Cultivation techniques

Where to start? There are a lot of mushrooms and there are many different growing techniques.

We provide you with detailed step-by-step information on various cultivation techniques.

Making a spore print

The best way to start the cultivating process is to get a good spore print. The spores are to the mushroom what seeds are to a plant. Hidden under the cap of a mushroom reside from hundreds of thousands to millions of spores that can all lead to the growth of new mushrooms.

Spores closeup

The making of a spore print has to be done in a sterile manner, so be sure to take all necessary precautions. To make a spore print you will need the following:

Sterilize the scalpel by heating it in the flame of a gasburner. Take it out of the flame when it starts to glow red and wait 20 - 30 seconds to let it cool off.

Sterile scalpel

Cut the cap from the stem at the highest possible point. Be sure not to touch the spores with the knife. Place the cap on the piece of paper with the spores facing down and cover it with a clean glass. In any case, the spores shouldn't be exposed to the outside air.

Cut hood

Hood on paper Print

After approximately 24 hours most of the spores will have fallen down. When the cap is still on the paper and you can see the spores have fallen around it that should be enough to continue. Remove the cap from the paper. Pick up the spore printed paper with a pair of tweezers and put it in a sealed bag. Small grip bags are excellent to use here. It is very important that the bags are 100% airproof. When placed in a cool and dark place, spore prints can be kept well for several years.

 

Hood on paper Print zipbag

Print in zipbag

Making a spore syringe

A spore syringe contains sterile water with hydrated spores in it. This spore water can be inoculated in the substrate of your choice. This way the chance that the spores will germinate is much higher.

Spore syringe

The reason that you have to hydrate the spores in water first is that they most likely will be too dry to germinate otherwise. To make a spore syringe you have to work under very sterile conditions, so take all necessary precautions. Try to work quickly, efficiently and with a steady hand when making a spore syringe. You can make several syringes from 1 print. It completely depends on the size and darkness of the spore print how much syringes you can make from it. From some prints only a few syringes can be made, while from other prints there can be made even 30 syringes from.

Print donker/licht

To make a spore syringe you will need the following:

The first thing to do is sterilize the water in a small bottle covered with tinfoil. The best way to go about this is to do it in a small flask. The narrow neck of the flask reduces the chance of contamination drastically.

Flask water

Place the bottle in the pressure cooker. When your pressure cooker has reached 15 psi, you can lower the gas. Let it all stay for about 30 minutes at 15 psi. After these 30 minutes you can turn the heat down and let it all cool off completely. This is very important. Don't be impatient, when the water is too hot the spores will not survive. Cooling off will take at least several hours.

Sterilise water

When the water has completely cooled off, you can do the next step. Sterilize the scalpel and your pair of tweezers by heating them with the gas-burner. Keep them in the fire till they start to glow red. Take them out of the fire and have them 20 or 30 seconds to let them completely cool off.

Sterilise scalpel

Remove the tinfoil from the flask with water and use the tweezers to take the sporeprint (partially) out of the bag. Scrape with the scalpel some spores from the print so they fall into the water.

Spore flask

The spore syringes can be made immediately. When doing the first syringe it's a good idea to fill it once or twice and then empty it again into the jar. The spores are then well divided over the water. When making larger amounts of spore syringes, it's wise to repeat this every few syringes.

Fill syringe Filled syringe

When the syringe is filled, it can not be used immediately. The spores need time to hydrate in the water. This will take at least 24 hours, but 2-3 days is even better (just roomtemperature or incubator).

Syringe in ziplock bag

You can also save them for a later moment. Spore syringes can be kept in the fridge at 2-4 °C for about 2-6 months. Save them in an airtight zip baggie.

