, Volume 79, Issue 1, pp 49–58 | Cite as

Interactive effects of Epichloë fungal and host origins on the seed germination of Achnatherum inebrians

  • Gensheng BaoEmail author
  • Meiling Song
  • Yuqin Wang
  • Kari Saikkonen
  • Hongsheng Wang


Cool-season grasses have developed a symbiotic relationship with Epichloë endophytes. In many environments, Epichloë endophytes have been shown to be mutualistic symbionts of plants by increasing the fitness of their host against abiotic or biotic stresses. The effects of Epichloë endophytes on other fitness-correlated plant characteristics are less intensively studied, and the results are usually variable and contradictory. In this study, we evaluated the effects of endophyte infection on seed germination in Achnatherum inebrians from four origins. Our results indicate that the germination rate of the seeds collected from alpine regions was higher at low temperatures than that of seeds with desert and arid grassland origins. By contrast, a higher germination percentage was detected in seeds with desert and arid grassland origins than in those with alpine origins in higher temperatures. Epichloë endophyte infection affects the cardinal temperatures of seeds from different origins. Endophyte-infected seeds have a lower base temperature and a higher ceiling temperature than their endophyte-free counterparts. The value of the base temperature was higher in seeds with alpine grassland origins than in those with desert and arid grassland origins. However, the ceiling temperature was higher in seeds with desert and arid grassland origins than in those with alpine grassland origins. Consequently, future experiments should consider the effects of endophytes on seed germination and seedling recruitment in suboptimal climatic conditions.


Endophyte Achnatherum inebrians Seed germination Habitat Temperature Thermal time model 



We would like to thank Prof. Xiaowen Hu and Carol C. Baskin for their constructive suggestions in interpreting the results of the study and Ph.D. students Xiuzhang Li and Xuekai Wei for providing technical support. This research was financially supported by the Natural Science Foundation of China (31660690, 31700098), the Program for Qinghai Province Thousand Talent Innovative Plan, Key Laboratory of Superior Forage Germplasm in the Qinghai Tibetan Plateau (2017-ZJ-Y12) and the Academy of Finland (grant nos. 295976 and 326226).

Supplementary material

13199_2019_636_MOESM1_ESM.docx (14 kb)
ESM 1 (DOCX 14 kb)
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13199_2019_636_MOESM3_ESM.jpg (940 kb)
Fig. S1 Molecular phylogeny derived from the maximum likelihood analysis of introns of the tefA gene from related Epichloë species and Achnatherum inebrians seeds originating from Dy, Td, Tl and Al. The tree was rooted with Claviceps purpurea 20.1 as the outgroup. The percentage of trees in which associated taxa are clustered together is shown next to the branches. The three gene copies in the hybrid endophyte, Epichloë coenophiala, are labeled with single-letter abbreviations of the extant Epichloë species related to its three ancestors. (JPG 939 kb)


