Application of Toxigenic Alternaria oxytropis to Soybeans and its Effect on Swainsonine Detection in Different Environments

  • Odeshnee MoodleyEmail author
  • Yue Sun
  • Frederick Leo Sossah
  • Makoto Kakishima
  • Igor N. Pavlov
  • Yu Li
  • Qi WangEmail author


Alternaria oxytropis is an endophytic fungus of locoweeds that synthesizes swainsonine toxin. In this work, we evaluated the effect of A. oxytropis on soybean seedlings and quantified swainsonine in different culture conditions. Soybean (Glycine max) seeds were co-cultured with A. oxytropis (at different concentrations of mycelial suspensions) in agar media and soil culture, and swainsonine was assayed using LC–MS/MS. The results showed evidence that A. oxytropis infected soybean seedlings produced detectable swainsonine in agar culture while the toxin was undetectable or below the detection limit (0.006% of swainsonine dry weight) in soil media even at higher concentrations of the fungus. These results suggest that swainsonine detection is highly dependent on culture conditions and that soybeans co-cultured with A. oxytropis in soil could potentially be used to limit toxin production.


Alkaloid toxin Alternaria oxytropis Soybean Swainsonine Locoweeds Toxin detection 



This work was funded by the Taishan Industry Leader Talent Project Fund (LJNY 201611) and the Program of Creation and Utilization of Germplasm of Mushroom Crop of “111 Plan” project (Grant No. D17014). We thank the following people for their contributions in this research study: Chunlan Zhang and Xiangyang Zhu.