Incubation conditions

Cultivation of Psilocybe cubensis on riceflour cakes

According to many the easiest method to cultivate the Psilocybe cubensis is the riceflourcake method. This method is also called the PF TEK, named after the person who invented it (Psilocybe Fanaticus). The substrate to use is a mixture of brown rice flour, vermiculite and water. In stead of brown riceflour you can also use ryeflower. For this method you will need the following: 

Cubensis on riceflour

The amounts for substrate ingredients are (by volume):

3.75 parts brown rice flour
10 parts vermiculite
5 parts water

Cake ingredients

Mix the dry components vermiculite and the brown rice flour very well. Add the water to it and mix it again. Fill the jars one by one with the substrate. Do this very loose; the mycelium needs space to develop itself. Make sure there is still 1 cm. remaining to the brim.

Ricecake mix Filling jar

Wipe the top clean with a tissue and fill the last centimeter with only vermiculite. This reduces the chance of contamination.

Clean jar Jar vermiculite

Close the jars with the lids and cover them with a large double layer of tin foil.

Holes in the lid Cake tinfoil

Now it is time to sterilize the substrate. Sterilize the jars at 15 psi for 30 minutes. Take the jars out of the pan after they have completely cooled down. Cooling the sterilized jars takes at least a couple of hours. Don't be too impatient. A substrate that is too warm can kill the mushroom spores and all preparations will be for nothing.

Sterilise cake

Before you inoculate your riceflourcakes, you have to sterilize the needle of your syringe first. Sterilize it by heating it with a gas burner until it starts to glow red. Let the needle completely cool down during 20-30 seconds.

After cooling down shake the syringe very well, so the spores will get divided in the syringe again. Remove the tinfoil and inoculate the culture medium with the spore syringe through the 4 holes in the lid against the glass of the jar. You should see the spore water running down. A total of 1-2 ml of spore water for every jar should be more then enough. After inoculation close the 4 holes again with some tape.

Inoculate cake Syringe glass

Tape hole Cake aluminium foil

Place the inoculated jars in a warm and dark place. The ideal temperature is 28-30°C. A cupboard above the fridge for example is a nice place. The fridge creates warmth at the back and heat always goes up. You can also use some heat elements. Now the work is done for a while, as the spores need time to germinate and develop themselves.

Incubation conditions

About 3 to 5 days after inoculating the jars, small white dots will appear. These are the mushroom spores that have germinated and grown into mycelium. After 2 to 4 weeks the riceflourcake will be totally white.

The next important action can begin. It's time to remove the substrate from the jar and place it into the fruiting conditions. The fruiting conditions mean that you expose the mycelium to fresh air, light and a lower temperature. The changing of these three elements will trigger the mycelium to form mushrooms. Place the riceflourcake upside down in your grow room and remove the jar. This can be done by (gently) tapping on the bottom. Finally place the box in a sunlit room. Make sure they are not in direct sunlight.

After 1 to 2 weeks the first pinheads will emerge. From this moment on growth can be very rapid. Within a few days the first mushrooms may already have reached their mature hood.

Casing layer

The casing layer is a layer of moist material that is put on top of the substrate with mycelium, before exposing this substrate to the fruiting conditions. The usage of a casing layer is highly recommended.

The riceflourcake method, is the only method where a casing layer is not been used. In other methods other ingredients will be used as substrates and these substrates will in a later moment be covered with a casing layer. This means that the substrate with mycelium itself will not be exposed directly to the outside air, which is the case with the riceflourcake method.

The casing layer makes sure the substrate doesn't dry out; it stimulates the formation of mushrooms and also serves as a water container for mature mushrooms.

There are many different recipes for casing layers. During time and experience a lot of different casing layers have proved to be very effective.

Some recipes are:

(both from Paul Stamets, The Mushroom cultivator, page 132)

We have had also very good results with the following casings:

and

Mix the dry components well together. Then steadily add the water and mix it well again.

Casing mix

Casing water

It is very important the casing layer will have the right degree of humidity. You have to go about this a little by instinct. A good way to check is to put a small amount of casing layer in your hand and squeeze. When the humidity will be right, a few drops of water will ooze through your fingers. If your entire hand gets wet, you should add some extra dry components. When there aren't any drops, you should add some water.

