Growing Gourmet Mushroomsfor Market

GOURMET
MUSHROOMS
FOR MARKET
A Complete Guide to Making a Living
Growing Gourmet Mushrooms at Home
Gary Heferle, MLS ASCP

Owner, Fresh from the Farm Fungi LLC
©Copyright 2022 by Gary Heferle – all rights reserved.
It is not legal to reproduce, duplicate, or transmit any part of this document in
either electronic means or printed format. Recording of this publication is
strictly prohibited.
2 Heferle/Growing Gourmet Mushrooms for Market

This Book is Dedicated to my loving wife Addie.
Without her enduring love, energy, and guidance, I would not be the man I am today or in the position
to write this book. Addie has been by my side every step of the way from day one and I express my full
gratitude and love in these words.
I also dedicate this book to my mother, father, family, friends, mentors, colleagues and the entire
mushroom community for the love and support that has been overwhelming from the start of my
mycology adventure.
“MUSHLOVE”
Heferle/Growing Gourmet Mushrooms for Market 3

INDEX:
Introduction 5
Part 1 – the beginnings 8

The journey 11
Part 2 – Mushroom Cultivation: 18
Mushroom life cycle 18
Anatomy of mushroom farm 22
Overview 22
Two areas of mushroom farm (Lab and Grow room anatomy) 22
Cultivation Processes: 34
Agar 35
Liquid cultures 51
Grain Spawn 55
Bulk Substrate 64
Fruiting 74
Harvest and storage 83
Distribution 84
Business Compliance: 85
Morchella cultivation research: 87
Breeding Mushrooms the new frontier: 89
Germination techniques 89
Isolation and Selection 90
Hyperbreeding 93
Epilogue/Conclusion: 95

Introduction
Thank you for purchasing a copy of this book! This information has helped many mushroom
With the dawn of internet technology and ease farmers start from scratch, scale up, and
of distributing eBooks, I am very grateful you succeed - the earlier you start cultivating, the
chose to support our efforts. This book will faster you will be able to serve your local
cover the basics on how to grow gourmet community! Enjoy this mushroom journey and
mushrooms at home and make a living! From prepare to immerse yourself in the wonderful
diving into cultivation methods, using today’s and fascinating world of fungi farming!
common procedures, and digging into some
real-life business strategies these words will
provide you with the knowledge you need to
start your own mushroom farm. Many books
cover the technical aspects of growing
mushrooms, but this will be a practical guide to
starting out, choosing a cultivation method
that will work for you, and providing key
discernments into best practices for new
businesses. From sourcing substrates and
navigating the tough world of Health
Inspectors, Finding Clients and Flowing Market
cycles, the goal is for the reader to have a
global understanding of the craft mushroom
industry so they can make better choices Everyone has a talent to add…
starting off. Utilizing simple liquid cultures
instead of investing in costly laboratory The magnificent aspect about the new age
procedures, finding the right niche in an craft mushroom industry is the untapped world
exploding industry, and creating a unique of potential innovation. Fungi Farming is a
presence in the mushroom community was relatively new endeavor in the West.
how we were able to grow our mushroom farm
from a 4’ x 4’ grow tent in our basement to a
manageable, sustainable commercial scale.

Most mushrooms in the United States are Therefore, the mushroom industry is looming
produced in Punnett Square Pennsylvania in with efficiency problems, supply sourcing
giant warehouses with easy access to peat and issues and many other niches where great
compost. This is slowly shifting. The new craft improvements can be made. You, as a fellow
mushroom industry is being developed from human have unique perspectives, experiences
grass roots movements and from people and ideas that can help move this flourishing
growing in urban settings, as well as DIYers industry forward. Humans being human is the
fruiting from their garages and basements in a oldest and truest form of innovation, so
work-from-home concept. hopefully this book can at the very least spark a
new thought that could lead to a breakthrough!
The mushroom family welcomes all – craftsmen,
home makers, plumbers, welders, artists,
scientists, citizens, chefs, every person young
and old! The mushrooms welcome all – help
spread the mycelium and bring your knowledge
and skillset to the table!

PART I: The Beginnings
There are many great technical mushroom-grow guides available and plenty of informative
videos on the innerwebs, so one might be curious why I decided to write another mushroom
grow guide. To this, I say “why not?” There are countless perspectives in this industry, and I am
going to provide guidance to growing mushrooms from a laboratory heavy, efficiency focused,
and “MacGyver” Do-it-yourself perspective which I believe led to the ability to grow my farm
from scratch without taking out any loans and risking the farm, literally. For those who want an
all-encompassing mycology manual, I recommend Paul Stamet’s books “The Mushroom
Cultivator” “Growing Gourmet and Medicinal Mushrooms” “Mycelium Running”, Tradd Cotter’s
book “Organic Mushroom Farming and Mycoremediation”, Peter McCoy’s book “Radical
Mycology” – all of these are great technical reads and helped to build the foundation of my
mushroom cultivation knowledge. For those of you looking for a down-to-earth guide with
practical tools around the house or workshop, this is your instruction manual.
“Poverty Playbook”
I will also cover a few shortcuts that I have used and have developed from other DIYers in the
field. Many of these shortcuts can save a lot of up-front capital which can buffer costly
mistakes during the learning period and early phases of your farm.
Solo Cup Agar: How to get 50 petri dishes for under $5

Sawdust Spawn: Working with local businesses and spreading the mycelium (for free)
Recycled Grain from Distillery: Free supplements = more mushrooms
Home Made Fruiting Chamber: Get your cycle timing down for pennies on the dollar

Solo cup agar dishes (shown in pictures above) were used early on in our journey to save on
laboratory costs. Local sawdust, which was used instead of grain spawn in the early days, is an
inexpensive alternative to hardwood pellets to learn for pennies on the dollar.
These first grow rooms were fashioned from reclaimed plastic and tents, two by fours,
humidifiers and lights all purchased locally on craigslist for pennies on the dollar.
Getting by without an industrial autoclave:
Since the beginning of this journey, I have contemplated the need for a large industrial
autoclave to scale sterilization. This is one of the major fulcrum points for many businesses in
the industry, but finding a viable alternative would save the operation many hours of costly
repairs, huge initial investments, and regulatory hurdles. To achieve larger scale production
without a costly autoclave was and is still my number one hitch. My first sterilizer was a steel
drum with a false bottom that I stacked on top of a turkey frier and covered with bricks in the
back yard. I quickly learned about the need for a more controlled process to achieve high

standards of sterilization after a whole
batch of 50, 5-lb bags contaminated with
mold a week into the first run. All that time
and energy wasted – but since then I have
developed a reliable system using
atmospheric steam sterilizers ran on
standard 120V power and a garden hose.
Thanks to Eric Myers and Bubba’s Barrels
for the at-home DIY innovation system
which brought about the next best thing to
an autoclave. There are now set-and-
forget systems that can help easily scale
this essential process. The main cost after
sterilization is labor (plenty of elbow
grease on-hand is a must for mushroom
farming, at least starting out).
The first year I spent a bit of time dialing in

sterilization times. My MMX (Master’s Mix
50:50 blends of sawdust and soy hulls)
bags that I use today are either sterilized in
a pressure cooker at 15PSI/250F for 90
minutes and cooled in front of a laminar
flow hood or steam sterilized at 200F for
20 hours and cooled in front of a hood. The
purchase of the “Bubbas Barrel”
significantly sped that process and now we
run 40, 5-lb bags in two barrels
intermittently to achieve a cycle of up to
200 blocks/week. Each block produces an
average of 1.2-2lbs per first flush. Plenty
of mushrooms for the community!
Master’s mix bags being sterilized in a 55 gal “Bubba’s Barrel” – this helped save 16 hours per
week in labor and freed up much needed time for operations early on.

The Journey
The journey of Fresh from the Farm Fungi LLC officially began in the summer of 2018, but the
prerequisites leading up to the launch started many years prior. For those of you who don’t
know, I began my career in Medical Laboratory Science. During my college years, I worked at a
blood bank in Buffalo, NY and began to get a feel for laboratory life. Long shifts with efficient
schedules were a great way to become indoctrinated to sterile environments, regular
cleaning/decontamination, and precise inventory tracking and environmental monitoring.
These were my duties then and became the foundations and disciplines that led to a successful
mushroom grow cycle. You will hear it again and again throughout this book that cleanliness is
at least 80% of growing high-quality mushrooms and I stand firm by this statement. The other
20% is really good time management.
Gary, the author, making karyotypes on clinical rotation at Genetics Lab

(Buffalo General Hospital) 2013.

After graduating from the State University of New York at Buffalo with my bachelor’s degree in
medical laboratory science, I was eager to explore the country for new opportunities with my
wife Addie (girlfriend at the time) and a mutual colleague from our years in college. We took a
late spring trip around the country in my Jeep Wrangler, camping at national parks, exploring
the famous Red Woods, Grand Canyon, Rocky Mountains and rolling planes of the Midwest. We
ended up staying in Glenwood Springs, CO for a few nights and fell in love with Colorado. My
wife and I immediately applied for open positions in the area. Luckily, she was able to transfer
positions with her current employer after attending a national training program and I was
accepted to take a position at a local tissue bank in Centennial, CO.
The new position was an entry level lab technician job which was great. Better yet, it was a
weekend shift which allowed me to explore the front range mountains on my days off. These
regular hiking sessions triggered my curiosity for mushrooms after one of my coworkers shared
some stories about his elk hunting trip that fall. It was an unsuccessful elk hunt, but he did
stumble upon a better haul – a large flush of bright orange Chanterelle mushrooms at an
undisclosed location not too far from Denver. These mushrooms were very fragrant, colorful
and grew in heaps, less than a couple hours’ drive from where I was living. I was intrigued and
was inculcated into a whole new world of mushrooms. These “wild crafted” and “gourmet”
mushrooms were much different than the portobella and cremini I was used to seeing in the
Western NY Italian/Polish Cuisine of my upbringing. From that point forward, hiking was ruined
for me (not really), and my eyes were glued to the forest floor. I soon noticed ropes of mycelium
clinging to debris, lots of exotic fungi (coral tooth, boletes, and the famous Amanita Muscaria)
on my subsequent hiking trips. I have not stopped exploring since.
Being the green thumb that I am, I decided to test my hand at growing my own mushrooms the
very next spring. I have always had a vegetable garden to tend since as far as I can remember
and just finished building some raised beds at our suburban split-level house in Aurora. We had
a decent compost pile going in our small, fenced in backyard so I thought that white buttons
would be an easy starter mushroom. Boy was I wrong! I purchased some “mushroom seeds”
from Amazon. A week later I had my very first “spawn”. It was a tiny bag filled with about two
dozen oats that were dried up and covered in white. I had no idea what I was doing so I planted
them in some potting soil in a Tupperware container and waited. And waited. Finally, some
new growth spurred! At last – Some white cottony growth began to show up on the surface of
the damp soil! This took my emotions to a whole new high, which was quickly destroyed a day or
two later. The white fluffy spots began to turn green… Trichoderma appeared, and my heart
was ripped in half. This is the classic roller coaster ride in mushroom cultivation. My first
attempt was a complete failure and waste of $15.99. I dumped the green matter in the compost
pile, gathered my gardening supplies and put growing mushrooms on the back burner.

A few months after that experience, I heard an amazing lecture by the Father of Fungi – Paul
Stamets on YouTube. It was a talk on how mushrooms could save the planet, and it was very
convincing. This was late Summer 2015 and was the final push of momentum I needed to get
the ball rolling and fall DEEP into the world of fungi.
After that first failed outdoor grow, I was determined to succeed in my next attempt. I had been
cultivating Cannabis for some time (It’s Legal in Denver) so I had enough equipment to set up a
cheap 4’ x 4’ grow tent. I highly recommend starting here. Known as a “martha” setup on most
forums – this small-scale tent setup is the perfect size to maintain humidity enough for a couple
pounds of mushrooms per month while maintaining enough fresh air to get decent fruits.
**POVERTY PLAYBOOK **
The fruiting room is one of the most important areas while cultivating mushrooms (second to
the laboratory in my opinion). The main objective is to control temperature, humidity and fresh
air exchange while maintaining complete sterility or as close to sterile as possible. This can be
done by building a super high-tech class IV BSL laboratory with space suits and HEPA filters
built into the ceilings, or - it can be achieved with a tent, some inline fans, and DIY air filters.
Both environments will produce mushrooms, and the disciplined efforts from the cultivator can
out-perform superior setups. Some degree of innovation is important to rig a decent fruiting
room as well as hours spent looking through wanted ads, craigslist, and next-door – all
important tools for a new cultivator. When just starting, use your imagination and spare some
free time to build a fruiting room from scratch. It will help you understand the fundamentals of
fruiting and prepare you for when you need to make larger investments and more permanent
structures.

First grow tents come online – most material was sourced locally second hand.
One summer day in 2017, my wife Addie happened to stumble on a grow kit that was left out on a
front porch from a Craigslist ad. There was a blurry dark picture, and the description was
basically “come pick up my spent mushroom block that I wasn’t able to care for”. I frantically
drove across town in our Subaru and rescued the oyster grow kit from the trash that afternoon.
This would be the perfect, low-cost experiment to dial in the “new” fruiting chamber which was
a repurposed hydroponic tent. I placed the mostly-dried out plastic bag that was packed with
mushroom mycelium-covered saw dust in the tent and waited for it to show some signs of life –
low and behold! I got my very first oyster mushroom! It was an amazing day -and cause for
celebration! I remember slicing that cool, plump, perfect little oyster mushroom into slices and
frying it up with some butter! That was one of the best mushrooms I had ever eaten to that day.
I immediately hopped on the “shroomery” forum, researched all the “teks” and in the meantime
gathered up some basic tools to “clone” my first strain. This is still the same “Italian oyster” that
we grow today, breed with regularly and it is a vigorous one!