  1. Alvarado V, Bradford KJ (2002) A hydrothermal time model explains the cardinal temperatures for seed germination. Plant Cell Environ 25:1061–1069Google Scholar
  2. Bacon CW (1993) Abiotic stress tolerances (moisture, nutrients) and photosynthesis in endophyte-infected tall fescue. Agric Ecosyst Environ 44:123–141CrossRefGoogle Scholar
  3. Bacon CW, White JF (1994) Microbial Endophytes. Marcel Dekker, New York, pp 341–388Google Scholar
  4. Bao GS, Saikkonen K, Wang HS, Zhou LY, Chen SH, Li CJ, Nan ZB (2015) Does endophyte symbiosis resist allelopathic effects of an invasive plant in degraded grassland? Fungal Ecol 17:114–125CrossRefGoogle Scholar
  5. Batlla D, Benech-Arnold RL (2014) Weed seed germination and the light environment: implications for weed management. Weed Biol Manag 14:77–87CrossRefGoogle Scholar
  6. Bradford KJ (2002) Applications of hydrothermal time to quantifying and modeling seed germination and dormancy. Weed Sci 50:248–260CrossRefGoogle Scholar
  7. Brem D, Leuchtmann A (2001) Epichloë grass endophytes increase herbivore resistance in the woodland grass Brachypodium sylvaticum. Oecologia 126:522–530CrossRefGoogle Scholar
  8. Bu HY, Du GZ, Chen XL, Xu XL, Liu K, Wen SJ (2008) Community-wide germination strategies in an alpine meadow on the eastern Qinghai-Tibet plateau: phylogenetic and life-history correlates. Plant Ecol 195:87–98CrossRefGoogle Scholar
  9. Charlton ND, Craven KD, Afkhami ME, Hall BA, Ghimire SR, Young CA (2014) Interspecific hybridization and bioactive alkaloid variation increases diversity in endophytic Epichloë species of Bromus laevipes. FEMS Microbiol Ecol 90:276–289CrossRefGoogle Scholar
  10. Chen L, Li XZ, Li CJ, Swoboda GA, Young CA, Sugawara K, Leuchtmann A, Schardl CL (2015) Two distinct Epichloë species symbiotic with Achnatherum inebrians, drunken horse grass. Mycologia 107:863–873Google Scholar
  11. Chen N, He RL, Chai Q, Li CJ, Nan ZB (2016) Transcriptomic analyses giving insights into molecular regulation mechanisms involved in cold tolerance by Epichloë endophyte in seed germination of Achnatherum inebrians. Plant Growth Regul 80:367–375CrossRefGoogle Scholar
  12. Cheplick GP (1997) Effects of endophytic fungi on the phenotypic plasticity of Lolium perenne (Poaceae). Am J Bot 84:34–40CrossRefGoogle Scholar
  13. Christensen MJ, Bennett RJ, Ansari HA, Koga H, Johnson RD, Bryan GT, Simpson WR, Koolaard JP, Nickless EM, Voisey CR (2008) Epichloë endophytes grow by intercalary hyphal extension in elongating grass leaves. Fungal Genet Biol 45:84–93Google Scholar
  14. Clay K (1987) Effects of fungal endophytes on the seed and seedling biology of Lolium perenne and Festuca arundinacea. Oecologia 73:358–362CrossRefGoogle Scholar
  15. Cochrane A, Hoyle GL, Yates CJ, Wood J, Nicotra AB (2014) Predicting the impact of increasing temperatures on seed germination among populations of Western Australian Banksia (Proteaceae). Seed Sci Res 24:195–205CrossRefGoogle Scholar
  16. Cook R, Lewis G, Mizen K (1991) Effects on plant-parasitic nematodes of infection of perennial ryegrass, Lolium perenne, by the endophytic fungus, Acremonium lolii. Crop Prot 10:403–407CrossRefGoogle Scholar
  17. Cui X, Graf HF (2009) Recent land cover changes on the Tibetan plateau: a review. Clim Chang 94:47–61CrossRefGoogle Scholar
  18. Ellis R, Butcher P (1988) The effects of priming and ‘natural’differences in quality amongst onion seed lots on the response of the rate of germination to temperature and the identification of the characteristics under genotypic control. J Exp Bot 39:935–950CrossRefGoogle Scholar
  19. Ellis R, Covell S, Roberts E, Summerfield R (1986) The influence of temperature on seed germination rate in grain legumes: II. Intraspecific variation in chickpea (Cicer arietinum L.) at constant temperatures. J Exp Bot 37:1503–1515CrossRefGoogle Scholar
