How to cultivate Flammulina velutipes

The cultivation of Flammulina velutipes or better known as Enoki or the Winter Mushroom, has a long history, especially in Asia, since its medicinal properties and nutritional values[1] (Fig. 1).

If you are interested in growing Enoki mushroom yourself, then you not only need to know the growing parameters but also how each one of them affects the quality and the yield.

And that is what this article is all about.

In order to achieve this, I analyzed and categorized all the scientific papers to find patterns and correlations for each growing parameter.

The graphical abstract of the nutritional values and biological activities of F. velutipes

Figure 1: The graphical abstract of the nutritional values and biological activities of F. velutipes[2]

Step 1: Culture media

The first category is the used culture media within these scientific papers.

The most used culture media were MEA[3] and PDA[4], but I also find media like MFM[5], MMM[6] and PDYA[7].

If you are new to this topic MEA stands for Malt Extract Agar, PDA for Potato Dextrose Agar, MFM for Mushroom Fermented Medium, MMM for Minimal Mushroom Medium and PDYA for Potato Dextrose Yeast Agar.

As the primary carbon source glucose was used.

While peptone, ammonium, and amino acids were used as the primary nitrogen sources[8].

If you know the composition of MEA and PDA, then you know that neither of these two contains these types of carbon and nitrogen sources.

In this regard, a GPA (Glucose Peptone Agar) would be, in my opinion, more suitable.

It could be found that adding phosphate, magnesium, iron, zinc, manganese, copper, cobalt, molybdian, calcium[9] as well as thiamine[10] to the agar improved the mycelium growth.

The pH value can range from 4.5 to 8.6[11], with an optimum pH value of 6[12],[13].

The optimum temperature is found to be 25 °C[14] (77 °F) with a range between 0°C to 34°C (32 to 93 °F) [15].

An interesting note is the age of the mycelium used for the inoculation. A study mentioned that the mycelium should not be younger than 5 days but not be older than 2 to 3 weeks[16].

Step 2: Substrate

The next step towards growing Flammulina velutipes is to find the right substrate mix. Table 1 summarized the main ingredients used in scientific articles.

The table shows not only the ingredients but more importantly their cellulose, hemicellulose and lignin content.

  Cellulose Hemicellulose Lignin
Sawdust 44 17 21
Wheat straw 50 24 14
Wheat bran 25 35 15
Rice straw 30 31 20
Rice bran 25 34 25
Cottonseed hulls 50 15 16
Soybean hulls 43 18 2
Corn cob 35 43 5

Table 1: Cellulose, hemicellulose and lignin content of different substrate compounds[17]

To these substrates, several supplements were added to increase the growth rate of the Enoki mushroom. Supplement mentioned in these articles were rice bran, wheat bran, sucrose, gypsum, lime, plaster, calcium superphosphate, corn powder, bean cake, CaSO4 and CaCO3.

The moisture content of the substrate ranged from 60 to 80 %[18],[19],[20],[21],[22], with most settling for 65 %.

The substrate is then filled into cropping containers such as plastic bags[23], 800 ml flask or 1,100 ml bottles[24],[25].

Slightly press down (if you are using bottles) the substrate to make it more compact.

Use for example a wooden stick and bore a hole in the center of the bottle.


In the following video I talk about a research project in which the authors could achieve a biological efficiency of 185 with the right substrate.

Step 3: Sterilization

After filling the cropping containers, the substrate is usually autoclaved at 121 °C (250 °F), 15 Psi for 1 hour[26] or if used less pressure at 95 °C (203 °F) for four hours[27].

Step 4: Inoculation

The only inoculation rate I could find was 10 %[28].

Pour the spawn in the hole (bottles) or on top of the substrate (bags).

Close the bottles with a lid (with filter) or seal the bag.

Step 5: Incubation

After the inoculation, the cropping containers were kept at a temperature of around 25 +/- 2°C (73°F)[29],[30],[31]. Some studies go even down to 18-20 °C (64 – 68°F)[32]. The temperature, though, should not reach more than 34 °C (93 °F) as the mycelium will die[33].

During this phase, no light is needed.

Step 6: Fruiting

After the cropping container is fully colonized, the surface is raked, and the temperature is dropped to 8 °C (46 °F)[34].

After the scratching of the surface, the bottles are placed upside down for a few days[35].

After the stem reaches 1-2 cm, at the top of the bottle, a paper was placed around the bottle to form a cylindrical shape[36]. This way, the oxygen level will be suppressed, and the Enoki will start stretching (elongating).

If primordium formation starts (stem: 1 to 2 cm) the temperature is raised to 15°C (59 °F)[37] with a range of 10 to 20 °C (50 to 68 °F)[38],[39],[40],[41],[42].

The humidity is kept between 60 % and 95 %[43],[44],[45],[46],[47]. After the stem reaches 2 cm, the humidity is adjusted to 75 to 80 %[48].

During the fruiting phase 1, the CO2 level is kept below 1,000-1,500 ppm. After the stem reaches 2 cm, some raising the CO2 level to 1,500-2,000 ppm[49] while others keep it below 1,000 ppm[50].

