Grow Diary - Wild Tricholoma Crassum (Berk.) (Rhino Feet Mushroom)

[Packdee]

On 16 May 2013 I found a thread in pantip.com web board asking to I.D. a flush of wild giant mushroom in her mango orchard.

I posted to tell her that it was the Tricholoma Crsassum (Berg.), an edible mushroom.

I also asked her to collect a mushroom for cloning.

Her house is near Prasat Phanom Wan an ancient Hindu temple in suburb Nakhonratchasima Province or it’s nick name is Korat.

Korat was an importance frontier city in North-East Plateau from Ayuthaya period to the begining of Ratanakosin period.

I live in Saraburi Province the central part of Thailand but fortunately on the part of Saraburi that next to Korat border.

My place is near the old rout that Ayuthaya and Ratanakosin moved her Army to control the North-East Plateau area and all area along the left bank of Mekong river.

I drove along the old rout to Huay Bong - Dan Khun Tot – Non Thai –

– Prasat Phanom Wan.

Finally I got a mushroom back to my LAB for cloning.

Day 7th after I laid a small piece of the T. Crassum tissue on PDA.

(To be continued.)

Edited May 30, 2013 by Packdee

[RedBullHorn]

Day 7th but the dates on the photos doesn't match, wrong date setting maybe ?

[Packdee]

Day 7th but the dates on the photos doesn't match, wrong date setting maybe ?

KHOP KHUN KRAP RedBullHorn. Last night I posted a wrong file.

And link to Army in English.

Army of Thailand in Haw wars (1875)

[sjefrie]

Col. Packdee
I read your reports about farming and i like them very much i also learn a lot of you
But maybe you can post youre historical pictures here
http://www.thaivisa.com/forum/forum/155-thailand-and-siam-history-photos/
I am sure they will appreciate very much by other members of Thaivisa

[Packdee]

Col. Packdee

I read your reports about farming and i like them very much i also learn a lot of you

But maybe you can post youre historical pictures here

http://www.thaivisa.com/forum/forum/155-thailand-and-siam-history-photos/

I am sure they will appreciate very much by other members of Thaivisa

Thank you sjefrie.

I have new place to post the historical pictures.

In this case I just follow our tradition to honor our ancestors who protected this land for us to do farming and ask them for help.

My target is to bring back the cloned mushroom to grow on the original ground in vicinity of Prasat Phanom Wan.

I vowed our ancestors help me to accomplish the mission.

I promise to offer them chicken, pork and Lao Khao.

Week 2 of the Tricholoma Crassum tissue on PDA.

Compare to day three of Straw mushroom tissue on the same batch of PDA.

One week of Milky mushroom tissue on the same PDA.

[Packdee]

Week 3 of the Tricholoma Crassum tissue on PDA.

Week 2 of Milky mushroom tissue on the same batch of PDA.

Compare to 10 days old of Straw mushroom.

[zeichen]

Do you find that potato agar works better than malt?

I haven't been able to find a supplier of agar especially premixed (saves time from boiling potatos, etc)

Those cultures look perfect. How many did you make? I usually make at least 10 different samples for a clone just incase of contaminations. I also like 3 partitioned peachtree dishes.

Do you make your slant master directly or do you first isolate the strains?

[Packdee]

Thank you Zeichen,

I am waiting some one to ask me about my PDA.
My PDA is not PDA. It is CWA (Coconut Water Agar)
1.5 liters of coconut water and one 25 gram pack of agar, that's all, no dextrose adding or PH adjusting. PH of coconut water exactly is 5.5.

The 25 and 50 gram pack of Food grade Agar can be found at almost every grocery shop in Thailand. A 25 gram pack costs about 50 B.
We use agar for making desert, the jelly like sweet with coconut milk or "วุ้น".

If I clone a rare mushroom I also make at least 10 different samples. In this case, the wild Tricholoma Crassum cloning, nine samples get contamination.
The pictures shown above I cloned mushrooms from fresh tissue.

Edited

One of the contaminated samples. Can you ID the contamination.
I heard that the germination of T. Crassum spores depend on some types of yeast.
I want to cultivate that yeast.