Test casing

Now the casing needs to be sterilized. Put the casing in a jar and sterilize it for about 50-60 minutes in the pressure-cooker at 15 psi. Then leave it to cool off until it has cooled down to room temperature. Casing material can also be pasteurized, 60-70 °C, for 1 hour. Some cultivators even choose to not sterilize it or pasteurize it at all.

Casing steriliseren

When the casing in the jar has completely cooled off, your casing layer is ready to be used.

Casing klaar

Psilocybe Cubensis on rye

Cultivating the Psilocybe cubensis on a substrate of rye, covered with a casing layer is one of the easiest and thus one of the most frequently used methods. All strains of the cubensis grow very well on a substrate of only rye covered with a casing layer.

Cubensis on rye

To cultivate cubensis on rye you need the following:

The amounts for substrate ingredients are (by volume):

1 part rye
0.7 - 0.9 part water

The rye, which will serve as substrate, has to be sterilized first. To sterilize rye you need to mix it with water. A often-used ratio is 1 part rye, 0.8 part water. When you use larger amounts of rye for in a filter bag less water is added otherwise it will become a sloppy substance. With only half filled standard jars you can use some more water. The general rule is: more rye is less water.

Rye ingredients

A full jar has to be sterilized at 15 psi for one hour. A full filter bag has to be sterilized at 15 psi for two full hours.

Keep in mind that the rye absorbs the water. So the rye grows during the process of sterilization. Especially when using jars to sterilize rye, it is important not to fill them up too much. The rye needs space. Also, it's good to have some space left in the jar so you can shake it well. When the rye has been sterilized and when it's still warm, it needs to be shaken well.

Rye in water Sterilising rye

This way the rye and the wet crumbs get the chance to mix about. After a good shake leave the rye to cool off, to about room temperature.

Sterilised rye Sterilised rye

When the sterilized rye has cooled off and has reached room temperature, it's ready to be inoculated. Open the jar and inject some spore water. Close the jar steady and quickly. After inoculation shake the jar or filterbag well, so all the spores are spread through the entire substrate. Basically your job is done for now. The spores need time to grow into mycelium. Put it away in a dark and warm spot of approximately 28-30 degrees C.

Inoculate rye Shake rye

Incubation conditions

Usually you will see small white dots appear after a few days. This is the start of the growth. After 2-4 weeks the entire substrate will be coloured white. The speed of growth of the mycelium is highly dependable on the warmth and the quantity of spores in the substrate. It can considerably improve the growing-speed to shake the jars well every 4-5 days. This keeps the rye and mycelium loose and crumbly.

Colonized jar Colonized jar

Colonized jar

Once the substrate is completely colonized, it's time to cover the mycelium with a casing layer. First shake the mycelium till it is completely loosened up. This can be a difficult task because of the rye that forms tough lumps of mycelium. Watch out not to break the jar.

Shake jar Mycelium shaken loose

Thoroughly clean the box you want to choose for growing the mushrooms. Open the jar and scatter the rye over the box. The next thing to do is to add the casing layer. The depth of the casing layer has to be around 1 - 1.5 centimeter. Take care that the casing layer is completely flat. Finally cover it with a lid.

Spread rye

Put the box back in the incubation room for a couple of days. You have to give the mycelium some time to grow through the casing layer. Make sure to close the box properly so the humidity remains as high as possible, close to 100 %.  

Incubation conditions

When you notice the white mycelium covering about 20-30% of the surface, it's time to expose it to the fruiting conditions. Do this by exposing it to light, fresh air and a lower temperature. After about a week the first pins will emerge from the casing. From this point the whole process will go considerably faster.

Psilocybe Cubensis on straw

Growing the Psilocybe cubensis on a substrate of straw is the best choice if you aim for growing at a somewhat larger scale, for it is easier to produce than a rye substrate. The straw delivers a better feeding ground for the Psilocybe cubensis than rye does; the mushrooms seem to grow larger and more numerous on straw.