There were many more moments and stories that lead to the startup of our farm. Spending
countless hours fixing up our first house led to the resourcefulness of building with what we
had. The experience working in the BSLIII lab trained me in the ways of aseptic techniques and
long, systematic tasks associated with growing living tissue. Extracting cells with careful and
methodical procedures was critical for establishing a foundation to preserve our mushroom
strains long term. I thank all my colleagues, supervisors, managers, and teachers for helping
me develop the discipline to perform sterile tasks fluidly and efficiently and I will do my best to
instill the same principles throughout this book.
It took about a year and a half to save the $15,000 seed money which in turn funded the
purchase of the flow hood and lab gear that drove Fresh from the Farm Fungi LLC out of the
starting gates! I am thankful for the opportunity and support that my wife gave me from the
very beginning and without her, we would not be in the position we are today, so I can write my
experiences and share them with the myco-community.
The day finally came when I quit my corporate gig, and I was on my own. Well, sort of. In the fall
of 2018, I was blessed by a friend, mentor, and fellow mushroom farmer Anthony DiFranco of
Niagara Falls Mushrooms. My Dad was introduced to Anthony through his work when Anthony’s
wife dropped by to sell some fresh mushrooms to the crew. Serendipity was in that moment
because my dad knew I had just quit my stable, corporate job to begin this new endeavor. I
called Anthony up one morning and was amazed by the amount of knowledge he spewed in the
first 20 minutes over the phone. I hung up, booked a flight back to Buffalo and visited his
mushroom farm the day before thanksgiving that autumn. My aunt, who is also an avid farmer
and teacher in upstate NY, gladly brought me to tour the mushroom farm. We explored two
separate grow houses, one was a traditional Agaricus setup with long metal trays stacked
vertically and hundreds of buttons flushing out of the peat moss. The second, and newly
constructed mushroom house contained over 10 varieties of “exotics” as he called them.
Hundreds of “blocks” were hanging from meat hooks, stacked on galvanized piping, and placed
in a cooler for later use. It was like walking into Wonderland. Pink, Gold, Blue mushrooms were
around every corner and the bright, clean environment didn’t smell musty or damp like most
people (including myself) imagined. I got to witness what a real high-quality farm was and got to
ask questions non-stop for 4 hours. It was the final thrust of motivation that pushed our
endeavor to the next level. The day after thanksgiving I hitched a ride back to Denver by helping
a longtime friend move across country that same weekend, got stuck in a snowstorm, and had
plenty of time to think about my next moves. That tour of Anthony’s mushroom farm will
forever be engraved in my soul. It was the solidification to the dream. From that point on I was
a mushroom farmer.

Touring Anthony DiFranco’s Niagara Falls Mushroom Farm, Niagara Falls, NY 2018.

First Flush of “Italian Brown Oyster Mushrooms” cloned from an abandoned grow
block from craigslist.

PART II: Mushroom
Cultivation
Mushroom Lifecycle:
To understand the entire cultivation process, one must have a firm comprehension of
the mushroom life cycle. Mushrooms are strange beings, but they, like the rest of the
living organisms on this planet have one goal in mind. To reproduce.
Spores released
Fruiting and
Spores germinate
creating spores
Pinning/Primordia
Hyphae mate to
form mycelium

Most mushrooms begin their life as a single, haploid spore. That is, a spore which contains half
of the genetic material it needs to produce a mushroom. The spore is manufactured and
released from tissue formed on the mushroom underside, through the pores of the mushroom,
or in some cases, from inside the mushroom when it is opened to the environment.
The spores use hydrophobic forces to project themselves into the atmosphere where wind
currents and gravity help to spread them throughout the nearby environment. Once the spore
lands on a surface and finds a suitable environment, it germinates (elongates into hyphae and
begins seeking nutrients). If, by a miracle, the germinated spore is close enough to another
spore that has germinated and is a compatible genetic partner, those spores will intersect,
(some form clamp connections) and they will fuse to form one whole, diploid strand called
mycelium. The genetic information from the two spores fuse together and the mycelium now
can produce viable fruits. Mycelium is a rope-like cellular conglomerate highway that transports
nutrients and genetic material throughout the organism. The mycelium will multiply and
consume its surroundings by secreting enzymes to break down larger molecules like lignans in
wood. Once the nutrients are digested, the mycelium absorbs the micronutrients through its
cell wall. This digestion, absorption and expansion takes place continuously until the substrate
(material that it is in contact with) is completely covered or “colonized”, or until the mycelium
meets another organism, or until it meets the proper parameters to start reproducing. Hyphae
begin to prepare for fruiting by forming dense structures where mushrooms will likely thrive. In
nature this occurs just after a heavy rainfall for most mushrooms. Often called “Pinning”, the
mycelium will form hyphal knots when conditions are ideal (high humidity with evaporation,
cooler temperatures, light). During this stage, the mycelium pumps water into the “pin” and it
quickly grows and becomes a sporocarp, ascocarp or what we consider as the “fruiting body”.
The mushroom then presents itself to the world in wonderful display which triggers insects,
animals, and humans’ senses. This often causes the mushroom to be disturbed, picked, eaten,
repositioned in a way to allow for the next generation of spores to be released into the universe
and start the cycle all over. This process happens continuously in nature. The fundamental goal
of a mushroom cultivator when growing mushrooms for market is to control this process in an
efficient and cost-effective manner.

Lions mane mushroom during first flush – excellent high yielding strain for commercial
cultivation.

Local Colorado oyster mushroom that was collected from nature, isolated, and fruited
in-house.

Mushroom Farm Design and
Operations
OVERVIEW:
The goal of the mushroom farmer is to replicate the ecological system in a way that guarantees
successful fruits using as little resources as possible to create an efficient and reliable method.
This can be accomplished in as small of a space as a clear plastic tote, to as large of a space as
a multi-acre plot or warehouse. This book focuses on the intermediate size mushroom farm and
so this should be kept in mind reading further. I began my farm inside a 4’ x 4’ hydroponic tent
and was able to produce upwards of 10 pounds per week in this space. At the time of writing
this book I am moving our operations into a new 1200 square foot grow space with room to
expand and am planning to implement the use of an acre of hillside forest, as well as raised
beds to stretch the substrate as far as possible. My advice for a new farmer is to grow within
your means, start small, and build on those successes.
ANATOMY OF A MUSHROOM FARM:
Grow space is valuable and should be carefully thought out before constructing your mushroom
farm. My rule of thumb for space allocation is 40% fruiting, 10% lab and 50% incubation,
processing, and storage. Incubation space is the most versatile and should be adjusted for
based on what mushrooms are in production. Processing includes sterilization of substrates,
mixing substrates, picking/packing mushrooms, and storing mushrooms (walk-in cooler).
Separate storage space is needed for supplies and bulk substrates (the materials used to grow
the mushrooms). If shiitake or other long-incubating mushrooms are being grown, then this
ratio should be adjusted.
Other factors to consider while planning your farm layout are power supply, water supply, HVAC
(fresh air exchange), insulation/temperature, cleaning methods, composting space, and
processing space. The environmental balance of the grow room requires substantial power for
air circulation, humidification, and temperature control. Make sure you prepare for high
humidity. For example, don’t use drywall in your fruiting room. Instead, use materials like mylar
or other waterproof layers to help in the long term. Think about your climate.

Mushrooms prefer cooler temperatures, high humidity, and clean environment. If you can, build
your farm in a shady, low spot on your property where mushrooms might normally thrive. This
will help with cooling costs in the hotter months of the year and allows the farmer to work with
the natural environmental conditions. Good insulation is key for temperature control. If you are
in a cooler climate, you may want to consider geothermal heating or other passive ways to
control temperature that will not be costly (i.e., heated slabs, carefully designed exterior
surfaces to collect solar heat). Lastly, think about the surrounding areas outside of the grow
space. There should be ample room to discard spent substrates without causing issue.
Compost piles can get quite large and may create unique odors. Grains left over from
cultivation attract birds and rodents. Keep this in mind so not to disturb the neighbors or
anyone down wind. A great solution is to use compost bins/tumblers. Not only do they contain
the substrate odors as they break down, keep out pests, but they also help speed up the
composting process incredibly.
As stated earlier, the goal of the cultivator is to take the mushrooms natural life cycle and
replicate it in a controlled setting to produce high quality mushrooms in the most efficient way
possible. To do this, there needs to be a basic understanding of workflow, cleanliness, and
culture maintenance at the very minimum. Furthermore, the most successful operations long
term, will have the ability to continuously refine cultures and produce highly selective
mushrooms with features chosen for by selective breeding. This would take place in the lab
(10% total space allocation should be plenty). This space must be protected from contaminants
and spores generated from the grow area. Once contamination takes foothold, it is nearly
impossible to eliminate it without shutting down operations and sterilizing the premises. The
grow space will get dirty from normal use and should be easy to clean and separated from the
lab. The cultivation area should therefore be divided into two main regions, the Lab, and the
Grow Space. The placement of these areas, the workflow through these areas and the
personnel chosen to operate in such work areas should be planned out with cleanliness in mind.

Region 1: The Lab
The lab is a designated area in the facility where

mushroom mycelium is maintained, replicated
and where the highest degree of sterility is
critical. Regular cleaning, thoughtful design and
use, and extra layers of protection play a critical
role in the mushroom lab. The main goal of the
lab space is to protect the sterility of the
mushroom mycelium and growing substrates.
This can be achieved by using aseptic
techniques, having proper controlled air flow,
implementing strict procedures, and
cleaning/disinfecting regularly.
**Poverty playbook ** Nowadays, grow tents are commonly used for a DIY lab and
can be great for learning the ins and outs of lab workflow, space design and air
flow. You can purchase a 10’ x 10’ grow tent at your local hydroponic store for a
couple hundred dollars and it is prefabricated to receive inline fans, hang lights,
and is completely sealed which makes it a great space for implementing positive
pressure.

Classic hydroponic grow tent setup. Can be used for lab and grow space – check
out our amazon affiliate page for our suggestions for grow equipment and lab gear.

Birds eye view of lab design (can use tent with inline fans and filters to achieve
positive pressure)

As you enter your lab space, you should consider the entrance to the doorway the highest risk
for contamination. Sticky mats on the floor, positive pressure, air locks, air showers, locker
rooms are all tools to help minimize the transfer of outdoor contaminants to the inside of the
lab. As you continue into the lab, there should be a separate area for materials, cleaning
supplies, lab supplies, clean lab coats/fresh garments, gloves, hair nets/face masks and other
PPE (personal protective equipment). This area is annotated as the lab’s “vestibule”.
The next area of the lab past the vestibule is the “wet bench” area. This is where procedures are
performed, spawn and grow blocks may be incubating there, and data collection/notebooks
should be stored. It is a clean workspace and must accommodate sterile processes either from
the use of a laminar flow hood or proper cleaning procedures. Fan Filter Units (FFU’s), UV lights,
bleach spray bottles, disinfectant wipes, stainless steel countertops, glassware, dishwashers,
sterile consumables etc. are all fundamental parts to the wet bench area.
The furthest and cleanest area of the lab should be the culture library/cryopreservation storage
freezer and any biosafety cabinets/flow hoods for culture work. This is the first line of defense
for keeping the library clean and healthy and is the most important part of the lab. Positive
pressure can be maintained for protection which is a constant flow of filtered air into the space
to ensure that contaminants do not enter this workspace.
Region 2: The Grow Space
The grow space can be defined as the region in which mushrooms are fruited
(sporocarps are produced here), processed and stored. This region needs to be able to
maintain proper environment (high humidity, temperature control, lighting, air
exchange) while maintaining cleanliness, easy access to harvesting mushrooms and
cold storage to maintain viability of the produce after it has been harvested for
distribution. The grow space should have enough room to move in and out with large
amounts of substrate bags or “blocks” and should have an efficient layout for optimal
performance. This area requires a decent amount of manual labor, so designing the
space efficiently can be critical for scaling up operations. Make sure floors are slip
proof, make sure any drains are protected with covers and can handle high amounts of
debris. Epoxy flooring is great for this application. Ceilings and walls should be water
resistant. Shelving should also be streamlined and have as few crevices as possible to
keep cleaning easy and prevent mold from growing in unwanted areas. Lights should
be waterproof; any electric access should also be rated for outdoor use.

Just imagine the interior of an industrial dishwasher – this would be the ideal grow
space scaled up – (minus the giant spinning blades, but maybe that would be a cool
concept).
Outside of the fruiting area, there should be access to clean countertops for
processing mushrooms during and after harvest. Some municipalities require
commercial kitchen status for this work to be performed so check with your local
health department authorities. There may be specific requirements like proper
plumbing, cleanliness standards and space design requirements which all benefit the
grower and consumer by ensuring the proper handling of food. Separate buckets for
waste are a good tool for the processing area. Large vacuum adaptors can be utilized
on 50-gallon drums with castors (imagine a large-scale shop vac). All of this will help
make cleaning more efficient and prevent the infestation of insects, molds, and other
unwanted intruders. Doorways should be sealed completely to prevent rodents from
entering the space. There are few things more annoying than walking into the storage
area to find millet spilled everywhere and an overfed chipmunk sleeping graciously on
top of its newly discovered lifetime supply of feed.

