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03-06-2022 23:11

Sergey Markov

I found small Lachnum on a stem of Filipendula ulm

05-06-2022 09:50

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Small Lachnum on Filipendula ulmaria
Sergey Markov, 03-06-2022 23:11
I found small Lachnum on a stem of Filipendula ulmaria.

Apothecia 0.2...0.4mm. Asci MLZ+. Croziers+. Hairs w/abundant crystals, not very firm attached to the tips, +-easily removed in the water mount. No VB's w/refractive contents in the hairs or paraphyses. Paraphyses far protruding. Sp. w/oil content, but not all.

Asci (living matherial) (43.3) 46.5 - 54.4 (58.2) × (4.1) 4.3 - 5.4 (6.5) µm

Sp. (not guaranteed mature) (7.1) 8.3 - 12.2 (14.1) × (1.4) 1.6 - 2 (2.1) µm

I think that this is L.morthieri according to the Zotto's key. But I doubt this choice because of too small apothecia, too large (including immature) spores and yellowish hymenium. Please help with identification.
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Sergey Markov, 04-06-2022 10:58
Re : Small Lachnum on Filipendula ulmaria
I found more adequate fruitbodies of this species in more wet place in one-two meters from previous find.

asci - (53.5) 57.7 - 65.7 (68.5) × (3.8) 4.2 - 5.2 (5.5) µm

sp. - (8.1) 9.8 - 14.9 (17.4) × (1.4) 1.6 - 2 (2.2) µm (w/oily content)

These specimens shows the near-same microscopy that doubts in the species identification. Spores are too big for L.morthieri, but asci are too small for L.nudipes.
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Ingo Wagner, 04-06-2022 13:20
Ingo Wagner
Re : Small Lachnum on Filipendula ulmaria
Hello Sergey!

In my opinion is your find Lachnum nudipes.
Why do you think that the asci too small for L. nudipes?

Greetings 
Ingo
Sergey Markov, 04-06-2022 13:30
Re : Small Lachnum on Filipendula ulmaria
Hello Ingo!

According to the H.O.Baral's key, published here earlier http://www.ascofrance.fr/forum/5566/lachnum , size of asci (combined from two measurement series - 46...65) and oil content of ascospores are more suitable for L.morthieri, but spore size are more suitable for L.nudipes...
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Hans-Otto Baral, 04-06-2022 16:54
Hans-Otto Baral
Re : Small Lachnum on Filipendula ulmaria
I assume you compared your dead (shrunken) asci with my living asci. And perhaps you mistook vacuoles in the spores for oil drops. I see only non-refractive vacuoles on your photos.

You would need to carefully mount in water and study living asci and spores.
Sergey Markov, 04-06-2022 20:14
Re : Small Lachnum on Filipendula ulmaria
Hello Zotto!

>>> I assume you compared your dead (shrunken) asci with my living asci.


There were guaranteed living asci. I mounted the squashed part of the living frb immediately after collection, no more than 10 minutes passed from taking a sample from the nature with part of Filipendula stem, and no more than 10-20 seconds after removing frb from the substrate. Asci and spores were mounted in SDS/Congo red.


At this time part of the stem with these specimens lives in the humid container. I now (some minutes ago) examined one new specimen (marked on the attached photo) in the water mount. Asci was observed with 60x lens, ascospores - with 100x oil immersion lens. It was 100% living and fresh specimen. This study shows that previous measurements were correct.


asci - (46.1) 46.9 - 53.1 (54.9) × (3.4) 3.8 - 4.9 (5.3) µm


ascospores (now measured bigger set of spores):
(7.7) 9.5 - 13.6 (16.8) × (1.2) 1.5 - 2.1 (2.4) µm
Q = (4.8) 5.5 - 7.7 (9.6) ; N = 87
Me = 11.6 × 1.8 µm ; Qe = 6.5


About oily content - i see that some spores contain refractive content, some other spores contains non-refractive vacuoles, still others contain none of that. But maybe I'm misinterpreting what I see.

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Hans-Otto Baral, 04-06-2022 20:45
Hans-Otto Baral
Re : Small Lachnum on Filipendula ulmaria
Sorry, but this is a so frequent mistake, that I explained it surely 10 times in this forum if not 20 times. Of course, I cannot expect that new users read my postings of 10 years ago.

You can have as fresh and living fungi as you want if your preparation methods are too rough. 100% for sure that your asci are dead. Your spores are alive, they resist your squashing technique because of their smaller size (width).

Lachnum species are difficult to treat under vital methods because of their thin stalk. Anyway it is possible. Perhaps you succeed to take out parts of the hymenium with a fine needle. Sections with a razor blade are much easier with sessile apothecia, which is the best method to keep the asci alive.

