A request and thoughts on Calycellina (Phialina) lachnobrachya

This characteristic leaf-inhabiting fungus is giving me trouble. The two last seasons I've been collecting "hairy" Helotiales and in most cases I've managed to retrieve a live culture on agar-media. Some genera are more reluctant than others, but with enough effort they usually make it. Still, lachnobrachya seems a bit special.

I have around ten collections of lachnobrachya, mainly from deciduous leafs (Betula, Quercus robur). They seem to shoot spores nicely on agar, spores germinate and an ok looking hypha appears on agar. But, once it gets through the extracting/PCR/sequencing protocol, the sequence is a contaminant, BLASTing to, say, near Dothideomycetes. Sure, leafs have a wide range of inhabitants and non-Helotialean fungi have a right to occupy leafs as well. But I need to change tactics. Other species in this group have been equally unsuccessful, but with n=1 or 2, it could be just bad luck.

The (Scandinavian) populations of lachnobrachya we have seen so far are usually small (<10 apos / leaf). Maybe you have it better in central Europe? I recently got Calycellina-like material from Rubén (Spain), a really wonderful collection of >100 apos, and some even shot spores. That was a peculiar fungus, but if any of you happens to run in to a handsome population of "ordinary" lachnobrachya (4 spored & whip-like hairs) I'd be more than happy to receive _a freshly dried specimen_ with preferably more than 30 apos. I can't promise much in return other than a warm thank you, a Christmas card and a round in a local bar once I get myself to some workshops.

Of course, if any of you have similar experiences I'd thrilled to hear. Or better, tell me how it should be done.

Getting there,

Timo

Hans-Otto Baral, 24-02-2017 09:59

Re : lachnobrachya

This is bad luck indeed, but the it is not the season now for this fungus. Did you ever try to take a sequence directly from the apos in such a case?

There is actually only C. lachnobrachyoides in my sequence collection, so this would be an urgent item.

Did any phialides appear in the culture? Calycellina should have such, at least some of the species.

Zotto

Kosonen Timo, 24-02-2017 12:53

Re : lachnobrachya

Yep, GenBank is lacking Calycellina -sequences. Looking forward in delivering atleast one. And yes, it's not the prime season. I'll need to repost this later during the season.

I'll check the cultures, but as I recall it, there were no conidia what so ever. but that would only be logical since I don't seem to have the right sequence. So, most probably something happens already when spores land on agar or immediately after. Somekind of facultative fungal "parasite" that dominates then on plates...

No dry extraction yet since all the populations are so small, theoretically possible of course.

T

 

Hans-Otto Baral, 24-02-2017 15:51

Re : lachnobrachya

Not only theoretically. It is actually possible to take a sequence from a single spore! At least if it is not too small. Phragmo-or staurosporous conidia have been sequenced and I saw it also for Nosema spores.

Our Orbilias are often only 0.5 mm or less, nevertheless you can take them with a needle in the hydrated state, including  the base with some algae and other fungi, and the result was often great.
 
Zotto

Kosonen Timo, 24-02-2017 16:00

Re : lachnobrachya

that's good news! It certainly looks like that I need to get into extracting from dry material for many species.

T