AIR INTAKE/TREATMENT EXIT
(filtering/humidity/fae) (FURTHEST
GROW ROOM DESIGN
FROM LAB)
(BIRDSEYE VIEW)
Intake O2:
should be
Floor elevated
drain in position
ENTRANCE
(LARGE DOOR
FOR EASY
TRANSPORT OF
GROW BLOCKS
Exhaust CO2 (near
floor/lower levels of
grow because CO2 will
fall) * NOTE: OVERALL
WORKFLOW: FROM FRONT OUT
NEGATIVE PRESSURE
BACK TO NOT CROSS-CONTAMINATE
Birds eye view of fruiting room and air flow – note pre-treatment of air and
direction of workflow from one side to other (avoid crossing workflow back
towards lab)

Grow room with ultrasonic misting humidification system and PVC racks. Note the small
amount of surface area exposed to allow for easy cleaning and quick access to mushrooms.
Thoughtful Design:
I cannot tell you how many times I have seen the proverbial mushroom startup begin by writing
out numbers on a napkin, working backwards from huge overhead costs of giant warehouse
spaces and highly speculated, inflated prices of fresh mushroom sales. 99% of these
endeavors end up crashing and burning and the other 1% usually pivot into something much
more manageable. If you are planning to make a lot of money in a short period of time,
mushrooms are not the industry for you. However, there are some keys to success and one
CAN ABSOLUTELY make an excellent standard of living off the profits from a mushroom farm.
This result can be accomplished by sticking to a solid plan, staying within the manageable limits
of production, and building on successes.
One of the first stages of mushroom farm preparation is to create your fruiting chamber. This
can be as fundamental as a clear bin that locks in humidity, has a lid to replace “fruiting blocks”
or units of myceliated-substrate that fruits the mushrooms. A bin (called a mono-tub, shotgun
fruiting chamber, etc.) should be large enough to fit a 5lb bag of sawdust (at least 5 gallons but
preferably the biggest you can find). I highly recommend starting with a grow kit so you can at
least see a mushroom start from pinning to harvest. So, step one, find a bin – step two, get a
grow kit and watch it grow in the bin. Very fundamental.

From this point forward, let your imagination come to life. Search craigslist, your local
hydroponic store or your grandma’s basement for the next size up container that you feel
comfortable filling with mushroom grow blocks. Keep in mind that you can produce multiple
flushes from one bag, they need to be disposed of eventually, and it needs to be easy to clean.
Some common fruiting chambers I have seen are shipping containers, old tractor trailers,
hydroponic tents, greenhouses, hoop houses, and old school buses. Make sure you check with
your local permitting office to see what requirements, if any, are necessary to consider your
space a legal, functional business. It is sad to see a new farm invest lots of time and money
only to get shut down from the health department because it was not up to code.
Basements can make a great starting point for a mushroom farm (this is where we began our
journey). However, make sure you take every precaution to design your grow properly or you
can damage the infrastructure of the house and end up losing out bigtime and creating a huge
problem. Many people have seen the meme image of a poorly kept decrepit house with oyster
mushrooms popping through the floorboards. Don’t be that person. If working in a basement, I
highly recommend using tents or creating rooms within rooms to mitigate humidity exposure
to the rest of the space. It can be done successfully.
After you have successfully fruited mushrooms, you can start innovating your grow space.
Adding volume to the room will help stabilize environmental factors (humidity, temperature,
fresh air exchange (FAE), lighting, and cleanliness). Larger volumes can be achieved using a
larger grow tent or building a separate room (this should be made somewhat temporary at first
so you can learn the ins and outs before investing a ton of money and time).
GROW ROOM BASICS:
There are important environmental factors that must be controlled in the grow room. These
are humidity, temperature, lighting, fresh air exchange (CO2 levels), and cleanliness.
Humidity – Humidity must be able to reach 99.9% in a relatively fast amount of time for
when mushroom blocks are first introduced to the fruiting room. This can be achieved
using an ultrasonic mister, pre-humidity chamber, high powered misters, hydrated
perlite or in desperate situations, soaking the floor with a garden hose. Humidity should
be controlled after temperature adjustments because dew point comes into effect and
improper air treatment can cause large amounts of condensation or inefficiencies.

Temperature – Temperature must be able to reach cooler temperatures (low 50s to
upper 60s) on demand and maintain stability within a few degrees to produce
environments to induce pinning. This can be done with A/C and heating, fans, thermal
piping in floor, and air recycling measures to help reduce costs of temperature control.
Lighting – Lighting must be bright enough to light the room for harvesting and cleaning
and set on a regular interval for mushroom cultivation. A 16 hour on-period works well
for most varieties but can be adjusted according to the work schedule of the farm.
Lighting can be implemented with waterproof LED’s which are low cost and low heat.
Mushrooms are phototropic so they respond to light but do not utilize light like plants do
for energy.
Fresh Air (CO2 levels) – Mushrooms inhale oxygen and exhale CO2 like us so keeping
oxygen levels high and exhausting CO2 as it builds up in the grow space is critical for
healthy fruits. My rule of thumb is to keep CO2 below 1000 ppm. This is done by
replacing the volume of air the room at specific intervals. The exhaust should be
located on the bottom of the room because CO2 weighs more than oxygen and will
accumulate near the floor. Intake should be placed near the ceiling or above the
mushrooms so the air can drop down towards the exhaust. Often, a perforated tube will
be used to distribute air directly in the center along a tapered ceiling. This creates an
ideal air current, and the newer hydroponic tents incorporate this air flow design by
having the extra head space above the canopy.
Cleanliness – Mushroom harvesting is messy. Regular and thorough cleaning must be

performed to keep up with debris falling to the floor and preventing contamination from
occurring and spreading. Grow rooms should be designed for efficiency of cleaning with
waterproofing, minimizing surface area, and reducing dead zones in the space that
stagnant air will build up and promote contamination. You can reduce liability of a
contamination outbreak by establishing multiple chambers. This also allows the grower
to have the ability to clean empty rooms without disrupting grow cycles while still
maintaining production in another room.
Humidity, temperature, lighting, fresh air exchange and cleanliness of the grow space must be
accounted for to succeed long term. If one of these factors is out of balance, the grow will
suffer, so carefully design your farm accordingly.

The finishing touches on preparing your
mushroom farm will be designing the
laboratory. This can be introduced gradually
as one gets more comfortable fruiting
mushrooms in the grow chamber. The lab
side of operations is more technical,
requires a larger investment and is a steeper
learning curve. In the early stages, a new
grower should focus efforts on creating an
environment suitable to maintain fruiting
and obtain consecutive “flushes”. As the
grower progresses, the laboratory can be
utilized to its potential, and this will take a
new level of discipline in cleanliness and
workflow. Of course, there are exceptions
to this, but most new growers buy spawn,
inoculate sterilized grow blocks, incubate
them, fruit them, and then repeat. This
requires a flow hood, a clean tabletop, and
enough space to store the required number
of mushroom blocks. Oh yes, and a lot of
labor. Mushroom farming is a labor of love if
you haven’t figured that out yet.

CULTIVATION PROCESSES:
The cultivation process can be divided into three main phases: Isolation, Expansion and
Fruiting. Isolation is the process of obtaining and protecting clean, healthy mycelium. This can
be done by purchasing already made cultures, cloning mushroom tissue, or starting from
spores. Expansion is the process from taking clean, pure culture and growing it out so it can
be commercially viable. There are many techniques to do this, but the golden standard is to
make grain spawn, use that to inoculate grow blocks and then repeat over and over to create a
continuous “grow cycle”. Be prepared to maintain this level of production so plan accordingly.
The final process, fruiting, is self-explanatory but also includes harvesting, packing, storage
and distribution.
There are many ways to begin the grow cycle production. As mentioned earlier, the easiest
way to start is with a grow kit. You can also purchase isolated cultures of mycelium (plate petri
dish, slant, liquid culture) as well as spores (prints, or spore syringe in solution). You can also
make your own spore print, clone tissue from healthy fruits, or use pieces of substrate with
healthy tissue and begin from there. All of these are viable options to begin your mushroom
cultivation journey. The most important aspect is to maintain clean cultures. This can be done
utilizing a very important tool in mycology – the agar petri dish.
Pholiota nameko and Stropharia rugusoannulata growing on agar petri dishes respectively. (you
can purchase our living mycelium cultures on our ETSY shop FRESHFUNGI)

Agar:
What is agar? Pronounced “Ah-gar” by some and “Ay-gar” by others, agar is a very important
tool for the mushroom lab. The history of agar goes back to early Japan as it has a long record
of use in cuisine from that region. It is derived from red algae and is a gelatinous substance
that melts and rehardens to a Jell-O-like consistency at room temperature. It has the
capability to hold large amounts of water and nutrients, is soft enough to cut easily and thus
makes a great matrix for bacteria, fungi, and yeast to grow and thrive. In the late 19th century,
it was discovered that agar could be used in vitro (in a petri dish) to support microbiological
life. Louis Pasteur discovered that bacteria could grow on the surface and form “colonies”
turning the invisible into visible. He observed these bacteria to help prove germ theory. Out of
this science emerged the process of aseptic techniques, the widespread use of agar in the
microbiology lab, and eventually the practice of mycology (the study of fungi).
Side Note: Aseptic Technique – With the discovery of germ theory, the immediate
implementation of a practice known as “aseptic technique” took hold in the microbiology world.
Aseptic technique is a fundamental and critical process by which the technician or cultivator
protects the material in use from foreign contamination. In the early days, microbiologists
used oil lamps and candles to create negative air currents around the area they were working.
Modern labs use highly sophisticated filters that push clean air in a uniform or “laminar”
direction over the workspace to protect the area from contamination. Contamination can be
bacteria, yeast, mold, other mushroom spores, chemicals, and debris from the surroundings. It
is critical to understand that to succeed in these delicate processes, one must be completely
aware of the surrounding organisms that are not visible with the naked eye. It took a long time
for humans to comprehend this phenomenon, and for the mushroom farmer, agar is a very
helpful tool to establish this mindset. An organism sluffed off a skin cell that fell because of
gravity onto the surface of an open plate culture, will not reveal itself until a few days’ time.
Because aseptic technique was not used correctly in this instance, this organism can hide out
of site expand itself in the next phases, grow out later after the damage has been done and
wreak havoc on an operation. I have witnessed the emergence of mold (the infamous
Trichoderma) days after inoculating hundreds of bags of sawdust only to go back to square
one. It will happen, but you as a cultivator need to understand “why” and the best way to do this
is by using agar.

Left: Agar being used to QC liquid culture to verify health and sterility.
Right: Various phenotypes of oyster mushrooms growing on agar – note that each variant has
their own unique characteristics.
The practical use of agar in a mushroom farm, stems from its ability to filter mycelial growth
into two dimensions (as opposed to three dimensions in the wild or in a liquid broth). When a
piece of mushroom tissue is in contact with agar, it grows along the surface, which makes it
easy for the cultivator to observe physical features. The health of the tissue and any other
organisms that might be present in that dish can also be observed easily. This is important
because it exposes contamination and allows for the farmer to make clear decisions at the
fundamental stage of growth for the mushroom. Agar is also a key tool for germinating spores,
breeding mushrooms, expanding mycelium, quality control testing and cloning copies of
desirable mushrooms that are in production or from the wild. This topic is often glossed over
so I will dive deep into the power of Agar.
Agar Preparation: Agar can be prepared from a powder form which is hydrated, dissolved in
solution, boiled, melted, sterilized, and then re-cooled to form the gel that is stored in a sterile
petri dish or slant.

Left: typical petri dish used for production – we use quick dissolve agar along with standard
size 100 x 15mm plates.
Right: Slant culture used for long term storage – note the depth of the nutrient media which
allows for continuous feeding during refrigerated periods of time.
**Poverty Playbook ** “Deli Dish”- some restaurant ketchup cups come nearly sterile
and can be used instead of the common petri dish. This saves significant amounts of
capital early on and allows one to learn with agar without a large cost. You can also use
jars or glass plates which are more eco-friendly options.

Step 1:
Choose your agar recipe. My favorite is the gold standard Malt Extract Agar (MEA) followed by
Potato Dextrose Agar (PDA). The recipe should include a sugar source and other additives to
allow for optimum growth of the fungi. For a list of agar links check out our amazon affiliate
page – we prefer to use the “quick dissolve” lab grade products from Hardy Diagnostics (no
affiliation there).
Step 2:
Weigh out agar in a clean bowl or weigh boat for appropriate volume to be prepared. (Typically,
15-20mL of liquid agar is aliquoted per dish/slant – 500 mL should pour about a full sleeve of 25
standard sized plates.)
Step 3:
Boil water in a clean pot (filtered, tap and distilled work best) and mix in agar until it is
completely dissolved. Agar should look viscous at this stage and will be extremely hot.

Step 4:
Transfer agar into media bottle, or appropriate autoclavable pouring vessel.
***Poverty Playbook*** You can save old jars, wine bottles, or other PP5 containers to
use for this stage. To see if a plastic container is suitable for sterilization, look on the
bottom of the container for the triangle graphic common for recyclables. The number
in the middle of this triangle should be 5 or greater to be autoclave safe.
Alternatively, you can pour agar out into jars, glass slants or glass petri dishes in 15-20mL
aliquots and sterilize. This is known as “pre-pour agar” and can be done without a flow hood.
Often these containers will contain higher amounts of condensation as a drawback, but this
will not harm the fungi.
Step 5:
Sterilize media in a pressure cooker or autoclave for 20 minutes at 15psi/250F. The pressure
cooker must hit 15psi to properly sterilize. Some “instapots” do not obtain this pressure so be
aware of the tool you are going to use. Start the timer once the sterilizer reaches pressure and
temperature.

