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RESEARCH ARTICLE
Contents Vol 59(3)

Detection and characterisation of novel fungal endophyte genotypic variation in cultivars of perennial ryegrass (Lolium perenne L.)

E. van Zijll de Jong A D E , M. P. Dobrowolski B D , A. Sandford C , K. F. Smith B D , M. J. Willocks C , G. C. Spangenberg A D and J. W. Forster A D F
+ Author Affiliations
- Author Affiliations

A Department of Primary Industries, Biosciences Research Division, Victorian AgriBiosciences Centre,La Trobe Research and Development Park, Bundoora, Vic. 3083, Australia.

B Department of Primary Industries, Biosciences Research Division, Hamilton Centre, Hamilton,Vic. 3300, Australia.

C New Zealand Agriseeds Limited, 2547 Old West Coast Road, RD1, Christchurch 8021, New Zealand.

D Molecular Plant Breeding Cooperative Research Centre, Australia.

E Present address: National Centre for Advanced Bio-Protection Technologies, PO Box 84, Lincoln University, Lincoln 7647, Canterbury, New Zealand.

F Corresponding author. Email: john.forster@dpi.vic.gov.au

Australian Journal of Agricultural Research 59(3) 214-221 https://doi.org/10.1071/AR07270
Submitted: 17 July 2007  Accepted: 13 November 2007   Published: 11 March 2008

Abstract

Alkaloids produced by the perennial ryegrass pasture and turf grass endophyte Neotyphodium lolii confer both beneficial (insect feeding deterrence) and deleterious (mammalian herbivore toxicoses) effects. Novel endophyte strains with altered and desirable in planta metabolic profiles have been introduced into cultivars by inoculation. The potentially severe effects of contamination with standard toxic endophytes provide an important incentive for quality control during varietal development. Genotyping with expressed sequence tag (EST)-derived simple sequence repeat (SSR) markers in order to monitor endophyte content was evaluated in this study. The NEA2 endophyte, described as producing low levels of lolitrem B and moderate quantities of ergovaline and peramine, has been deployed in the cultivar (cv.) Tolosa and transferred to other proprietary varieties by crossing or controlled inoculation. In planta genotypic analysis of cv. Tolosa-derived plants revealed the presence of 2 distinct NEA2 genotypes (A and B). Variable proportions of NEA2 A and B were observed at different stages of cultivar multiplication, and NEA2 B was present in inoculated varieties. Several accessions also revealed the presence of a third genotype (C), identical to standard toxic types. Alkaloid measurement in plants containing individual A and B endophytes identified distinct profiles which collectively account for the average NEA2 profile. The C variant is apparently a contaminant arising from seed or adventitious seedlings during varietal multiplication or incomplete removal of endophyte from seed before inoculation. SSR-based genotyping provides an efficient means to continuously assess endophyte prevalence and identity in pasture grass breeding programs.

Additional keywords: EST-SSR, alkaloid, quality control, livestock toxicity, breeding program.


Acknowledgments

This work was supported by funding from the Victorian Department of Primary Industries and the Molecular Plant Breeding Cooperative Research Centre. The authors thank Gavin Kearney for assistance with statistical analysis of data, and Prof. Michael Hayward for careful critical assessment of the manuscript.


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