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Copyright © 2018 I. G. Safonov (IGSafonov)
Copyright © 2018 I. G. Safonov (IGSafonov)

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Thanks, Alan
By: I. G. Safonov (IGSafonov)
2018-11-23 22:52:42 EST (-0500)

Yes, I will append them to the end of the most recent FASTA file. Will take some time for me to retrieve the data from a bunch of files. Some of these TEF-1s are already on MO, others I haven’t attached yet because I wanted to work on my discussions, which hasn’t happened yet.
It don’t know why GenBank hasn’t published any of them yet — I submitted the data 6 months ago! It was a lot of work because I had to manually find all the introns and do the tedious protein translation… I need to give GB a nudge one of these days.

Sure
By: Alan Rockefeller (Alan Rockefeller)
2018-11-23 22:40:35 EST (-0500)

These trees are super easy to make, go ahead and edit the FASTA I sent you a few minutes ago and add the TEF1 sequences. If you don’t want to publish the sequences yet we’ll just show people the tree.

Thanks Alan — this looks great!
By: I. G. Safonov (IGSafonov)
2018-11-23 22:21:11 EST (-0500)

These several known genera really stand out as monophyletic clades. Note that the topology of Neoboletus is the most complex, as expected, but it still looks reasonable. B. huronensis and N. venenatus are buried inside Neoboletus.
Now it would be interesting to include all of my TEF-1 sequences of eastern red-poreds that should clade in Neoboletus into this tree. I have GB accessions nos. for these, but they haven’t been published yet. What say you, Alan?

New TEF1 Tree
By: Alan Rockefeller (Alan Rockefeller)
2018-11-23 22:06:34 EST (-0500)

Here is a TEF1 tree that Igor and I worked on. Igor removed a lot of extraneous sequences and cleaned up the names, making it a whole lot more readable.

https://images.mushroomobserver.org/huronensis-tef1.jpg

DNA results discussion
By: I. G. Safonov (IGSafonov)
2018-11-22 17:03:31 EST (-0500)

Results:
> A contiguous nrLSU sequence of 937 bps was obtained from this material and uploaded to this observation. The read starts in the beginning of the ITS4 primer and ends at the LR5 primer. There are no ambiguous characters. The 856 bps fragment (the region between ITS6R and LR5 primers) was BLASTn-ed, and the annotated GenBank-derived phylogram was posted here.
> A contiguous TEF-1-alpha sequence of 954 bps was obtained from this material and uploaded to this observation. The region between and including the EF1-983F and EF1-1567R primers is 610 bps long. There is a single ambiguity, “R”. This exact fragment was BLASTn-ed, and the annotated GenBank-derived phylogram was posted here.
Discussion:
As can be seen from the two phylograms, B. huronensis and N. venenatus are genetically related, which makes good sense in light of their morphological likeness. In fact, this was originally surmised by Nagasawa, who described B. venenatus in 1996.1
In 2015, B. venenatus was officially transferred to Neoboletus.2 Subsequent to this, Neoboletus was collapsed into Sutorius3. However, this synonymy has not been universally accepted, and questions still remain regarding the monophyly and taxonomy of Neoboletus s. lat.
Still, as it now stands, the yellow-pored and bluing huronensis and venenatus appear to be related to a large and diverse clade of red-pored boletes that define and delimit – both morphologically and genetically – the current generic concept of Neoboletus more than to any other known group or clade within the Boletaceae.
References:
1 Nagasawa, E., “A new poisonous species of Boletus from Japan” — Rep.
Tottori. Mycol. Inst.
33:1–6 (1996)
2 Wu et al., “Four new genera of the fungal family Boletaceae” — Fungal Diversity: 10.1007/s13225-015-0322-0, 18 (2015)
3 Wu et al., “One hundred noteworthy boletes from China” — Fungal Diversity 81: 145 (2016); https://www.researchgate.net/...

The pictures really do look like the asian venenatus
By: Scott Pavelle (Scott Pavelle)
2018-11-20 16:11:53 EST (-0500)

I have to question the Neoboletus genus, however. The morphology is just too different. Especially that weird texture! My guess – and it’s little more than that – is that full genome testing will ultimately find some set of genes that move huronensis and venenatus into their own, distinct category.