Edited June 14, 2013 by Packdee

[zeichen]

I will definitely try you CWA. I have never used food grade agar. Do you find that they gel strong enough? I usually can keep a lab grade agar master culture in storage for almost 2 years. I tend to repropogate every 6 months to a year, if it is a good strain though.

I definitely haven't seen that pick contamination before. I usually get the green, and sometimes a scary black one. I wonder if it was carried in with the mushroom or is it a common contamination for you? Try turning your flow hood or hepa filter on 1 hour before working and as you know never wear the same clothes from outside. Lucky you got one good culture from those 10. Would have been sad to loose the whole batch.

I lost a batch of lions mane to contamination. I forgot to wash my glasses and I touched them for while switching plates. I know a very novice move but it happens. Costly mistake because I bought the spores and they were expensive.

I am kind of interested how hard/easy it is to get samples or innoculate more cultures from using the bottles. I use petri dishes and very easy to open lid and take slices. But since bottles are almost free would save a lot, but not worth it if it adds extra difficulty to getting or growing samples.

[Packdee]

This FAO paper describes every steps of small bags tek of mushrooms growing.

http://www.fao.org/docrep/004/ab497e/ab497e07.htm

PDF version

ftp://ftp.fao.org/docrep/fao/004/ab497e

[Packdee]

Comparing the progress of mycelia of different mushroom tissue in same batch of CWA (Coconut Water Agar)

From left; 4 weeks Tricholoma Crassum, 3 weeks Colocybe Indica, 17 days Volvariella volvacea (Pady rice straw mushroom)

I decided to multiply the T. Crassum to ten bottles of CWA and some bags of grain.
I prepare CWA from coconut water. The fruit should not be too young or too old.

Recipe: coconut water 1.5 liters + agar powder 25 grams (food grade agar is available in 25 gram pack and 50 gram pack).
Boil coconut water, mix well agar power with cold water in separate bowl and pour into boiling CW, keep stirring.
Use a syring to fill the bottles 30 CC each.

Normally PH of CWA is 5.5. No need to be adjusted for mushroom tissue cultivation.

[zeichen]

Those are perfect cultures. No small satelites, contamination, growth spreads out quite full, only see one or two segments also so the strain looks pure. That is really good for one cycle. I have found that sometimes I need to make 2-3 generations before I get something that pure.

If we had a forum member of the year. I would definitely vote for you. You add so much knowledge and kindness to these forums. Thanks again for your wonderful contributions.

I still cannot find a decent pressure cooker here for cheap and wonder if there is a way to sterilize the agar in the bottle that is low tech but still effective.

Edited June 17, 2013 by zeichen

[sjefrie]

Thanks again Col. Packdee for the info

[Packdee]

Week 5 of wild Tricholoma Crassum culture in CWA.
It’s time to transfer to more new CWA and the directly to the grain to shorten time.

Grain spawn preparation.

I grow sorghum for my own purpose to make sure the grain is free from pesticide.

Soak the grain over night.

Precook the grain with rice cooker.

Spread the grain on a sieve allow it to dry by air to suitable moisture content.

Fill the grain into bottles or pp bags.
Sterilize at 15 psi or 121 degrees C for 30 - 45 minutes.

[zeichen]

I noticed that there are a few different sectors on that culture. Are you going to test different strains or just cut sections and propagate your grain?

Is sorghum the cheapest grain in Thailand or is it just most effective?

[Packdee]

I noticed that there are a few different sectors on that culture. Are you going to test different strains or just cut sections and propagate your grain?

Is sorghum the cheapest grain in Thailand or is it just most effective?

In this case I just cut sections and propagate to more CWA bottles and some pieces will be transfered directly to the grain bottles.

I think that the culture from one piece of fresh tissue should be one strain.

If spores germnations we have to test different strain before release for comercial use.

Corn is the cheapest grain here.

We can use corn or rice to make grain spawn, the result is same.

The round grain of sorghum helps to pour grains into the bottle neck of plastic bags easily and quickly.

