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Helotium schimperi
Nina Filippova, 03-02-2013 19:50
Good time),

I've compared this specimen with the description in the paper Redhead, Spicer (1981, Discinella schimperi). It seems fits well. Only spores in mine some smaller (measured in alive apos), and the host is another (Sphagnum magellanicum). Yet i was not able to observe medullary excipulum (probably  since apothecia small and it is not very developed).


Apothecia turbinate, translucent, white to pink, to 0,5 mm wide, smooth, sited at leaves (phyllids) in capitula of S. magellanicum.


Excipulum from textura prismatica (oblita?), with dextrinoid reaction in MLZ (KOH), cells at the base larger , more ellipsoid (20 x 10), narrower to the edge, end cells obtuse, cohered (15 x 5); receptacle attaches to the substrate by the bunch the hyphae from cylindrical cells (15 x 5); asci clavate, with pronounced clamp, with CR+ content, with amyloid pore, 85 (78-93) x 10,5 (9,2-11); spores fusoid, with obtuse ends, with amorphous oil content in fresh apothecia, with several medium and small guttules in KOH, mature spores 1-2 septate, septum thin, scarsely seen, content CR+, 15,3 (13,7-17,2) x 5,3 (4,8-5,8) (Q=,9; N=16, spores from alive apth.); paraphyses cylindrical, slightly bulged in different parts, segmented, branched in 2-4 parts or unbranched, 1,5-3 mk broad.


Collected 06.08.2012, N61,063892° E69,455695°, ombrotrophic bog.

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Hans-Otto Baral, 03-02-2013 21:10
Hans-Otto Baral
Re : Helotium schimperi
Hi Nina

this is great!! I once saw Discinella schimperi when I was in Lapland, thanks to Marie Davey from Finland, who said that it exclusively grows on Sphagnum squarrosum. I made the atached spore photos which show these striking refractive vacuoles in the living spores which I can also see in your pics in water.


What is not the case in your specimen: the asci always contained 4 large and 4 smaller spores:


"Always 4 large spores with VBs and 4 smaller spores without VBs in each living ascus. Large sp. *16-19 x 5.3-5.7 µm, fusoid, containing very elongate 1-2refractive large VBs, const. 2 per spores."


so maybe we have different species?


Zotto

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Nina Filippova, 04-02-2013 08:51
Re : Helotium schimperi
Hi, Zotto,

there are no close species described (?) - as i could see from Redhead's paper and IF - i checked Discinella spp., but it is difficult to check all Helotium spp. Yet i am not imformed at all in this group, since it is difficult to do conclutions. Do you think it may worth to describe sp. nov.? There are about 10 apos in the collection, i could microscopy once more to get clear picture (if you want, point to the features which should be checked more precisely). All previous micro pictures are there: http://www.flickr.com/photos/bog-fun/sets/72157631521757540/?

Hans-Otto Baral, 04-02-2013 10:16
Hans-Otto Baral
Re : Helotium schimperi
Hi Nina

H. schimperi was described by Nawashin near Moscow, on Sphagnum squarrosum.

No mention of heterospore asci, the drawing shows  5 or 6 spores in the living ascus, all containing these large vacuoles. Spore size 18-21 x 5-6, so your spores look shorter. Asci in Nawashin are 90-100 x 10-13 µm (alive), not clearly larger than yours (in dead state).

I do not see strong evidence for a different species. 

The generic placement. I see some similarity with Pezoloma, and a sequence analysis (Stenroos et al. 2010) show it to be near that genus, but also near Mitrula and Epiglia.

We could genetically show (unpublished) that Discinella and Pezoloma are very close, so that genus is not bad for schimperi.

Zotto
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Nina Filippova, 04-02-2013 15:55
Re : Helotium schimperi
Thank you, Zotto.

It will be interesting to compare D. schimperi  from the both sphagna. The ecology of these two species quite different, S. squarrosum is minerotrophic and common in fens and bogged forests, and S. magellanicum is ombrotrophic and inhabits bogs. But i did not look at S. squarrosum, though it is common there as well.
Hans-Otto Baral, 04-02-2013 16:12
Hans-Otto Baral
Re : Helotium schimperi
Yes, maybe! What I failed to look is for the croziers. Yours has them very clearly thanks to your good pics, also your photo of the apical rings I like very much. Sometimes nvery closely related species differe herein.

This type of ring is similar as in Ascoocoryne or Peziloma but very different from Hymenoscyphus (Helotium).

Zotto
Hans-Otto Baral, 08-02-2013 11:56
Hans-Otto Baral
Re : Helotium schimperi
Seppo Huhtinen just answered me the following:

I am very surprised about D. schimperi. It is very specific, hyphae always in one special cell at shoot apex, always on S. squarrosum.

Would be nice to know whether such a cell is present in magellanicum and whether the species is behaving as in S. squarrosum.