Left: 15PSI/250F is the point at which sterilization begins, set timer when pressure cooker
reaches this stage.
Right: Pressure cooker cooling in front of laminar flow to prevent contaminants from entering
system during cooling period.
Step 6:
Allow pressure cooker/autoclave to cool back down to atmospheric pressure. Remove media
bottles and place in front of a sterile flow hood - you can move the pressure cooker and place it
in front of the flow hood before removing the lid to prevent contaminants from entering the
now sterilized media as a vacuum will form from the cooling liquid. Some pressure cookers
prevent this phenomenon using backflow preventers so make sure you understand your
sterilizer.

Step 7:
Allow liquid agar to cool to around 130F (agar will start to solidify around 120F) and pour into
sterile dishes, tubes, or jars. *Use aseptic techniques to guarantee the sterility of the final
product*. THIS SHOULD BE PERFORMED IN A SEPARATE CLEAN AREA IN FRONT OF A FLOW
HOOD, INSIDE A STILL AIR BOX, GLOVE BOX, OR IN SOMEWAY TO MITIGATE CONTAMINATION.
Above: Media Bottles cooling before being used to pour (use laser thermometer to measure
temperature of agar as it cools to not contaminate freshly sterilized media).
Below: Technician pours petri dish and slant using aseptic technique (does not place hand over
dish/slant or in front of air flow to prevent contaminants from entering the system).

Step 8:
Allow agar to solidify in the vessel which should take around an hour. **Pro Tip** condensation
will occur when the outside air is cooler than the air inside the vessel. To mitigate this, the lab
should be warmer (70s), the agar could be poured as cool as possible before solidifying, and the
dishes could be cracked open in front of the flow hood as they cool off to allow escape of
steam.
Above: Agar cooling in front of flow hood – will become gelatenous in an hour or so.
Step 9:
Allow agar to remain unused for 72 hours or so to verify that the process was performed
correctly. This helps to ensure sterility prior to use. Plates should be free from growth until use.
Step 10:
To store, place in a sterile sleeve or bag, wrap with parafilm or allow to remain in front of a flow
hood at room temperature. This will keep for a couple months. For longer storage, place in a
refrigerator agar-side up to reduce evaporation. Eventually the agar will dry out into a thin film
which renders the media useless.

Below: Black Charcoal Yeast Agar – provides beautiful contrast of mycelium growing and
provides a buffer for pH swings during growth stages.
Malt extract agar – Coloring with food dye can help to distinguish variations of concentrations
of sugars for experimentation. Try to mix up your recipes to tailor towards your own strains.
Save sleeves and keep sterile so they can be

placed over cooled plates for storage and
protection.

Agar Recipes:
In general, Agar recipes contain three main components. The agar-agar
which provides the physical gel matrix for the substrate. Sugars and
minerals for the macro and micronutrients that feeds the mycelium in
vitro. My rule of thumb is about 12-14g of agar-agar per L and 20-24g of
nutrients per L.
(PER L)
Malt Extract Agar (MEA) 12g Agar, 24 g malt extract
Potato Dextrose Agar (PDA) 12g Agar, 14g dextrose, 8g potato flakes
Light Malt Extract Agar (LMEA) 12g Agar, 16g malt extract, 4g potato flakes
V9 Agar 1L V8 fruit juice, 8g potato flakes, 12 g Agar-agar
Black Charcoal Yeast Agar (BCYA) 12g Agar-agar, 20 g malt extract, 4g

nutritional yeast, 4g potato flakes, 3g activated charcoal
Water Agar 1L water, 20g agar agar
Grain Water Agar 1L grain water, 14g agar-agar
Dichloran Rosebengal agar with chloramphenicol (DRBC): 15g agar, 10 g

glucose, 1mL dichloran, .1g chloramphenicol .5mL Rosebengal 5g peptone,
1g potassium phosphate, .5g magnesium sulphate heptahydrate
Antibiotic agar (novobiocin) 20g agar, 10g sucrose, 10g lactose, 5g peptone
3g yeast extract, 20mg novobiocin

AGAR PROCEDURES:
In general, agar procedures should take place in your designated lab. They should be done using
aseptic techniques in an organized fashion in front of a laminar flow hood or still air box on a
clean surface. Stainless steel, glass, or granite counters should be used because they can be
wiped clean with alcohol or bleach. Consider your surroundings. You don’t want a shelf of plants
above your workstation that could be dropping mold spores onto your petri dish. Make sure the
lab is cleaned thoroughly before using agar. Start with wiping the ceiling, furthest inside the
lab, the walls from top to bottom, and work your way back towards the entrance of the lab. Any
debris in the air will fall downward because of gravity and if the petri dish is open, and an
organism lands on the surface it will start to grow and render that media useless. You can even
place open plates around the lab, see what grows and use that data to gather more information
on where some weak spots of your lab might be. The rest of the dust and debris will fall to the
floor, so make sure it’s a clean floor preferably concrete or laminate that can be mopped easily
with bleach or other disinfectants.
Wear a lab coat, scrubs, or special garments that are designated for lab procedures. These add
an extra layer of protection between your skin and the surface of the agar. Gloves, hair nets,
surgical masks, Tyvek sleeves, Tyvek suits, pressurized space suits with internal filters are all
additional layers of protection but ultimately it is up to the cultivator to utilize proper
movements, placement of tools and protection of the sterile materials to successfully perform
the following procedures.
While working with tools, make sure to not touch them to dirty surfaces. Use a tool stand to
rest tools on while they are not in use. Be aware of any “fomites”, or vectors that could carry
potential organisms into the workspace. The most common and dirtiest fomite is your cell
phone. It’s best to leave that out of the lab or disinfect it before beginning your work. The tool
of choice for most cultivators is the scalpel. You can purchase pre-sterilized blades that are
used for surgery and other aseptic processes on our amazon affiliate page. I recommend the
#11 blade as it is very sharp and easy to cut into mushroom tissue. Gloves should be washed
frequently with 70% isopropyl alcohol and breathing should be controlled when working in the
lab. Talking, singing, or breathing heavily in the direction of the sterile work field can release a
slew of organisms, so wearing a mask and avoiding these actions will help mitigate
contamination. If you drop a tool on the table, sterilize it and start over. If your hand slips and
gently touches the surface of the plate or the interior of the lid, start over. Aseptic technique is
the most difficult discipline of mushroom cultivation and takes a lot of self-awareness and
accountability on the side of the cultivator. The best way to learn is to make mistakes, think
about what went wrong and try to mitigate those actions in the future.

Nowadays, recording yourself on video doing work can be extremely valuable because if
something goes wrong you can go back, watch the procedure, and try to pinpoint any errors
that might have caused the issue. Understand that most, if not all the problems in the
mushroom growing system, can be traced back to the cultivator. If you see contamination, it is
most likely your fault. Swallow your pride and figure out what happened. Now, onto the fun
part!
Lab space cleaned and ready for use – note the gloves, lab coat, mask, sterilization pouches,
placement of cultures and slants for transfers. All work performed in front of laminar flow helps
to mitigate contamination.
Germinating spores: One of the fundamental procedures for utilization of agar is the
germination of spores. This is critical to the understanding of aseptic technique as one will
discover the nuances of working with microorganisms quickly on a petri dish with very little
cost relative to the entire operation. There are a few technical options for germinating spores
that will be described in the breeding section of this book, but the basic idea is that the
cultivator is taking special care to place microscopic viable cells onto the surface, allowing
them to germinate in a timely fashion while eliminating competition that would be present in
the wild. This can be done on water agar if the starting material is dirtier, or it can be done on
most nutrient agar types with the understanding that there will be a high chance of
contaminants present. Frequent observations and quick isolation will be necessary for the
next steps to isolate the mushroom tissue so be prepared with additional plates on-hand.

Spores germinating on agar petri dish. Each colony represents a new genetic variety and are
numbered as they appeared on the dish. To target haploid colonies, aim for one spore per dish,
this can be accomplished using serial dilutions (explained in breeding section).
Cloning mushrooms: Cloning mushrooms is the process of taking a sterile piece of tissue and
allowing it to recover and continue to grow on agar. Mushrooms are amazing creatures in that
they can reproduce an entire copy of the organism by separating tissue from any region and
allowing that to grow (except for gill or pore tissue because it contains spores which would not
be an exact copy). Tissue should be taken from the center of the mushroom where the tissue is
sterile, healthy and should be done using aseptic technique, so the cultivator doesn’t transfer
unwanted organisms to the agar along with the mushroom. To do this, carefully split the
mushroom in half ensuring that you do not touch the clean, split region. Use your clean hands
to split the mushroom because cutting it with a knife or scalpel blade could transfer the
organisms from the exterior to the area where you are trying to extract clean tissue. Use a
fresh, sterile blade for this procedure. Flame sterilizing a blade can work but this causes
deformities in the blade and greatly reduces its sharpness. Think about the best area to take a
sample from. If there are worm holes or dark, squishy regions, avoid taking tissue from these
areas. Cut into the tissue using a gentle sawing motion. You can create 2 semicircular cuts or 3
separate pyramid-shaped cuts so that all the cuts overlap, and the tissue can be easily removed
using the tip of the scalpel blade. Carefully transfer the tissue to the surface of the agar and
observe for growth. If the tissue turns brown or black, it most likely is contaminated and should
be discarded. It is important to take many samples at first to ensure you get a clean healthy
clone.

Sequence of cloning mushroom – open sterile tissue (near stem for this oyster), cut and place on
agar dish. Note the 3 pieces of tissue on the dish – more copies ensure higher success rates.
Left: Contaminated clone of an oyster mushroom – to get clean mycelium, one should collect
the next sample from a new dish or carefully take from a contaminated plate in the clean white
region of the mycelium. Avoid cutting or digging out contaminants – it will spread like wildfire
through the lab.

Right: Clean healthy mycelium taken from clone. Note the white healthy periphery of the
colony, dark white center, and uniformity. This is an ideal culture to transfer.
Cloning mycelium: Once you have established healthy mycelium from a clone or from spores,
you can clone mycelium onto other petri dishes or slants for long term storage. This is another
fundamental procedure for mushroom cultivation. Known as “agar transfers” one can replicate
hundreds of copies of the same mushroom for production, storage, distribution, and
observation of growth. It only takes one cell to colonize an entire petri dish so small segments
of mycelium can be transferred over and over to new containers. Slants are tubes with deeper
volumes of agar settled at an angle - ideal for long term storage. Transfer the healthiest
mycelium to slants so they can be capped and sealed in a refrigerator for months or deep frozen
in cryopreservation freezers for years.
Sequence of transferring a single spore colony to a new plate where it will grow out void of
competition.
Cleaning up mycelium: If you observe unhealthy growth or any other organisms growing on the
dish, the mycelium should be quickly transferred onto a new, clean dish. This should be done
with care, as opening a contaminated dish is a liability for that unknown organism to spread
rapidly throughout the lab. Transfer clean mycelium from the leading edge of the colony. This
is best practice because the center of the mycelium may have overgrown the contaminant in
question, thus hiding it from sight. The leading edge is easy to observe clearly and thus will be
less likely to hide other contaminants that could get transferred over to the new plate. Another
tactic that I discovered to mitigate contamination is a technique from Andrew Reed at “Mossy
Creek Mushrooms”. By implementing cuts on the surface of the agar in the outer regions of the
plate, it forms barriers for bacteria that mushroom mycelium can cross. The idea behind this is
that the mycelium can jump across the cut and outpace any contaminants that would grow
down into the cut. This is a cool way to utilize a third dimension on the surface of the agar and I
was blown away when I first saw this practice in use. It is much better than utilizing antibiotics
in my opinion and should be utilized instead of antibiotics because antibiotics can create
resistant bacteria if overused.

Utilizing a third dimension with the cuts in the agar give the mycelium an edge over potential
bacteria or yeast contaminants.
Quality Control: Agar can be used as a tool for quality control by the mushroom farmer. To
troubleshoot issues, one must have a clear understanding of where a problem (like
contamination) occurred. By using agar as a “timestamp” of cleanliness, one can discover where
a procedure went wrong. For example, if a cultivator just purchased a liquid culture from a
reputable vendor, inoculated a grain jar and watched as it became contaminated with
Trichoderma, they might be quick to blame the distributor for a poor product. If, on the other
hand, the cultivator had inoculated a petri dish with that same culture at the same time and
discovered that the petri dish came out clean without any signs of Trichoderma, they would be
able to conclude that either the sterilization process or the inoculation into the grain was
performed inadequately. That brings us to the next topic – liquid culture.

Liquid Culture:
Liquid culture syringes being packaged and sent for distribution. Because of the highly
nutritious nature of liquid cultures, every batch is quality controlled on agar to ensure sterility.

The advent of the liquid culture has changed the game for new mycologists. Liquid culture
allows for mitigation of contamination during grain spawn production and is significantly
cheaper than doing agar procedures. Prior to liquid culture (LC) one would need to grow out
mycelium on agar petri dishes, segment mycelium using aseptic techniques with an open plate
and inoculate grain spawn one wedge at a time. As you will find out in the grain spawn segment,
this can quickly become a tedious task requiring lots of focus and time with a large risk for
contaminants to enter the system. Liquid culture eliminates this amount of focus and energy
because it reduces the liability of contamination down to the opening of the needle and the
cleanliness of an injection port. However, in the example of the contaminated spawn jar
described in the quality control section, it is very possible that the injection port contained
spores from Trichoderma on the surface which were carried into the bag because of poor
practices. It isn’t 100% fool proof but if you take the time to understand the system, it is a
remarkable innovation. Liquid culture can be 100% effective if the needle is kept sterile, the
injection port is cleaned correctly with alcohol and the culture was clean to begin with.
Apart from the great reduction in contamination risks, liquid cultures can also greatly increase
the efficiency of spawn production. Because a liquid is a 3-d matrix compared to agar (2-d
matrix), the mycelium can rapidly expand and colonize a broth in a matter of days, or hours in
some extreme cases. This time saving process quickly adds up and frees up the cultivators’
time to focus on other tasks which are extremely important early on.
With every great innovation, there is a cost, and for liquid culture, the major drawback is the
fact that it can hide contamination very easily. For this reason, it is very important to perform
quality control on every batch of LC. This can be done by taking a sample from the stock
solution with a syringe, placing a drop onto agar and watching for abnormalities. Another quick
screen is the “turbidity” test. Any bacteria will quickly outpace mycelium and the solution will
turn cloudy. If a liquid culture broth looks cloudy, smells pungent or is frothy, it is a good
indication that something entered the system and should be tested or discarded. Apart from
this, there is no way to know if the sample is pure or not, so it should always be tested on agar.

