27:3–9.

    Article  Google Scholar 

  11. Kobayashi, T. 2011. Regulation of ribosomal RNA gene copy number and its role in modulating genome integrity and evolutionary adaptability. Cell. Mol. Life Sci. 68:1395–1403.

    PubMed  Article  CAS  Google Scholar 

  12. Li, Y. Z. and Nan, Z. B. 2007a. Symptomology and etiology of a new disease, yellow stunt, and root rot of standing milkvetch caused by Embellisia sp. in Northern China. Mycopathologia 163:327–334.

    PubMed  Article  Google Scholar 

  13. Li, Y. Z. and Nan, Z. B. 2007b. A new species, Embellisia astragali sp. nov., causing standing milk-vetch disease in China. Mycologia 99:406–411.

    PubMed  Article  Google Scholar 

  14. Marsh, C.D. 1909. The locoweed disease of the plains. USDA Bureau of Animal Industries Bull. 112.

  15. McLain-Romero, J., Creamer, R., Zepeda, H., Strickland, J., and Bell, G. 2004. The toxicosis of Embellisia fungi from locoweed (Oxytropis lambertii) is similar to locoweed toxicosis in rats. J. Anim. Sci. 82:2169–2174.

    PubMed  CAS  Google Scholar 

  16. Molyneux, R. J. and James, L. F. 1982. Loco intoxication: Indolizidine alkaloids of spotted locoweed (Astragalus lentiginosus). Science 216:190–191.

    PubMed  Article  CAS  Google Scholar 

  17. NOAA 2006a. Climatological Data. Annual Summary. Colorado 2006. 111(13)

  18. NOAA 2006b. Climatological Data. Annual Summary. New Mexico 2006. 110(13)

  19. NOAA 2007a. Climatological Data. Annual Summary. Colorado 2006. 112(13)

  20. NOAA 2007b. Climatological Data. Annual Summary. New Mexico 2007. 111(13)

  21. NOAA 2008a. Climatological Data. Colorado. February 2008. 113(02)

  22. NOAA 2008b. Climatological Data. Colorado. May 2008. 113(05)

  23. NOAA 2008c. Climatological Data. New Mexico. February 2008. 112(02)

  24. NOAA 2008d. Climatological Data. New Mexico. May 2008. 112(05)

  25. Oldrup, E., McLain-Romero, J., Padilla, A., Moya, A., Gardner, D., and Creamer, R. 2010. Localization of endophytic fungi in locoweed seed and influence of environmental parameters on a locoweed in vitro culture system. Botany 88:512–521.

    Article  CAS  Google Scholar 

  26. PARKER, J. 2008. Effects of insect herbivory by the four-lined locoweed weevil Cleonidius trivitattatus (Say) (Coleoptera: Curculionidae), on the alkaloid swainsonine in locoweeds Astagalus mollissimus and Oxytropis sericea. MS thesis. New Mexico State University, Las Cruces.

  27. Patterson, P. E. 1982. Loco, la yerba mala. Rangelands 4:147–148.

    Google Scholar 

  28. Pfister, J. A., Stegelmeier, B. L., Gardner, D. R., and James, L. F. 2003. Grazing of spotted locoweed (Astragalus lentiginosus) by cattle and horses in Arizona. J. Anim. Sci. 2003:2285–2293.

    Google Scholar 

  29. Pryor, B. M., Creamer, R., Shoemaker, R. A., McLain-Romero, J., and Hambleton, S. 2009. Undifilum, a new genus for endophytic Embellisia oxytropis and parasitic Helminthosporium bornmuelleri on legumes. Botany 87:178–194.

    Article  CAS  Google Scholar 

  30. Ralphs, M. H., James, L. F., Nielsen, D. B., and Panter, K. E. 1984. Management practices reduce cattle loss due to locoweed on high mountain range. Rangelands 6:175–177.

    Google Scholar 

  31. Ralphs, M. H., Graham, D., Molyneux, R. J., and James, L. F. 1993. Seasonal grazing of locoweeds by cattle in northeastern New Mexico. J. Rangeland Manage. 46:416–420.

    Article  Google Scholar 

  32. Ralphs, M. H., Pfister, J. A., Welsh, S. L., Graham, J. D., Purvines, J., Jensen, D. T., and James, L. F. 2003. Locoweed population cycles. Rangelands 25:14–18.

    Google Scholar 

  33. Ralphs, M. H., Creamer, R., Baucom, D., Gardner, D. R., Welsh, S. L., Graham, J. D., Hart, C., Cook, D., and Stegelmeier, B. L. 2008. Relationship between the endophyte Embellisia spp. and the toxic alkaloid swainsonine in major locoweed species (Astragalus and Oxytropis). J. Chem. Ecol. 34:32–38.

    PubMed  Article  CAS  Google Scholar 

  34. Rasmussen, S., Parsons, A. J., Bassett, S., Christensen, M. J., Hume, D. E., Johnson, L. J., Johnson, R. D., Simpson, W. R., Stacke, C., Voisey, C. R., Xue, H., and Newman, J. A. 2007. High nitrogen supply and carbohydrate content reduce fungal endophyte and alkaloid concentration in Lolium perenne. New Phytol. 173:787–797.

    PubMed  Article  CAS  Google Scholar 

  35. Stegelmeier, B. L., James, L. F., Gardner, D. R., Panter, K. E., Lee, S. T., Ralphs, M. H., Pfister, J. A., and Spraker, T. R. 2005. Locoweed (Oxytropis sericea)-induced lesions in mule deer (Odocoileius hemionus). Vet. Pathol. 42:566–578.

    PubMed  Article  CAS  Google Scholar 

  36. Vallotton, A. D. and Sterling, T. M. Sterling. 2007. Physiology, growth and swainsonine content response to water deficit stress in Astragalus mollissimus and Oxytropis sericea. Paper presented at the 47th Annual Meeting of the Weed Science Society of America in Houston, TX.

  37. White, T.J., Bruns, T., Lee, S., and Taylor, J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. pp. 315–352 in: PCR protocols: a guide to methods and applications. Edited by M.A. Innis, D.H. Gelfand, J. Sninsky and T.J. White, Academic Press, San Diego, CA.

  38. Williams, M. C., James, L. F., and Bond, B. O. 1979. Emory milkvetch (Astragalus emoryanus var emoryanus) poisoning in chicks, sheep, and cattle. Amer. J. Vet. Res. 40:403–406.

    PubMed  CAS  Google Scholar 

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Acknowledgements

We thank Dr Jose Valdez-Barillas and David Graham for their guidance during field seasons, and Dr Steve Hanson, Dr Champa Sengupta-Gopalan, and Dr Tracy Sterling for allowing the use of their lab facilities and equipment. We acknowledge USDA Special Grant # 59-5428-1-327 and the New Mexico State University Agricultural Experiment Station for supporting this work.

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Correspondence to Rebecca Creamer.

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Achata Böttger, J., Creamer, R. & Gardner, D. Seasonal Changes in Undifilum Colonization and Swainsonine Content of Locoweeds. J Chem Ecol 38, 486–495 (2012). https://doi.org/10.1007/s10886-012-0120-z

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Keywords

  • Undifilum
  • Quantitative PCR
  • Swainsonine