How they got there and what they branched off from… That is the real question that DNA will have to unravel. I suppose it might be the Neoboletus group but somewhere there’s a genetic left turn…

LSU Tree
By: Alan Rockefeller (Alan Rockefeller)
2018-11-20 15:28:06 EST (-0500)

I just made a tree which includes the new B. huronensis sequence:

https://images.mushroomobserver.org/neoboletus.jpg

It appears about 3/4th of the way towards the bottom, just above Xerocomus.

Hello, Linas
By: I. G. Safonov (IGSafonov)
2018-11-20 13:24:44 EST (-0500)

I was unaware you sequenced 174643 because, as you say, you got the material directly from Dave. I don’t have my own voucher of huronensis yet. So far I saw this species only once (2014 NEMF in ME) and at that time I had no plans to start my own herbarium.

I have ITS and LSU for …
By: Linas Kudzma (baravykas)
2018-11-20 12:53:25 EST (-0500)

https://mushroomobserver.org/174643 , which are identical. Have you seen these Igor? I think I got this sample directly from Dave not through you.

Linas

If there is a toxic clade in Neoboletus
By: Alan Rockefeller (Alan Rockefeller)
2018-11-20 11:12:47 EST (-0500)

That might be a good place to draw the genus line. I like the idea of having toxic and edible genera.

PGW,
By: I. G. Safonov (IGSafonov)
2018-11-20 01:09:59 EST (-0500)

This is based on one collection, but there is no question about the identity of this material, as the rare B. huronenesis is a very distinct species with no lookalikes (unless you consider B. pallidus as one). And no, B. huronensis has not been looked at by anyone phylogenetically — it missed all the recent Boletaceae studies.
The DNA evidence points in the direction of red-pored taxa ca. Neoboletus s. lat.
Yes, it is probably premature to draw larger conclusions, but not for the reason you state. The thing is, Neoboletus is probably polyphyletic and shouldn’t be lumped with Sutorius s. str.

Re: Hot off the press — DNA solves the mystery of B. huronensis
By: Peter G Werner (pgwerner)
2018-11-20 00:49:03 EST (-0500)

Is this based just on one collection, or is this based on a larger study of Boletus huronensis material? If it’s the former, it’s probably premature to draw the larger conclusion.

I’m very surprised B. huronensis hadn’t already been looked at phylogenetically.

obsi.
By: Robert(the 3 foragers) (the3foragers)
2018-11-19 09:07:25 EST (-0500)

These observation is been found in same spot as observation 250775.

Hot off the press — DNA solves the mystery of B. huronensis
By: I. G. Safonov (IGSafonov)
2018-11-18 19:10:05 EST (-0500)

Both nrLSU and TEF-1 sequences derived from this material indicate that B. huronensis is a member of Neoboletus (=Sutorius sensu Wu et al.). Specifically, TEF-1 data indicate that huronensis is closely related to the poisonous Neoboletus venenatus. Googling venenatus (as Boletus or Neoboletus) revealed that is actually close morphological lookalike of B. huronensis!!! For example, see http://toolate.s7.coreserver.jp/....
Neoboletus venenatus grows in association with conifers in Abies, Picea and Tsuga.

May be
By: Robert(the 3 foragers) (the3foragers)
2018-09-13 08:57:08 EDT (-0400)

I will send for LSU and TEF-1 as well

The ITS sequence…
By: I. G. Safonov (IGSafonov)
2018-09-12 16:10:13 EDT (-0400)

…matches KY826031 (Boletales sp. voucher B1178) and KY826065 (Boletales sp. voucher B1998) from Canada, sans a few ambiguities and deletions in the homopolymeric “G” motif near the 5’ end. The next hit is only 91% similar and belongs to a red-pored bolete.
Clearly, ITS is stripped of any useful phylogenetic information in this case, and serves as a simple genetic barcode, i.e., the pictured mushroom has this sequence. This result is perfectly in line with the unique morphological gestalt of B. hurnenesis that precludes assignment to any known genera. B. huronensis likely represents a new monotypic genus in the Boletaceae.