Turbidity test: note the center liquid culture: Graphics cannot be discerned behind
the middle bottle while the one on the right can clearly be seen through and the
letters are clear. The middle jar is likely contaminated.
Now that we talked about the powerful attributes of liquid cultures and the major drawback, we
can cover how to prepare and grow one out. There are many complicated recipes out there, but
for me, the standard 4% Honey water works best. You can also use Karo corn syrup which is
usually filtered and clear so it is a good starting point for new mycologists. If you use honey, be
aware that some raw or natural honey will have debris. If this bothers you, get Bjorn’s Colorado
Honey from Boulder. Pontus is a great multi-generational beekeeper and takes great care to
provide a pure, natural filtered honey. This is my go-to nutrient source for LC.
Prepping liquid culture is easy, simply add 4% weight/vol of honey/karo or other pre-mix to
water, mix thoroughly, place a stir bar, glass marble, or stainless-steel screw into the jar and
sterilize for 20 minutes at 250F/15PSI. Once the stock solution has been sterilized, let it cool in
front of a flow hood or in a still air box. It is important to let it sit at room temperature for a few
days prior to use. This is another form of quality control because if there is any growth prior to
inoculating the mycelium of choice, then it can be verified that the sterilization was done
incorrectly. If you inoculate the mycelium too quickly and it contaminates, you will not be able
to tell if it was from the syringe, from the original solution being not sterile or from the actual
injection process. Patience is key here.

Once you have a clean and sterile solution, now you can introduce the mycelium. If you have a
flow hood, you can cut a piece of clean mycelium from a plate, carefully crack open the lid of the
liquid culture and aseptically transfer a piece of agar into the solution. This is the riskiest way to
inoculate a liquid culture but can be done correctly with proper aseptic technique. You can also
use a needle to take a small biopsy of the mycelium from the agar using liquid to fill a hole in the
dish and scraping mycelium into the hole and aspirating into the syringe. Then inject into the
fresh broth. You can also expand a current syringe by injecting into new broth. Get creative, use
your imagination, and bring your own skills to the table.
After the mycelium begins to grow, observe for any abnormal coloration, smells, or cloudiness.
Once again, you will need to test on agar to verify the sterility and health of the liquid culture.
The only way to know for sure is to test. If the plate comes out clean and the mycelium looks
healthy, you are ready to move onto grain spawn.
Observing healthy growth of liquid culture.

Grain Spawn:
Local oyster mushroom mycelium leaping off agar onto oats
After a culture has been cleaned up, expanded, and stored on agar (or LC), the next step in the
journey to producing delicious mushrooms is to grow it on grains. This achieves a few things.
One, it brings the mycelium out of the two-dimensional surface of agar or small container of
sterile liquid media and brings the mycelium to a three-dimensional matrix with air spaces and
various complex nutrients. Two, it allows for efficient expansion of mycelium into a larger bulk
substrate once the mycelium is fully colonized in the grain spawn. Three, it provides some
amount of nutrients and water to the overall mushroom substrate. There is also a risk for
contamination when using grain spawn so proper aseptic technique must be followed when
performing these procedures.

What Is Grain Spawn?
One of the challenges of producing high yielding mushroom blocks is the balance between
nutrient content, moisture content, and the speed at which the mycelium colonizes the
substrate. If the nutrient content in the final substrate is too high, contaminants can quickly
take over and out compete the mushroom. If the Moisture content is too low, then growth will
be stalled and if it is too high, water pools up in the grow block and allows for anaerobic bacteria
to out compete the mushrooms as well. The perfect balance between nutrition, moisture
content and the acceleration of growth from the mushroom mycelium can be achieved using
grain spawn. Grain spawn is simply a nutritious vehicle for mushroom mycelium to go from in
vitro, to bulk substrate efficiently in a way where optimum growth can happen. Grains are used
because they naturally can soak up a lot of water inside the grain while protecting the nutrients
from contaminants because of its natural shell. Mycelium can break through the shell
surrounding the grains using enzymes and access the water and nutrients more effectively than
contaminants such as bacteria. This gives the mushrooms an advantage in performance, gives
the mushrooms adequate nutrition and water, and in the process of inoculations, it spreads the
mycelium evenly throughout the bulk substrate to increase colonization speeds.
Grain spawn inoculated with agar wedge – note the placement along the glass for visibility.

Types of grain spawn:
Different types of grains (left to right) Millet, Corn, Rye, Oats, Wheat – Top is hydrated.
Various grains can be used for the purpose of creating grain spawn.
Oats: My favorite of grains for starting commercial production is oats. Because of the price
and resistance to contamination, it makes a great option for beginners. Oats have a harder,
leafy shell than most grains so they are more resilient to contamination outbreaks. The
downside is that they hold less moisture, so it takes a more resilient strain to jump off agar onto
oats.
Wheat and Rye Berries: Wheat and Rye can also be used for grain spawn. These two grains are
very great at holding water; however, they are sensitive to overhydration and can create a
sticky mess if overcooked. This can lead to contamination, so err on the dryer side when
working with Wheat and Rye.
Popcorn: Popcorn is a cheap and effective grain to be used for grain spawn. It has moderate
difficulty hydrating but holds water well once hydrated. It has larger cornels than all other
grains which makes it easy to mix but also, it has less inoculation points so it may take longer to
colonize than other grains.
Milo, Millet and Quinoa: These grains are very small in comparison to the other grains described
and will have many inoculation points when inoculated into a bulk substrate. For this reason,
they can colonize substrates rapidly. On the downside, the smaller grains hold less moisture,
and are susceptible to over hydration. There is a fine balance when it comes to cooking Milo,
Millet and Quinoa.
***Poverty Playbook *** Sawdust spawn: Sawdust can also be used as a “grain spawn”.
The downside is that it does not hold as much moisture as traditional grain spawn. This
can be corrected by adding a small amount of wheat bran. Sawdust is very inexpensive,
can be selective for mushroom mycelium over other contaminants and is a great way to
learn the in between steps without losing your shorts.

Left: Sawdust spawn – locally sourced hardwood sawdust growing local oyster mushroom.
Right: Lions Mane Mycelium leaping off agar onto oat sawdust – note the spindly mycelium.
Grain Spawn Prep: The goal for preparing effective grain spawn is to hydrate every grain to the
maximum water holding capacity while leaving the outer shell of the grain intact and relatively
dry for the introduction of mycelium. The target for water content should be around 60%. This
can get tricky especially with the smaller grains like millet that will hydrate more rapidly than
the larger grains like corn.

Cold hydration: One way to hydrate grains is to pour them into a bucket, cover adequately with
water and let them soak overnight. This method requires very little energy but does take a long
time and risks some of the grains sprouting. I usually do this at the end of the day and process
them first thing the next morning to prevent the grains from sprouting. Once the grains have
soaked in the water, they should be expanded. There are a couple of options at this point. You
can use a sieve and pour off the remaining water and then scoop the now hydrated grains into
jars or bags for sterilization. You can also bring them up to a boil at this point and quickly take
off the heat and pour off the excess water. The heat at this point is not for killing
contamination. It is to generate steam and create a dryer exterior surface on the grains which
helps prevent pooling in the container. Once the grains have been transferred to their
appropriate vessel (glass jar with lid or unicorn grow bag with 0.2-micron filter patch for grain
spawn) they can now be sterilized. Sterilization times vary with the volume of the load, but you
want to ensure that every grain reaches 250F/15PSI for at least 20 minutes. This means that for
the grains in the center, it can take longer to heat. My rule of thumb for grain spawn is to
sterilize for an hour for 20 lbs of grains (4, 5lb bags will fit in a 23qt. pressure cooker). Let the
pressure cooker cool slowly in front of a flow hood, and carefully remove the grain spawn. If lids
are loose, tighten the lids and place in front of the flow hood to cool. As the grains cool, they will
create a vacuum which can pull in contaminants, so I strongly recommend using a flow hood
from this point forward. If you are using unicorn bags, make sure you don’t touch the inside of
the bag when you are removing from the pressure cooker. The bag will naturally create a
vacuum if folded correctly and will seal itself until use.
Simmer hydration: If you are in a time crunch, you can also use the simmer preparation
technique. The purpose of this method is to speed up hydration time and at the same time it
makes a dryer spawn because the heat from the simmering produces steam which evaporates
quickly from the outer shell of the grain. Simply fill a kettle with grains, cover adequately with
water, bring the grains and water up to a boil. Turn off heat and cover for about an hour, making
sure you don’t see any grains exploding. Aim for 60% moisture content (this can be
approximately calculated by dividing the weight of the hydrated grains into the weight of the
dry grains and multiplying by 100 – there will be a deviation if the starting grains were moist, to
correct this dry them in an oven before weighing them for this equation). Once the grains have
hydrated for about an hour, remove them from the kettle and pour off the hot water.

***POVERTY PLAYBOOK *** The grain water that is poured off from this hydration
makes a great nutrient source for agar. Simply mix with 14g/L and sterilize for a cheap
and effective agar recipe. Now that the steam has evaporated the water from the
outside of the grains, you can load into the jars or bags and sterilize for about an hour at
250F/15PSI.
No Soak, No Simmer Hydration: This technique is the trickiest preparation for grain spawn and
needs to be adjusted each batch because of the initial moisture content of every batch of grain
will be slightly varied. It is the fastest preparation. Simply place a specific volume/mass of
grains in the jar or bag dry. Pour a specific amount of water into the bag or jar and sterilize. As
the grains sterilize, they will hydrate and expand so account for this extra expansion in the jar or
pressure cooker if using bags. As a rule of thumb, I start with about 2L of grains and 1.5L of
water. Do a small test batch first and adjust from there. I have saved significant amounts of
time doing this once it got dialed in.
Inoculations: After the grains have been hydrated, sterilized, and cooled, they can then be
inoculated with mycelium. It is a good idea to leave a “blank” jar or bag aside during this process
to quality control the sterilization times. Ideally, that jar or bag should remain sterile indefinitely
until mycelium is introduced. If the grains contaminate, you can go back and adjust your
sterilization times. If the grains remain clean, then you know you did your work correctly.

Grain jars cooling in front of laminar flow hood – these grains were hydrated, sterilized and now
are ready for inoculation.
Inoculations should be done in front of a laminar flow hood. Carefully open the jar or bag in the
direction of the air current and carefully cut a wedge of agar, place over the opening and let
gravity drop the myceliated wedge into the new food source. You can also inoculate the grains
with a liquid culture. Mix the culture syringe, clean the injection port with alcohol and carefully
inject the needle into the port. 1-2 cc is plenty of volume to get the grain spawn started but
adjusting the volume of liquid culture can help speed up colonization. Just note that as you add
more liquid inoculum, it will affect the overall moisture content of the grains. You can adjust
the hydration prior to this point to allow for more LC to be inoculated at this stage, thus
decreasing colonization times. It’s a balancing act.
Once the mycelium has been inoculated, close the lid, or seal the bag with a heat sealer. Mix the
jar/bag carefully and keep the agar wedge in view if you used agar so that you can observe
growth. The mycelium should begin “leaping” onto the grain after a couple days. Once the
mycelium has fully covered some grains, you can shake the bag and break up the grains to
evenly distribute the mycelium. This can cause some delays in growth as the mycelium
recovers but ultimately can speed up colonization overall because the mycelium will be more
evenly distributed. During this period, observe your grain for abnormal growth, slimy or greasy
spots which could indicate contamination. If the grain spawn stalls out or stops growing,
chances are that the grains were not hydrated correctly or there is a contaminant. If all goes
well and the inoculation was clean, the grains should be fully colonized in about 10 days to 2
weeks. From here one can inoculate more grains, test on agar to ensure sterility or move on to
inoculate bulk substrates.

Completely colonized grain spawn jar ready to be inoculated onto bulk substrates. One half pint
jar is sufficient to colonize one 5 lb grow block.
Bulk Substrate:
Bulk substrate – The “nutrient sponge” for growing mushrooms.
After grain spawn, mushroom mycelium should go into its final substrate, which is designed to
hold water, provide the adequate nutrients to fruit, and prevent contaminants from attacking
the mycelium, especially once it is exposed to a non-sterile fruiting chamber. For most culinary
mushrooms, hardwood substrate works adequately. Oak is the preferred hard wood, but I have
also used elm, maple, cottonwood, ironwood, (walnut – should be avoided because of
antimicrobial properties) cherry and other local timber with great results. Softwoods like pine
contain antimicrobial resins and can inhibit the growth of the mushroom. Straw, Coco Coir,
Peat Moss, and other faux composts are also common substrates to grow on. They hold large
amounts of water and are resistant to contaminants.

