Production of loline alkaloids by the grass endophyte, Neotyphodium uncinatum, in defined media

Jimmy D. Blankenship, Martin J. Spiering, Heather H. Wilkinson, Franklin F. Fannin, Lowell P. Bush, Christopher L. Schardl

Research output: Contribution to journalArticlepeer-review

108 Scopus citations


Lolines (saturated 1-aminopyrrolizidines with an oxygen bridge) are insecticidal alkaloids produced in symbioses of certain Epichloë (anamorph-Neotyphodium) species (fungal endophytes) with grasses, particularly of the genera Loliun and Festuca. Prior to the present study, it was unknown whether lolines were of plant or fungal origin. Neotyphodium uncinatum, the common endophyte of meadow fescue (Lolium pratense = Festuca pratensis) produced loline, N-acetylnorloline, and N-formylloline when grown in the defined minimal media at pH 5.0-7.5, with both organic and inorganic nitrogen sources and sugars as carbon sources. In contrast, lolines were not detected in complex medium cultures. GC-MS and 13C NMR spectroscopic analyses confirmed the identity of the alkaloids isolated from the defined medium cultures. Lolines accumulated to ca. 700 mg/l (4 mM) in cultures with 16.7 mM sucrose and 15-30 mM asparagine, ornithine or urea. Kinetics of loline production and fungal growth were assessed in defined medium with 16.7 mM sucrose and 30 mM ornithine. The alkaloid production rate peaked after the onset of stationary phase, as is common for secondary metabolism in other microbes.

Original languageEnglish
Pages (from-to)395-401
Number of pages7
Issue number3
StatePublished - Oct 1 2001

Bibliographical note

Funding Information:
We thank Dr. Mark S. Meier and Mr. John Layton for their expert advice on 13 C NMR analysis, and we thank Dr. Harold H. Burton, Dr. Peter A. Crooks and Dr. Xiaochen Wei for their expert advice on GC–MS. Capable assistance was kindly provided by Mr. A.D. Byrd, Ms. J.L. Dueñas, Ms. D.W. Franks, Mr. W. Hollin, Ms. A.C. Mallory, Ms. S.A. Mathews, and Ms. S. Van Orden. This research was supported by National Science Foundation grant IBN-9808554 to C.L.S. and L.P.B., and the United States Department of Agriculture NRI grant 96-35303-3578 to H.H.W.


  • 1-Aminopyrrolizidines
  • Clavicipiteae
  • Epichloë endophyte
  • Festuca pratensis
  • Fungal fermentation
  • Loline alkaloids
  • Lolium pratense
  • Meadow fescue
  • Neotyphodium uncinatum
  • Poaceae

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Plant Science
  • Horticulture


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