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

doi:10.1016/S0031-9422(01)00272-2

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

Plant Pathology

Research output: Contribution to journal › Article › peer-review

108 Scopus citations

Abstract

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 language	English
Pages (from-to)	395-401
Number of pages	7
Journal	Phytochemistry
Volume	58
Issue number	3
DOIs	https://doi.org/10.1016/S0031-9422(01)00272-2
State	Published - 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.

Keywords

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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@article{f217bf069f6c418892bebaacf818bd30,

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

abstract = "Lolines (saturated 1-aminopyrrolizidines with an oxygen bridge) are insecticidal alkaloids produced in symbioses of certain Epichlo{\"e} (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.",

keywords = "1-Aminopyrrolizidines, Clavicipiteae, Epichlo{\"e} endophyte, Festuca pratensis, Fungal fermentation, Loline alkaloids, Lolium pratense, Meadow fescue, Neotyphodium uncinatum, Poaceae",

author = "Blankenship, {Jimmy D.} and Spiering, {Martin J.} and Wilkinson, {Heather H.} and Fannin, {Franklin F.} and Bush, {Lowell P.} and Schardl, {Christopher L.}",

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. Due{\~n}as, 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.",

year = "2001",

month = oct,

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doi = "10.1016/S0031-9422(01)00272-2",

language = "English",

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pages = "395--401",

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T1 - Production of loline alkaloids by the grass endophyte, Neotyphodium uncinatum, in defined media

AU - Blankenship, Jimmy D.

AU - Spiering, Martin J.

AU - Wilkinson, Heather H.

AU - Fannin, Franklin F.

AU - Bush, Lowell P.

AU - Schardl, Christopher L.

N1 - 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.

PY - 2001/10/1

Y1 - 2001/10/1

N2 - 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.

AB - 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.

KW - 1-Aminopyrrolizidines

KW - Clavicipiteae

KW - Epichloë endophyte

KW - Festuca pratensis

KW - Fungal fermentation

KW - Loline alkaloids

KW - Lolium pratense

KW - Meadow fescue

KW - Neotyphodium uncinatum

KW - Poaceae

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U2 - 10.1016/S0031-9422(01)00272-2

DO - 10.1016/S0031-9422(01)00272-2

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AN - SCOPUS:0035479608

SN - 0031-9422

VL - 58

SP - 395

EP - 401

JO - Phytochemistry

JF - Phytochemistry

IS - 3

ER -

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

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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