Orange on Ulota crispa - Forum ASCOFrance

Edvin Johannesen, 05-01-2019 14:59

Re : Orange on Ulota crispa

The collector has sent med these two photos. I see these in the microscope, but apparently not inside asci. I am uncertain whether these are paraphyses or conidia of some sort.

Lothar Krieglsteiner, 05-01-2019 15:03

Re : Orange on Ulota crispa

Hello Edvin,

whatever this is - no apothecia.

Oddly enough: there is no Octospora known from Ulota by now (as far as I know).

Best regards, Lothar

Hans-Otto Baral, 05-01-2019 15:19

Re : Orange on Ulota crispa

Could you please test the asci with Lugol? Are there any paraphyses? And did you see the excipulum?

Edvin Johannesen, 05-01-2019 15:20

Re : Orange on Ulota crispa

Hi Lothar,

Actually I was in doubt whether to call them apothecia or not. They are rather shapeless.

I also found that there were no ascomycetes known on Ulota in Hansruedis review from 1988. But much has happened since then ...

Thanks.

Edvin Johannesen, 05-01-2019 15:49

Re : Orange on Ulota crispa

I will check what you ask for, Hans-Otto.

Lothar Krieglsteiner, 05-01-2019 16:06

Re : Orange on Ulota crispa

Sorry, I was too fast - and did not see the foto of the ascus first - and thought the second fotos (the very long spores, which more likely are conidia) would be the spores of the fungus.

So - this could be apothecia. Maybe of a species of Octosporella or something similar.

Best, Lothar

Edvin Johannesen, 05-01-2019 16:43

Re : Orange on Ulota crispa

I am beginning to doubt that the spores and ascus I submitted initially, actually belong here. I am struggling to find further asci and spores. What I do see is tufts/bundles of what looks like paraphyses. I doubt they are spores, since they appear to break loose with a hyaline cell at the break-off point.

Here are a few images of these "paraphyses" and a few from "excipulum". Tissue is partially gelatinised. It is very difficult to make clean sections.

Hans-Otto Baral, 05-01-2019 17:44

Re : Orange on Ulota crispa

I cannot help at this point.

Edvin Johannesen, 05-01-2019 18:00

Re : Orange on Ulota crispa

That's ok. Thanks anyway!

Björn Nordén, 07-01-2019 12:50

Re : Orange on Ulota crispa

could it be a dried up mycomycete?

Björn Nordén, 07-01-2019 12:51

Re : Orange on Ulota crispa

myxo not myco

Edvin Johannesen, 07-01-2019 13:09

Re : Orange on Ulota crispa

No, it's definitely a fungus. I am looking at muscicolous myxomycetes virtually daily. It's interesting to note that this fungus grows in the middle of the apical growth point on virtually every branch of the moss.

Björn Nordén, 07-01-2019 14:38

Re : Orange on Ulota crispa

Ok, we can barcode it if you like.

Best

Björn

Edvin Johannesen, 07-01-2019 14:41

Re : Orange on Ulota crispa

That would be interesting! I will drop it off at NINA :-) Thanks.

Lothar Krieglsteiner, 07-01-2019 15:00

Re : Orange on Ulota crispa

I also do not believe in a myxo - if there is a bet between you I go with Edvin.

Best, Lothar

Martin Bemmann, 07-01-2019 15:29

Re : Orange on Ulota crispa

The long "spores" in the second post could be cyanobacteria (Oscillatoria or something similar).

regards

Martin

Edvin Johannesen, 07-01-2019 15:54

Re : Orange on Ulota crispa

I do see the similarity in shape to some Oscillatoria species. However these filiform "spores" are hyaline without any sign of photosynthesising structures. And they clearly belong to the fungus (see latest photos). Thanks.

Martin Bemmann, 07-01-2019 15:56

Re : Orange on Ulota crispa

At least on my screen the contents of them seems to be green. But this is probably due to a colour abrerration of the photographs.

Björn Nordén, 07-01-2019 15:56

Re : Orange on Ulota crispa

New phylum!? : )

Edvin Johannesen, 07-01-2019 16:26

Re : Orange on Ulota crispa

Cyanoplasmodium? :-)

Jokes aside - I take your point, Martin. The images you are referring to were taken by the collector and sent to me, but I have assumed that they are the same structures which I observed. In which case the greenish colour is an artifact.

