It’s been a few weeks since I started my experiment in growing a bioluminescent mushroom and I thought it’d be a good time to give an update on my progress.
Not everything has gone according to plan, but that is to be expected for a DIYer working in a largely uncontrolled environment. In spite of the challenges, I was able to successfully capture a glowing petri dish and my friends, it was well worth the effort.
Update on the Goals of the Experiment
Let me review the goals and briefly update you on where I’m at with each one:
Replicate results from published research on the optimal growing conditions for maximizing luminosity of Panellus stipticus
I can say definitively that I was unsuccessful in replicating the two published research papers in their entirety.
Here’s a summary of what I’ve observed:
- The growth of my cultures have taken 4x longer than published
- I’ve been unable to see any light emitted from the liquid cultures
- The MEA petri cultures grew quicker and do emit light
* Prepare and inoculate PG (potato glucose) liquid culture
* Test my new custom lids with self-healing injection ports
* Test which jar labels survive the pressure-cooker sterilization
* Calibrate my cheap pH meter found on Amazon
All of these goals were successful! I made PG liquid culture, although as mentioned above, the expected growth rates did not match the published research.
My new custom lids with injection ports and 2-ply breathable micropore tape appear to be working with one caveat: when I shake the jars to mix and breakup the growth, sometimes a little bit of culture seeps through the tape. This is also problematic when I draw a sample from the culture with a syringe. The needles I use aren’t very long and after a good shake, much of the culture floats to the top requiring that I tilt the jar on the side while I draw in a sample. If I’m not careful, I’ll tilt the culture jar so far that it can leak from the breathing port covered in the micropore tape.
As for the labels, both the tape and marker labels survived the pressure cooker. While I prefer the label that is written directly on the jar with a marker because it’s less work to add and remove (it wipes off easily with a little alcohol), it’s prone to smudging with all of the handling that happens during a grow cycle. The label written and affixed to tape takes longer to make but is generally more durable over time.
As for pH, I did calibrate my cheap meter with the reference solutions and use it to create cultures at varying pH levels around the published ideal of 3-4. But, I’ve spoken to many folks who strongly recommend using a better meter as these cheap devices are prone to unreliable readings. Unexpectedly, the jars where I did not do any pH adjustments appear to be showing the fastest colonization. These were also the cultures jars that showed a baseline reading of 7.3 – 7.6 pH.
What Might Account for the Differences in Results
I’m obviously not working under highly controlled laboratory conditions, but I have a hunch about what might be causing some of the variations in my results vs the published research.
- My colonization temps have been about 10 degrees cooler.
- I used a cheap, probably faulty, pH meter.
- I adjusted the pH only before sterilization and not after.
- pH was just adjusted using lime juice concentrate as opposed to HCL (hydrochloric acid) and NaOH (sodium hydroxide). Maybe the citrus concentrate had other compounds that interfered with growth.
Quick Note on Tracking the Experiment
I originally intended to track all of the experiment data in a structured and organized way, first starting with a spreadsheet and then working my way into something I could more easily update and share on the site.
Unfortunately, I still haven’t found a great solution. I don’t know yet how to properly quantify growth in a liquid culture given the manner in which it grows. I also don’t have meter to measure luminosity. So even if I had a good way to track and share the experiment data, it wouldn’t be so useful without these two important variables. Nevertheless, resolving these challenges are on my roadmap to sort out.
Now, onto the fun stuff – photo updates!
I’ve decided to remove these members from the experiment. 1-A is definitely contaminated and the rest are either showing little to no growth or questionable growth altogether.
This experiment required that I create a liquid culture for the first time. Based on everything I’ve read, healthy fungi liquid cultures should be crystal clear with the exception of clearly differentiated fungal growth suspended in the solution.
If we compare these two jars, it’s clear to me that 1-A is definitely not healthy and is contaminated.
Let’s take a look at the remaining healthy liquid cultures.
Note how all jars show nice clear liquid substrate and coagulated clumps of the Panellus stipticus culture hanging out.
OK – now, recall from that first part of this experiment that I added some members: some grain jars inoculated from healthy cultures and a couple of plates. Let’s take a quick peak at how those are coming along.
As you can see, we have some decent growth in these jars. The grains I’m using here are what I have available in my area: winter rye berries.
Note how the jar says 5mL on 6/1 and then +10mL on 6/8. That is because I added extra culture after a week of not seeing any growth. My advice is don’t do this! I was impatient and not only did I lose the data from the 6/1 inoculation, I introduced another opportunity for contamination.
Luckily, this jar is looking good! This species exhibits a light cottony wispy-like growth on the grain.
The Challenge of Capturing Bioluminescent Light
There’s a lot of information on the net and many cool pics of bioluminescent mushrooms, but much of it is conflicting. Some things say the LC culture glows, some say it doesn’t. Same for cultures on solid mediums. Some say the glow can be viewed with the naked eye.
I’ve heard some people say that the glow is so bright, it can illuminate a hallway enough to act as a nightlight.
In my experience, the glow is quite dim. It was not until I created a dark room in my closet and stood in it at night in complete darkness that my eyes adjusted enough to see a subtle glow from one of the petri dishes.
The night I discovered that the petri dish was glowing, I was ecstatic. I called my wife, Cat, into the closet and we just stared in awe of one of natures gifts. It was incredible.
Once I knew that the dish was glowing, I became semi-obsessed with taking a decent photo. First, I tried taking the photo with a paid iPhone app that provides fine-grained manual controls not available in the native app. This is the best I could achieve using an iPhone 11.
If you look closely, you can see a little bit of light but the overall image quality is terrible. The high ISO generates a lot of noise.
So I switched over to an old DSLR with a 50mm lens and started working on which manual settings would give me a better pic. Here’s what the first round of attempts looked like:
When I saw these, boy was I excited! For the first time, you could clearly see the green glow of the petri dish with no image processing.
But I didn’t stop there. I knew that it could get better. Specifically, the light was still so faint and blurry.
A little more trial and error, spending upwards of two hours in a dark closet, anxiously awaiting the result of a new round of manual settings, when finally, a wonderful preview comes across the display:
The photo was taken with a Canon EOS Rebel T3i with a 50MM lens, manual settings of ISO 3200 f/1.8 with a 30-second exposure time.
I’m absolutely thrilled to have successfully cultured and captured a glowing mushroom on camera. I’m moving forward with fruiting this culture and capturing more amazing shots while also expanding my learning of science, mycology and experiment replication.
If you have any thoughts about how to address some of the challenges I mentioned above or you want to jshare any feedback, drop me a comment below or on the EverymanBio instagram page.
See you soon!