The air exchange is therefore set to about 2-4 per hour[51].

The light during the primordia formation is raised to 20-50 lx[52],[53]. Some setting the duration to 0.5 hours/day at approximately 300 lx[54].

Day to mature (DTM)

With this setup, the whole mushroom cycle takes according to Stamets between 22 and 31 days. While it takes about 14 to 18 days to fully colonize a bag, the pinhead formation needs roughly 3 to 5 days and the flushing 5 to 8 days[55].

Shelf life

After this brief introduction, let’s take a look at different parameters and how they impact the yield as well as the quality of Flammulina velutipes.

Winter mushrooms have approximately 14 to 20 days at 1°C (34 °F) a good shelf life. If the temperature is raised to 6 °C (43 °F), the shelf life drops to 10 days. At 20 °C (68 °F), the shelf life is about 2 to 3 days[56].

Mycelium

The first parameter or ingredient I want to look at is the nitrogen sources. As mentioned in the beginning, peptone, ammonium, and amino acids are the way to go. If we take a look at figure 2, we can see the effect of different nitrogen sources on the growth of Flammulina velutipes.

While none of the other nitrogen sources could beat the control (which is a mixture of peptone and yeast extract), peptone and yeast extract alone was among the four best nitrogen sources.

This means that adding yeast extract and peptone to your culture media will increase the growth of the mycelium.

Effect of different nitrogen sources on the growth and crude polysaccharide extract of F. velutipes

Figure 2: Effect of different nitrogen sources on the growth and crude polysaccharide extract of F. velutipes 6 (control: peptone + yeast extract)[57]

The other side of the coin is the carbon sources. Here glucose is the way to go as described earlier and in figure 3. All other carbon sources had a smaller effect on the growth of the Winter mushroom.

Effect of different carbon sources on the growth and crude polysaccharide extract of F. velutipes

Figure 3: Effect of different carbon sources on the growth and crude polysaccharide extract of F. velutipes 6[58]

And finally, an interesting finding was the influence of the C:N ratio on the growth rate of Flammulina velutipes (Fig. 4). According to this study, the higher the C:N ratio, the higher the growth rate. But overall, there is almost no influence at all.

Correlation between C-N ratio and mycelium growth rate of Flammulina velutipes

Figure 4: Correlation between C:N ratio & mycelium growth rate (mm/day) of Flammulina velutipes[59]

Results

With that information in mind, we can now talk about the yield you can expect from growing Enoki mushrooms. If the conditions are right, the potential yield is around 160-220 g per 800 ml bottle with a maximum of 600 g per 800 ml bottle[60]. To understand this broad range, let’s analyze different parameters.

One of the main factors is the substrate composition. In figure 5, several different mixtures were analyzed, and their influence on the yield measured. It could be shown that the yield ranged from almost 2 g/bottle up to 84 g/bottle.

The study used poplar sawdust as the main ingredient which was supplemented either with rice bran, wheat bran, cattle manure or a mixture to the supplements.

The best result with 84 g/bottle was achieved by mixing poplar sawdust and rice bran in a ratio of 10:3 (Fig. 5, substrate no. 2).

Effect of different substrate mixtures on the yield of Flammulina velutipes

Figure 5: Effect of different substrate mixtures on the yield of Flammulina velutipes[61]

Why this is the case that can be figured out by looking at the influence of the nitrogen content (Fig. 6). As the figure shows, the higher the nitrogen content, the higher the yield.

The total nitrogen content for substrate number 2 was 0.82 %.

Influence of the total nitrogen content on the yield of Flammulina velutipes

Figure 6: Influence of the total nitrogen content on the yield of Flammulina velutipes[62]

As we talked already about the influence of the carbon source, let’s have a look at the influence more closely. In figure 7, we can see the correlation between sugar content and the yield. As the figure indicates, the higher the sugar content, the higher the yield.

For our substrate (No. 2) the sugar content was about 19 %.

Influence of the total sugar content on the yield of Flammulina velutipes

Figure 7: Influence of the total sugar content on the yield of Flammulina velutipes[63]

Besides these two ingredients (nitrogen and carbon), adding phosphate (Fig. 8) or potassium (Fig. 9) to the substrate can improve the yield.

While phosphate in high doses reduces the yield (Fig. 8), adding potassium to the substrate increases the yield (Fig. 9).

The phosphate content for substrate number 2 was 1,107 ppm.

Influence of the P2O5 content on the yield of Flammulina velutipes

Figure 8: Influence of the P2O5 content on the yield of Flammulina velutipes[64]

The potassium content for our substrate (No. 2) 23 me/100g.

Influence of the potassium content on the yield of Flammulina velutipes

Figure 9: Influence of the potassium content on the yield of Flammulina velutipes[65]

The next study from 2014 showed similar results (Fig. 10 and Fig. 11). The authors compared different substrates and measured their effect on the biological efficiency of Enoki mushrooms.

The best result (B.E. 185 %) was achieved by mixing paddy straw (PS) and palm empty fruit bunches (EFB) at a ratio of 25:75.

