Continuous Liquid Culture for Arbuscular Mycorrhizal Fungi

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Solution Overview

Problem

Current methods for the in vitro cultivation of arbuscular mycorrhizal fungi (AMF) are inefficient, costly, and prone to contamination, limiting the mass production of AMF material for industrial use due to the obligate biotrophic nature of AMF and the complexity of existing cultivation techniques.

Innovation Solution

A continuous liquid culture system for AMF-colonized root material that allows for cost-efficient and scalable production by maintaining high colonization rates and spore yields, using root material capable of building a branched secondary root system, which can be easily handled and stored, and does not require complex bioreactors or active aeration.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional substrate culture or conventional ROC is used for AMF cultivation, then colonization can be achieved, but productivity and scalability are markedly limited

Engineering Contradiction:
ImprovecolonizationVSAvoidmass production
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The patent applies liquid culture medium instead of solid substrate to cultivate AMF-colonized root material. The liquid medium allows for better mixing, oxygen distribution, and scalability in bioreactors, thereby increasing productivity while maintaining colonization effectiveness.

Inventive Principle:
Principle #29Pneumatics and hydraulics

Solution Approach 2:

The patent changes the physical state of the culture medium from solid (conventional ROC) to liquid, and optimizes parameters such as aeration rate, agitation speed, and nutrient composition to enable both high colonization rates and scalable mass production.

Inventive Principle:
Principle #35Parameter changes

2Productivity

If complex bioreactors with active aeration are used, then productivity increases, but device complexity and cost increase

Engineering Contradiction:
Improveproduction yieldVSAvoidbioreactor complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent employs root material capable of autonomous growth and branching in liquid culture without requiring complex bioreactor systems. The roots self-organize and form branched secondary root systems that naturally support AMF colonization, eliminating the need for sophisticated aeration and mixing devices.

Inventive Principle:
Principle #25Self-service

Solution Approach 2:

The patent uses simple, inexpensive liquid culture systems that can be easily prepared and disposed of, replacing complex, expensive bioreactors. The method achieves high productivity through optimized liquid culture conditions rather than through complex equipment.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

3Reliability

If conventional ROC techniques are used, then AMF colonization is achieved, but contamination risk and operational complexity increase

Engineering Contradiction:
ImprovecolonizationVSAvoidcontamination resistance
Core Design Contradiction:
ReliabilityVSEase of operation

Solution Approach 1:

The liquid culture system allows for better control of sterile conditions through fluid handling techniques. The liquid medium can be sterilized more effectively and maintains better sterility during cultivation compared to solid substrates, reducing contamination risk while preserving colonization effectiveness.

Inventive Principle:
Principle #29Pneumatics and hydraulics

4Productivity

If root material capable of branched secondary root system is used, then productivity and ease of handling improve, but selection of suitable root material becomes more specific

Engineering Contradiction:
Improvespore yieldVSAvoidroot material selection
Core Design Contradiction:
ProductivityVSAdaptability or versatility

Solution Approach 1:

The patent selects specific root materials with the particular quality of forming branched secondary root systems. This local quality (branching capability) is identified as critical for success in liquid culture, allowing the system to achieve high productivity while maintaining ease of handling and storage.

Inventive Principle:
Principle #3Local quality

Data Source

PatentEP3038456B1System and methods for continuous propagation and mass production of arbuscular mycorrhizal fungi in liquid culture
Publication Date: 2018.03.21 SYMPLANTA GMBH & CO
  • EP3038456B1 patent drawingFigure 1A~2
  • EP3038456B1 patent drawingFigure 3~4
  • EP3038456B1 patent drawingFigure 5~6

AI summary

The present invention relates to a method and a system for the production of arbuscular mycorrhizal fungi (AMF). More specifically, the invention described herein relates to a method and a system for the continuous in vitro culture of arbuscular mycorrhizal fungi, preferably including steps of harvesting and/or passaging. Further, the invention relates to culture conditions suitable for said continuous cultivation method and system. Another aspect of the invention is a sterile inoculum comprising AMF-colonized root material and other propagules, like AMF spores. Yet another aspect of the invention relates to the use of AMF-colonized root material for continuous production of arbuscular mycorrhizal fungi material in liquid culture medium.