Nina Filippova, 05-03-2013 13:44
Re : Helotium schimperi
Here is what i was able to understand now about the mode of parasitism. Surely, the fruitbodyes are the tip of the iceberg there ).

Apothecia attaches to upper part of leaves (phyllida) by the weft of cylindrical hyphae.  Hyphae radiate from stipe base, several go down to reach the base of the leaf. Other hyphae spread in leaf tissue, they seen inside cells, along and penetrating green (chlorophyllous) cells, often in places of cell contacts.


I was not able to trace all way hyphae does from leaf tip to it's base and further down to base of branch (where mentioned mucilaginous cells are placed). Separately, i looked these cells in capitula. Hyphae were spread there abundantly as well. Part of them were inside cells. Others were free, in bundles (2-3), ending with higly branched entangled agglomerages. Some of these were seen branching close to muciaginous cells. Mucilagenous cells with such agglomerages outside also were seen, as well as single hyphae attached as cap to it's tip and penetrationg inside.


There was only one capitula which i was studying (unfortunatelly i separated branches with apothecia right after collection, being not informed that other parts may be important). Mounts were not very clear.


But it seems close to what was described on S. squarrosum after such short examination. How to interpret all this... i don't know.

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Hans-Otto Baral, 05-03-2013 16:37
Hans-Otto Baral
Re : Helotium schimperi
Good photos and observations! So these are clear haustoria inside the necrobiotic (or dead?) cells of the Sphagnum. Your mucilaginous cells I do not understand, do they belong to the moss?

Zotto
Nina Filippova, 05-03-2013 17:54
Re : Helotium schimperi
Ok, now i don't have clear answer ). I relied on desciription in Redhead and Spicer (1981), they mention "mucilaginous cells" which are infected by fungus. They show these cells in the pictures (Fig. 6-8 there) and i copied their notices.

But now i opened a book on Sphagnum biology and see, these balls must be antheridia actually. Antheridia "nestled among the leaves near the tips of the capitulum branches". So, it is antheridia infected, probably.


What is mucilaginous cells, then? - haven't found anywhere in books about sphagnum. There is a citation in Bryophyte biology (2009): "Juvenile leaves hide delicate filaments that line the insertion of the leaf. These hairs originate from the leaf initial and secrete a mucilage of polysaccharides that may be essential in preventing the delicate growing apices from dehydrating".


I have not understand the description by Nawaschin yet, since it is in German and Google translation worked quite littered. May be, there will be more clarity.

Hans-Otto Baral, 05-03-2013 18:26
Hans-Otto Baral
Re : Helotium schimperi
From a diagrammatic illustration of Sphagnum reproduction I see that an antheridium is a multicellular organ which produces inside the sperms. So I think that these mucilaginous cells are something different, or it may be a very young antheridium.

Zotto
Chris Yeates, 01-07-2013 13:09
Chris Yeates
Re : Helotium schimperi
I have downloaded the protologue for this species (as in the images Zotto has posted above) and produced this PDF which may be of use
Chris

PS - when downloaded it looks fine - honestly!

Nina Filippova, 11-07-2013 19:17
Re : Helotium schimperi
Hello,

Another collection of D. schimperi has showed up here. But now it parasiting on another sphagnum - S. papillosum (also ombrotrophic bog, 40 km from previous collection).

The measurements of asci, paraphyses and spores coinside with previous collection.

Apothecium (single apos collected) turbinate,  650 mk high, 900 mk broad, whitish, watery.
Asci clavate, with crozier, with euamyloid cylindrical pore, 83.2–113.5 x 9.2–13.4; paraphyses cylindrical, rarely branched, 2 broad, with pale round to elongated vacuoles; spores ellipsoid, with pale irregularly oils at the ends, 14.7 (13–17.7) x 4.8 (4.4–5) (n=12).?
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Hans-Otto Baral, 11-07-2013 21:10
Hans-Otto Baral
Re : Helotium schimperi
Very good! The distinctly smaller spores (Nawaschin: 18-21 x 5-6) with much less refractives vacuoles, consistently 8-spored asci together with Sphagnum species other then S. squarrosum actually point to a different species.
Zotto
Nina Filippova, 12-07-2013 05:18
Re : Helotium schimperi
Interesting! I will try to collect more representative collection then.
Nina Filippova, 13-07-2013 08:30
Re : Helotium schimperi
There was luck with the collection, and now the are about 20 apothecia, which should be usefull for descripiton. The attacking hyphae were seen very well, they are abundant in leaf cells (piercing through walls of photosinthetic cells); and bunches of them descending along branches; antheridia (= mucilaginous cells) was empty (don't contained any hyphae) at this stage of life cycle.?
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Jan Eckstein, 07-10-2022 14:28
Re : Helotium schimperi
Hello,

I would like to add to this old thread because I found a similar fungus but on a further peatmoss (Sphagnum teres).