Effect of different substrates on the biological efficiency of Flammulina velutipes

Figure 10: Effect of different substrates on the biological efficiency of Flammulina velutipes[66]

As already shown, the higher the C:N ratio, the higher the yield (Fig. 11).

Correlation between C-N ratio & biological efficiency of Flammulina velutipes

Figure 11: Correlation between C:N ratio & biological efficiency of Flammulina velutipes[67]

Grow factors

Temperature

Now let’s talk a little bit about the influence of the temperature. As mentioned during the fruiting phase the temperature is around 15 °C (59 °F).

But what will happen if we increase or decrease the temperature?

This question was answered in another study. The authors found that reducing or increasing the temperature by 5 °C (41 °F) will decrease the mycelium growth rate by up to 10 % (Fig. 12).

The effects of culture room temperature on mycelium growth of Flammulina velutipes

Figure 12: The effects of culture room temperature on the mycelium growth of Flammulina velutipes[68]

But the temperature does not only impact the mycelium growth rate but also the quality of the mushroom itself.

In an in-depth study, the authors found out that be lowering the temperature, all parameters (thickness, diameter, and length) will be reduced (Fig. 13).

But that does not have to be a bad thing. Depending on your region, your customer prefers only a specific size of the Winter mushrooms. You, therefore, can adjust the properties by adjusting the temperature.

Change of morphological characteristics of fruit-body on winter mushroom of white-line by the different temperature

Figure 13: Change of morphological characteristics of fruit-body on Winter mushroom of white-line by the different temperature[69]

Moisture

One of the most adjusted parameters is the moisture content of the substrate. How it will influence, the mycelium growth rate was part of the same study, which measured the influence of the temperature.

The study changed the moisture content from 55 % to 70 % in 5 % increments. The authors found out that the highest mycelium growth rate was achieved at a moisture content of 65 % (Fig 14).

The effects of the moisture content of the culture medium on mycelium growth of Flammulina velutipes

Figure 14: The effects of the moisture content of the culture medium on mycelium growth of Flammulina velutipes[70]

Humidity

Besides the moisture content, the humidity level within the grow chamber plays as you already know an important role. The same study mentioned earlier investigated the influence of the humidity level on the growth rate of the mycelium.

The authors found out that the mycelium growth rate was the highest if the humidity level is set to 75 % (Fig 15). Higher or lower will reduce the mycelium growth rate by around 15 %.

The effects of culture room humidity on mycelium growth of Flammulina velutipes

Figure 15: The effects of culture room humidity on mycelium growth of Flammulina velutipes[71]

Now I want to hear from you:

Are you growing Flammulina velutipes yourself?

If yes, what are your experiences with it?

If no, did you get excited growing Enoki mushrooms while reading my article?

Let me know by leaving a quick comment.

Literature

Osman

Tang

Harith

Chang

Yun

Stamets

Sharma

Chang-Sung

Kwon-Il

Hiramori


[1] Osman 2014

[2] Tang 2016

[3] Harith 2014

[4] Chang 1995

[5] Osman 2014

[6] Yun 2015

[7] Stamets 1983

[8] Sharma 2009

[9] Sharma 2009

[10] Sharma 2009

[11] Chang 2004

[12] Harith 2014

[13] Sharma 2009

[14] Sharma 2009

[15] Chang 2004

[16] Sharma 2009

[17] Own table based on Salihu 2015, Kasmani 2011, Aranata 2019, Cantero 2014, Xiao 2001, Ravindra 2006, Demirbas 2004

[18] Harith 2014

[19] Sharma 2009

[20] Chang-Sung 2010

[21] Stamets 1983

[22] Chang 2004

[23] Harith 2014

[24] Kwon-Il 2014

[25] Kwon-Il 2017

[26] Harith 2014

[27] Chang 2004

[28] Sharma 2009

[29] Harith 2014

[30] Stamets 1983

[31] Chang-Sung 2010

[32] Chang 2004

[33] Sharma 2009

[34] Harith 2014

[35] Sharam 2009

[36] Harith 2014

[37] Harith 2014

[38] Zhao 1989 in Sharma 2009

[39] Kwon-Il 2017

[40] Chang-Sung 2010

[41] Chang 2004

[42] Stamets 1983

[43] Harith 2014

[44] Zhao 1989 in Sharma 2009

[45] Kwon-Il 2017

[46] Chang-Sung 2010

[47] Stamets 1983

[48] Kwon-Il 2017

[49] Kwon-Il 2017

[50] Stamets 1983

[51] Stamets 1983

[52] Stamets 1983

[53] Stamets 1983

[54] Hiramori 2017

[55] Stamets 1983

[56] Tang-Xiang 2001 in Sharma 2009

[57] Osman 2014

[58] Osman 2014

[59] Own figure based on Harith 2014

[60] Stamets 1983

[61] Chang 1976

[62] Chang 1976

[63] Chang 1976

[64] Chang 1976

[65] Chang 1976

[66] Own figure based on Harith 2014

[67] Own figure based on Harith 2014

[68] Kwon-Il 2017

[69] Chang-Sung 2010

[70] Kwon-Il 2017

[71] Kwon-Il 2017

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