Expressed sequence tags and genes associated with loline alkaloid expression by the fungal endophyte Neotyphodium uncinatum

Martin J. Spiering; Heather H. Wilkinson; Jimmy D. Blankenship; Christopher L. Schardl

doi:10.1016/S1087-1845(02)00023-3

Expressed sequence tags and genes associated with loline alkaloid expression by the fungal endophyte Neotyphodium uncinatum

Martin J. Spiering
, Heather H. Wilkinson
, Jimmy D. Blankenship
, Christopher L. Schardl

Plant Pathology

Producción científica: Article › revisión exhaustiva

46 Citas (Scopus)

Resumen

Loline alkaloids (LA), which are 1-aminopyrrolizidines with an oxygen bridge, are produced by Epichloë (anamorph = Neotyphodium) species, endophytes of grasses. LA are insecticidal, thus, helping to protect host plants from insect herbivory. The objective of this study was to identify genes associated with LA biosynthesis. Suppression subtractive hybridization PCR was used to isolate transcripts up-regulated during LA production in cultures of Neotyphodium uncinatum. Subtracted cDNAs were cloned and a λ-phage cDNA library from an LA-expressing N. uncinatum culture was screened with subtracted cDNA. In BLAST searches, several cDNAs identified had sequence similarities to aspartate kinases and another with O-acetylhomoserine-(thiol)lyase. Differential expression of these two genes in LA-producing cultures of N. uncinatum was confirmed and in a survey of 23 isolates from 21 Neotyphodium and Epichloë species these two genes strictly correlated with LA production. These findings open up the possibility for detailed studies on genes and enzymes involved in LA production.

Idioma original	English
Páginas (desde-hasta)	242-254
Número de páginas	13
Publicación	Fungal Genetics and Biology
Volumen	36
N.º	3
DOI	https://doi.org/10.1016/S1087-1845(02)00023-3
Estado	Published - 2002

Nota bibliográfica

Funding Information:
This work was supported by the US National Science Foundation Integrative Plant Biology Program 9808554 to C.L.S. and US Department of Agriculture NRICGP grant 9901343 to H.H.W. We thank Ms Rebecca R. Singleton for assistance with the library screening and cDNA sequencing. We also thank Dr. Christina D. Moon and Mr. Kelly D. Craven for providing endophyte genomic DNAs and Mr. Alfred D. Byrd and Mr. Walter Hollin for technical assistance. This is contribution 02-12-49 of the Kentucky Agricultural Experiment Station, published with the approval of the Director.

Financiación

This work was supported by the US National Science Foundation Integrative Plant Biology Program 9808554 to C.L.S. and US Department of Agriculture NRICGP grant 9901343 to H.H.W. We thank Ms Rebecca R. Singleton for assistance with the library screening and cDNA sequencing. We also thank Dr. Christina D. Moon and Mr. Kelly D. Craven for providing endophyte genomic DNAs and Mr. Alfred D. Byrd and Mr. Walter Hollin for technical assistance. This is contribution 02-12-49 of the Kentucky Agricultural Experiment Station, published with the approval of the Director.

Financiadores	Número del financiador
US Department of Agriculture NRICGP	9901343
US National Science Foundation	9808554

ASJC Scopus subject areas

Microbiology
Genetics

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10.1016/S1087-1845(02)00023-3

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@article{15145b6c71a14ceca8b7739ef77f91c5,

title = "Expressed sequence tags and genes associated with loline alkaloid expression by the fungal endophyte Neotyphodium uncinatum",

abstract = "Loline alkaloids (LA), which are 1-aminopyrrolizidines with an oxygen bridge, are produced by Epichlo{\"e} (anamorph = Neotyphodium) species, endophytes of grasses. LA are insecticidal, thus, helping to protect host plants from insect herbivory. The objective of this study was to identify genes associated with LA biosynthesis. Suppression subtractive hybridization PCR was used to isolate transcripts up-regulated during LA production in cultures of Neotyphodium uncinatum. Subtracted cDNAs were cloned and a λ-phage cDNA library from an LA-expressing N. uncinatum culture was screened with subtracted cDNA. In BLAST searches, several cDNAs identified had sequence similarities to aspartate kinases and another with O-acetylhomoserine-(thiol)lyase. Differential expression of these two genes in LA-producing cultures of N. uncinatum was confirmed and in a survey of 23 isolates from 21 Neotyphodium and Epichlo{\"e} species these two genes strictly correlated with LA production. These findings open up the possibility for detailed studies on genes and enzymes involved in LA production.",

keywords = "1-aminopyrrolizidines, Aspartate kinase, Homocysteine synthase, Loline alkaloids, Neotyphodium uncinatum, Suppression subtractive hybridization",