The structure of bulk substrates should be diverse and allow for adequate gas exchange
between particles while simultaneously holding enough water to produce mushrooms. Unlike
traditional farming, where water is irrigated into the soil, water is forced into the bulk substrate
one time, prior to inoculation and must remain in the substrate by being placed in a humid
environment through the duration of the fruiting process. (One exception is shiitake which can
be rehydrated by submersion after a tough layer has formed on the surface of the mycelium).
The high humidity of the fruiting environment reduces evaporation and maintains the water
content in the substrate until the mushroom can push that water into its cells and blow up the
fruiting body like a water balloon. The more water holding capacity, the higher yields in general.
Hand-mixed blend of hardwoods getting scooped into unicorn grow bags prior to sterilization.
Each bag or “grow block” contains 5 lbs of hydrated sawdust substrate – ideal food source for
gourmet mushrooms.

Top left (sawdust produced from a “wood wiz” CNC machine) – ideal medium size texture
Bottom left (sawdust “curls” produced from a wood planer) – good large size texture
Right (sawdust from sander) – great small size texture

Dry bagging vs wet mixing:
There are two progressions of thought when it comes to preparing bulk substrates. One, is
aliquoting out the dried ingredients into mushroom bags, and storing them until use. This
creates a uniform product ready to be hydrated and sterilized. Think “pancake batter in the
box”. The other method would require a bulk mixer or “ribbon mixer”, and the substrate would
be hydrated before being aliquoted into bags. Think “cement truck”. The second is more
common commercially and saves time but requires more capital, equipment, and storage space
for larger volumes of bulk substrates.
For learning, I believe it is important to mix small batches of substrate to understand the water
holding capacity of various materials. Hardwood Fuel Pellets have become the golden standard
for small scale craft mushroom production and are great because of their consistency, but one
should understand the scope of possible water holding capacities to really see the value that
pellets have provided the industry.
“Master’s Mix”: Credit to Earth Angel Mushroom’s owner TR Davis, Master’s Mix is a revolutionary
substrate because of its water holding capacity and perfect blend of nutrients to support a
wide variety of mushrooms. Master’s mix is 50:50 blend of oak fuel pellets (used to fuel wood
burning stoves and very accessible from hardwood stores) and Soy Hull Pellets (a common feed
that is made from the hulling of soybeans) – an agricultural byproduct.
Bulk Substrate Prep:
To prepare a masters mix bag, simply put equal amounts of HWFP and SHP into the mushroom
grow bag, hydrate to 60% moisture content and sterilize. We use 5lb “14A unicorn” grow bags
from Unicorn Bags for our bulk production, but it is common to use 10lb bags as well. This just
comes down to space, variety, and the strength of the grower (mushroom blocks get heavy
when you shake them during inoculations). We use volumetric measurements for the pellets
and find that this was consistent enough – 1L HWFT, 1L SHP, 1.8-2L Water per 5lb bag.
Once your mushroom block has been prepared, it must be sterilized. Sterilization times depend
on the overall mass of the batch being processed. I started by sterilizing 4 blocks at a time for
90 minutes in a 23qt presto pressure cooker. I now use the famous “Bubba’s Barrels” and pack
40 5lb blocks per run and let them cook overnight.

Grow blocks cooling off after being sterilized in a pressure cooker for 90 minutes at 250F/15PSI.
Inoculations: Once the bags have been properly sterilized, carefully remove them from the
cooker and place in front of a flow hood to cool. If using unicorn bags, they will naturally create
a seal as the bag compresses from cooling. Like in the rest of the phases of processing, you
should leave behind a bag to prove that the sterilization run was performed correctly. The bag
should remain free of growth if it sits to cool in front of a flow hood.

Grow blocks ready for inoculation – they have been cooling overnight in front of the flow hood.
Once the bags have been cooled, they can be inoculated with grain spawn, sealed with a heat
sealer or clip, mixed thoroughly, and moved into incubation. The whole purpose of the grain
spawn is to spread the mycelium evenly in three dimensions throughout the substrate to speed
up colonization times. Grain spawn should be carefully poured into each bag (about 1 pint per
10lb bag is sufficient or ½ pint per 5lb bag. Inoculation volumes can be adjusted to speed up or
slow down colonization times. This is critical for establishing a routine grow cycle). Aseptic
technique is important during this stage. Avoid using extra tools like spoons or scoopers if
possible. These are a potential “fomite” and can be excluded with good pouring practice. If
using a bag, sterilize the outside of the bag with isopropyl alcohol and use a sterile blade to cut
a corner of the bag open, or use scissors that have been sterilized and are specific to this
procedure. Use the hole in the corner like piping frosting and inoculate the bags without
touching the interior. Make sure you fill your bags with as much clean air from the flow hood as
possible before heat sealing as this will make mixing much easier. Mix the bag thoroughly
making sure to spread the grains evenly throughout the substrate.
Inoculation sequence being performed in front of a laminar flow hood.
***Poverty playbook *** Instead of using grow bags, try using 5-gallon buckets with 1/2-
inch holes drilled into the sides or other reusable plastic containers to fruit. There will
be a learning curve during this period, and it might help to reuse containers at first. The
drawback is that they will need to be cleaned thoroughly with bleach and the substrates
will not be completely sterile, but you can replace the master’s mix with pure sawdust
or straw which is a more selective media. Simply hydrate the sawdust or straw in a
clean area and layer intermittently with grain spawn in the bucket (think lasagna
layers). Cover the bucket with the lid and wait for the substrate to colonize and you will
get fruits in no time. This works best with oyster mushrooms and is a low cost, low risk
way to watch grain spawn colonize a substrate. You can also use spent coffee grounds
as a supplement. Use your imagination and bring your skills to the table.

Once your bags are inoculated, sealed, and mixed, place them on a sturdy shelf or rack and
observe their growth regularly. Look for abnormalities or foul odors which could indicate
contamination. If a bag is suspect, pull it from the shelf and place it in quarantine. If it continues
to lag, discard the bag. Incubation should take place in a clean environment and should be
monitored for temperature. As the bags begin to colonize, they release heat so spacing is
important in this stage. The exothermic reactions of mycelium consuming the substrate
creates an environment where competition flourishes, therefore aseptic technique is so
important during the inoculations. The bag should be fully colonized in about 10 days to 2
weeks. If growing shiitake, there should be a second phase of colonization where the block
grows a thick brown skin or “bark”. This could take an additional 4-6 weeks. Once the bags are
colonized, they can be moved into the cooler, damp fruiting area to begin their final stage of
producing mushrooms.
Grow blocks ready for inoculation – they have been cooling overnight in front of the
flow hood.

Bulk substrate bags fully colonized and ready to go into fruiting.
*Side note* During the time of writing this book, I am experimenting with smaller scale
Agaricus spp. cultivation. You can grow Agaricus mushrooms in bags like the way that
is described but it requires a casing layer of nutrient deprived substrate about an inch
thick after inoculation and a more finicky pasteurization of the substrate compared to
the sterilization process described.

New variety of golden oysters fully colonized and ready to go into fruiting – note how
the bag inflated during colonization.

FRUITING:
Mushrooms fruiting on the shelf of this home-made fruiting chamber – environment is dialed in.
Fruiting fungi is by far the most satisfying step in the whole cultivation process. Watching your
own mushrooms reveal their identity to the world after imagining them developing from
microscopic spores, carefully feeding them the right nutrients and water, caring for them for
weeks, and finally seeing your efforts payoff is extremely rewarding. There are some key
environmental factors that the cultivator can control which will maximize the yield, quality, and
consistency of fruits from every grow block produced and we will cover them in this section.

Environmental factors:
Humidity – Once again, I stress that humidity levels must be able to be quickly and easily
controlled. Ranges are from 99% for pinning to 80% rH just before harvest. The common
system I use implements ultrasonic misters, humidstats (humidity controller) and inline fans.
The mister sits inside the bin which is filled with water through a float valve to maintain the
level of water. Inline fans push the humidified air from the inside of the bin out a pipe into the
grow space. The humidstat keeps control of the humidity digitally by turning fans on and off
when the humidity rises and falls. The exhaust runs continuously outside of the building to
create overall negative pressure.

Temperature – Temperature should be cool and manipulated prior to humidification. If the air
is cooled after it is humidified, dewpoint fluctuations can cause the water to condense on the
walls and floor and it will become a vicious cycle of humidifying and cooling the room which is a
significant waste of energy.
Fresh Air Exchange – (FAE) should be able to maintain a level of CO2 below 1000 ppm.
Mushrooms exhale CO2 and inhale O2 and when they are deprived of oxygen, mutations and
less-than-ideal mushrooms form. Exhausts should be placed near the floor because CO2 falls
and intake/fresh air should be expelled above the mushrooms because the cooler, humid air will
fall towards the floor as well. A systematic circulation of fresh air can be achieved in the grow
room with careful planning, placement of intake fans and circulation of air using fans and
dampers. Continuous monitoring of CO2 with a meter is important because once mushrooms
develop elongated, they will not correct themselves even when fresh air is corrected.
Lighting – Lighting should be sufficient to read this text. Lighting is less important than other
factors but should be able to be controlled for times during harvest. I recommend at least 12
hours of light and rest times to give the mushrooms a proper growth cycle. Blue spectrum light
should be considered for more temperamental strains like cordyceps militaris.

Evaporation – Evaporation occurs when the bags are opened or sliced and the moisture from
the substrate leaves the surface and enters the grow room. Evaporation can be controlled by
the relative humidity of the room and the timing of the bag slice. In nature, mushrooms emerge
after a rainstorm. Cooler temperatures, higher humidity and the evaporative effects all
contribute to pinning, so this should be considered by the cultivator as well.
Fruit body development – As the mushrooms emerge, they will be extremely delicate. Even a
single water droplet landing on a baby pin set can have detrimental effects on the mushroom
later. Those pins can develop mutated fruits, or abort all together. Careful design of the
shelving can prevent water droplets from hitting baby pin sets. Reducing the humidity as the
mushroom matures can also make the fruits better quality. If the humidity is too high just
before harvest, the mushrooms will be too mushy and suffer a shorter shelf life because of this.
Therefore, it is best to taper off humidity just prior to picking.

Spore dispersion – Mushrooms should be harvested just before spores are dropped. However,
this is sometimes not the case and spores will be released readily into the grow room. This can
cause problems with spore accumulation in the ventilation, it can also cause health risks for the
cultivator, so proper cleaning and PPE should be implemented to handle the spore load of the
operation. Fans should have easy access for cleaning and replacing. I like to say that “one is
none, and two is one” when it comes to inline fans. I like to keep a spare on hand so that when
spore loads build up, the swift replacement can keep oxygen flowing and the dirty fan can be
thoroughly cleaned and dried for replacing the next time.
Initiating pinning: When mushrooms fruit in the wild it is usually after a significant rainstorm.
Think about the factors that happen when it rains. First, the temperature of the air drops and
wind might pick up a bit. Next, the rain comes, soaks the ground for a while and begins to
absorb into the earth. Trees, bark, and other foliage quickly soaks up any access moisture and
evaporation soon begin to occur. The sky eventually clears (if you live in Colorado, it’s usually
soon) and the sun heats up the soil and increases evaporation. Micro-climates near the soil and
forest floor begin to form and mushrooms soon appear. To get the mushrooms to produce their
wonderful fruits, this sequence must be mimicked in the grow room.
There are a couple approaches for this process. If growing Agaricus mushrooms, large wooden
boxes like raised beds can be inoculated, cased with peat moss, and then kept cool and moist
long enough for mushrooms to fruit. Fan speeds are controlled at either end to implement
evaporation. This is an efficient process. For most gourmet mushrooms however, they prefer
to grow on hardwood and for this style of operation, bags are the preferred method. There are
two ways to fruit mushrooms from bags: top fruiting and side fruiting. Both can be initiated by
carefully and thoughtfully slicing the bag open with a clean blade or knife.
Top fruiting mushrooms like these Pioppino initiate pinning towards the opening of the bag.

Side pinning was induced on these Beech mushrooms by slicing the bag open – this allows for
evaporation and air exchange to occur on the surface of the mycelium and results in mushroom
formation. Note rubber bands keeping the bag firm against the substrate, this helps to prevent
pinning inside the bag.

Agaricus mushrooms fruiting from long beds (left). Pioppino mushrooms fruiting from hardwood
grow block (right).
Top Fruiting vs Side Fruiting:
Top Fruiting: Like Agaricus mushrooms fruiting out of the soil, some mushrooms (the typical
“stem and cap” like Pioppino, Chestnut and Nameko) like to grow upwards in a linear direction
towards the light source. To induce pinning, small cuts in the top seam of the bag can be made
at specific intervals and eventually the top of the bag can be cut and removed to simulate the
excessive rate of evaporation. A rubber band can be placed just below the surface of the
substrate sealing most of the colonized block from excess evaporation. This allows the
evaporation to be concentrated near the top surface to produce strong pin sets. There are
ways to extend the life of the bag as well. For example, if you cut the bags down the gusset
seams on all four corners and carefully fold the “flaps” that were just created, they can be folded
back over once the mushrooms were harvested. This will protect the mycelium during its
“resting” period and will eventually allow the mushroom block to pin once more with careful
humidity control. This process can be repeated over and over until the mushroom weakens and
produces less mushrooms or gets contaminated. I have generated up to 5 flushes from a single
bag before and some people claim to get 7 or 8 – that is really increasing the ROI!