First, you have to prepare the spawn. You begin with inoculating a jar with rye, just the same as you would make the jars for the cultivation of mushrooms on only rye. When this jar with rye is fully colonized we are not going to use it as substrate. However, we are going to use this jar with mycelium on rye to inoculate a bag with straw. When you distribute colonized rye throughout straw, the mycelium will develop further throughout the straw. When you use mycelium this way to inoculate another substrate it's called spawning. In this method the colonized rye will serve as spawn for straw. For a standard filterbag with straw you need about 2 jars of spawn.

First the straw needs to be chopped to a maximum of 5 cm. If the straw is too long the mycelium will have difficulties with colonizing the straw. When the straw has been chopped to the right length, it is time to soak it in water. This can be done by putting the straw in a large plastic bag and fill this with water. Watch out with this, it can be become easily a mess. Let this stand for at least a few hours, although some cultivators prefer to let it soak for even 2-3 days. After the soaking the straw has to be drained very thoroughly. Finally fill a filterbag with the soaked/drained straw. Then the filterbag with straw has to be sterilized (15 psi, 3 hours) or pasteurized (60-70 °C, 2 hours). Either way, don't close the filterbag with an impulse sealer yet, the spawn has still to be distributed throughout the bag at a later moment.

When the sterilized or pasteurized straw is completely cooled off, it's time to divide the spawn over the bag of straw. Before you inoculate the straw with the spawn, carefully shake the spawn in the jar so the chunks of mycelium break apart. The straw can now be inoculated with the spawn. Open the filter bag with the straw and pour the mycelium in the bag. Close the bag immediately with a good sealing. It's important to evenly divide the spawn over the straw, to avoid the substrate from not colonizing completely. Especially with a mass of moist straw in a filterbag this can prove to be be a troublesome job.

Straw

Finally place the full filterbag in the incubationroom. Again, let time do its work and after 2 to 4 weeks, the straw will be fully colonized with mycelium. Now it's time to prepare the final phase. Take a large clean box, spread out the colonized straw with a height of 7-10 cm and put a casing layer on it. Thoroughly cover the box with plastic foil. Place the box in the incubationroom again, and give the mycelium time to form a strong network through the casing-layer. After a couple of days, when the mycelium is clearly visible through the casing-layer, it can be exposed to fruiting conditions. Under normal conditions the first pinheads will show up after 7 -10 days.

Incubation

Growing Panaeolus cyanescens

Panaeolus cyanescens and the Panaeolus tropicalis are grown almost exactly in the same way. And this is not that different from growing the Psilocybe cubensis either.

However the Panaeolus cyanescens and the Panaeolus tropicalis are in many ways much more vulnerable than the Psilocybe cubensis. Vulnerable in a sense that these 2 types need more attention during the preparation and maintenance of their ideal growing circumstances than the Psilocybe cubensis does. You can also see on the mycelium that these types are more vulnerable. Their mycelium is normally less rhizomorph than the mycelium of the Psilocybe cubensis.

When growing the Psilocybe cubensis it's sometimes possible to deviate from the ideal conditions and still get a fine harvest. When growing the Panaeolus cyanescens and the Panaeolus tropicalis, the delivery of ideal conditions may be the difference between a good harvest of no harvest at all.

Required items:

The first thing you must have for the growing of these mushrooms is strong spawn. Rye is the best basis for spawn. It's wise to inoculate the rye with a strong single strain on agar instead of a spore syringe.

When the spawn is totally colonized, it's time to prepare the substrate.

The substrate consists of (in volume):
10 parts of straw 
4 parts of cow manure
3 parts of vermiculite
3 parts of water

Mix the dry components in a pot and add the water afterwards. Mix everything very well.

Next sterilise the substrate in a filter bag. Don't seal the bag yet, because you still need to add the spawn to it later on. After it's cooled down you can spread the spawn in the filter bag with straw. For a normal filter bag a pot with spawn is enough. The spawn must first be well shaken to get loose. After this you can scatter the spawn in the filter bag and seal the bag immediately after this.

It's important to spread the spawn equally on the whole substrate to prevent it from colonizing unevenly or not colonizing at all. Try to do this by shaking the bag. Especially with a humid mass substrate in a filter bag this can sometimes be difficult.