  20. Ellis R, Simon G, Covell S (1987) The influence of temperature on seed germination rate in grain legumes: III. A comparison of five faba bean genotypes at constant temperatures using a new screening method. J Exp Bot 38:1033–1043CrossRefGoogle Scholar
  21. Estrelles E, Güemes J, Riera J, Boscaiu M, Ibars AM, Costa M (2010) Seed germination behaviour in Sideritis from different Iberian habitats. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 38:9–13CrossRefGoogle Scholar
  22. Faeth SH, Helander ML, Saikkonen KT (2004) Asexual Neotyphodium endophytes in a native grass reduce competitive abilities. Ecol Lett 7:304–313CrossRefGoogle Scholar
  23. Fenner M, Thompson K (2005) The ecology of seeds. Cambridge University Press, CambridgeCrossRefGoogle Scholar
  24. Fortunel C, Paine CET, Fine PVA, Kraft NJB, Baraloto C (2014) Environmental factors predict community functional composition in Amazonian forests. J Ecol 102:145–155CrossRefGoogle Scholar
  25. Fuchs B, Krischke M, Mueller MJ, Krauss J (2017) Plant age and seasonal timing determine endophyte growth and alkaloid biosynthesis. Fungal Ecol 29:52–58CrossRefGoogle Scholar
  26. Gratani L (2014) Plant phenotypic plasticity in response to environmental factors. Advances in Botany 2014:1–17CrossRefGoogle Scholar
  27. Gundel PE, Maseda PH, Ghersa CM, Benech-Arnolo R (2006a) Effects of the Neotyphodium endophyte fungus on dormancy and germination rate of Lolium multiflorum seeds. Austral Ecol 31:767–775CrossRefGoogle Scholar
  28. Gundel PE, Maseda PH, Vila-Aiub MM, Ghersa CM, Benech-Arnold R (2006b) Effects of Neotyphodium fungi on Lolium multiflorum seed germination in relation to water availability. Ann Bot-london 97:571–577CrossRefGoogle Scholar
  29. Gundel PE, Zabalgogeazcoa I, De Aldana BV (2011) Interaction between plant genotype and the symbiosis with Epichloë fungal endophytes in seeds of red fescue (Festuca rubra). Crop Pasture Sci 62:1010–1016CrossRefGoogle Scholar
  30. Gundel PE, Martínez-Ghersa MA, Ghersa CM (2012) Threshold modelling Lolium multiflorum seed germination: effects of Neotyphodium endophyte infection and storage environment. Seed Sci Technol 40:51–62CrossRefGoogle Scholar
  31. Hardegree SP (2006) Predicting germination response to temperature. I. Cardinal-temperature models and subpopulation-specific regression. Ann Bot-london 97:1115–1125CrossRefGoogle Scholar
  32. Hesse U, Schöberlein W, Wittenmayer L, Förster K, Warnstorff K, Diepenbrock W, Merbach W (2003) Effects of Neotyphodium endophytes on growth, reproduction and drought-stress tolerance of three Lolium perenne L. genotypes. Grass Forage Sci 58:407–415CrossRefGoogle Scholar
  33. Hu XW, Zhou ZQ, Li TS, Wu YP, Wang YR (2013) Environmental factors controlling seed germination and seedling recruitment of Stipa bungeana on the loess plateau of northwestern China. Ecol Res 28:801–809CrossRefGoogle Scholar
  34. Hu XW, Fan Y, Baskin CC, Baskin JM, Wang YR (2015) Comparison of the effects of temperature and water potential on seed germination of Fabaceae species from desert and subalpine grassland. Am J Bot 102:649–660CrossRefGoogle Scholar
  35. Iannone L, Pinget A, Nagabhyru P, Schardl C, De Battista J (2012) Beneficial effects of Neotyphodium tembladerae and Neotyphodium pampeanum on a wild forage grass. Grass Forage Sci 67:382–390CrossRefGoogle Scholar
  36. ISTA (2009) International rules for seed testing, 2009th edn. International Seed Testing Association, SwitzerlandGoogle Scholar
  37. Kauppinen M, Saikkonen K, Helander M, Pirttilä AM, Wäli PR (2016) Epichloë grass endophytes in sustainable agriculture. Nat plants 2:15224CrossRefGoogle Scholar
  38. Li CJ, Nan ZB, Paul VH, Dapprich PD, Liu Y (2004) A new Neotyphodium species symbiotic with drunken horse grass (Achnatherum inebrians) in China. Mycotaxon 90:141–147Google Scholar
  39. Li CJ, Gao JH, Nan ZB (2007) Interactions of Neotyphodium gansuense, Achnatherum inebrians, and plant-pathogenic fungi. Mycol Res 111:1220–1227CrossRefGoogle Scholar