  1. Ayolagha GA, Peter KD (2013) Effect of remediation on growth parameters, grain and dry matter yield of soybean (Glycine max) in crude oil polluted ultisols in Ogoni land, South Eastern Nigeria. Afr J Environ Sci Technol 7:61–67. Google Scholar
  2. Brundrett M, Bougher N, Dell B, Grove T, Malajczu N (1996) Isolation of fungi and production of inoculum. In: Lynch P (ed) Working with mycorrhizas in forestry and agriculture. Pirie Printers, Canberra, pp 249–253Google Scholar
  3. Bultman TL, Aguilera A, Sullivan TJ (2012) Influence of fungal isolates infecting tall fescue on multitrophic interactions. Fungal Ecol 5:372–378. CrossRefGoogle Scholar
  4. Cook D, Shi L, Gardner DR, Pfister JA, Grum D, Welch KD, Ralphs MH (2012) Influence of phenological stage on swainsonine and endophyte concentrations in Oxytropis sericea. J Chem Ecol 38:195–203. CrossRefGoogle Scholar
  5. Cook D, Grum DS, Gardner DR, Welch KD, Pfister JA (2013) Influence of endophyte genotype on swainsonine concentrations in Oxytropis sericea. Toxicon 61:105–111. CrossRefGoogle Scholar
  6. Cook D, Gardner DR, Pfister JA (2014) Swainsonine-containing plants and their relationship to endophytic fungi. J Agric Food Chem 62:7326–7334. CrossRefGoogle Scholar
  7. Cook D, Gardner DR, Lee ST, Pfister JA, Stonecipher CA, Welsh SL (2016) A swainsonine survey of North American and taxa implicated as locoweeds. Toxicon Google Scholar
  8. Cook D, Donzelli BGG, Creamer R, Baucom DL, Gardner DR, Pan J, Moore N, Krasnoff SB, Jaromczyk JW, Schardl CL (2017) Swainsonine biosynthesis genes in diverse symbiotic and pathogenic fungi. G3 Genes Genomes Genet 7:1791–1797. Google Scholar
  9. Creamer R, Baucom D (2013) Fungal endophytes of locoweeds: a commensal relationship? J Plant Physiol Pathol 1:1–4. Google Scholar
  10. Dorling PR, Huxtable CR, Colegate SM (1980) Inhibition of lysosomal α-mannosidase by swainsonine, an indolizidine alkaloid isolated from Swainsona canescens. Biochem J 191:649–651CrossRefGoogle Scholar
  11. El-Azouni IM (2008) Effect of phosphate solubilizing fungi on growth and nutrient uptake of soybean (Glycine max L.) plants. J Appl Sci Res 4:592–598Google Scholar
  12. European Commission (2002) Decision (2002/657/EC) of 12 August 2002 implementing council directive 96/23/EC concerning the performance of analytical methods and interpretation of results. Off J Eur Commun 8–36Google Scholar
  13. Gardner DR, Cook D (2011) A Comparison of alternative sample preparation procedures for the analysis of swainsonine using LC–MS/MS. Phytochem Anal 22:124–127. CrossRefGoogle Scholar
  14. Gardner DR, Molyneux RJ, Ralphs MH (2001) Analysis of swainsonine: extraction methods, detection, and measurement in populations of locoweeds (Oxytropis spp.). J Agric Food Chem 49:4573–4580. CrossRefGoogle Scholar
  15. Gelfand I, Robertson GP (2015) A reassessment of the contribution of soybean biological nitrogen fixation to reactive N in the environment. Biogeochemistry 123:175–184. CrossRefGoogle Scholar
  16. Hamayun M, Khan SA, Khan AL, Afzal M, Lee IJ (2012) Endophytic Cephalotheca sulfurea AGH07 reprograms soybean to higher growth. J Plant Interact 7:301–306. CrossRefGoogle Scholar
  17. Hamayun M, Hussain A, Khan SA, Kim HY, Khan AL, Waqas M, Irshad M, Iqbal A, Rehman G, Jan S, Lee IJ (2017) Gibberellins producing endophytic fungus Porostereum spadiceum AGH786 rescues growth of salt affected soybean. Front Microbiol 8:1–13. CrossRefGoogle Scholar
  18. Klypina N, Pinch M, Schutte BJ, Maruthavanan J, Sterling TM (2017) Water-deficit stress tolerance differs between two locoweed genera (Astragalus and Oxytropis) with fungal endophytes. Weed Sci 65:626–638CrossRefGoogle Scholar
  19. Latch GCM (1994) Influence of Acremonium endophytes on perennial grass improvement. N Z J Agric Res 37:311–318. CrossRefGoogle Scholar
  20. Latch GCM (1998) Grass endophytes as a model. Sydowia 50:213–228Google Scholar
  21. Liu B, Yan H, Wang B (2011) Breeding report of new soybean cultivar ‘Jiyu 86’. J Jilin Agric Sci 36:11–12Google Scholar
  22. Lu H, Quan H, Ren Z, Wang S, Xue R, Zhao B (2016) The genome of Undifilum oxytropis provides insights into swainsonine biosynthesis and locoism. Sci Rep 6:30760. CrossRefGoogle Scholar
  23. Lu H, Quan H, Zhou Q, Ren Z, Xue R, Zhao B, Creamer R (2017) Endogenous fungi isolated from three locoweed species from rangeland in western China. Afr J Microbiol Res 11:155–170. CrossRefGoogle Scholar
  24. Molyneux RJ, James LF (1982) Loco intoxication: indolizidine alkaloids of spotted locoweed (Astragalus lentiginosus). Science 216:190–191CrossRefGoogle Scholar
  25. Molyneux RJ, James LF, Ralphs MH, Pfister JA, Panter KE, Nash RJ (2004) Polyhydroxylated glycosidase inhibitors from poisonous plants of global distribution: analysis and identification. In: Colegate SM, Dorling PR (eds) Plant-associated toxins, agricultural and phytochemical aspects. CABI, Wallingford, pp 107–112Google Scholar
  26. Molyneux RJ, Lee ST, Gardner DR, Panter KE, James LF (2007) Phytochemicals: the good, the bad and the ugly? Phytochemistry 68:2973–2985. CrossRefGoogle Scholar
  27. Noor AI, Nava A, Cooke P, Cook D, Creamer R (2018) Evidence for nonpathogenic relationships of Alternaria section Undifilum endophytes within three host locoweed plant species. Botany 96:187–200CrossRefGoogle Scholar
  28. Pryor BM, Creamer R, Shoemaker RA, McLain-Romero J, Hambleton S (2009) Undifilum, a new genus for endophytic Embellisia oxytropis and parasitic Helminthosporium bornmuelleri on legumes. Botany 87:178–194. CrossRefGoogle Scholar
  29. Puteh AB, ThuZar M, Mondal MMA, Abdullah NAPB, Halim MRA (2013) Soybean [Glycine max (L.) Merrill] seed yield response to high temperature stress during reproductive growth stages. Aust J Crop Sci 7:1472–1479Google Scholar
  30. Ralphs MH, Creamer R, Baucom D, Gardner DR, Welsh SL, Graham JD, Hart C, Cook D, Stegelmeier BL (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. CrossRefGoogle Scholar
  31. Rodriguez RJ, White Jr JF, Arnold AE, Redman RS (2009) Fungal endophytes: diversity and functional roles. N Phytol Google Scholar
  32. Rufty Jr TW, Raper Jr CD, Jackson WA (1981) Nitrogen assimilation, root growth and whole plant responses of soybean to root temperature, and to carbon dioxide and light in the aerial environment. N Phytol 88:607–619CrossRefGoogle Scholar
  33. Schulz B, Boyle C (2006) What are endophytes? In: Sieber TN (ed) Soil biology: microbial root endophytes. Springer, Berlin, pp 1–13CrossRefGoogle Scholar
  34. Thies JE, Singleton PW, Bohlool BB (1995) Phenology, growth, and yield of field-grown soybean and bush bean as a function of varying modes of N nutrition. Soil Biol Biochem 27:575–583. CrossRefGoogle Scholar
  35. Wang Q, Nagao H, Li YL, Wang HS, Kakishima M (2006) Embellisia oxytropis, a new species isolated from Oxytropis kansuensis in China. Mycotaxon 95:255–260Google Scholar
  36. Waqas M, Khan AL, Hamayun M, Kamran M, Kang SM, Kim YH, Lee IJ (2012) Assessment of endophytic fungi cultural filtrate on soybean seed germination. Afr J Biotechnol 11:15135–15143. CrossRefGoogle Scholar
  37. Waqas M, Khan AL, Hamayun M, Shahzad R, Kang SM, Kim JG, Lee IJ (2015) Endophytic fungi promote plant growth and mitigate the adverse effects of stem rot: an example of Penicillium citrinum and Aspergillus terreus. J Plant Interact 10:280–287. CrossRefGoogle Scholar
  38. Woudenberg JHC, Groenewald JZ, Binder M, Crous PW (2013) Alternaria redefined. Stud Mycol 75:171–212. CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Odeshnee Moodley
    • 1
    • 2
    Email author
  • Yue Sun
    • 1
  • Frederick Leo Sossah
    • 1
  • Makoto Kakishima
    • 1
    • 3
  • Igor N. Pavlov
    • 4
  • Yu Li
    • 1
  • Qi Wang
    • 1
    Email author
  1. 1.Engineering Research Centre of Chinese Ministry of Education for Edible and Medicinal FungiJilin Agricultural UniversityChangchunChina
  2. 2.South African Department of AgricultureForestry and Fisheries (DAFF)PretoriaSouth Africa
  3. 3.Life and Environmental SciencesUniversity of TsukubaTsukubaJapan
  4. 4.V.N. Sukachev Institute of Forest SB RASKrasnoyarskRussia

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