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PMID: 15654104 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Gene clusters for insecticidal loline alkaloids in the grass-endophytic fungus Neotyphodium uncinatum.

Genetics ·Vol. 169 ·No. 3 ·2005-03-00 ·Pages 1403-14

Spiering MJ, Moon CD, Wilkinson HH, Schardl CL

Abstract

Loline alkaloids are produced by mutualistic fungi symbiotic with grasses, and they protect the host plants from insects. Here we identify in the fungal symbiont, Neotyphodium uncinatum, two homologous gene clusters (LOL-1 and LOL-2) associated with loline-alkaloid production. Nine genes were identified in a 25-kb region of LOL-1 and designated (in order) lolF-1, lolC-1, lolD-1, lolO-1, lolA-1, lolU-1, lolP-1, lolT-1, and lolE-1. LOL-2 contained the homologs lolC-2 through lolE-2 in the same order and orientation. Also identified was lolF-2, but its possible linkage with either cluster was undetermined. Most lol genes were regulated in N. uncinatum and N. coenophialum, and all were expressed concomitantly with loline-alkaloid biosynthesis. A lolC-2 RNA-interference (RNAi) construct was introduced into N. uncinatum, and in two independent transformants, RNAi significantly decreased lolC expression (P < 0.01) and loline-alkaloid accumulation in culture (P < 0.001) compared to vector-only controls, indicating involvement of lolC in biosynthesis of lolines. The predicted LolU protein has a DNA-binding site signature, and the relationships of other lol-gene products indicate that the pathway has evolved from various different primary and secondary biosynthesis pathways.

MeSH Terms
Alkaloids/genetics Amino Acid Sequence Ascomycota/genetics Cloning, Molecular Conserved Sequence DNA Primers Enzymes/genetics Genome, Bacterial Genome, Fungal Insecticides Molecular Sequence Data Multigene Family Plasmids Poaceae/microbiology Polymerase Chain Reaction RNA Interference RNA, Fungal/genetics Restriction Mapping
Chemicals
Alkaloids DNA Primers Enzymes Insecticides RNA, Fungal loline
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Spiering Martin J
Department of Plant Pathology, University of Kentucky, Lexington, 40546-0312, USA.
Moon Christina D
Wilkinson Heather H
Schardl Christopher L
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Article Info
Journal
Genetics
Abbr.
Genetics
ISSN
0016-6731
DOI
10.1534/genetics.104.035972
Published
2005-03-00
Epub
2005-00-16
Pages
1403-14
Language
English
Region
United States
NLM ID
0374636
PMCID
PMC1449547
Subset
IM
Databases
GENBANK
AY723749, AY723750, AY724686
External Links
PubMed source DOI: 10.1534/genetics.104.035972 PMC Full Text View journal details
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