Notes:

apothecia on capitulum of Sphagnum teres, arising between appressed leaves of young branches, white or faintly pink, to 0.8 mm in diameter, shortly stalked, hymenium first concave then flat and becoming slightly convex, excipulum small, of the same colour as hymenium
paraphyses filiform, straight, rarely branched, 1.6-2.5 µm wide below, 2.0-3.5 µm wide above, walls slightly wavy
asci cylindrical, K/I+, small tholus, 8-spored but often only 4 (or 2) spores reach maturaty, 4(-6) spores aborted
ascospores ellipsoid, one-celled, 13-17 x 5-6 µm, to 20 x 6 in two-spored asci, several large dropdlets 1-4 µm in diameter (weakly refractive) and several small droplets c. 0.5 µm in diameter (strongly refractive); ascospores germinating shortly after discharging, germinates spores sometimes with one or two septa
hyphae on surface of Sphagnum leaves going down to the tiny spaces between young leaves, squeezing into the leaf axils of very young leaves near the shoot apicies, often growning around the axillary hairs but without entering these cells;
no infection seen, hyphea remain outside the Sphagnum cells only rarely growing through hyalocysts (dead cells)

Are there any new devopments on this topic? I can send some apothecia for studying or sequencing, if someone is interested. There are about 30 infected Sphagnum capitulums.

With best wishes

Jan
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Hans-Otto Baral, 07-10-2022 16:02
Hans-Otto Baral
Re : Helotium schimperi
Interesting collection! I wonder whether two the large drops in the spores disappear in KOH like in schimperi. Maybe it is schimperi woith a higher lipid content? Or is the spore size deviating? Asci always with 4 spores aborting?
Jan Eckstein, 10-10-2022 16:38
Re : Helotium schimperi
Dear Zotto,

most asci with four aborting spores, sometimes six. However this wasn't so obvious in the very fresh state but obvious after one week in the refrigerator.

The droplets don't dissolve in KOH.

It looks fairly similar to Helotium schimperi, but I couldn't find the charateristic infection on the muscilaginous cells (axillary hairs) which should be easy to observe according to Redhead and Spicer. I looked in at least ten Sphagnum shoots but all axillary hairs were without infection. Unfortunately, I don't found other infected cells either. Additionally, the spore size seems smaller although its variable.

With best wishes

Jan
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Hans-Otto Baral, 10-10-2022 20:25
Hans-Otto Baral
Re : Helotium schimperi
I have no idea about the infection organs in this species, but I see very clearly that the two large refractive vacuoles in the spores lose their refractivity in KOH while the LBs resist. So in this point there is no difference to what I so far saw in H. schimperi.

I did not compare measurements, I suppose this will be a difficult thing, often a species shows some variation.
Jan Eckstein, 11-10-2022 08:22
Re : Helotium schimperi
Dear Zotto,

thank you very much for your comments!

As conclusion, the specimen can be called Helotium schimperi or is very close. Sphagnum teres is closely related to Sph. squarrosum (same section) after all.

I will try to obtain ITS and LSU sequences of my find if I can find a lab.

The locality is a spring mire in the Thuringian fores at about 700 m asl.

More finds should be possible in the future.

With best wishes

Jan
Hans-Otto Baral, 11-10-2022 09:34
Hans-Otto Baral
Re : Helotium schimperi
This would be very good to try a sequence. D. schimperi is badly sequenced. Only two ITS sequences exist: M168_Discinella schimperi (T. Laukka, but only 5.8S in GenBank, not sure why the entire ITS was not uploaded) and Discinella "schimperi"_Kh-4517_(30.08.2014, N. Filippova, not in GB). The two strongly differ, but Nina's was morphologically clearly different from schimperi, so is something else.
Zdenek Palice, 11-10-2022 23:06
Zdenek Palice
Re : Helotium schimperi
Hallo to all of you,

I collected this species in the Czech Republic in N  Bohemia - Jizerske hory Mts (Isergebirge) on Sphagnum centrale (I am not completely sure with ID, but I hope it is). It was sampled in autumn 2018 on an open raised bog at altitude of almost 1000m, few months ago I sent the specimen to Zuzana Sochorova for revision for sure

best Zdenek
Georges Greiff, 12-10-2022 10:14
Re : Helotium schimperi
I collected D. schimperi on Sphagnum squarrosum a month ago. It also had asci with 2 types of spores that both get to maturity. 4 larger ones each with 2 obvious refractive vacuoles and 4 smaller ones without vacuoles. The infected mucilage cells with fungal infection "caps" as in Redhead & Spicer paper were abundant on the leaves at the shoot apices. Perhaps because of this, the plants were somewhat atypical in appearance, more like S. fimbiratum macroscopically.

