Jump to content
The Corroboree
spooge

subaeruginosa phenotypes that were DNA tested and the results

Recommended Posts

Holy shit that's awesome work mate!!!

  • Like 2

Share this post


Link to post
Share on other sites

You know you're a sickcunt. Nice display pic too.

  • Like 1

Share this post


Link to post
Share on other sites

I said GODAMN! Hot shit Spooge, and an awesome resource!:o

 

*Come to think of it I'll keep that mob in mind for next season... which is pretty much now through next week if your a SE QLDian :lol:

Edited by Responsible Choice
grammar
  • Like 4

Share this post


Link to post
Share on other sites

Great work Spooge! The one with chantarelle-looking gills is amazing!

  • Like 1

Share this post


Link to post
Share on other sites
2 hours ago, DualWieldRake said:

What did you test em for?

Maybe too much visual variation between various isolated patches of subaeruginosa in his local area?
You'd be wanting to know what the fuck your picking etc, hearing stories that Psilocybe Cyanscens (not referring to pan cyan) might be in Aus etc and picking things that don't quite look exactly like they should, gill variances/cap shape 'n structure. I'd be wondering myself...  Or wondering if you've discovered a new mushroom.

You know the stories of Urban Victorian (childrens playground chipped beds etc) subs being twice as strong as fkn wild forest subs. Also having difference in gill/cap structure and fruiting in busy clusters vs lone soldiers etc...    People theorize that the chipped victorian urban sub could infact be or have strong  'AZURESCENS'  dna present.

Hearing what spooge found are like 99% identicle to Psilocybe Cyanscens is pretty amazing, I think its been stated in literature online before though.

Its like when Gondwana land split apart cyancens myc was on our Aussie cut of the earth and as we floated south-east and away from other shit the Cyanscens over millions of years slowly developed into our southern hemisphere continental Subaeruginosa mushroom phenotype we have today (clearly with other forms/subspieces etc floating around in certain isolated areas).  And the north American cut of the earth floated north-west and their Cyanscens myc slowly developed into the northern hemisphere 'Azurescens' phenotype known today...  With plenty of Psilocybe Cyanscens still around various parts of the planet too, seems different phenotypes have developed and peeled off that original species. I could just be off my head rambling way off track, but it is certainly froth worthy shit to think about.

It wasn't GC/MS testing or anything was it? Did you get a psilocybin percentage out of any of the tests done spooge?   Nice work again mate.  :worship:

  • Like 5

Share this post


Link to post
Share on other sites

I wanted to have these mushrooms tested as they are different from the run of the mill differening phenotypes amongst subaeruginosa that occur where I am, we get all kinds of subs, white gills, odd caps, weird stypes. 

 

Though I had the 3 samples tested I'm being told by a leading Psilocybe mycologist that I have at least 2 different species. I'm told also that I should have had ITS sequences done as they are more variable and that there are ITS sequences of p. subaeruginosa on Genbank to refference. Hopefully this guy can find the sample of the crazy gills I sent him so he can do the ITS sequences, if not I will get more samples this season and get them ITS sequenced. 

 

 

  • Like 3

Share this post


Link to post
Share on other sites

Did your mycologist contact encourage you to get the same DNA tests for future finds, or will you use the ITS sequencing alone next time?

 

Share this post


Link to post
Share on other sites

Thats awesome, very cool info. And how are you finding subs this early in the year, with such high temps and low rainfall? :blink:

Share this post


Link to post
Share on other sites

^ They were last years mushroom samples mailed off shortly after picking I'm guessing. Picked in winter 2017. Right now is too early for wild subs(?), a whole season too early. Must of taken like 8+ months to arrive at Pablo's Lab in Spain & for them to do the tests & forward the info onto Spooge.

I'm wondering how the fuck u keep mushies fresh 'n viable for testing going in the mail from Aus to Spain... lol...  Can they DNA test a dried specimen?

 

Edited by Skellum
paragraph structure
  • Like 1

Share this post


Link to post
Share on other sites

I reckon if it keeps raining consistently for the next few weeks we're about a month off sub season in the mountains. But it's been hell dry since June 2017, so who knows what will happen. It could go either way. 

There's a known sub spot where someone spread a lot of azurescens LC, I wonder if the wild population swapped any genes...

I didn't even bother heading into the forests last year as it had been so crazy dry. I feel a mission coming on in April! 

  • Like 2

Share this post


Link to post
Share on other sites

Nice work spooge! Those are some funky fungi indeed.

 

On 3/8/2018 at 8:13 AM, spooge said:

I encourage people to do this, pay a couple bucks and get your mushrooms tested

So how much did it cost you to have each of these samples tested? I found their pricing page, but am still a bit unclear on which items you chose and what you would do differently next time. Just to give us a rough idea of the cost, waiting time, and level of sample-prep needed from our end - a little more info on this would be great!

