Treehouse Farm

Cannabis Cooperative on Moku o Keawe in Wao o Kele

AChE Inhibition – Biochemistry Informing Cannabis Microbiology

Animated gif of the top of a bright green cannabis plant in a golden ray of sunshine.

A very short study on an attempt to convert a research paper on cannabis treatments for Autism into a method for producing an AChE inhibiting terpene profile.

In a new paper published in the Nature Journal, Dr.Bonnie and their team at cannaformatics evaluated Cannabis-Responsive salivary biomarkers to measure the impact of medical cannabis treatment in children with Autism Spectrum Disorder (ASD).

I thought, hmmm, I wonder if I can translate that positivity into microbial content my plants might use to to produce a terpenoid profile higher in AChE inhibitors. Put another way, I became interested in learning how this new information can be translated into soil health.

For example, I want to encourage my cannabis plants to learn from and ingest any specific microbes that will result in higher acetylcholinesterase inhibition in humans. Would the result be measurable in CBG for example?

I asked ChatGPT to read Dr. Bonnie’s paper and this paper on “Food-derived Acetylcholinesterase Inhibitors as Potential Agents against Alzheimer’s Disease” as sort of control for AChE. This can of course introduce variables that I can easily mistake for facts. So I read up on the topic so I could have a base for working with ChatGPT which summarized the research thusly:

Translating the findings on acetylcholinesterase inhibition into soil health would involve understanding the symbiotic relationships between cannabis plants and soil microbes. Certain microbes can help the cannabis plant produce phytochemicals that may contribute to acetylcholinesterase inhibition. By enriching the soil with these specific microbes, you might be able to cultivate cannabis with enhanced therapeutic properties.

So I asked ChatGPT, “which specific microbes and the source of those microbes could help me alter my soil.”?

ChatGPT replied, “here’s how you can modify your soil to encourage the cannabis plant to produce phytochemicals that may contribute to acetylcholinesterase inhibition.” Btw, I’m dropping the quotes and I promise not to use the word thusly again in this post. I got into the strange habit when writing prompts. 🤢🤮

Specific Microbes and Their Sources:

  1. Mycorrhizal Fungi: These fungi form a symbiotic relationship with plant roots and can help in the uptake of nutrients. They are known to stimulate the production of secondary metabolites in plants.
    • Sources: Commercial mycorrhizal inoculants are available, or you can introduce them naturally through forest soil.
  2. Trichoderma: This is a genus of fungi that can promote plant growth and also increase the plant’s resistance to diseases.
    • Sources: Available as commercial biofertilizers and biopesticides.
  3. Rhizobacteria: Certain strains of rhizobacteria like Pseudomonas and Bacillus have been known to induce systemic resistance in plants and could stimulate the production of beneficial compounds.
    • Sources: Can be isolated from healthy plant roots or purchased as commercial inoculants.
  4. Azotobacter: Nitrogen-fixing bacteria that can also produce natural plant growth regulators.
    • Sources: Available in commercial biofertilizers.

Steps to Amend the Soil:

  1. Soil Testing: Conduct a comprehensive soil test to understand its current structure, pH, and nutrient levels.
  2. Inoculation: Introduce the selected microbes into the soil. This can be done through commercially available products or natural sources as mentioned above.
  3. Organic Matter: Add organic matter like compost, which serves as a food source for microbes.
  4. Monitor and Maintain: Regularly check soil pH and moisture levels to ensure they are optimal for both the cannabis plants and the microbial community.
  5. Regular Testing: Perform plant tissue tests and recheck soil health to monitor the effectiveness of your microbial amendments.

By creating a more biologically active soil, you’re not just feeding the plant; you’re also encouraging a microbial community that can help the plant produce specific phytochemicals, potentially increasing its therapeutic effects.

I did also ask for more information about Specific Microbes and Their Sources including ideal pH levels and other environmental conditions such as pests, and molds that must be controlled for the soil altering experiment to be a success, please.

