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I will report back here.
I’m sorry to hear about your difficulties with spore measurements. I’m glad I’ve been able to reduce some future grief.
Yes, I still have the .ab1 file. Unfortunately the sequencing reaction was a complete failure and had “NNNNN” in the margin of the Chromas histogram but I will still send the file to you.
Thanks for the tip about the spore measurements I wish I would have known that sooner. It would have made spore comparison much simpler.
Yes, I would like to add your material (or a portion of it) to the herbarium here in Roosevelt.
You should round your spore measurements to a single digit to the right of the decimal point. Two place accuracy is not possible with light microscopy.
Did you retain the unsuccessful .ab1 file or files?
Amanita “sp-lavendula01” produces incoherent .ab1 sequences for nrITs…for the most part.
However, the 3’ terminal motif of 18S (nrSSU) is present (GGATCATTA-3’) slong with the a few coherent characters at the 5’ end of ITS1.
Similarly, near the opposite [<—EDIT] end of the “failed sequence” you will find a few coherent characters of ITS2 followed by a coherent beginning of the 28S (nrLSU) sequence…i.e. a 5’-terminal motif of that sequence (5’-TTGACCTCAAATCA).
In just those two fragments of the proposed barcode (along with the adjacent motifs from 18S and 28S), there is enough data to separate sp-lavendula01 from sp-lavendula03. [<—EDIT]
You’re absolutely correct that the two taxa (up to the present date) do not have known characters that permit their separation without DNA sequencing.
If you can send me what you have of the raw data (.ab1 files), I will see if I can determine which taxon you have. If not, I will see if we can get a sequence from the dried material. We are really finished with data collection for the lavendula-related project; however, if you have the patience to wait a few months (our sequencing queue is long), I will see what I can find out for you.
I hope that you have the .ab1 files. This has a chance of producing a meaningful result sooner. I’m very curious to know if you can see the two motifs in your data.
I wanted to let you know, I have a duplicate of this specimen that I did not turn in with my project. Do you want it to add to your collection?
I think it may be Amanita sp-lavendula03 because I don’t recall the stipe being easily compressed and I think it has a faint radish-potato-like odor. Its difficult because the descriptions for A.sp-lavendula03 and A. sp-lavendula01 seem so similar.
When I did the DNA sequencing it was one of several (including “A.Peckiana” 189471 from another post ) that were unsuccessful. I’m really curious of what you would get with the DNA.
Nobody has done a study, even a brief one. Did you, or can you, record the results in photos? That would be cool, and your specimens should then be sequenced so that we know which species you saw fluoresce.
I’ve known several people who tried fluorescence as a way of adding a character for determining mushrooms quickly. However, I don’t think that a systematic collection and publication of results ever happened. Maybe that was because of doubt about determinations or because too many mushrooms produced the same results. You’d probably want to know what frequency(ies) of light were produced by the reaction to UV…maybe what spectra were produced. It would take a little organization and some care in designing the lab set-up.
thanks to MO and Patrick Harvey. Any of the three could be in your area.
The true lavendula has spores that are smaller and have a higher length:width ratio than do the other two. We can tell them apart genetically. Bacterial cloning (there’s something to google) using E. coli was necessary to separate the nrITS sequences from the hybrid; however, despite the apparent chaos of an uncloned ITS from the hybrid, you can still tell the three species apart based solely on the tiny fragments of recognizable “proposed barcode” that can be obtained from the hybrid. It just happens that the three taxa are different in those tiny fragments. A chance occurrence.
The refrigerator is not cold enough. [EDIT] You have to collect them after a freezing rain or a hard frost. At the Cape Cod NEMF foray of 2009 we had several consecutive days with temperature nearly constant at 0-1 degrees C with freezing rain. Someone collected very large buttons of one of the three taxa (I’ll look it up in a minute…the hybrid…“sp-lavendula01”). The collections was composed of very large buttons like spherical amethysts…deep, gorgeous purple.
Some how I came away without a photograph. I can’t understand it. Rats.
Thank you for the links and info. When I found them what was very apparent to me is that they seemed to glow florescent yellow (with a hint of green). They are dry now and are a very pretty golden yellow. I don’t recall them ever turning lavender. Even after I had put them in the refrigerator till I had time to Identify them. Just going by pictures I’d say Amanita sp-lavendula03 looks the most similar. Whether or not that is correct, I have no idea! I’ll add some photos of the dried specimens.
You can take a look at these three temporary codes with attached descriptions:
http://www.amanitaceae.org?Amanita%20sp-lavendula01 – a species with a complex of many different “proposed barcode” genes
http://www.amanitaceae.org?Amanita%20sp-lavendula02 – the true Amanita citrina f. lavendula
http://www.amanitaceae.org?Amanita%20sp-lavendula03 – a non-hybrid taxon with spores very similar to that of the hybrid taxon.
From the genes we know there are three (at least) in eastern North America. They all will turn lavender or amethyst or a similar color if they get cold enough. The European species of Schaeffer apparently doesn’t occur in N.Am.
Created: 2014-11-27 15:05:42 PST (-0800)
Last modified: 2014-11-30 07:51:09 PST (-0800)
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