  40. Liu W, Liu K, Zhang CH, Du GZ (2011) Effect of accumulated temperature on seed germination: a case study of 12 Compositae species on the eastern Qinghai-Tibetan of China. Chinese Journal of Plant Ecology 35:751–758 (in Chinese with English abstract)Google Scholar
  41. Ma MZ, Christensen MJ, Nan ZB (2015) Effects of the endophyte Epichloë festucae var. lolii of perennial ryegrass (Lolium perenne) on indicators of oxidative stress from pathogenic fungi during seed germination and seedling growth. Eur J Plant Pathol 141:571–583CrossRefGoogle Scholar
  42. Madej CW, Clay K (1991) Avian seed preference and weight loss experiments: the effect of fungal endophyte-infected tall fescue seeds. Oecologia 88:296–302CrossRefGoogle Scholar
  43. McCulley RL, Bush LP, Carlisle AE, Ji H, Nelson JA (2014) Warming reduces tall fescue abundance but stimulates toxic alkaloid concentrations in transition zone pastures of the U.S. Front Chem 2:1–14CrossRefGoogle Scholar
  44. Morse L, Faeth SH, Day T (2007) Neotyphodium interactions with a wild grass are driven mainly by endophyte haplotype. Funct Ecol 21:813–822CrossRefGoogle Scholar
  45. Novas MV, Gentile A, Cabral D (2003) Comparative study of growth parameters on diaspores and seedlings between populations of Bromus setifolius from Patagonia, differing in Neotyphodium endophyte infection. Flora 198:421–426CrossRefGoogle Scholar
  46. Oberhofer M, Güsewell S, Leuchtmann A (2014) Effects of natural hybrid and non-hybrid Epichloë endophytes on the response of Hordelymus europaeus to drought stress. New Phytol 201:242–253CrossRefGoogle Scholar
  47. Pańka D, West C, Guerber C, Richardson M (2013) Susceptibility of tall fescue to Rhizoctonia zeae infection as affected by endophyte symbiosis. Ann Appl Biol 163:257–268CrossRefGoogle Scholar
  48. Parmoon G, Moosavi SA, Akbari H, Ebadi A (2015) Quantifying cardinal temperatures and thermal time required for germination of Silybum marianum seed. The Crop Journal 3:145–151Google Scholar
  49. Pérez-García F, Hornero J, González-Benito ME (2003) Interpopulation variation in seed germination of five Mediterranean Labiatae shrubby species. Isr J Plant Sci 51:117–124CrossRefGoogle Scholar
  50. Phartyal SS, Thapliyal RC, Nayal JS, Rawatm MMS, JoshiI G (2003) The influences of temperatures on seed germination rate in Himalayan elm (Ulmus wallichiana). Seed Sci Technol 31:83–93CrossRefGoogle Scholar
  51. Pinkerton B, Rice J, Undersander D (1990) Germination in Festuca arundinacea as affected by the fungal endophyte, Acremonium coenophialum. In: Proceedings of an international symposium on Acremonium/grass interactions. New OrleansGoogle Scholar
  52. Saikkonen K, Faeth SH, Helander M, Sullivan T (1998) Fungal endophytes: a continuum of interactions with host plants. Annu Rev Ecol Syst 29:319–343Google Scholar
  53. Saikkonen K, Ion D, Gyllenberg M (2002) The persistence of vertically transmitted fungi in grass metapopulations. P Roy Soc B-Biol Sci 269:1397–1403Google Scholar
  54. Saikkonen K, Wäli P, Helander M, Faeth SH (2004) Evolution of endophyte-plant symbioses. Trends Plant Sci 9:275–280CrossRefGoogle Scholar
  55. Saikkonen K, Phillips TD, Faeth SH, McCulley RL, Saloniemi I, Helander M (2016a) Performance of endophyte infected tall fescue in Europe and North America. PLoS One 11:e0157382CrossRefGoogle Scholar
  56. Saikkonen K, Young CA, Helander M, Schardl CL (2016b) Endophytic Epichloë species and their grass hosts: from evolution to applications. Plant Mol Biol 90:665–675CrossRefGoogle Scholar
  57. Schardl CL (2001) Epichloë festucae and related mutualistic symbionts of grasses. Fungal Genet Biol 33:69–82CrossRefGoogle Scholar
  58. Schutte BJ, Regnier EE, Harrison SK, Schmoll JT, Spokas K, Forcella F (2008) A hydrothermal seedling emergence model for giant ragweed (Ambrosia trifida). Weed Sci 56:555–560CrossRefGoogle Scholar
  59. Shiba T, Arakawa A, Sugawara K (2015) Effects of alkaloids from fungal endophytes in grass–Epichloë associations on survival of the sorghum plant bug (Stenotus rubrovittatus). Grassl Sci 61:24–27CrossRefGoogle Scholar
