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Influence of Epichloë coenophiala, a seed-borne endophyte, on the micropropagation of tall fescue

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Abstract

Improving micropropagation success rate is important since this technique allows the development of other biotechnological tools, such as genetic transformation, in vitro polyploidization, or endophyte inoculation. This is particularly relevant for grasses, as they are traditionally considered recalcitrant to plant tissue culture. Grasses of the subfamily Pooideae establish mutualistic associations with grass endophyte fungi of the genus Epichloë. These associations are specific, as each host species is associated with one or few Epichloë species. Lolium arundinaceum (Schreb.) Darbysh. is a perennial C3 species, native to Europe, which has become an important cool season forage crop worldwide. This grass usually establishes beneficial symbioses with E. coenophiala, but these benefits depend on both plant and fungal genetics and environmental conditions. Micropropagation protocols have been published for L. arundinaceum, but the influence of Epichloë endophytes was never examined. The aim of this manuscript was to study the effect of E. coenophiala on different micropropagation stages of L. arundinaceum and growth of the regenerated plantlets. Association with E. coenophiala (E+) increased callus induction, enhanced their proliferation, and increased shoot regeneration in L. arundinaceum. However, the biomass of plantlets regenerated from E+ seeds was lower than from non-associated seeds, both in vitro and ex vitro, and the in vitro multiplication rate also decreased. Contrary to other endophyte Epichloë-host associations with Lolium multiflorum and Bromus auleticus, the association with E. coenophiala did not improve tall fescue micropropagation. We can conclude that the effects of Epichloë endophytes on micropropagation and growth of their host differ for each endophyte-host association.

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Funding

This work was supported by funding from Agencia Nacional de Promoción Científica y Tecnológica (PICT Joven 2016-0487, PICT 2016-0877, PICT 2016-0980, PICT 2018-4436), CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina) Grant PIP 11220150100956CO, and from Universidad de Buenos Aires UBACyT (20020150100067BA and 20020170100598BA).

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Authors and Affiliations

  1. Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Biodiversidad y Biología Experimental (DBBE), Laboratorio de Cultivo Experimental de Plantas y Microalgas, No. 68, Ciudad Autónoma de Buenos Aires, Argentina

    J. J. Regalado González, V. Berdion, V. E. Tossi & S. I. Pitta-Alvarez

  2. CONICET - Universidad de Buenos Aires, Instituto de Micología y Botánica (INMIBO), Ciudad Autónoma de Buenos Aires, Argentina

    J. J. Regalado González, V. E. Tossi, L. J. Iannone, S. I. Pitta-Alvarez & M. V. Novas

  3. Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Biodiversidad y Biología Experimental (DBBE), Laboratorio de Micología y Fitopatología, No. 69, Ciudad Autónoma de Buenos Aires, Argentina

    V. Berdion, L. J. Iannone & M. V. Novas

Authors
  1. J. J. Regalado González
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  2. V. Berdion
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  3. V. E. Tossi
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  4. L. J. Iannone
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  5. S. I. Pitta-Alvarez
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  6. M. V. Novas
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Contributions

RJJ designed and carried out the in vitro micropropagation protocol and wrote the manuscript. BV carried out in vitro micropropagation and plantlet acclimatization. TVE supervised plantlet acclimatization and ex vitro growth analysis and critical revision of the manuscript. ILJ provided and characterized the starting plant material and critical revision of the manuscript. PASI designed the in vitro micropropagation protocol and supervised the English edition of the manuscript. NMV supervised and coordinated the research and wrote the manuscript.

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Correspondence to J. J. Regalado González or M. V. Novas.

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Fig. S1

Callus induction in Lolium arundinaceum seeds: A) associated with Epichloë coenophiala B) not associated with Epichloë coenophiala. Calli obtained from Lolium arundinaceum seeds: C) associated with Epichloë coenophiala D) not associated with Epichloë coenophiala. Shoots regenerated from callus obtained from F. arundinacea seeds: E) associated with Epichloë coenophiala F) and not associated with Epichloë coenophiala. (PNG 53368 kb)

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Regalado González, J.J., Berdion, V., Tossi, V.E. et al. Influence of Epichloë coenophiala, a seed-borne endophyte, on the micropropagation of tall fescue. Mycol Progress 19, 1301–1308 (2020). https://doi.org/10.1007/s11557-020-01627-y

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