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Case Study: Advancements in Biostimulant Antifungal Mechanisms
Project Duration: October 2024 – December 2025

Project Overview

Our team undertook a project to investigate the antifungal properties of a proprietary biostimulant designed for agricultural applications. The objective was to determine whether the biostimulant could consistently suppress economically significant fungal pathogens and to uncover the biochemical and microbial mechanisms underlying its activity. The project aimed to address batch-to-batch variability, identify pathogen-specific efficacy, and provide a mechanistic understanding to guide reproducible, targeted use as a biofungicide.

Challenges and Scientific Uncertainties

The test biostimulant is a complex, multi-component product derived from cold-burst extraction and a 42-day fermentation of plant exudates. Unlike conventional single-compound antifungals, the product contains diverse metabolites and microbial communities. This complexity created several uncertainties that standard testing methods could not address:

  • Significant variability in antifungal performance between production batches, potentially due to differences in metabolite composition, microbial populations, or their interactions.

  • Unknown spectrum and selectivity of antifungal activity across different plant pathogens.

  • Unclear mechanisms of action, which could involve direct antifungal metabolites, antagonistic microbes, soil microbiome modulation, or synergistic interactions between these factors.

Existing literature and quality control methods were insufficient to resolve these uncertainties, requiring novel experimental designs and mechanistic investigations.

Best Farming achieves at least a 4x return on
their R&D re-investments from SR&ED tax credits,
made available through its partnership with Digitize.

Research and Work Performed

To address these uncertainties, the team conducted a series of targeted studies:

1. Batch Consistency Testing
  • Adapted the Agar Well Diffusion Assay for the biostimulant.
  • Tested six production batches (140–640) against Escherichia coli ATCC 2592
  • Observed batch-dependent antifungal activity: only batches 440, 540, and 640 showed measurable inhibition, confirming that batch-specific factors s
2. Pathogen-Specific Screening
  • Tested active batches against 15 agriculturally relevant fungal pathogens, including Fusarium graminearum, Rhizoctonia solani, and Sclerotinia sclerotior
  • The biostimulant selectively inhibited Fusarium graminearum, Fusarium avenaceum, Septoria sp., Mycosphaerella pinodes, and Leptosphaeria maculans, while other pathogens showed no inhibition
  • Results defined a precise spectrum of activity for targeted application strate
3. Mechanistic Investigation
  • Untargeted metabolomic profiling identified over 12 bioactive compounds with antifungal properties, including 3-phenyllactic acid, azelaic acid, s
  • Microbial isolation identified Paenibacillus polymyxa JB0501-1 as a contributor to antifungal activity. Both its cell-free supernatant and fermented broth strongly inh
  • Practical assays on contaminated barley seeds confirmed effective suppression of funga
  • The bacterial isolate also exhibited plant growth-promoting traits, including nitrog
Throughout the project, over 150 laboratory assays, metabolomic analyses, and microbial isolations were conducted, with meticulous documentation of batch numbers, assay conditions, inhibition zones, and chromatographic profiles.


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Scientific and Technological Advancements

The project achieved several key advancements:
  • Established a reproducible framework for assessing complex, multi-component biostimulants, addressing batch variability and emergent biological effects.
  • Defined the pathogen-specific antifungal spectrum, enabling targeted crop protection strategies.
  • Identified the chemical and microbial mechanisms contributing to antifungal activity, revealing synergistic interactions between metabolites and microbial populations.
  • Confirmed multifunctional benefits of the microbial isolate, including plant growth promotion.
  • Provided data to improve fermentation protocols, standardize production, and optimize biostimulant efficacy.
This work provides a solid scientific foundation for the development and application of complex, fermentation-derived biostimulants, advancing both agricultural technology and our understanding of multi-component antifungal systems.

Best Farming Technologies has partnered with tax-incentive specialist Digitize to ensure they capture all eligible benefits available through the Canadian government’s Scientific Research and Experimental Development (SR&ED) program. Each year, the Canadian government provides billions of dollars in tax incentives to support companies that invest in innovation

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