Edited June 25, 2013 by Packdee

[zeichen]

Nice. The left bottle looks like rice with husk on, do you cook that like the sorgum or just soak? Are you testing to see which one cultures better?

[Packdee]

Nice. The left bottle looks like rice with husk on, do you cook that like the sorgum or just soak? Are you testing to see which one cultures better?

There are many straw mushroom farms in my area.

I am trying to be a staw mushroom spawn supplier.

Normally the compost waste cotton and bean shell are used to be substrate for straw mushroom spawn.

Most of straw mushroom spawn suppliers inoculate the fully colonized substrate into new sterilized substrate bag.

I cloned fresh tissue of straw mushroom fresh tisue into rice grain directly (left bottle).

Straw mushroom mycelium grew very fast in rice grain and produced good fruit in mushroom bed.

But my substrate, the rice grain, costs much higher than the cotton waste and bean shell.

I am looking for other cheap material to be substrate for straw mushroom growing.

I pre-cook the grain and dry by air to get proper moisture content before filling into the bottles and sterilize them.

The other way, without pre-cooking, by soaking the grain to get 70% more weight then fill the grain into the bottles.

[zeichen]

Yeah that has been my problem also is finding the right grain that is cheapest for grain spawn and also substrate materials for growing beds. Keeping costs down is the hardest thing to making it worth it.

Cloning straight into the grain, you must have perfectly sterile conditions. I don't know many that have had success doing that. You are really perfecting your skills.

I would think that the natural rice in the husk would be a lot cheaper than other things just because of the amount that is grown here.

I would think that manure and chopped straw with a little EM and lye would be a common substrate for straw mushroom beds. I wouldn't think that cotton was that available. What bean shells? Soybean?

Again thanks again Pacdee for your wonderful contributions to the farming forum. If we had a poster of the year, you would be my choice.

[Packdee]

Straw mushroom spawn is made from cotton waste and soybean shell or greenbean shell compost.
Long time ago we used horse manure and lotus seed shell compost.

The substrate for growing this mushroom we can use variety of farming waste that contain more cellulose than lignin.
The mushroom growers in my area use skin of cassava root.
It is the waste of starch factory, so it is very cheap just the cost of loading and transportation.

[Packdee]

The mycelium of wild tricholoma Crassum in CWA was transfered into ten CWA bottles and ten grain bottles.

During waiting the grain to be fully colonized I would like to show how to make straw mushroom grain spawn.

Prepare the sterilized grain in PP bag.

Select fresh straw mushroom from a farm or from a flea market.
The good one should have complete cover tissue, no crack to reveal the cap.

Working in a sterile condition.
Peel out the cover without touching the inner tissue.
Cut a piece of tissue with a sterilized needle.
Lay it on the grain or cover with the grain in the bag.

3 hours, 24 hours, 48 hours.

[Packdee]

72 hours

[Packdee]

The mycelium fully colonizes 500 grams grain within 5 days.

Tomorow I will spawn it into a portion of the mushroom bed being pasteurized today.

I expect that within ten more days I will get the fruiting like this.

[IsaanAussie]

Col Packdee,

Do you sell straw mushroom grain spawn yet ?

[Packdee]

IsaanAussie

Not yet.
I am testing to substitute the grains for waste cotton and bean shell compost.

Paddy straw mushroom spawn is a fast money making.

The mushroom grower pay for the spawn more than substrate (in my area they use the waste from tapioca starch factory).

The rotation for a new corp of mushroom is every six weeks.

There are about 12 mushroom houses in a farm to keep earning money up to 5k B every day (whole sale for 100 kg/day)

Normally spawn suppliers use waste cotton and bean shell compost.
I cannot find enough amount of waste cotton at this time.

http://www.phetphichit.com/manufacturing.php

Edited July 9, 2013 by Packdee

[zeichen]

Is the cotton waste that cheap?

I would think that the rice would be cheapest at about 6-8 baht a kilo. Some people have had success with dried and shredded coconut husk for certain types of mushrooms if that is cheaper and available to you.