Now let time do its work. Put the filter bag in the incubator with a temperature of 28 - 30 °C and after two to four weeks the substrate will be totally colonized with the mycelium.

Now the last phase has began. Get a big, clean bag, spread the colonized substrate in it and put the casing layer on top. Close the pot well again with plastic foil. Place the pot again in the incubation space and give the mycelium time to create a strong network through the casing layer. After a couple of days, when the mycelium is undoubtedly visible through the casing layer, it can be placed in fruiting conditions.

The fruiting conditions are:
- (indirect) sun light
- 24 - 25 °C
- air humidity 95 - 100%

After a week the first pinheads will appear. From now on it's really important that the mushrooms get enough fresh air (min. 4 - 5x a day). The air humidity can go slightly down to 90 - 95%.

Growing psilocybe Azurescens (outdoor)

These 3 mushrooms grow in the colder and wetter places on Earth. Therefore they are no good to grow indoors. This means that growing these mushrooms is not for everybody. You must live in a place where autumn is cold and wet and besides you need to find a location where you can lay the bed and where it is safe throughout the year.

We are thinking of a bed in the garden (or woods) of 80 * 80 * 10 cm.

These mushrooms come up in the autumn and in the beginning of the winter. The whole process from preparing the spawn until the mushrooms come out is a process that will take very long. If you want to harvest these mushrooms in the autumn, you actually must start at the beginning of the year (Jan/Feb).

The growing process normally has 3 phases: - preparing the spawn - growing the mycelium outdoors - growing the mushrooms

First you must prepare the spawn. You do not need that much: 300 to 400 ml. is enough. Rye is the best basis for spawn and the mycelium of these mushrooms grows best in the incubator with a temperature of 20 °C.

When the rye is totally colonized, it is time for the next step in preparing the spawn. The Psilocybe azurescens, cyanescens and baeocystis love woodchips. The colonized rye will also be helpful in colonizing a part of the woodchips with mycelium.

It does not really matter what kind of woodchips you use. In general the mushrooms grow very well on almost all kinds of woodchips. We use the ones that you can buy in the pet shops to use as underground for your pet house.

Before you inoculate the woodchips with the colonized rye, you must first soften them by soaking them. Fill up 2 pots of 1000 ml for 60% with small woodchips and totally cover the woodchips with water. Let them soak for 48 hours. After 48 hours let the 2 pots with the woodchips leak out in a strainer. Do this very thoroughly. Fill the 2 pots again with the prepared woodchips and close the pots with a lid with a filter. The pots must be now sterilized for 1 hour on 15 psi. Let the pots quietly cool down afterwards.

If the pots have cooled down entirely, you can start to inoculate with rye. Divide de rye over the 2 pots. After inoculation you must give the 2 pots a good shake to spread the rye. After that, place the pots in the incubator (18 - 20 °C.).

After 2 up to 4 weeks the pots have been entirely colonized. From time to time shake the pots well to speed up the process. Once this has happened, it is time to prepare for the open air.

For an outside 'bed' of 80 * 80*10 cm. you need approximately 10 liters of woodchips. It is all right if these woodchips are a bit larger than the ones you used for making the brood. Put these 10 liters in a garbage bag and add plenty water. Let this now soak for 24 hours. After these 24 hours the bag must leak out entirely. Make some small holes in the bag tot make that happen. And cut off, after a while, the corners on the bottom of the garbage bag. Wait till there is no more water left.

Put your boots on! Go outside and dig a hole in the ground of your desired size. It is smart to choose a spot with much shade. Scatter a layer of 4 - 5 centimeters woodchips on the floor. Take the 2 pots with brood (loosened up by shaking) and scatter these over the complete bed. Cover this again with a few centimeters of wet woodchip. The bed is almost ready now. Irrigate the bed and cover it with a garbage bag. This makes the bed the ideal environment for the mushrooms, nice wet and dark. You can keep the bag in its place by laying some stones on the edge.