Perhaps half of the spores have no oil inside but only vacuoles (transparent regions), whereras the others have minute refractive drops outside the vacuoles which might be oil drops.
Sergey Markov, 04-06-2022 21:37
Re : Small Lachnum on Filipendula ulmaria
Thank You very much for your attention, Zotto!

I found several long asci in a new mount, without squashing the sample, with division of it's upper layer with hymenium into small pieces with a very thin needle (0.1mm).

Do I understand correctly that the presence of some oil content in the spores is not important in this case, and these samples are L. nudipes?

PS Previous mounts were from squashed (not very strong) sections made by razor blade.
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Hans-Otto Baral, 04-06-2022 21:49
Hans-Otto Baral
Re : Small Lachnum on Filipendula ulmaria
Hmm, again I see only dead elements. Did you ever look in my article in Mycotaxon, 1992, on vital taxonomy? In this forum you can find numerous examples of living asci, how they look, very different from dead asci.

For comparing spore contents we must try to disregard spores other than mature in the definition I gave in 1992: i.e. spores that were freshly ejected. And they should be alive, this is better because oil drops tend to fuse to larger aggregations when killed or in the process of dying.

My impression is that those with minute granules are in some way altered, though I am not sure. In any case, this is only a slight difference. Important is that you did not confuse vacuoles with oil drops, which sometimes occurs.
Sergey Markov, 04-06-2022 23:36
Re : Small Lachnum on Filipendula ulmaria
Thank You very much again!

>>>>Did you ever look in my article in Mycotaxon, 1992, on vital taxonomy?


Yes, I often look at this article when I study ascomycetes under a microscope.
But in this case, all the requirements of the preparation were met. The material is fresh and not dehydrated (not even a little), there was no extensive squashing. In the last preparation no squashing at all was.
I thought that here the contents of the asci in the form of a large number of small and medium VB's, behind which the ascospores are almost not visible (new photo from the last mount with 100x immersion lens), indicates that the asci are alive (as in the paraphyses, as on fig.33 in the article mentioned). They should not have died, especially in the last careful mount.
For comparsion - microphoto #2 in Congo red/SDS of dead asci from the same frb - VB's in the asci are dissolved, and some asci clearly shows ascospores inside.


But it turns out that I'm wrong? In this case, I can't understand why asci are died in the last and pre-last mount and paraphyses not.


 


PS - previous photo was with 40x lens whos resolution can't clearly show small VB's as discrete objects.

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Hans-Otto Baral, 05-06-2022 11:37
Hans-Otto Baral
Re : Small Lachnum on Filipendula ulmaria
You used the term VBs for contents of the ascospores? Or asci? Or paraphyses? In Lachnum they occur only inside paraphyses, but L. nudipes does not have any.

What can be seen inside your paraphyses is dead granular plasma.

Your asci show the typical papillate apex of dead Lachnum asci, with swollen apical ring. In IKI this would be more clear.

Generally it is possible that your collection experienced some unfavourable events in the field. It can be that apothecia look externally sound but contain no living elements.

Regarding asci it is not difficult to recognize the living state, especially when they are mature: The spores are crowded in the upper half, whereas yours are spread across the ascus.
Sergey Markov, 05-06-2022 15:43
Re : Small Lachnum on Filipendula ulmaria
Thank You again!

>>> You used the term VBs for contents of the ascospores? Or asci? Or paraphyses?


I used the term "VB" in general for any content of vacuoles in all tissues, disregarding of their content type (refractive/non-refractive, lipidic, e.t.c.)


>>> Generally it is possible that your collection experienced some unfavourable events in the field. It can be that apothecia look externally sound but contain no living elements.


Highly likely this is my case. Temperature stress could be another cause of death besides death in the habitat. Being taken from a lying stem from under a layer of last year's leaves in a very wet place in dark dense thickets at a temperature of 12-14C (as surface groundwater), they were placed in a container with wet cotton, and transported for 10-20 minutes at a temperature of 22-24 C (like air). Usually samples of such small fungi, being taken in a wet habitat, not removed from the substrate and placed into in a moist container, continue to mature and begin to sow spores. But in this case, the fruiting bodies really stopped growing, which indicates their death.
I almost always study microscopic features of only fresh living material, and very rarely exsicates. I considered these samples to be a priori alive, and for me this is the first such case for more than 10 years. For me, the study of dead material is unusual. That's why I'm confused.


In any case, thank You very much again for your attention, now everything is clear and I should remember this case.

Hans-Otto Baral, 05-06-2022 16:57
Hans-Otto Baral
Re : Small Lachnum on Filipendula ulmaria
Then you cause confusion. If you read my paper you will learn that LBs are easily recognized by resisting in KOH, unlike VBs. LBs are not vacuolar. I introduced these term with reason.

Non-refractive, normal vacuole possesses every cell, unless the cell's vacuoles are all refractive, and only the latter deserve the name VBs (refractive vacuolar bodies).

Since I do not know your other research on fresh fungi, I cannot comment on your last statement.