Are you doing grain to grain transfers after your first colonization or putting tissue culture directly in each and every bag? I would think that g2g would be easier and faster if you have a culture master.

Where are you getting the pvp bags?

What kind of autoclave set up are you using for mass production? You had a picture of a large beast but you said that your wife won't let you use it because of fuel consumption.

I was looking into using a dual 55gallon drum system. The bottom filled with water and heated and the second attached with pipes to fill with steam and a release valve. http://www.mushroomvideos.com/55-Gallon-Drum-Sterilizer

But I would like to be able to do more than 100 bags a day. and since pasteurizing takes longer time, it uses more fuel. I think that if I could pasteurize about 1k bags a day in one unit would be more cost efficient.

Figure you would need about 20x 500gm spawn bags per 5x8 meter hut?

So if these farmers are doing 10 huts or so every 6-8 weeks, 100kilos of grain spawn at even 40 baht a 500gm bag would make a pretty good profit. (not sure the going rate for straw mushroom spawn but I figure it is probably higher than 40)

8k baht per farm that you sell to every 2 months minus production cost, even using rice, pvp bags and fuel for pasteurizing, you would still make around 6k.

Get 10 farms and you are pulling in 60k every 2 months. But then you will need to hire more help which would add to your costs.

Overall it seems that supplying farmers is lower cost with higher profit margin.

Soon all the farmers will stop growing mushrooms and start making spawn. Then no one will be there to buy the spawn.

[Packdee]

Only five days the mycelium almost fully colonize the pasteurized compost.

Next step is spaying water on the fully colonized substrate pretending to kill the mycelium with the water.
Before we used a flame torch to burn the mycelium around the bed.

Once the mycelium is going to die it starts to produce fruiting and produce spores for it's surviving.
That is the life cycle of cultivated paddy rice straw mushroom.

@zeichen
We can use other material to make straw mushroom spawn but I grow sorghum so I try to use it first.

For straw mushroom, putting tissue culture directly in each and every bag or to PDA and the first generation on PDA to each bag is the best way to do.
Some supplier buy reliable spawn from other and transfer spawn to spawn.

Normally the local mushroom growers who make the substrate bags for sale will order large amount of the bags and we can buy from them.
Or you can order here http://anonbiotec.gratis-foros.com/f1-forum (You can ask any question about mushroom growing here, Dr.Anon he was a mushroom expert working for FAO in South Africa countries for many years. He has a vast collection of mushrooms strain)

I added an outside steamer using rice husk as fuel to my large autoclave in order to save LPG. It is OK and I am allowed to use it.

To pasteurize about 1k bags we use a thick tarpaulin to cover the piled up racks of the substrate bags.
Steamer may use rice husk to be fuel if available.

I am planing to use solar energy as a heat source.
Only making a collector like a green house and some focusing reflectors to increase the temperature.
Then I can pasteurize my substrate bags up to 90 degrees C for at least 6-8 hours.

There still be some secret for making good reliable straw mushroom spawn, so not any one can do it.
Once the mushroom growers do not get good product from our spawn they never come back again.

Edited July 10, 2013 by Packdee

[Packdee]

My steamer is working to heat the large size autoclave to pasteurize straw. (lower than 100 degrees C for six hours)
For sterilizing the grain it is used with an electric blower to provide more heat.
After purging air out a LPG stove may be used to help bringing hot steam pressure to 15 psi.

[Packdee]

After the grain bottles of Tricholoma Crassum was fully colonized I multiplied the grain spawn into 400 grams grain bags.
I simply poured the fully colonized grains into the sterilized 400 gram grain bag in sterile condition.

Now the 400 gram grain bags are fully colonized.
Next step, I need pasteurized substrate to be spawned.

Edited August 5, 2013 by Packdee

[Packdee]

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Last week I did mass casing the fully colonized wild Tricholoma Crasssum by peeling the bag out
and placed many blocka in the same bed next to my giant Milky mushroom bed.
Hope that the wild Tricholoma Crassum will give the fruit larger than the Milky.

(The size of mushrooms are exaggerated by wide angle lense)

Edited September 12, 2013 by Packdee