author = "Spiering, \{Martin J.\} and Wilkinson, \{Heather H.\} and Blankenship, \{Jimmy D.\} and Schardl, \{Christopher L.\}",

note = "Funding Information: This work was supported by the US National Science Foundation Integrative Plant Biology Program 9808554 to C.L.S. and US Department of Agriculture NRICGP grant 9901343 to H.H.W. We thank Ms Rebecca R. Singleton for assistance with the library screening and cDNA sequencing. We also thank Dr. Christina D. Moon and Mr. Kelly D. Craven for providing endophyte genomic DNAs and Mr. Alfred D. Byrd and Mr. Walter Hollin for technical assistance. This is contribution 02-12-49 of the Kentucky Agricultural Experiment Station, published with the approval of the Director.",

year = "2002",

doi = "10.1016/S1087-1845(02)00023-3",

language = "English",

volume = "36",

pages = "242--254",

number = "3",

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T1 - Expressed sequence tags and genes associated with loline alkaloid expression by the fungal endophyte Neotyphodium uncinatum

AU - Spiering, Martin J.

AU - Wilkinson, Heather H.

AU - Blankenship, Jimmy D.

AU - Schardl, Christopher L.

N1 - Funding Information: This work was supported by the US National Science Foundation Integrative Plant Biology Program 9808554 to C.L.S. and US Department of Agriculture NRICGP grant 9901343 to H.H.W. We thank Ms Rebecca R. Singleton for assistance with the library screening and cDNA sequencing. We also thank Dr. Christina D. Moon and Mr. Kelly D. Craven for providing endophyte genomic DNAs and Mr. Alfred D. Byrd and Mr. Walter Hollin for technical assistance. This is contribution 02-12-49 of the Kentucky Agricultural Experiment Station, published with the approval of the Director.

PY - 2002

Y1 - 2002

N2 - Loline alkaloids (LA), which are 1-aminopyrrolizidines with an oxygen bridge, are produced by Epichloë (anamorph = Neotyphodium) species, endophytes of grasses. LA are insecticidal, thus, helping to protect host plants from insect herbivory. The objective of this study was to identify genes associated with LA biosynthesis. Suppression subtractive hybridization PCR was used to isolate transcripts up-regulated during LA production in cultures of Neotyphodium uncinatum. Subtracted cDNAs were cloned and a λ-phage cDNA library from an LA-expressing N. uncinatum culture was screened with subtracted cDNA. In BLAST searches, several cDNAs identified had sequence similarities to aspartate kinases and another with O-acetylhomoserine-(thiol)lyase. Differential expression of these two genes in LA-producing cultures of N. uncinatum was confirmed and in a survey of 23 isolates from 21 Neotyphodium and Epichloë species these two genes strictly correlated with LA production. These findings open up the possibility for detailed studies on genes and enzymes involved in LA production.

AB - Loline alkaloids (LA), which are 1-aminopyrrolizidines with an oxygen bridge, are produced by Epichloë (anamorph = Neotyphodium) species, endophytes of grasses. LA are insecticidal, thus, helping to protect host plants from insect herbivory. The objective of this study was to identify genes associated with LA biosynthesis. Suppression subtractive hybridization PCR was used to isolate transcripts up-regulated during LA production in cultures of Neotyphodium uncinatum. Subtracted cDNAs were cloned and a λ-phage cDNA library from an LA-expressing N. uncinatum culture was screened with subtracted cDNA. In BLAST searches, several cDNAs identified had sequence similarities to aspartate kinases and another with O-acetylhomoserine-(thiol)lyase. Differential expression of these two genes in LA-producing cultures of N. uncinatum was confirmed and in a survey of 23 isolates from 21 Neotyphodium and Epichloë species these two genes strictly correlated with LA production. These findings open up the possibility for detailed studies on genes and enzymes involved in LA production.

KW - 1-aminopyrrolizidines

KW - Aspartate kinase

KW - Homocysteine synthase

KW - Loline alkaloids

KW - Neotyphodium uncinatum

KW - Suppression subtractive hybridization

UR - https://www.scopus.com/pages/publications/0036346292

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U2 - 10.1016/S1087-1845(02)00023-3

DO - 10.1016/S1087-1845(02)00023-3

M3 - Article

C2 - 12135580

AN - SCOPUS:0036346292

SN - 1087-1845

VL - 36

SP - 242

EP - 254

JO - Fungal Genetics and Biology

JF - Fungal Genetics and Biology

IS - 3

ER -

Expressed sequence tags and genes associated with loline alkaloid expression by the fungal endophyte Neotyphodium uncinatum

Resumen

Nota bibliográfica

Financiación

ASJC Scopus subject areas

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