Specific Microbes Ideal Conditions Sources Pest/Mold Control
Mycorrhizal Fungi Ideal pH: Slightly acidic to neutral (pH 5.5–7) Commercial mycorrhizal inoculants or forest soil Generally resistant to many soil pathogens but avoid fumigants that can kill beneficial fungi.
Trichoderma Ideal pH: Neutral to slightly alkaline (pH 6–8) Commercial biofertilizers and biopesticides Trichoderma itself is used as a biopesticide and is generally resistant to molds.
Rhizobacteria (Pseudomonas, Bacillus) Ideal pH: Neutral (pH 6.5–7.5) Healthy plant roots or commercial inoculants Some strains produce antibiotics that control soil-borne pathogens.
Azotobacter Ideal pH: Slightly acidic to neutral (pH 5.8–7) Commercial biofertilizers Generally not susceptible to most common soil pests and molds.

Environmental Conditions:

  • Soil Moisture: Overly wet conditions can lead to mold growth and may drown beneficial microbes. Aim for moist but well-drained soil.
  • Temperature: Most beneficial microbes thrive at moderate temperatures (15–30°C or 59–86°F), similar to cannabis plants.
  • Organic Matter: This serves as a food source for microbes, so consider adding compost or well-rotted manure.

Pest and Mold Control:

  • Biological Controls: Consider introducing predatory insects or other biopesticides to manage harmful pests.
  • Cultural Practices: Proper plant spacing, and adequate air circulation can reduce the chances of mold and pest infestations.
  • Chemical Controls: If you must resort to chemical pesticides, opt for those labeled as safe for beneficial microbes.

Here are some additional details about the potential safety concerns for each of the specific microbes mentioned:

Mycorrhizal Fungi

  • Safety Concerns: Generally considered safe for plants and humans. However, some people with compromised immune systems might be more susceptible to fungal infections.
  • Precautions: Use commercial inoculants from reputable sources and avoid fumigants that can kill beneficial fungi.


  • Safety Concerns: While Trichoderma is used as a biopesticide, it can sometimes cause “green mold” disease in mushrooms. It’s not generally harmful to humans or other plants.
  • Precautions: Be cautious if you’re also cultivating mushrooms. Always follow the manufacturer’s guidelines when using commercial biofertilizers or biopesticides that contain Trichoderma.

Rhizobacteria (Pseudomonas, Bacillus)

  • Safety Concerns: Some strains of Pseudomonas and Bacillus can be pathogenic to humans or animals. However, the strains used in agriculture are generally considered safe.
  • Precautions: Use strains that are proven to be non-pathogenic and adhere to guidelines when applying these bacteria. Always buy from reputable sources.


  • Safety Concerns: Generally considered safe and is used in the production of various biofertilizers. It is non-pathogenic to humans and animals.
  • Precautions: Always use commercial products that have been tested for safety and efficacy.

Environmental Conditions & Pest/Mold Control

Even beneficial microbes can become a concern if environmental conditions are not properly managed. Overly wet conditions can lead to mold growth, and incorrect application of pest and mold controls can harm these beneficial microbes.

  • Soil Moisture: Ensure the soil is well-drained to avoid creating a breeding ground for harmful molds.
  • Temperature: Maintain moderate temperatures for the optimal growth of both plants and beneficial microbes.
  • Pest and Mold Control: Choose products that are labeled as safe for beneficial microbes and always follow application guidelines.

By adhering to best practices and guidelines, you can minimize any potential risks associated with using these microbes.

In general, the microbes mentioned—Mycorrhizal Fungi, Trichoderma, Rhizobacteria (Pseudomonas, Bacillus), and Azotobacter—are often compatible and can coexist in the same soil ecosystem without posing a threat to each other. However, interactions can be complex and sometimes depend on specific strains and environmental conditions.

more specific details on how these microbes might interact with each other:

Mycorrhizal Fungi and Other Microbes

  • Interaction with Trichoderma: Generally compatible. Some Trichoderma strains even have a synergistic relationship with mycorrhizal fungi, helping them to establish more effectively in the soil.
  • Interaction with Rhizobacteria: Mostly synergistic. Certain rhizobacteria can stimulate the growth of mycorrhizal fungi.
  • Interaction with Azotobacter: Generally compatible and can coexist without any issues.

Trichoderma and Other Microbes

  • Interaction with Mycorrhizal Fungi: As mentioned, generally compatible and sometimes even synergistic.
  • Interaction with Rhizobacteria: Generally compatible but depends on the strains. Some Trichoderma strains may have antibiotic properties.
  • Interaction with Azotobacter: No known negative interactions.

