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Endophytes and related methods

a technology of endophyte and ergot alkaloids, applied in the field of endophytic fungi, can solve the problems of reducing herbivore feeding, toxic to grazing animals, limiting usefulness, and low inoculation frequency of endophyte metabolites,

Inactive Publication Date: 2013-04-25
AGRI VICTORIA SERVICES PTY LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patent is about a method to create new strains of a beneficial endophyte called Neotyphodium lolii, which is found in grasses like perennial ryegrass. These new strains can have better abilities to protect the grass from disease and improve its growth in different conditions. The method involves using X-ray mutagenesis to modify the genes in the endophyte and create new variants with improved properties. This would make these new strains more stable and compatible with different grass host genotypes, and would also eliminate the production of certain toxins. Overall, this patent describes a way to create new and improved endophyte strains for grasses that would be better suited for agricultural applications.

Problems solved by technology

Other metabolites produced by the endophyte, lolitrems and ergot alkaloids, are toxic to grazing animals and reduce herbivore feeding.
Difficulties in artificially breeding of these endophytes limit their usefulness.
For example, many of the novel endophytes known to be beneficial to pasture-based agriculture exhibit low inoculation frequencies and are less stable in elite germplasm.

Method used

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  • Endophytes and related methods
  • Endophytes and related methods
  • Endophytes and related methods

Examples

Experimental program
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Effect test

example 1

Identification of Novel Endophytes

[0293]A collection of 244 perennial grass accessions was assembled for the discovery of novel endophyte strains. The collection targeted accessions from the Northern Mediterranean and Eastern Europe for endophytes that lack lolitrems, as well as accessions from the Middle East, the proposed centre of origin of perennial ryegrass and N. lolii.

[0294]Genotypic analysis of endophyte content was performed across a total of 189 accessions. From each accession 1-5 plant genotypes were analysed for endophyte. Endophyte incidence was low, with endophyte detected in 51% of accessions. Endophyte was consistently detected (with ≧10 SSR markers) in 77 of the accessions.

[0295]Endophytes representing five different taxa were detected across the 77 accessions with 18 SSR markers used to investigate endophyte diversity in perennial ryegrass (FIG. 2). N. lolii was predominant, occurring in 63 accessions. Also detected, although less common, were LpTG-2 and putativel...

example 2

Candidate Endophytes

[0301]Candidate endophytes for further study were chosen on the basis of their genetic identity and metabolic profile. Host-endophyte combinations producing significant amounts of lolitrem B were eliminated, as the ryegrass staggers syndrome produced by this alkaloid is the most important limitation for livestock production.

[0302]The candidate endophyte NEA10 (originating from Spain) was identified as a novel genotype in this analysis with an unknown toxin profile. Its genetic identity is a unique N. lolii strain. Following in planta metabolic profiling analysis, candidate endophyte NEA10 was found to produce ergovaline and peramine, and not lolitrem B.

[0303]The candidate endophyte NEA11 (originating from France) was identified as a novel genotype in this analysis with an unknown toxin profile. Its genetic identity is a unique LpTG-2 strain. Following in planta metabolic profiling analysis, candidate endophyte NEA11 was found to produce ergovaline and peramine, a...

example 3

Methodologies for Endophyte Characterisation

Endophyte Isolation

[0306]Novel candidate endophytes were isolated from their host plant to establish an in vitro culture. Following isolation, the genotype of each endophyte was confirmed by SSR analysis to ensure a high level of quality control prior to inception of isogenic inoculations.

Establishment of Meristem Cultures for a Diverse Perennial Ryeqrass Host Panel

[0307]A set of cultivars representing elite germplasm were obtained, including forage and turf types. Meristem cultures from different cultivars were established to evaluate and compare the phenotypic properties of novel endophyte strains in diverse isogenic host backgrounds. Embryogenic genotypes were identified for each of the cultivars through callus induction and proliferation. Subsequent regeneration of embryogenic genotypes identified primary tissue culture responsive (pTCR) genotypes for each of the cultivars. The number of pTCR genotypes with regeneration frequencies ran...

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Abstract

The present invention relates to a method for identifying and / or characterising an endophyte strain, said method including providing a plurality of samples of endophytes, subjecting said endophytes to genetic analysis, subjecting said endophytes to metabolic analysis and selecting endophytes having a desired genetic and metabolic profile.The present invention also relates to novel endophytes having a desired toxin profile wherein the endophyte produces significantly less toxic alkaloids compared with a control endophyte such as standard toxic (ST) endophyte; and / or significantly more alkaloids conferring beneficial properties compared with a control endophyte such as ST endophyte.The present invention also relates to endophyte variants having a desired genetic and metabolic profile, wherein said endophyte variants possess genetic and / or metabolic characteristics that result in a beneficial phenotype in a plant harbouring or otherwise associated with the endophyte variant. Preferably said endophyte variants are generated by polyploidisation or induced chromosome doubling.

Description

STATEMENT OF RELATED CASES[0001]This application is a continuation in part of PCT / AU2011 / 000020, filed Jan. 7, 2011, which claims priority from Australian Patent Application filed Jan. 7, 2010, and Australian Patent Application 2010902821, filed Jun. 25, 2010, and also claims priority from Australian Patent Application Nos. 2012902275 and 2012902276, filed Jun. 1, 2012. All of these applications are incorporated herein by reference in their entirety.FIELD OF THE INVENTION[0002]The present invention relates to endophytic fungi (endophytes), including modified variants thereof, and to nucleic acids thereof. The present invention also relates to plants infected with endophytes and to related methods, including methods of selecting, breeding, characterising and / or modifying endophytes.BACKGROUND OF THE INVENTION[0003]Important forage grasses perennial ryegrass and tall fescue are commonly found in association with fungal endophytes.[0004]Both beneficial and detrimental agronomic propert...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12Q1/68A01H5/10A01H5/12A01H6/46
CPCC12Q1/6895C12N1/14C12Q2600/156A01H17/00A01H5/12A01H6/46A01H5/10A01H15/00
Inventor SPANGENBERG, GERMAN CARLOSGUTHRIDGE, KATHRYN MICHAELAFORSTER, JOHN WHITESAWBRIDGE, TIMOTHY IVORLUDLOW, EMMA JANE ISOBELKAUR, JATINDERROCHFORT, SIMONE JANERABINOVICH, MAIA ANDREAEKANAYAKE, PIYUMI
Owner AGRI VICTORIA SERVICES PTY LTD
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