Endophyte Strain Selection Using Genetic and Metabolic Analysis

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Solution Overview

Problem

Current methods for breeding and identifying endophytes, particularly Neotyphodium endophytes, face challenges such as low inoculation frequencies, instability in elite germplasm, and difficulties in delivering desired traits, limiting their agricultural applications.

Innovation Solution

A method involving genetic and metabolic analysis of endophytes using SSR markers and nucleic acid profiling to select and characterize endophyte strains with desired traits, including the use of mating-type genes and hybridization for molecular breeding, and the identification of novel endophyte strains like E1 with high inoculation success and stability.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If traditional breeding methods are used for endophytes, then natural association is maintained, but inoculation frequency is low and stability in elite germplasm is poor

Engineering Contradiction:
Improvestability in elite germplasmVSAvoidinoculation frequency
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The patent replaces traditional mechanical/natural breeding methods with molecular biology techniques including PCR-based marker-assisted selection, DNA sequencing, and genetic transformation. This substitution enables precise identification and selection of endophyte strains with desired traits, significantly improving both inoculation frequency and stability in elite germplasm while maintaining natural association characteristics.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent creates molecular copies of endophyte DNA through PCR amplification and sequencing to identify and select specific strains. This copying process allows rapid multiplication and distribution of successful endophyte strains without relying on slow natural propagation, thereby increasing inoculation frequency while maintaining strain stability.

Inventive Principle:
Principle #26Copying

2Measurement precision

If molecular analysis methods are implemented, then endophyte identification and selection precision is improved, but method complexity and cost increase

Engineering Contradiction:
Improveendophyte identification precisionVSAvoidmethod complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent divides the complex molecular analysis process into discrete, manageable segments: DNA extraction, PCR amplification of specific markers, gel electrophoresis, sequencing, and data analysis. This segmentation makes the complex methodology more systematic and reproducible, reducing the barrier to entry while maintaining high identification precision for endophyte strains.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent introduces molecular markers (such as SSR markers and DNA sequences) as intermediaries between the endophyte organism and the selection process. These markers serve as detectable proxies that simplify identification and selection, reducing the direct complexity of analyzing entire genomes while maintaining precise endophyte characterization.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Reliability

If endophyte strains with desired metabolic profiles are selected, then agricultural performance is improved, but the breeding process becomes more time-consuming

Engineering Contradiction:
Improveagricultural performanceVSAvoidbreeding process duration
Core Design Contradiction:
ReliabilityVSLoss of time

Solution Approach 1:

The patent performs preliminary molecular characterization and metabolic profiling of endophyte strains before the actual breeding process. By pre-screening for desired traits such as stress tolerance, pest resistance, and metabolic profiles using PCR markers and DNA sequencing, the process eliminates time-consuming trial-and-error testing later, significantly reducing overall breeding duration while ensuring agricultural performance.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent implements continuous molecular monitoring and selection throughout the breeding process using PCR-based markers and DNA sequencing. This continuous action allows real-time tracking of endophyte strain characteristics, ensuring desired traits are maintained and improved without interruption, thereby reducing the time required compared to traditional intermittent selection methods.

Inventive Principle:
Principle #20Continuity of useful action

Data Source

PatentEP3011828B1Endophytes and related methods
Publication Date: 2018.12.05 AGRI VICTORIA SERVICES PTY LTD
  • EP3011828B1 patent drawingFigure 1
  • EP3011828B1 patent drawingFigure 2A
  • EP3011828B1 patent drawingFigure 2B

AI summary

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.