Rhizobacteria (Pseudomonas, Bacillus) and Other Microbes

  • Interaction with Mycorrhizal Fungi: Generally synergistic. Some rhizobacteria can stimulate the growth of mycorrhizal fungi.
  • Interaction with Trichoderma: Generally compatible, although some strains of rhizobacteria might produce substances that could inhibit fungal growth.
  • Interaction with Azotobacter: Generally compatible and can even have synergistic interactions where both benefit.

Azotobacter and Other Microbes

  • Interaction with Mycorrhizal Fungi: No known negative interactions.
  • Interaction with Trichoderma: No known negative interactions.
  • Interaction with Rhizobacteria: Generally compatible and can sometimes be synergistic.

Important Notes

  • Strain Specificity: It’s crucial to note that the interactions can be strain-specific. Different strains of the same type of microbe might have different interactions with other microbes.
  • Environmental Factors: Soil pH, moisture, and temperature can influence microbial interactions. Ensuring ideal conditions for each microbe will likely reduce any potential for negative interactions.

Please consult with experts and refer to scientific literature for the most accurate information on microbial interactions, especially in Hawaii. We don’t need people altering our environments. If you don’t have sufficient containment measures, consider focusing your search on finding IMOs as replacements. I’ll put something together to refocus on IMOs in another post for those who are considering introducing multiple types of microbes into the same environment.

Here’s some methods to effectively introduce beneficial microbes into your cannabis garden:

Bacillus Strains

Methods of Application

  1. Soil Inoculants: Bacillus spores can be mixed directly into the soil or growing medium before planting.
  2. Foliar Spray: Liquid formulations can be sprayed directly onto the leaves.
  3. Drench: Liquid or water-soluble granules can be applied to the soil as a drench.
  4. Compost Tea: A microbial-rich compost tea can also be brewed and applied either as a foliar spray or soil drench.

Ideal Conditions

  • pH: Neutral to slightly alkaline (6–8)
  • Temperature: Moderate (15–30°C or 59–86°F)


  • Seedling Stage: To establish a healthy root microbiome.
  • Vegetative Stage: To promote plant growth.
  • Before Flowering: To ensure the plant is strong before putting energy into flower production.

Lactobacillus (e.g., Lactobacillus acidophilus)

Methods of Application

  1. Fermented Plant Extracts: Lactobacillus can be cultured in a carbohydrate-rich medium like rice wash or milk and then mixed into compost teas.
  2. Soil Drench: The fermented liquid can be used as a soil drench.
  3. Foliar Spray: Can be sprayed on leaves but is less commonly used in this way for plants.

Ideal Conditions

  • pH: Slightly acidic to neutral (5.5–7)
  • Temperature: Slightly cooler (10–25°C or 50–77°F)


  • Seedling Stage: To help establish initial soil microbiota.
  • Stress Periods: During transplanting or other stress events to help improve resilience.

Important Considerations

  • Compatibility: Both Bacillus and Lactobacillus can generally coexist, but it’s crucial to monitor for any unforeseen interactions.
  • Local Conditions: Given that you’re in Hawaii, consider the local climate and soil conditions when applying these microbes.
  • Organic Matter: As someone interested in sustainability, incorporating organic matter like compost can serve as a food source for these microbes.

By tailoring the introduction of these beneficial microbes to the specific needs of your cannabis plants and local conditions, you can better promote plant health, improve yields, and manage pests sustainably.

Type of Microbe Methods of Application Ideal Conditions Timing
Bacillus Strains
  • Soil Inoculants
  • Foliar Spray
  • Drench
  • Compost Tea
  • pH: Neutral to slightly alkaline (6–8)
  • Temperature: Moderate (15–30°C or 59–86°F)
  • Seedling Stage
  • Vegetative Stage
  • Before Flowering
Lactobacillus (e.g., Lactobacillus acidophilus)
  • Fermented Plant Extracts
  • Soil Drench
  • Foliar Spray
  • pH: Slightly acidic to neutral (5.5–7)
  • Temperature: Slightly cooler (10–25°C or 50–77°F)
  • Seedling Stage
  • Stress Periods

The microbes mentioned—Bacillus strains, Lactobacillus strains, Mycorrhizal Fungi, Trichoderma, Rhizobacteria (Pseudomonas), and Azotobacter—are generally ubiquitous and can be found in various soil environments around the world. However, the specific strains and their abundance can vary depending on local soil conditions, climate, and land use. Given Hawaii’s unique ecosystem, it’s possible that some strains may be less common or not naturally occurring there.