Side Fruiting: Some tree dwelling mushrooms (Oysters, Lions Mane, Turkey Tail, Coral Tooth)
prefer to fight gravity the long way and fruit parallel to the earth. This “side fruiting” strategy
produces more shelf-shaped mushrooms or longer teeth in the case of lions mane. I like to
think of the fruiting block like a sponge. In this way, the mushroom bag “squeezes” out the fruits
of the mushrooms. One large cut will “squeeze out” one large cluster and many small cuts placed
uniformly around the bag will produce many small clusters totaling relatively the same yield.
This might be important when deciding what the customer prefers as a final product. Are you
serving chefs who may want big meaty oyster caps in one large chunk or are you serving small
variety mixes at the farmers markets where small little clusters are preferred? This is where
you can get creative. Experiment with different cuts and different end products for your
customers.
Left: top fruited beech mushrooms. Right: side fruited golden oysters.
Cleanliness, moisture retention, shiitake rehydration:

The biggest differentiator of mushroom quality in my opinion stems from the cleanliness of the
operations. This includes the growing room. The best way to maintain cleanliness is to have a
system down for regular cleaning. I think, at a minimum, you should be wiping surfaces and
mopping floors every other day and deep cleaning the grow room between every batch of
mushrooms. It is important to keep any spore load or mold outbreak from building up because
once contamination enters the system, it is nearly impossible to mitigate without shutting
down operations and doing a thorough deep clean. Spores fall on the surfaces, and mold
attracts insects which create an even more difficult situation. Insects can spread
contamination rapidly and attack healthy mushrooms directly. You can monitor insects with
yellow sticky pads and bug zappers, but it is best to eliminate the source which is from unkempt
grow rooms.

I think the best strategy for deep cleaning is using high water pressure and vacuums in
combination with FDA and Locally approved disinfectants. Cleaning must be done when
mushrooms are not present. This is best practice and makes cleaning much easier of a task.
Between flushes, mushrooms can be kept outside of the grow space if moisture can be
retained (in a walk-in cooler, or I like to rubber band them and place them on racks to pull out
healthy blocks for cleaning cycles). Another good strategy for shiitake mushrooms especially is
to hose them between flushes or even “dunk” them in clean water to allow them to absorb water
and swell for the next flush. This can be done by placing a heavy steel grate or brick on top of
the mushroom block and submerging it in water for a few minutes depending how dry the block
was initially. The mycelium will swell up and the block can be placed back in the fruiting room
for the next flush. This usually can stretch a block for another week or so, usually hosing keeps
the outer layer which can be prone to drying out healthy. This can also extend the life of the
block.
HARVESTING and STORAGE:
Once the mushrooms reach maturity, it is crucial to pick, pack and refrigerate in a timely
manner. Mushrooms will continue to grow until they are placed into a refrigerator and reach a
temperature between 37-40F. Timing during harvest is critical to procure the highest quality
mushrooms. Along with timing, moisture control, the proper storage materials and
considerations for customers is critical as well.

Mushrooms should be harvested just before the spores drop. This is important because once
the spores start to drop, the quality of the mushrooms degrade quickly. Spores are a perfect
food source for contaminants like yeast and bacteria and once they begin to fall on the fresh
tissue, the mushrooms start to degrade. Some mushrooms (like Pioppino, Nameko and
Chestnut) have veils that protect the spores until they are fully mature for dispersion. A good
indicator is to watch the veil and harvest just before they unfurl.
For Mushrooms like Oysters and Shiitake, the ideal time to harvest is before the cap unfurls
upward. This is a good indicator for when the spores will drop. Keep an eye on the outer rim of
the cap. They naturally grow outward and thin out. Keep in mind what your customers are
looking for – this is the time when art meets science, and the cultivator can put their final
touches on the masterpiece of their fungi.
DISTRIBUTION:
When it comes to distribution of mushrooms, timing, consistency, and velocity of mushroom

sales must all be considered for every farm’s unique situation. Some farms choose to distribute
directly to consumers. This creates the largest margins and is a great starting point because
you can develop strong relationships, get direct feedback and tailor directly to individuals
needs. We prefer to sell our mushrooms at local farmers markets along with other locally
produced foods and goods because it allows us to provide the most value to our customers.
There is a lot of educating about mushrooms early on, so the one-on-one conversations at
farmers markets help to spread the word and can be a great way to network. We have met so
many great mushroom lovers, from chefs to farmers, children to adults, being at the Cherry
Creek Farmers Market has been an enjoyable way to spread our product across our community!
Another approach to distribution is going to chefs and restaurants. This requires a larger supply
and consistency. Check with your local health department and food safety regulators to
understand what might be required to sell your products at this wholesale level.
A third approach for distribution is to wholesale to grocery stores and other food distributors.
This is feasible for larger scale farms and comes with a price. To stay competitive, prices need
to be kept low, which leads to tighter margins and less room for error. I recommend new farms
to stay away from wholesale until an efficient process is established and consistent results are
being obtained week in and week out. People eat A LOT of mushrooms!

Business compliance for a
mushroom farm:
Two caveats before diving into this topic.

❶ Compliance varies greatly from region to region so this is more of a generic
overview that can be applied across the board.
❷ This is coming from a food compliance point of view with a bias toward food
safety and working with the CDPHE here in Colorado.
I like to begin thinking of business compliance from a larger scope (like the FDA) and shrinking
that focus down to the individual business.
Federal and State Compliance: Check with FDA for current regulations on farming practices,
storage, processing, and distribution. Generally, as volume of production on your farm
increases, liability and levels of regulation increase as well. There are certain thresholds of
mushroom production that require higher levels of compliance at the federal level. Sometimes,
state regulations supersede federal regulations, especially for medium to small scale farms, so
contacting the local state authorities to verify what measures must be accommodated for local
food health safety is a great starting point. At our farm, the CDPHE (Colorado Department of
Public Health and Environment) oversees all food manufacturing regulations and compliance.
Calling them directly and establishing was our first step. From there, other agencies like the
Department of Agriculture, helped us to establish best practices. There are free resources and
annual meetings held by these agencies to give new farmers a chance to develop their own
compliance plans. I highly recommend starting here, or else seeking out another farmer in your
area and figuring out what they needed to have in place for their farm to be compliant. Farmers
markets are great for networking and forming trusted partnerships with fellow producers.
Usually, more established farmers are open to collaborating and will have the latest information
when it comes to compliance standards in your area.

Once federal and state regulations are met, you can seek out more localized authorities for any
County/Township regulations. In some cases, there may be zoning restrictions for agricultural
production or waste issues. Mushroom farms can produce a lot of compost with spent
substrates, so there may be regulations that should be followed when designing the scale and
location within your county or township. Building permits and zoning are controlled at this
level, so make sure that your farm is up to code (safety, size, number of parking spaces,
frequency of customers, deliveries etc.) all must fall within the scope of what your county
allows. For us, we needed to be very clear when applying for our farm’s permits. This helped us
really figure out the scope of what we were aiming to accomplish and made sure we had the
proper paperwork from the county so that we could begin operations.
Once you have received the proper permits, ensured that your farm is zoned correctly, followed
all state and federal authorities’ guidelines for food production, then you should consider the
protection of your business and business assets. This would include insurance, accounting, and
tax accreditation. Business liability insurance is a must have for attending most farmers
markets, but it is a good idea to have at least enough coverage to protect your business from
worst-case scenario events like food allergies and food-born illnesses. You can find small
business insurance online now and another good resource is from fellow producers at your local
food markets. You may even get a better rate from referrals or grouping together with other
farms or coalitions.
Accounting, like in all businesses, is very important for a mushroom farm. I recommend using
software like QuickBooks or Microsoft excel to keep track of all expenses. Opening an S-Corp or
LLC is a good way to establish your farm under its own identity and adds a layer of protection
from personal assets. A separate business account should be used to keep business expenses
separate from personal expenses. I recommend finding a good accountant to help you work out
the details of your operation. Make sure you keep up with taxes (quarterly or annually depending
on your local regulations).
These are all general ideas for business compliance and protecting your assets. I recommend
hiring an attorney to go over any liabilities that you might incur on your farm and protect
yourself with insurance. Mushroom farming is like all businesses, so good bookkeeping and
compliance with current authorities is extremely important for the long-term success of your
farm. I know many farms that operate without following any of these recommendations, and I
have also seen farms get shut down by local health departments which is a sad sight to see.
Don’t lose all your hard-earned money and make sure your “ducks are in a row” before you start.
That is my recommendation.

Morchella Cultivation
Research
We are now fully existing in the age of the internet. The internet is a great tool for research and
appreciatively, people have been slowly revealing the successful morel projects throughout the
past half century or so. With the first brief description that I came across appearing in Tradd
Cotter’s book “Organic mushroom farming and mycoremediation”, and after hearing his keynote
lecture in Telluride, I caught the itch of morel cultivation curiosity. Recent developments in
China have been outlined in the famous paper “Artificial cultivation of true morels: current
state, issues and perspective” (June 2017) since the release of the American Patent “Cultivation
of Morchella” patent number 4,866,878 (September 1989). Even more recently, “The Danish Morel
Project” has caught a wave in the mycology community describing in detail, with plenty of solid
evidence, how they succeeded in creating and implementing a commercially viable process.
From my research, there are some key aspects of cultivating morels (Allegedly – as I have yet to
produce a significant fruiting body at the date of writing this). From my understanding, genetics
is the most important factor, followed by substrate composition, nutrient balance,
environmental conditions and swings between dryness, necessary freeze/thaw period and a
period of extremely high moisture content that maintains constant water transfer properties.
Also, conidia formation is an important factor that is not completely described but is key to
fruitbody production.
My latest experiment is underway at both of my properties in Colorado. I have developed a

small patch for this spring’s monsoons in Denver, which is my more technical operation, and
have a few smaller patches in the hillside of our property in Sedalia. Both methods are based off
the Chinese method and a method established by Blue’s Best Mushrooms, a successful
commercial scale farm in Iowa.
First, substrate must be prepared. Morels are surprisingly fast growers and can be cultured like
other mushrooms in agar. They are unique in that they will eventually fade into a beige or tan
color, and form sclerotia on the surface of the agar, so fresh tissue or mycelium that has been
refrigerated works best for expansion. Keep in mind, the mushroom needs to be planted in the
fall prior to fruiting that spring (a mistake I made in my first experiment). Furthermore, patches
may take a few years to establish.

Once the culture has been expanded on agar, it can be transferred to grain. I used oats in my
latest experiments, but any grain should work, if the genetics of the mushroom are high quality
and have key features for forming sclerotia and conidia. Transfer the mycelium onto grain and
move onto a hardwood bulk substrate like other gourmet mushrooms. I also read that you can
supplement the bulk substrate with pine bark, but this is speculation. Once the mushroom
colonizes the block, this is where the differences occur from standard production. An incision
can be made, or many holes poked evenly on one side of the bag. This side where the incision
was made, or holes were poked should be placed down on the soil where the mushroom patch
will be grown. Choose a well shaded area that receives good moisture. The idea is that the
mycelium will jump off the hardwood substrate (imagine this block as the tree reservoir normally
storing the mushroom mycelium in the wild) and will grow outwards from the bag into the soil.
This nutrient gradient will cause the mushroom to form conidia, a secondary reproductive stage
that is unique to some mushroom species. Once conidia have formed, the mushroom will start
to go into dormancy. The mushroom should be covered with leaves, shade cloth and a wire
mesh to protect from harsh winter environments and keep moisture levels high, the wire is to
protect from rodents.
In the following spring, carefully observe for sclerotia development. These will be hard nodules
in the surrounding soil and are the beginnings of what will eventually become the fruiting morel
mushrooms. If you see sclerotia, maintain moisture content in the soil around 60% until you see
pins form. A heavy flush of water is recommended in some procedures, this may help induce
pinning. Carefully monitor pins as they are extremely sensitive and keep the moisture content
around 60% to keep the mushrooms from drying out (this is the hardest part in Colorado). With
a little bit of luck, you might establish a little morel patch! Do your own research, this is one of
the more experimental projects on our farm but also one of the ways to really get to know
mushrooms up close and personal.

Breeding Mushrooms –
The new frontier:
As I develop my process for cultivation, I believe that there is a significant portion of fungi
farming that is being greatly overlooked. This overlooked frontier is in strain performance and
breeding. My philosophy on this topic stems from my experience in the commercial Cannabis
industry. The saying goes that “you can have the best setup in Colorado, but if you don’t have the
genetics, you will not be able to compete”. I believe this holds true in mushroom farming as well
which is why I have spent about 10% of my earned capital on developing curated, craft
mushroom genetics over the past 3 years. The best setup in the world will not be able to sustain
itself long term without high quality genetics. It’s half the equation.
DNA:
Fungi are like all living creatures in that they carry an elaborately constructed protein that
encodes for every function of the organism known as Deoxyribonucleic acid (DNA). Fungi are
more complex than other microorganisms especially the Basidiomycota. They are more closely
related to us as humans than plants genetically and therefore are extremely complex. Fungi have
large regions of highly repeated code which can adapt over time according to the mushroom’s
environment, food source and spontaneous mutations that occur from continuous replication.
Therefore, growing mushrooms from spores can be so exciting. Spores from a mushroom
represent a genetic lottery, some spores are more selected for than others, but overall, there is a
standard curve of trait variations that can be separated out and implemented with careful
selection.
Thinking back to the lifecycle of the mushroom, one can recall that each spore may contain half
of the genetic information required to produce viable fruits. This single “haploid” spore isolate
culture can be systematically crossed with other haploids to produce very well-defined
offspring.
This is the basic principle of the following breeding tactics, but this should not be thought of as a
boundary to what may be possible as far as genetic manipulation and careful selection of traits.
It’s the first step, or the first advancement beyond the natural selection process in the wild.

