Just curious if you ever sequenced “sp-N50”. If so, what were your findings?

placed in Phalloideae. I did have difficulty convincing myself that the spores in https://mushroomobserver.org/242125 (which looks like a dead ringer for Alexey’s), were not at least weakly amyloid.

…of “sp-57” also supports placement into sect. Phalloideae.
There are three ITS accession in GenBank, at least one of which is a full-length sequence. A BLAST of KX061520 (with SSU/LSU ends trimmed off) gives A. levistriata as the closest relative at 97.6%, followed by pseudogemmata, hesleri, zangii, franzii, and ballerina. The remaining names on this 100-strong hit list are all taxa in Phalloideae.
With regard to the above named entities, in their multi-locus analysis of the Amanitaceae Cui et al. (2018) write:
“It is noteworthy that A. ballerina, A. franzii, A. hesleri, A. levistriata, A. pseudogemmata and A. zangii, grouped with high support with species of the sect. Phalloideae, and occupied a position basal to the rest of the section as two subclades (A. hesleri-A. zangii subclade and A. pseudogemmata-A. levistriata-A. ballerina-A. franzii subclade) [(Cai et al. 2014; Figs. 1b, S11]. Because the six species in these two subclades were significantly different morphologically from the remaining species of sect. Phalloideae, with the characters discussed in the commentary on Amanita sect. Phalloideae, they should eventually be treated formally as subsections of this section, in order to highlight the morphological and chemical diversity. However, formal descriptions of the subsections are not made here because more extensive sampling of Amanita species is necessary for a better resolution of the internal topology of sect. Phalloideae (Tulloss et al. 2016).”

We were able to obtain over 2,050 characters beginning within the nrSSU gene, extending through nrITS and containing a large portion of the nrLSU gene.

This species is genetically distant by a minimum of 5.5-6% from the top 27 matches in a BLAST run. This indicates it is not closely related to any sequence posted to GenBank. Common properties of taxa with sequences listed in the top the top 27 include membership in the morphologically defined Phalloideae and the presence of amatoxins in their fruiting bodies.

It should be emphasized that the similarity was judged based on the nrLSU component of our composite sequence, which is possibly misleading. When we compare our data to data sets restricted to nrITS, we get 100% match to the sequences we have posted for Amanita “sp-57.” This entity has inamyloid spores. In the past we have judged it to belong in section Amanita.

Consequently, we have situation that is a difficult one. The use of BLAST with relation to two commonly used “ID” genes would place the species in two different subgenera.

Situations such as this are motivators for experimentation with singly occurring, protein-coding genes over the whole of the genus Amanita.

I have a New Jersey collection of what I identified as “sp-57” last year, obs 323946. If memory serves, this is not the first time our club members have collected this taxon in this part of the Garden State; however, we haven’t seen it anywhere else…
I looked for the glass slide with the spore print, but couldn’t find it. I could take a tissue sample and look at the spores in Melzer’s to check for amyloidity. Also, I should consider getting my “sp-57” sequenced…
Ron, the spores of your TX collection, obs 242125, do look vaguely amyloid. I think this makes some sense in light of the recent Amanita study of Cui et al. (2018) that puts A. levistriata and the similar looking, albeit all white, Asian A. ballerina (see the link below) into the basal position of section Phalloideae (!).
https://www.researchgate.net/...

https://mushroomobserver.org/242125 but there was some question as to if it is a match to A. “sp-57”

R

Thank you for sending this material.

Very best,

Rod