Here’s how to assess (😁🤣) the microbial diversity in your local soil:

Soil Testing

  1. Basic Soil Test: A basic soil test can provide you with information on pH, nutrient levels, and organic matter, which indirectly indicate microbial activity.
  2. Microbial Soil Test: More advanced tests can directly assess microbial populations, including beneficial bacteria and fungi. These tests often require specialized labs that can perform DNA sequencing or plate culturing.

On-site Observations

  1. Plant Health: Healthy plants are often a good indicator of a balanced soil microbiome.
  2. Smell and Texture: Healthy soil often has an earthy smell and crumbly texture, both of which suggest good microbial activity.

Scientific Literature

  • Research papers and agricultural reports specific to Hawaii could provide insights into the native microbial flora.

Consulting Local Experts

  • Contact the local agricultural extension services or UH College of Tropical Ag or Community College who might provide expertise.

DIY Methods

  1. Compost Teas: You can create your own compost teas tailored to your needs, using locally sourced compost to capture native microbial communities.
  2. Microbial Inoculants: If you find that certain beneficial microbes are lacking, commercial microbial inoculants can be used.

Critical Things to Consider

  1. Native vs. Introduced: Hawaii has unique native flora and fauna, so introducing non-native microbes should be done cautiously to avoid disrupting local ecosystems.
  2. Climate: Hawaii’s climate can influence microbial activity, so consider the moisture and temperature needs of any microbes you’re looking to introduce.
  3. See number one.

By taking a systematic approach, we can better understand the microbes in your local soil and make informed decisions about which beneficial microbes to introduce for optimal plant health. Let’s take a closer look at AChE Inhibiting Terpenes.

Terpenes and AChE Inhibition

Terpenes are aromatic compounds found in various plants, including cannabis. Some terpenes have shown promise in inhibiting acetylcholinesterase (AChE), although most of this research is preliminary and not specific to cannabis strains.

  1. Pinene: Alpha and beta-pinene have demonstrated AChE inhibitory activity in some studies. They are commonly found in many cannabis strains.
  2. Limonene: This common cannabis terpene has also shown some promise as an AChE inhibitor in non-cannabis studies.
  3. Caryophyllene: Known for its spicy, peppery aroma, caryophyllene has also shown some AChE inhibitory effects in preliminary studies.

More in this research paper: Inhibitory Effects of Cannabinoids on Acetylcholinesterase and Butyrylcholinesterase Enzyme Activities

The authors (Tess Puopolo, a , b Chang Liu, a Hang Ma, a , b ,* and Navindra P. Seeram a , b ,**) wrote, “In summary, several cannabinoids exhibited moderate inhibitory effects against the activities of cholinesterases including AChE and BChE. In addition, enzyme kinetic studies suggested that CBD is a competitive inhibitor of both AChE and BChE. Further mechanistic studies using molecular docking methods explored the binding capacity between cannabinoids and AChE and BChE proteins. Findings from the experimental- and computational-based assays suggested that cannabinoids are more selective for the inhibition of AChE, which shed light on the development of cannabinoids-based treatment for neurodegenerative diseases but further cells- and animal-based studies are warranted to evaluate cannabinoids’ efficacy.”

Cannabinoids and AChE

  1. CBD (Cannabidiol): Some research suggests that CBD may have neuroprotective effects, although evidence for AChE inhibition specifically is limited.
  2. THC (Tetrahydrocannabinol): THC primarily interacts with the endocannabinoid system and its role in AChE inhibition is not well understood.

Strain Specificity

Given the complexities of cannabis phytochemistry and the lack of targeted research, it’s difficult to recommend specific strains for AChE inhibition. Strains high in pinene, limonene, or caryophyllene might be more likely to have AChE inhibitory effects based on the terpenes they contain, but this is speculative.

Research Gaps

It’s important to note that much of the research on terpenes and AChE inhibition has been conducted in vitro or in animal models, not in humans. The concentrations used in these studies may also not be reflective of the concentrations one would get from consuming cannabis.