The best time of the year to make such an outside bed is around March/April. Your work is down for now. During spring and summer the mycelium needs enough time to build a firm network thorough the whole bed of woodchips. Verify, however, on a regular basis if the bed still has a nice humid condition. Spray the bed with water if nescessary.

At the beginning of September the weather becomes colder and wetter: the ideal circumstances for the growing mushrooms. Remove the garbage bag and give the bed, if needed, a good irrigation to keep it humid. You will probably see a number of insects and other small beasts walking around in and on your bed. Do not worry; these small creatures will not damage your work. They do not have an influence on the quality of the bed or the growing of your mushrooms, except in extreme cases (plagues etc.) I would almost say: give them the space and let them enjoy the mycelium...

It is difficult to indicate when the mushrooms will rise. This is entirely dependent on the specific weather of that particularly autumn. You can expect them from mid- September up to beginning December. Care at all times that the bed will not dry out.

It can be that in one autumn several flushes will rise. After the last flush you can simple leave the bed. The mycelium should not have too much trouble surviving the winter and giving flushes the next year. It is nevertheless wise to spread out, at the beginning of a new spring, a fresh layer of woodchips on the bed.

Growing truffles (Philosopher Stones)

The Psilocybe mexicana and the Psilocybe tampanensis are two mushrooms which are known for their sclerotia-forming. Sclerotia (plural of sclerotium) are hard pieces of mycelium which the mushroom grows throughout the substrate. These hard pieces mycelium have a much better resistance against bad influences that could cause delays in growing than normal (soft) mycelium. They serve thus as protection against a too high or too low temperature, dehydration and a too high air humidity. The sclerotia are looking a bit like walnuts.

The sclerotia of the Psilocybe mexicana and the Psilocybe tampanensis contain a much higher concentration psilocybine then the mushrooms themselves. This makes that most people only grow the sclerotia instead of the mushrooms. 'Truffles' they call this kind of mushroom-less mycelium, or 'Philosopher Stones'. The 'A'-strain of the Psilocybe mexicana is commonly known as the mushroom which forms most of the sclerotia. Growing sclerotia is not that different from growing mushrooms. While growing mushrooms you only go a few phases further in the life circle of the mycelium. The Psilocybe mexicana and the Psilocybe tampanensis are growing bests on grass seed. A lot of different types of grass seed of several brands have conducted to a good result. Rye or white rise can also be used.

Required substrate (in volume):

Sterilize the substrate in a filter pocket, filter box or glass jar with a self built-in filter. Filter boxes and glass jar must be sterilized for 1 hour at 15 psi. A full filter pocket for 3 hours. If the substrate has cooled down entirely, you can start to inoculate it with a sporesyringe or mycelium on agar. Naturally we prefer a strong single strain on agar. Shake the pocket or pot well to divide the spores all over the substrate. After inoculation you can put the substrate aside in the incubator. The mycelium grows best in the dark at a temperature between 21 and 25 °C. After 2 to 4 weeks, and a couple of good shakings now and then, the substrate will have entirely colonized. You should now decide whether you want to grow the mushrooms or sclerotia.

Growing the sclerotia

When choosing this option, you are basically done 'gardening'… The only thing that counts now is that you need to be patient. You can just leave the substrate where it is (still at a temperature between 21 - 25 °C). Also important from now on is not to shake the substrate anymore.

The substrate will quite rapidly start its forming of sclerotia. Slowly the sclerotia will grow into large, hard pieces with a yellow-brownish color. This sclerotia-growing last about 3 up to 4 months. After this period they will not grow any bigger

Growing the mushrooms

If you choose to grow mushrooms instead of the sclerotia, for instance because you wish to make spore prints, you will have to take it some steps further. If the substrate has been entirely colonized and has not yet started its sclerotia-growing, you need to add a 'casing-layer' to the substrate. These mushrooms perform best on a casing-layer of peat, vermiculite and calcium carbonate.

Shake the bag or jar to loosen up the mycelium. Scatter the mycelium in a clean barge and cover it with a casing-layer of 1 to 1.5 cm thick. Put the lid on the barge or cover the whole thing with foil. After all this, the barge must be placed back in the incubator for some days.