 

On 3/12/2018 at 11:17 AM, spooge said:

I'm told also that I should have had ITS sequences done as they are more variable

I'm confused - I thought this was what you'd done! The files are labelled "1F +ITS4". Or did you just mean using a more specific ITS primers? I gather that ITS1/ITS4 combo is kinda general (the "universal fungal primers") - and others may work better with particular fungal families or testing/sampling conditions. I guess if there's an existing database of reference samples then you'd want to match their methods so you got comparable results (maybe?)... but what are those? And how important are the differences?

  • Like 3

Share this post


Link to post
Share on other sites

I have been meaning to make this comment for a little while.

 

but i am very concerned with the idea that specimens are leaving the country and not being deposited with local herbariums.

 

Its important that we support local opportunities for research, and while taxonomic work on Australian Psilocybe species is not likely to happen for a while, when someone from australia is ready to be able to it would be nice if there was a decent amount of samples ready to begin work.  I dont like the idea that overseas researchers are taking possible opportunities from local researchers.

 

In terms of getting DNA testing or sequencing done I appreciate that we have to go overseas, all uni's do the same anyway,  but is important that the results are lodged with genbank so they become available then to researchers.

  • Like 2

Share this post


Link to post
Share on other sites
On 12 March 2018 at 11:20 AM, Wile E. Peyote said:

Did your mycologist contact encourage you to get the same DNA tests for future finds, or will you use the ITS sequencing alone next time?

 

 

The international mycologist is very busy, I get responses to my messages Wile, still waiting to hear if the sample I sent him first before alvalab has been found. This is why I went with alvalab as when you pay for a service it usually gets completed a bit faster. I had let this go and was content with the 98% from alvalab and 99% from a local mycologist who viewed the results also, Then i sent the results to the international guy and it's all in the air again as he is quite sure just from external characteristics that I have at least 1 different species that may not be a sub in the 3 samples.

 

On 14 March 2018 at 9:31 PM, Anodyne said:

Nice work spooge! Those are some funky fungi indeed.

 

So how much did it cost you to have each of these samples tested? I found their pricing page, but am still a bit unclear on which items you chose and what you would do differently next time. Just to give us a rough idea of the cost, waiting time, and level of sample-prep needed from our end - a little more info on this would be great!

 

I'm confused - I thought this was what you'd done! The files are labelled "1F +ITS4". Or did you just mean using a more specific ITS primers? I gather that ITS1/ITS4 combo is kinda general (the "universal fungal primers") - and others may work better with particular fungal families or testing/sampling conditions. I guess if there's an existing database of reference samples then you'd want to match their methods so you got comparable results (maybe?)... but what are those? And how important are the differences?

 

Was around 80eu to have the 3 samples tested Anodyne, you pay for the services you require from a list. Took around 6 weeks including post time of 7-10 days to get the results back and then a futher week as I asked for another test to be done to tryn narrow down the extra 2% thing.

 

Im confused as well man. Tryn to get this done so I have a definitive answer on at least one of the samples is frustrating. I've no idea but I think I need a result that says 100% like the first sample in the pics, then I will be happy. Mushroom tests prolly don't work like that, who knows...... 

 

On 15 March 2018 at 11:16 AM, obtuse said:

I have been meaning to make this comment for a little while.

 

but i am very concerned with the idea that specimens are leaving the country and not being deposited with local herbariums.

 

Its important that we support local opportunities for research, and while taxonomic work on Australian Psilocybe species is not likely to happen for a while, when someone from australia is ready to be able to it would be nice if there was a decent amount of samples ready to begin work.  I dont like the idea that overseas researchers are taking possible opportunities from local researchers.

 

In terms of getting DNA testing or sequencing done I appreciate that we have to go overseas, all uni's do the same anyway,  but is important that the results are lodged with genbank so they become available then to researchers.

 

Im all for keeping the things we find here, here. Prob is though the whole Psilocybe mushroom thing here is so covert. Even transporting the sample or samples to a herbarium one would risk prosecution, prob can get an official letter etc, Pablo offers to upload the data for a fee to Genbank which I will prob do, once I have a definitive answer one way or the other on what the 2 samples are that are still in question.

 

This season I will collect samples of the 2 odd subs and have them tested agian, I may go with a couple labs and get the full range of tests done so the data is there for everyone to analyse. Both these mushroom grow in quite isolated places, in pine coups, that were once bush.

 

 

 

 

Edited by spooge
  • Like 4

Share this post


Link to post
Share on other sites
2 hours ago, spooge said:

i will happily supply samples to who ever finally does choose to do the work on subaeruginosa that desperately needs to be done.