To carefully separate spores and get them to germinate for the selection process, one must
prepare agar media as described in the lab cultivation section of this book. Agar is a two-
dimensional filter that allows for clear observation of germination, growth, and cross
compatibility of different fungal spores. Agar must be prepared aseptically to create an
environment void of any competition. This allows for the spores to germinate without any
competition which will benefit the mushroom cultivator.
The first consideration during the breeding process is the concentration of spores that will be
planted or “inoculated” on the nutrient agar plate. Spores will germinate microscopically at
various intervals and will immediately start seeking compatibility to mate with other spores.
This is important because if the proximity of other spores is close, then spores will mate before
they can be separated. This isn’t necessarily a problem if the cultivator is trying to produce
random pairings or phenotypes, but to systematically breed and maintain haploids with very
specific traits that could otherwise be covered with dominant traits, one needs to be able to
germinate, separate, and analyze a single haploid spore colony. Ideally, one should target
germination of one spore per plate to obtain a pure haploid colony. This can be done by using a
microscope and carefully selecting an individual spore physically with a loop or scalpel blade, or
it can be done by a systematic process of dilutions (called “serial dilutions”) so that one spore
can be collected in a specific volume of diluent. The physical separation can also be achieved
using various streak techniques that help spread the spores on the agar surface to help the
breeder isolate a single haploid colony. These techniques require the use of a sterile loop or
plate spreader and the careful aseptic techniques of the cultivator.
Making a Spore Solution (mixture):
Starting with a spore print, one should consider how it was prepared. Some prints are done on
sterile glass slides in front of a laminar flow hood and taken from a mushroom that was carefully
elevated above the surface of the glass so no contaminants can enter the system. Some prints
are formed in the middle of the forest on a piece of tin foil that was dug out of the picnic basket
from earlier that day. Either way, clean colonies can be isolated. The ideal spore print would
come from a sterile mushroom and collected in vitro or in front of a flow hood. I prefer to take
my prints in sterile petri dishes or used dishes that have been cleaned and disinfected because
they store nicely and make an easy vessel for spore dilution and collection.
To transfer spores into solution, you need to have sterile water ready. It doesn’t have to be
distilled water, but it should be drinkable water that has been pressure cooked or autoclaved in
a jar with an injection port and syringe filter to easily draw up with a syringe. The syringe should
be sterile and the ones with leur locks are preferred because the needle can be capped and then
stored for long term viability.

First, using a self-healing injection port, sterile syringe, and sterile needle, take up a syringe full
of sterile water. Cover the spores with the sterile water and gently agitate the dish while being
careful not to spill the water. Suck up the spore solution by placing the tip of the needle into the
thin layer of spore water and drawing up with the plunger. Cap the needle carefully and label
with the species name, date, and other critical information. You will need to make this label
clear because all spores look relatively the same once diluted in a syringe. Now you have
hydrated spores ready for inoculation.
If the spore syringe is highly concentrated, spores will be visible to the naked eye and will start
to clump in one area of the syringe. If you can visibly see clumps of spores in the syringe, it
should be diluted. Diluting the spores does a few things: it extends the use of the syringe, it
helps to disperse the spores to make for more isolated germination, and it may dilute out any
contaminants present in the solution.
From the concentrated spore solution, there are a few options for inoculating spores for
germination. You can put the spores directly onto a sterile substrate, you can inoculate them
onto the surface of the agar and let them germinate in place, or you can put them on agar,
spread them in a technical way and then let them germinate. All these techniques are viable
options, and each procedure has its pros and cons.
Direct Spore Inoculation:
If you are confident that your spore prints were prepared aseptically, then you can inoculate
directly onto agar. This can be done by scraping spores right off the print with a sterile loop,
blade, or swab. Take the sterile tool, carefully scrape spores off the surface of the print. Some
spores may stick to the surface, some will attach themselves to the tool and will leave an area
void of spores on the spore print. It only takes one microscopic cell to begin.
Once the spores are on the sterile tool, carefully lift the lid of the dish, scrape the tool gently
across the surface and place the lid back on the dish. From here, a few techniques can be
utilized to help further separate the spores. This is known as “streaking”. You can streak spores
in one direction to make a line. This is helpful because it allows for more sectors to emerge than
if all the spores were kept in one small point. To separate spores even further, one can streak in
multiple directions. Three-part streaks accomplish the most dilution by carefully dragging
spores across the surface to achieve isolated colonies. Start with one area off-center. This is
the initial inoculation point. Spread spores in that region with a sterile loop and flame in-
between spreading. Take the loop and gently pull spores from the first region to the next region

sterilizing the loop with a flame between streaks. Repeat this until you have a final target region
that single colonies will emerge from. It may be helpful to mark the area of inoculation on the
backside of the plate with a marker. Label the dish and wait patiently for the spores to begin to
germinate. Older spores can take a couple weeks to emerge, fresh prints can germinate spores
in a few days. The spores will start to germinate and produce hyphae. They will begin to emerge
where the sterile tool met the surface of the agar. Keep in mind the concentration of the spores
that were inoculated onto the dish. Highly concentrated spores will most likely create many
diploid colonies in a small region. These colonies will “sector” out to reveal numerous
phenotypes or different genetic combinations that can be isolated at the peripheral edges.
Diluted or spread spores can be isolated as haploids and placed on their own separate dishes
for later.
Hydrated or not:
If using a sterile spore syringe, the spores will be hydrated prior to inoculation. This primes the
spores for germination and can speed the process. Instead of directly spreading the spores, a
drop or two of spore solution should be used as the starting point. Spores will be diluted and
more likely to create separate colonies. To reduce clumping, a surfactant like tween 20 can be
utilized. Use lab-grade surfactants so that they can be added to the final spore dilution without
causing contamination.
Spore Isolate selection:
Once clean spores have started germinating, continuous observation is critical. I like to
number the colonies as they emerge on the back of the petri dish using a marker. Watch for any
abnormal growth that could indicate contamination. This would include blotchy areas, greasy
spots, small uniform circular colonies can also be contaminants. The challenge is in isolating
pure mycelium. The ability to identify this confidently will come with experience.
After a target number of colonies or different sectors from one colony emerge, they should be
transferred to their own plates to grow them out and observe for characteristics. If colonies
are presumed to be haploid isolates, store them on slants so they can be retrieved in the future
once characteristic phenotypes are traced back to progenitor haploids. If sectors are selected
and isolated, they can be grown out and fruited for selection. Each “sector” should produce
unique phenotypes. Each haploid must be systematically introduced to another haploid for
compatibility. Compatible haploids will fuse together to form viable mushroom tissue.
Incompatible colonies will form a zone of inhibition and compete on the dish.

Crossing haploid isolates.
Using a mathematical permutation, take each haploid and carefully place on one side of the
dish. Multiple haploids can be placed on the same dish, however, careful timing and constant
observation should be done to catch the mycelium at the points where they fuse because one
colony can quickly outpace the rest and cause confusion. Mushrooms do not stop growing for
the cultivator, so have a few spare plates ready for transfers. Once strains have been crossed,
you can move them onto grains or liquid culture and fruit. Each cross will produce a unique
phenotype. If there is a common trait that emerges, work backwards, and figure out what
haploid carried that trait. That single isolate can then be used to systematically cross in the
future. This is the most technical approach to breeding that I utilize and has led to some
vigorous commercial strains.
PFtek, Multi-spore liquid cultures (hyperbreeding) and Tissue Culture Multi-plex wells:
There are other, less-technical ways to breed mushrooms. One technique, often used for
psilocybe mushrooms is the “Pf tek”. Psilocybe finaticus on the online forum “the shroomery” is
credited with this ingenious approach for growing out mushrooms from spores. The method is
performed by inoculating a spore syringe into a carefully procured all-encompassing media
known as the “brf cake”. This media is a mixture of vermiculite, and brown rice flower, and
hydrated and sterilized like grain spawn. The “Brf cake” contains the right nutrients and the
right matrix to germinate spores at various locations when injected with a few ccs from a spore
syringe. The whole procedure can be found on “the Shroomery”.
The benefit of this process is that it takes spores and allows them to compete invitro and
eventually fruits a mushroom. The theory is that the mushroom produced contains the genes
to outcompete the rest of the paired mycelium in the media, and if cloned, can create a reliable
strain for commercial production. This process has produced a few good strains (pioppino,
chestnut, nameko) for our own farm and should be utilized to obtain high quality, commercial
varieties.
Another technique that I derived, “hyperbreeding”, is another fast-breeding method but

requires a high degree of aseptic technique. You can find the complete details in my YouTube
video “hyperbreeding mushrooms” and this applies to most mushrooms that can grow in liquid
culture. The idea is to take clean spores, let them germinate and cross in solution and then take
that multi-spore solution and inoculate grains to fruit. This will generate the strongest variety
from competition much like the pfTek method.

The latest technique being explored currently is from the use of tissue culture multiplex wells
and utilizing serial dilutions. If the cultivator can dilute the spores out to a solution that
contains one spore per drop, spores can be divided out uniformly on a multiplex plate
containing agar. One spore from one variety can be systematically crossed with a single spore
from another variety in each well and evaluated under a microscope for confirmation. This
process is highly technical but can save a significant amount of space and can produce many
crosses very quickly. All the compatible crosses can then be grown out to fruit and selected
for.
Mushrooms are relatively easy to genetically manipulate because the DNA is accessible through
a single cell layer that encompasses the mycelium. Plasmid induction, Di-Mon Mating, and
Crispr Cas9 are all techniques to horizontally transfer genetic material from one mushroom to
another in real time. These techniques are a bit more technical and require some more
materials but ultimately can create a wider variety of phenotypes without waiting for the
subsequent generation. If performed routinely and systematically, this can quickly change the
landscape of mushrooms being offered to market. I believe in the next few decades, genetic
technologies like Crispr Cas9 will be affordable for the every-day mushroom farm, and this is
the most exciting time to start getting a firm grasp on these technologies. Perhaps we will
start seeing blue lions mane mushrooms and polka dotted oysters in the not-so far distant
future!

Epilogue/Conclusion
I hope this book has provided you with the information you need to get
started on your own mycological journey. The first step of mushroom
farming is to get a plan together. Outline your market, set a target
production, and work backwards. Start small, learn fast and build on your
success. This is what we did to gain momentum in our local community.
We look forward to watching this industry evolve. Take your knowledge,
apply it to these principles and Grow More Mushrooms! MUSHLOVE

Bibliography
Below is a list of references used for the basis of this book and for your further
reading:
Cotter, T. (August 18, 2014). Organic Mushroom Farming and Mycoremediation: Simple to
Advanced and Experimental Techniques for Indoor and Outdoor Cultivation. Liberty, South
Carolina: Chelsea Green Publishing.
McCoy, P. (January 1, 2016). Radical Mycology: A Treatise On Seeing And Working With Fungi.
Portland, OR: Chthaeus Press.
Stamets, P. (December 1, 1983) The Mushroom Cultivator: A Practical Guide to Growing

Mushrooms at Home. Olympia, WA: Agarikon Press
Stamets, P. (October 25, 2000) Growing Gourmet and Medicinal Mushrooms. Olympia, WA. Ten
Speed Press
Stamets, P. (October 1, 2005) Mycelium Running: How Mushrooms Can Help Save the World.
Olympia, WA: Ten Speed Press
Journal References:
Bart P.S. Nieuwenhuis ⇑, Sil Nieuwhof 1, Duur K. Aanen . (November 2013). On the asymmetry
of mating in natural populations of the mushroom fungus Schizophyllum commune . Fungal
Genetics and Biology, 56 (2013) 25-32.
Ellingboe, Albert H, and John R. Raper. (2016 September, 7) The Buller Phenomenon in
Schizophyllum Commune: Nuclear Selection in Fully Compatible Dikaryotic-Homokaryotic
Matings. American Journal of Botony.(2016) 454-459
Qizheng Liu, Husheng Ma, Ya Zhang & Caihong Dong (2017): Artificial cultivation of true
morels: current state, issues and perspectives, Critical Reviews in Biotechnology, DOI:
10.1080/07388551.2017.1333082

Acknowledgments
I would like to thank all the members of the mycological community for the support received
during the emergence of Fresh from the Farm Fungi. There are many open sources of
information that helped me to gain the perspectives and insights to write this book.
Specifically, Mossy Creek Mushrooms, which helped me to standardize substrates by strongly
suggesting the use of hardwood fuel pellets over the curated blend of sawdust I was utilizing
early on. This freed up substantial amounts of time and created a more stable production.
Thanks Andrew and the rest of the Mossbacks for helping us gain more time! Another thanks to
Myers Mushrooms which exposed me to atmospheric steam sterilization and the Bubba’s
Barrel. This innovation and piece of equipment freed up even more time to allow for innovation
in breeding practices which has further pushed the envelope of efficiency on our farm. Lastly, I
would like to acknowledge my friend and colleague Zach from Mushroom Cult. During the lulls in
the offseason, the cultivation classes that we developed in house and curated over the years
has fueled my own continuous expansion of knowledge, curiosity, and innovations and the farm
would not be where it is today without carefully thinking through the processes and teaching
them to other people month after month. Thanks Zach and all the students (over 400 people
taught at the time of this writing).

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GOURMET

Gary Heferle, MLS ASCP

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Part 1 – the beginnings 8

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Solo Cup Agar: How to get 50 petri dishes for under $5

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Getting by without an industrial autoclave:

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The first year I spent a bit of time dialing in

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Gary, the author, making karyotypes on clinical rotation at Genetics Lab

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ANATOMY OF A MUSHROOM FARM:

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The lab is a designated area in the facility where

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Region 2: The Grow Space

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GROW ROOM BASICS:

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Cleanliness – Mushroom harvesting is messy. Regular and thorough cleaning must be

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Transfer agar into media bottle, or appropriate autoclavable pouring vessel.

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Save sleeves and keep sterile so they can be

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Malt Extract Agar (MEA) 12g Agar, 24 g malt extract

V9 Agar 1L V8 fruit juice, 8g potato flakes, 12 g Agar-agar

Black Charcoal Yeast Agar (BCYA) 12g Agar-agar, 20 g malt extract, 4g

Water Agar 1L water, 20g agar agar