Notes: > A family of four specimens growing on the grassy margin of a trail under pitch pine and black jack oak saplings in the North Gate section of FPP near Bertha’s canal.
> This is the same mysterious bolete originally reported in obs 102459 and obs 143371.
> Spore measurements and pix will be posted later.
DNA Sequencing Discussion:
In the past I tried to get a nrITS sequence for obs 143371 (the original holotype, obs 102459 went to Dr. Roy Halling in 2012), but the effort was thwarted by a repeating nucleotide fragment that caused the ensuing sequence to be non-specific. It was explained to me that since such motifs can vary in length and occur more that once (naturally, it doesn’t have to be the same nucleotide), the scrambling renders the sequence unreadable. The only way to overcome this intra-genomic variation problem is to run sequences for at least two collections (in case of one repeating motif) or to do cloning, the latter being expensive and labor intensive.
Unfortunately, the intra-genomic variation problem occurred with this collection for both nrITS and nrLSU!!! Apparently this annoying problem is a feature of this bolete.
The central portion of the LSU sequence is unreadable due to intra-genomic variation, and only two front/rear partial sequences (419 and 915 nucleotides long, respectively) are available. Neither produced a hit in GenBank.
However, reconstruction of a contiguous nrITS sequence from the various overlapping fragments that came from the two collections became possible (the sequence had only one repeating motif, “AAAAAA”). This full-length ITS sequence did not produce a match in GenBank.
So, this DNA saga ends here. Getting Tef1 or RPB sequences for this sp. nov. would not make sense now.
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Created: 2014-09-19 16:22:21 CDT (-0500)
Last modified: 2015-05-16 21:29:12 CDT (-0500)
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