^ I need pounds.
Lots of work to be done....    :wink:

  • Like 2

Share this post


Link to post
Share on other sites

Citizen science at its best, and done without a vested interest.

 

Spent your own coin and risks.... Buy this bloke a drink:wink:

  • Like 6

Share this post


Link to post
Share on other sites
On 3/16/2018 at 9:11 AM, spooge said:

Im all for keeping the things we find here, here. Prob is though the whole Psilocybe mushroom thing here is so covert. Even transporting the sample or samples to a herbarium one would risk prosecution, prob can get an official letter etc, Pablo offers to upload the data for a fee to Genbank which I will prob do, once I have a definitive answer one way or the other on what the 2 samples are that are still in question.

 

This season I will collect samples of the 2 odd subs and have them tested agian, I may go with a couple labs and get the full range of tests done so the data is there for everyone to analyse. Both these mushroom grow in quite isolated places, in pine coups, that were once bush.

 

I have been thinking about this.

 

I would think that if you had collected a few specimens, dried them out appropriated, and then written up a herbarium style label, with species name, approximate location and habitat details etc., such that the herbarium could then create its own label with sufficient data, you would have a sufficient case to claim its for academic purposes.

 

You have already paid to have the specimens tested, it would be a shame that the data were not available to others.

 

i think i have made the point before though that I really dont think we will have a sufficient answer till we are doing full genome sequences at a decent depth so that someone can do some proper bioinformatics on specimens.  untill then every bit helps so you are to be congratulated on your efforts.

  • Like 2

Share this post


Link to post
Share on other sites
On 3/15/2018 at 11:46 AM, obtuse said:

but i am very concerned with the idea that specimens are leaving the country and not being deposited with local herbariums.

 

Its important that we support local opportunities for research

 

 

Yup, this /\/\/\/\/\

 

I'm not sure if there is some process involved in herbarium/ info submission which would give preference to the first institution to log the data- I did bump up against something of the kind a few years back but didn't pursue the specifics.

 

But the grumpy taxonomist ( is there any other kind? ) at the other end gave me a stern reminder that we need to remember to submit first to Australian institutions if we want them to get recognition and funding.

 

Which is totally fair enough IMO

 

Alvalabs in Spain do a great job of the sequencing for a reasonable price, but I think there are now local services which offer identical services at similar prices.

  • Like 2

Share this post


Link to post
Share on other sites

I've tried for years to get local help and always find closed doors. It's the reason things get sent overseas. If local institutions and enterprises want to open some doors then I will support them. Until that time I will deposit samples in the proper state herbariums and continue to send samples out to be sequenced.

 

I'd love to know of any local places offering services, if you know.

  • Like 1

Share this post


Link to post
Share on other sites

I've often theorized aussie subs are actually Cyanscens due to forestry links between australia and west coast USA and the fact subs are almost always found in proximity to Pinus plantations and indigenous australians having no history of use. The variation between the species could be drift and local adaption over the relatively short period of logging in australia. 

 

Unless someone finds evidence to the contrary, I am not yet convinced subs are endemic to Australia.

Edited by botanika
  • Like 2

Share this post


Link to post
Share on other sites

Botanika, i think you and i have discussed this idea before.  I am very open to to this idea and would love to investigate it further.  on one hand i suspect Psilocybe subaeruginosa may represent a very fast evolving species that has taken rapidly to Australian eucalyptus woodlands.

 

or on the other, if it werent for Psilocybe azurescens, i would suspect it could be the case the other way.  maybe it did but P. azurescens represents a speciation event outside the time frame of at least colonial movment

 

And then there is the very close genetic similarity between P.subaeruginosa and P. weraroa, and thats where things get a little confusing, as it has been suggested that P. subaeruginosa evolved from P. weraroa.  additionally P. subaeruginosa and P. cyanescens are damn near identical genetically.

 

i think i have to review the literature again.

 

do you have thoughts?

Edited by obtuse
  • Like 1

Share this post


Link to post
Share on other sites

The 99% similarity to P. cyanescens needs to be considered in the context of which locus you got sequenced, and there seems to be some confusion about that. The letter says TEF, while the chromatograms say ITS4. As mentioned, ITS is ideal for this, as it is the most widely used locus in molecular species delimitation. Species-level difference in ITS is typically 3 - 5% depending on the taxon. 

 

Can you upload or PM a text file of the sequences? This is the easiest way to work with the information. I'd be happy to help you make the most of this. 

 

  • Like 3

Share this post


Link to post
Share on other sites

what about P tasmanica? Is the jury out on that yet?Easily the best mushroom I've ever talked to in my life....

Share this post


Link to post
Share on other sites

Create an account or sign in to comment

You need to be a member in order to leave a comment

Create an account

Sign up for a new account in our community. It's easy!

Register a new account

Sign in

Already have an account? Sign in here.

Sign In Now

×