  60. Song ML, Chai Q, Li XZ, Yao X, Li CJ, Christensen MJ, Nan ZB (2015a) An asexual Epichloë endophyte modifies the nutrient stoichiometry of wild barley (Hordeum brevisubulatum) under salt stress. Plant Soil 387:153–165CrossRefGoogle Scholar
  61. Song ML, Li XZ, Saikkonen K, Li CJ, Nan ZB (2015b) An asexual Epichloë endophyte enhances waterlogging tolerance of Hordeum brevisubulatum. Fungal Ecol 13:44–52CrossRefGoogle Scholar
  62. Steinmaus SJ, Prather TS, Holt JS (2000) Estimation of base temperatures for nine weed species. J Exp Bot 51:275–286CrossRefGoogle Scholar
  63. Tadych M, Ambrose KV, Bergen MS, Belanger FC, White JF (2012) Taxonomic placement of Epichloë poae sp. nov. and horizontal dissemination to seedlings via conidia. Fungal Divers 54:117–131CrossRefGoogle Scholar
  64. Tobe K, Zhang LP, Omasa K (2006) Seed germination and seedling emergence of three Artemisia species (Asteraceae) inhabiting desert sand dunes in China. Seed Sci Res 16:61–69CrossRefGoogle Scholar
  65. Trudgill D, Squire G, Thompson K (2000) A thermal time basis for comparing the germination requirements of some British herbaceous plants. New Phytol 145:107–114CrossRefGoogle Scholar
  66. Vázquez de Aldana BR, Gundel PE, García Criado B, García Ciudad A, García Sánchez A, Zabalgogeazcoa I (2014) Germination response of endophytic Festuca rubra seeds in the presence of arsenic. Grass Forage Sci 69:462–469CrossRefGoogle Scholar
  67. Wäli PR, Helander M, Saloniemi I, Ahlholm J, Saikkonen K (2009) Variable effects of endophytic fungus on seedling establishment of fine fescues. Oecologia 159:49–57CrossRefGoogle Scholar
  68. Wang MY, Liu W, Liu K, Bu HY (2011) The base temperature and the thermal time requirement for seed germination of 10 grass species on the eastern Qinghai-Tibet plateau. Pratacultural Science 28:983–987 (in Chinese with English abstract)Google Scholar
  69. Welty RE, Craig AM, Azevedo MD (1994) Variability of ergovaline in seeds and straw and endophyte infection in seeds among endophyte-infected genotypes of tall fescue. Plant Dis 78:845–849CrossRefGoogle Scholar
  70. White Jr JF, Cole GT (1985) Endophyte-host associations in forage grasses. III. In vitro inhibition of fungi by Acremonium coenophialum. Mycologia 77:487-489Google Scholar
  71. Zhang XX, Fan XM, Li CJ, Nan ZB (2010a) Effects of cadmium stress on seed germination, seedling growth and antioxidative enzymes in Achnatherum inebrians plants infected with a Neotyphodium endophyte. Plant Growth Regul 60:91–97CrossRefGoogle Scholar
  72. Zhang XX, Li CJ, Nan ZB (2010b) Effects of cadmium stress on growth and anti-oxidative systems in Achnatherum inebrians symbiotic with Neotyphodium gansuense. J Hazard Mater 175:703–709CrossRefGoogle Scholar
  73. Zhang XX, Li CJ, Nan ZB, Matthew C (2012) Neotyphodium endophyte increases Achnatherum inebrians (drunken horse grass) resistance to herbivores and seed predators. Weed Res 52:70–78CrossRefGoogle Scholar
  74. Zhao XY, Ren JZ, Wang YR, Li YM (2005) Germination responses to temperature and moisture in seed from three species of Caragana. Acta Botan Boreali-Occiden Sin 25:211–217 (in Chinese with English abstract)Google Scholar
  75. Zhou LY, Li CJ, Zhang XX, Johnson R, Bao GS, Yao X, Chai Q (2015) Effects of cold shocked Epichloë infected Festuca sinensis on ergot alkaloid accumulation. Fungal Ecol 14:99–104CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Gensheng Bao
    • 1
    • 2
    Email author
  • Meiling Song
    • 1
    • 2
  • Yuqin Wang
    • 1
    • 2
  • Kari Saikkonen
    • 3
  • Hongsheng Wang
    • 1
    • 2
  1. 1.State Key Laboratory of Plateau Ecology and AgricultureQinghai UniversityXiningChina
  2. 2.Key Laboratory of Qinghai-Tibet Plateau Forage Gerplasm ResearchAcademy of Animal and Veterinary Medicine, Qinghai UniversityXiningChina
  3. 3.Biodiversity UnitUniversity of TurkuTurkuFinland

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