Discovery and characterisation of novel asexual Epichloë endophytes from perennial ryegrass (Lolium perenne L.)
Jatinder Kaur A D E , Piyumi N. Ekanayake A D E , Pei Tian A B C D , Eline van Zijll de Jong A D , Mark P. Dobrowolski B D , Simone J. Rochfort A C E , Ross C. Mann A C E , Kevin F. Smith B C D E , John W. Forster A C D E , Kathryn M. Guthridge A D E and German C. Spangenberg A C D E F
+ Author Affiliations
- Author Affiliations
A Department of Economic Development, Jobs, Transport and Resources, Biosciences Research Division, AgriBio, the Centre for AgriBioscience, 5 Ring Road, Bundoora, Vic. 3083, Australia.
B Department of Economic Development, Jobs, Transport and Resources, Biosciences Research Division, Hamilton Centre, Mount Napier Road, Hamilton, Vic. 3300, Australia.
C School of Applied Systems Biology, La Trobe University, Bundoora, Vic. 3086, Australia.
D Molecular Plant Breeding Cooperative Research Centre, Victorian AgriBiosciences Centre, La Trobe Research and Development Park, Bundoora, Vic. 3083, Australia.
E Dairy Futures Cooperative Research Centre, AgriBio, the Centre for AgriBioscience, 5 Ring Road, Bundoora, Vic. 3083, Australia.
F Corresponding author. Email: german.spangenberg@ecodev.vic.gov.au
Crop and Pasture Science 66(10) 1058-1070 https://doi.org/10.1071/CP14332
Submitted: 24 November 2014 Accepted: 18 May 2015 Published: 30 September 2015
Abstract
Asexual fungal endophytes of the genus Epichloë form mutually beneficial associations with cool-season pasture grasses such as ryegrasses (Lolium spp.). Alkaloid production by the fungus confers both beneficial (deterrence of invertebrate herbivory) and detrimental (toxicity to mammalian livestock) attributes. A few novel strains with desirable metabolite profiles have been advanced into commercial production by inoculation of perennial ryegrass cultivars. In the present study, an integrated process for discovery of novel endophytes based on exploitation of genotypic information has been designed and implemented. A survey of genetic diversity was performed on a large-scale, customised germplasm collection (containing 244 accessions) in order to identify previously uncharacterised endophyte genotypes. Preliminary qualitative metabolic profiling in the endogenous genetic background permitted elimination of undesirable combinations, and definition of a subset of priority candidates. A novel method was developed for inoculation of endophytes into meristem-culture-derived callus tissue of single genotypes from multiple perennial ryegrass cultivars, in order to allow isogenic comparisons with respect to both host and endophyte genotype. Beneficial toxin profiles were confirmed for associations formed with the grass genotypic panel, and semi-quantitative metabolite analysis provided evidence for genotype-specific effects of both host and genotype on levels of alkaloid production. Vegetative stability was also assessed over both shorter and longer terms. A final set of three prioritised candidates was obtained, two of which (belonging to the known taxa E. festucae var. lolii and LpTG-2) produce the alkaloids ergovaline and peramine. The third candidate endophyte, belonging to a putative novel taxon, solely produces representatives of an additional alkaloid class, the epoxy-janthitrems.
Additional keywords: alkaloid, genetic diversity, isogenic, meristem culture, PNT, symbiosis, vegetative stability.
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