Quote:

Paecilomyces hepiali Chen (strain CS-4) is probably the most popular commercial strain used to produce most of the mycelial products sold in the world marketplace as O. sinensis. It used to be called Cordyceps sinensis (strain CS-4). The strain CS-4 was isolated by the Chinese Academy of Sciences in 1972 from specimens found in Qinghai province, central China. It was tested for its medical potential, and by 1988 it had been approved in China as a medicine under the name Jin Shui Bao. At present there is a lot of reported evidence that CS-4 is not a correct anamorph of O. sinensis but belongs to a different fungus, Paecilomyces hepiali Chen.

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So, what was previously called the CS-4 strain of Cordyceps sinensis is really Paecilomyces hepiali, which been found to be an endoparasite of Cordyceps sinensis.

What is really interesting here is that one of the "Cordyceps" strains that was used for hybridization in the Holliday paper was CS-4 (note: it is written as CX4 in the paper, but that is clearly a typo as it is written CS4 here).


So, I would suggest that what was isolated is not a "hybrid" of two Cordyceps sinensis strains, but rather Cordyceps sinensis parasitized by Paecilomyces hepiali.

I also want to make it clear that I have no reason to question that the paper was written in good faith, or that the authors isolated a different kind of mycelium, or that the metabolite profile of the isolated mycelium was different. In fact, many microorganisms produce more and different metabolites when stressed. However, I think it is clear that the isolated mycelium is not a hybrid, and that the snake venom method for hybridization is not valid.

Quote:

Hybrid definition is - an offspring of two animals or plants of different races, breeds, varieties, species, or genera.
Hybrid | Definition of Hybrid by Merriam-Webster

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Peter65199 said:

I have never tried using rattlesnake venom so cannot say 100% but that first paper you quoted seems entirely feasible. Besides the proteolytic enzymes you mentioned the venom in question also contains chitinase.

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Peter65199 said:

@Pinback I meant to add above look into PEG mediated somatic fusion, could have potential?????


Hi Inski! Maybe we are using the same word for different things, the definition I am using is;

Quote:

Hybrid definition is - an offspring of two animals or plants of different races, breeds, varieties, species, or genera.
Hybrid | Definition of Hybrid by Merriam-Webster

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But this is just semantics... Inski is correct in biology you are taught that you can have intra or inter hybrids, both are by definition hybrids.

@Pinback, I sincerely apologize I also can not find what I was looking at when I said Crotalus Atrox has chitinase in its venom. I have papers that document other snakes that have it. But I dont know why I was under that impression, I think I was confusing it with the somewhat chitinolytic vipericidin in Crotalus durissus terrificus(wrong rattlesnake)? As you said it does not matter that much anyway since chitin only makes up small percentage of the fungal cell wall(similar to lignin in wood, a structural polymer). And as you said the venom is proteolytic, and could degrade/inhibit the vast majority of the cell wall anyway allowing creation of protoplasts and exchange.

If the paper was flawed you would hope that the peer reviewed journal in which appeared would have picked it up (but it happens sometimes), but why then did the researchers in the paper above even attempt this, quote from paper;

Quote:

The present study is based on the study conducted by
Holliday et al. (2004) using Snake venom for somatic fusion of C. sinensis and the developed strains
were reproducible strains and showed improved quantitative bioactive compounds. A similar effective
somatic fusion was observed in the present study using abrin, a plant poison.

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Quote:

Abrin was extracted by soaking 10 g pulverized seeds of Abrus precatorius in PBS (pH 7.7)
with 5 % NaCl for overnight at 4°C. The homogenate was filtered and centrifuged at 10,000 rpm for 30
min at 4°C.The crude abrin was dialyzed against distilled water against 10 mM Tris-HC1 pH 7.7 for 24
h each. The dialyzed solution was centrifuged at 15, 000 for 20 min, and the partially purified abrin was
used for hybridization.

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Snake Venom Into Hybrids <- Link to post.
This is an image he posted;

Quote:

I first learned of rattlesnake venom from attending a medicinal mushroom conference in 2005. John Holliday did a presentation on it. Here's a link to a paper he wrote. John Holliday is director of research at alahomedicinals.com and there's lots more info on his website.
RR
http://www.nwbotanicals.org/nwb/lexicon/hybridcordyceps.htm

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Quote:

Search this forum for the Redboy thread(s). I had some 20 year old redboy spores that simply would not germinate, no matter what I tried. After placing the non-viable spores on venomated agar for a couple of weeks, I released monokaryotic PR mycelium to crawl over the dish. Low and behold, when the monokaryotic mycelium reached the area where the spores were near the edge, two separate rhizomorphic strains emerged. Somehow the snake venom allowed the PR mycelium to access the nucleus of the spores for mating. I had tried this without the venom to no avail. The very first time venom was used, the result was dikaryotic mycelium, and the redboy strain was back as a 50/50 cross with PR. The experiment was repeated with the offspring, resulting in 75% Redboy(perhaps? I'm not a geneticist), which was then released to the community.
RR

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Quote:

John Holliday IS a real mycologist.  Write him to ask for more info if you wish.  He owns www.alohamedicinals.com

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@Pinback; I hope you dont take this an attack on you, its far from it! I actually respect your style, it would serve you very well when researching. I like people who dont just take things at face value, and dig deeper. Anyway, I hope we can work together on anything one of us may be having trouble with in the future....

Quote:

Potato lectin: a modular protein sharing sequence similarities with the extensin family, the hevein lectin family, and snake venom disintegrins(platelet aggregation inhibitors).

"Finally, all plant chitin-binding domains examined bore a remarkable sequence similarity, particularly in the spacing of Cys residues, to the disintegrins (platelet aggregation inhibitors) which occur in crotalid and viperid snake venoms."

https://www.ncbi.nlm.nih.gov/pubmed/8054990

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Crotalus Atrox venom profile;


Lysing the cell wall;

C.Atrox venom is has a high proteolytic activity compared with other snakes in general, these degrade and inhibit proteins. Since the chitin content of a fungal cell wall(in our case) is from 10% to 20%, which means 80% to 90% of the cell wall containing proteins and glucans are vulnerable. The snake venom disintergrins above bind chitin and act like some plant lectins which have some agglutination properties (agglutination is not fusion, but in a lot of cases once the polarity issues are overcome fusion can then proceed). There are also host of other enzymes I could get into like the metalloproteinase(but alas my time has run out).

Peter65199 said:

And when Pinback said it he doubted the hybridization would work, and Holiday was not a real mycologist RR replied;

Quote:

John Holliday IS a real mycologist.  Write him to ask for more info if you wish.  He owns www.alohamedicinals.com

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