When the mycelium is clearly visible through the casing-layer, you can place the whole cluster in the fruit forming conditions. For the Psilocybe mexicana and the Psilocybe tampanensis these conditions do not differ that much from those of other types of mushrooms. For the Psilocybe mexicana and the Psilocybe tampanensis these conditions mean that they must be exposed to fresh air, (indirect) sunlight, air humidity of 95% and a temperature of 20 °C or a even a little bit higher.

After 1 up to 2 weeks the first pinheads will appear. These pinheads will develop  into adult mushrooms in 4 to 7 days.

Truffle Growkits

A quick and easy method of growing sclerotia is by making use of a pre-inoculated and colonized truffle grow box. These are glass jars filled with a substrate of grass seed that is already colonized with a truffle forming psilocybe strain. All you need to do with such growkits is wait for the truffles to grow. When they are large enough to be harvested, the jar can be opened and the truffles can be harvested and cleaned.   

Multispore and single strains

Multispore and single strains

A lot of people that start growing mushrooms start running into the same questions and problems. After a number of successes growing, a lot of people wonder why mushrooms of the same 'flush' don't grow equally. They all seem to have their own natural rhythm. Also, there are large differences in the way the mushrooms look. Some are big and healthy, but not all mushrooms grow up to adulthood.

 

The most important reason for this is that most home growers inoculate their substrate with a spore syringe. Inoculation with a syringe is called "multi-spore inoculation."

Every spore-syringe contains thousands, maybe even millions of spores, from the same spore print. All these spores, in theory they can all germinate into a unique genotype with their own characteristics. So, when inoculating a substrate with a spore syringe, a large number of different genotypes end up in the substrate. Every unique genotype that grows to be a mushroom has its own characteristics when it comes to speed of growth, size and looks. This explains the unregular flushes and why one mushroom can look much nicer and healthier than the other.

The more experienced grower therefore looks for the stronger genotypes and inoculates his substrate with a so-called "single strain" or "pure strain". To understand this, first you need to know more about how you can separate the strong genotypes in the mycelium from the weak genotypes.

When you've inoculated the substrate with a spore-syringe and large parts of the substrate are colonised, often you can distinguish two different kinds of mycelium. One kind looks somewhat fluffy/down-like; This is called Tomentose Mycelium. The other has a wiry-type structure; this is called Rhizomorph Mycelium. This is the kind we are looking for. Rhizomorph Mycelium has a larger potenty to grow into nice, healthy mushrooms than Tomentose Mycelium.

To make sure that only rhizomorph mycelium will develop, it's possible to take a part of rhizomorph mycelium from a colonised substrate. This part of mycelium can serve as the inoculant for another container of sterilised substrate. If you took a good part with a strong genotype of the mycelium, within the new substrate will mostly grow healthy and strong mycelium.

But when a container is largely colonised, it can be very difficult to select the best mycelium from it. The risk of contamination is very large; so it's not advisable to try.

Growing the mycelium on agar in petri dishes first offers a solution for that problem. The agar media is not a fertile soil for growing mushrooms, but the mycelium develops very nicely on it. Therefore mycelium that grows in Petri dishes can only be used to inoculate a substrate where it can develop further.

Before we'll continue with why and how agar can be used, we'll show how you can make this cultivation media from agar agar.

Throughout time it has appeared that there are many different recipes that can all lead to a good result. These agar media recipes can be bought, ready to use, in local stores or through the internet. It is also not that difficult to make these recipes yourself. 

Mix the dry components well together and add the water to it. It is best to do this in special 'laboratory-bottles', the ones with the long narrow necks. These bottles are easy to use and have a narrow neck which reduce drastically the risk of contamination. Of course you can also use a different kind of jar or glass.

Cover the bottle well with a piece of tinfoil and mix it again thoroughly. Place the bottle in the pressure-cooker and sterilize the agar media for approximately 40 minutes.