Practical Implications

If you’re interested in experimenting with cannabis cultivation for this purpose, you may want to consider strains known to be high in the terpenes listed above. However, this would be more of an exploratory endeavor than one based on solid scientific evidence at this point.

If you are interested in cannabis cultivation and soil microbiology, you could potentially explore how different soil microbes influence the terpene profile of cannabis plants, although this would be a complex and long-term research project.

I hope this provides a thorough overview of what is currently known about cannabis components and AChE inhibition.

A few things you can do to follow-up / fact-check this post:

  1. Paper in Nature Journal: Read the original research. Search academic databases or directly go to Nature’s website to check.
  2. AChE Inhibiting Terpenes: Research papers and more information in pharmacological journals or databases like PubMed.
  3. Microbes and Terpenes: Learn exactly how specific microbes influence the terpene profile of plants.
  4. Safety Concerns: Academic papers, government advisories, and environmental studies could be good sources for safety data on these microbes to research BEFORE introducing them into a local ecosystem like Hawaii.
  5. Microbe Interactions: Scholarly articles and studies on soil microbiology would likely have information on the compatibility or interaction between different microbes.
  6. Ideal Conditions for Microbes: Academic journals and agricultural extension publications often have detailed information on the ideal growing conditions for various microbes.
  7. Microbial Strains in Hawaii: Since you’re in Hawaii, local agricultural extension services, university departments related to agriculture or environmental science, and even local experts could be excellent resources for this.
  8. Soil Testing Methods: Soil science textbooks, agricultural extension publications, or academic courses could have this information. Consulting with local experts could also be valuable.
  9. Local Expertise: Given your involvement in sustainability and community education, you might have contacts who could confirm the availability of local resources.

Steps to Explore

  1. Literature Review: Start by looking into existing research on how soil microbes influence terpene production in plants, not just cannabis. This will give you a foundational understanding.
  2. Identify Target Terpenes: Given that certain terpenes like pinene, limonene, and caryophyllene have shown some promise in AChE inhibition, these could be your target terpenes.
  3. Soil Testing: Before introducing any new microbes, get a baseline soil test to understand its current microbial community, pH, and nutrient levels.
  4. Microbe Selection: Choose microbes that are known to influence the production of your target terpenes. Mycorrhizal fungi, Trichoderma, and certain rhizobacteria are often cited for their role in enhancing terpene profiles.
  5. Inoculation: Introduce these microbes into your soil using commercial inoculants or other methods.
  6. Control Group: Maintain a control group of cannabis plants without the microbial amendments for comparison.
  7. Monitoring: Regularly monitor soil conditions, plant health, and terpene profiles through lab tests.
  8. Data Analysis: Compare the terpene profiles of the control group and the test group to assess the impact of the microbial amendments.
  9. Adjustments: Based on your findings, you may need to adjust the microbial community, soil conditions, or other variables.


  1. Local Ecology: Given that you’re in Hawaii, be cautious about introducing non-native microbes that could disrupt local ecosystems.
  2. Long-term Study: This would be a long-term project requiring multiple growth cycles to get reliable data.
  3. Expert Consultation: Consult with soil microbiologists, cannabis experts, and possibly even neuroscientists who specialize in AChE for a well-rounded approach.
  4. Regulatory Compliance: Make sure to comply with all local and state regulations regarding cannabis cultivation and scientific research.
  5. Documentation: Keep thorough records of all procedures, observations, and test results. This can not only guide your ongoing work but also serve as valuable data for any future publications or presentations you may wish to make.

Originally found on Dr. Bonnie’s Instagram: @bonnigoldsteinmd


Puopolo T, Liu C, Ma H, Seeram NP. Inhibitory Effects of Cannabinoids on Acetylcholinesterase and Butyrylcholinesterase Enzyme Activities. Med Cannabis Cannabinoids. 2022 Apr 19;5(1):85-94. doi: 10.1159/000524086. PMID: 35702400; PMCID: PMC9149358.

Disclosure: I’m barely smart enough to put this post together and I rely heavily on preconceived notions and bias to try to understand if ChatGPT is lying. While I’m certain this information is going to be helpful it may just as likely become a buried artifact at a time when we only knew one half of one percent of that there is to know about soil microbiology. Remember to scrutinize the credibility of your sources, especially when considering scientific or health-related claims. Do your own research.