Luis Quijada would be interested in these finds.

All the best,
George
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Hans-Otto Baral, 12-10-2022 10:25
Hans-Otto Baral
Re : Helotium schimperi
very typical spores!
Jan Eckstein, 16-10-2022 11:50
Re : Helotium schimperi
Dear all,

a further fresh specimen showed that fresh asci mostly contain 8 equally sized spores. I have the impression that the cell content is different from George's collection on Sphagnum squarrosum.

Is there any chance to get sequences from specimens on Sph. squarrosum?

Jan
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Georges Greiff, 16-10-2022 11:57
Re : Helotium schimperi
Hi Jan,

That is quite striking and certainly looks different to mine.

I have material on S. squarrosum but not the capacity to sequence it myself at this point. I know Luis Quijada is interested in this group, maybe he will be able to sequence some. I think he is on holiday now but I will email him.

In your fresh material were you able to find where the hyphae localise? I was thinking a character of this Sphagnicolous group was the distinctive infection on the mucilage cells but perhaps not.

All the best,
George
Jan Eckstein, 16-10-2022 12:16
Re : Helotium schimperi
Dear George,

the infection also puzzles me. I observed lots of hyphae and mucilage cells but couldn't find an infection.

With best wishes

Jan
Hans-Otto Baral, 16-10-2022 12:27
Hans-Otto Baral
Re : Helotium schimperi
Jan, your new sample was on S. squarrosum? Which mounting medium you used for the last two pics?

The spores look as if they are without vacuolar bodies, although Nina's sample on Sphagnum magellanicum with 8-spored asci also has in fact distinct but low-refractive VBs.

Perhaps this is only variation of a single species.

Zotto
Jan Eckstein, 16-10-2022 13:34
Re : Helotium schimperi
Dear Zotto,

the last specimen was also on Sphagnum teres only from another site.

with best wishes

Jan
Georges Greiff, 02-11-2022 21:22
Re : Helotium schimperi
Hi Jan,

I wonder if this one from Sunday on S. papillosum is related to your second collection. It seems to fit Helotium vasaense / Hymenoscyphus vasaensis quite well with spores falling within the range given by Dennis (1962: British Helotiales). Seems to also sometimes have weakly refractive vacuoles in the spores so perhaps close to H. schimperi.

Bests,

George
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Jan Eckstein, 06-11-2022 13:03
Re : Helotium schimperi
Dear George,

great, this looks exactly like my fungus. Hymenoscyphus vasaensis is an old name (Peziza vasaensis P. Karst. [as 'varsaënsis'], Not. Sällsk. Fauna et Fl. Fenn. Förh. 10: 150 (1869)). Do you know if ever H. vasaensis was compared with H. schimperi? Please try to find sufficient material for a DNA study.

With best wishes

Jan
Georges Greiff, 06-11-2022 16:58
Re : Helotium schimperi
Hi Jan,

Despite searching, I only found one apothecium last week at the site. That said, the apo is quite large. I can try to get sequence data from half of it and still have a bit left.

I have the real H. schimperi as well so I will try with both. Morphologically, they are quite similar. I have not yet tried to look for the infection in this one though.

Dennis (1962) said that H. vasaensis was closer to H. schimperi than to anything else he had seen. I agree. In Stenroos et al. (2010), they show a few Hymenoscyphus spp. from Sphagnum as sister to H. schimperi. It is likely they also collected this vasaensis-like one as it does not seem to be uncommon, generally. I recently saw images of an identical collection from Bosnia & Herzegovina.

Bests,
George
Elisabeth Stöckli, 07-11-2022 11:52
Re : Helotium schimperi
Bonjour,

Voici également une récolte sur Spaghnum (pâturage boisé à 1'000m), apothécies 0.3 - 0.7 mm de diamètre, blanchâtres/roses, stipitées, Ascospores 4 - 5.25 µm, hyalines, non septées, lisses. Asques 93 - 105 x 10 - 13 µm, avec crochet, J+, contenant 8 spores (moins avec l'âge). Paraphyses droites, hyalines.

Elisabeth

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Georges Greiff, 07-11-2022 15:32
Re : Helotium schimperi
Wow, great documentation, Elisabeth! Nice to see the refractive vacuoles in the spores and also a shot of the hyphae.

Best wishes,
George
Jan Eckstein, 07-11-2022 15:48
Re : Helotium schimperi
Dear Elisabeth,

thank you for this nice documentation! For me it looks like the same as my and George's collections. The Sphagnum on your image should be S. palustre. However, in my collections only Sph. teres was infected even when Sph. palustre was growing nearby.

With best wishes
Elisabeth Stöckli, 07-11-2022 22:34
Re : Helotium schimperi
Bonsoir Georg et Jan,

Merci pour vos commentaires.

Elisabeth