When the pressure has completely left the pot, you can remove the bottle from it. The agar media is close to boiling, so be careful. Boiling agar can cause nasty burns.

It is now important to pour the agar into the dishes at the right moment. If you wait too long, the agar starts to clot. This means it's not easy pourible anymore. This situation is not very handy.

If the agar is too hot when you pour it into the dishes, this causes another unpleasant situation. It will start to condense within the Petri dish. Condense is harmful to the growth of mycelium and it also prevents you from seeing what is going on inside the dish.

The right temperature for the agar to be poured is about 40-50 degrees Celsius. When you're able to hold the bottle for about 10-12 seconds without it being painfully hot, then you know the temperature is right

Place the dishes next to each other in piles of about 10 or 20 pieces and remove the tinfoil from the bottle of agar. Take the whole pile and pour the agar in the lowest dish, then continue to work upwards. Let the dishes sit for a while so the agar can clot. When it is completely clotted, the dishes are ready to use. 

Agar is very susceptible to contaminations. Especially at this stage it is very important to work very clean and with a steady and efficient hand.

Now you know how you can make agar media, we can continue with explaining the advantages of the usage of this agar media in Petri dishes.

When your Petri dishes with agar media are ready, you can inoculate them with some spores. Take the print out of the zip bag with a sterilized tweezers, open de Petri dish and scrape some spores with the sterile scalpel so they fall on the agar media on several places. Close them quickly again and place them in the incubation room at 28 - 30 C.

After a few days the spores will germinate and the growth of white mycelium will be visible. Because the mycelium on agar media grows in a flat, 2-dimensional way, it's possible to extract a part of mycelium with a scalpel and place it in a new dish with agar. So cut out one or more of the germinated spores and place each of them in the middle of a new dish. Close the dish quickly.

After a few days you can already distinguish the stronger and the weaker parts in the structure of the growing mycelium. Look if there are any sectors of mycelium which are rhizomorphic.

If you've made the right selection of mycelium, new mycelium will grow stronger and larger, compared to the first dish you used.

This process can be repeated a number of times. After a while you will notice that the mycelium doesn't grow in "sectors" anymore. Now you have your own "pure" or "single" strain. From this single strain, you can inoculate numerous containers substrate or spawn by cutting a small part of mycelium from the the petri dish, and putting this in the new substrate/spawn. In these jars there will be a development of rhizomorph mycelium of 1 specific genotype. All the mushrooms that develop from this strain will all have the same properties in terms of growth and looks.

The procedure of moving parts of mycelium cannot be repeated too often. If you select sectors too often, after some time degeneration will set in. The mycelium in the agar dishes will then produce more down-like/fluffy sectors again. Degeneration has to be prevented.

In general, it's advised to make no more than 3 parts of rhizomorph mycelium from the same mother culture to petri dishes. 5 times is the absolute maximum for most kinds.

Mushroom cloning

Cloning a mushroom is a very good way to preserve the genetic blueprint of that same mushroom. You can clone a mushroom by making a so-called "tissue culture". Tissue cultures are made with the help of agar media. A little bit of flesh from the inside of the mushroom has to be placed on agar media in a Petri dish. Around this tissue from the mushroom will develop mycelium. This mycelium will have the same genetic pattern as the mother mushroom. 

A tissue culture has to be made within 48 (but preferably 24) hours after the mushroom has been harvested. 

Basically, you can take tissue from any part of the mushroom. But the tissue must be taken from the inside of the mushroom, for this part is practically sterile.

You can extract a part of tissue by breaking the stem or the hat in 2. Use a sterile knife or scalpel to remove a part of the mushroom's inside. A part of 2 by 2 mm is enough. Never touch the outside of the mushroom with the knife, the outside is not sterile and contact with it will most certainly lead to contamination. 
 

 

Place this piece of tissue in a petri dish with agar as quickly as possible. The risk of contaminating is very large with this method. The chance of succeeding will be larger if you inoculate a number of dishes with a part of tissue. 
 

It could be possible that there is a necessity for selecting a stronger sector of mycelium and taking it to a new dish for further growth.