Georges Greiff, 07-01-2019 21:51

Re : Orange on Ulota crispa

Hello,

Interesting find. I have been finding other ascomycetes on Ulota bruchii and U. crispa agg. but I have not seen this one. I will keep an eye out.

Georges

Viktorie Halasu, 07-01-2019 23:03

Re : Orange on Ulota crispa

Hello,
I wonder if this collection could be related to Belonium coroniforme (in Zotto's folder under Belonioscyphella here: https://drive.google.com/drive/u/0/folders/0B5SeyOEkxxZhaG11dlBYTDZtREk ). The spore shape and contents and apothecium structure look similar to me, however spore size is different and no septa there.
Viktorie

Edvin Johannesen, 08-01-2019 10:38

Re : Orange on Ulota crispa

Very interesting, Viktorie. I think this is the same, or something very close to the fungus displayed in the folder you are referring to. Look at the pictures added on 22. May 2012! Let me quote the comments by Zotto (I assume) in the folder:

"Helotiales, 18.V.2012-2a.jpg" 2012-05-24, on Orthotrichum striatum, leg. Carlos Sánchez. Inoperculate asci are RR, with croziers, eight-spored, up to 200 x 17. Paraphyses and ectal excipulum are also hemiamyloid! Medulla is strongly gelatinized and exudates carotenoids everywhere. There are also carotenoids in paraphyses and ectal excipulum. Spores from scale 46-55 x 5.1-5.6 µm, 7(-13) septate, which germinate or form conidia? (I don't think so). Flesh composition is different, not Belonioscyphella. --- 1) I was wrong, spores only get 6-8 septated. I was confused because in some spores oil contents looked as septa. 2) Hemiamyloid reaction is not restricted to ectal excipulum and paraphyses, it is also present in the medulla. Even some spore walls react blue.3) All the apothecia is more or less gelatinized. This makes it difficult to section. Maybe gel gives the hemiamyloid reaction?4) Again not sure about microconidia on spores. Probably they just only germinate as you think. Maybe I confuse microconidia with plenty of ¿bacteria?5) Some of these ¿bacteria? react blue in IKI.Please see footnotes of the pics attached to understand what I´m talking about.After comparing with your Naeviopsis, it fits very good except for the lack of hemiamyloid reaction.
"Helotiales, 18.V.2012-4k, ectexc.jpg" ectexc fide Raul
"Helotiales, 18.V.2012-4l2, ectexc.jpg" ectexc?"

Regarding Belonium coroniforme, it was described on Orthotrichum. The collector of the present fungus now tells me that is may well be Orthotrichum on which it grows. I don't know mosses well enough to decide.

I will try once again to find ascospores in the material. And maybe will will have its DNA sequenced in the near future.

Edvin Johannesen, 08-01-2019 12:15

Re : Orange on Ulota crispa

Finally I found an apothecium with a ripe hymenium. The spores I have measured this time are ca. 30-45 x 4-5 µm, asci are ca. 110-140 x 10 µm (all measures in CR in water). Ripe spores germinate at both ends. It seems they are mostly 7-septate. Asci 8-spored. Asci I believe have croziers (?). The photos are taken at 1000x (oil) - scale intervals ca. 1 µm. There is little doubt in my mind that this is the same fungus as was displayed in Zotto's folder from May 2012.

Björn Nordén, 08-01-2019 12:16

Re : Orange on Ulota crispa

Very cool fungus, i wonder were it should end up, it is not anything like Belonium in current circumscription as i can understand it, but the name seems to fit well! Yes, let's barcode it!

Edvin Johannesen, 08-01-2019 12:17

Re : Orange on Ulota crispa

And a few more.

Martin Bemmann, 08-01-2019 12:22

Re : Orange on Ulota crispa

Wonderful discovery Viktorie!

@Edvin: How about the IKI-reaction of ascus-tips? The one in Zotto's folder reacts strongly hemiamyloid. But Rehm's original description sais: "J-".

Regards

Martin

Edvin Johannesen, 08-01-2019 12:30

Re : Orange on Ulota crispa

I tried to put the second half of the apo in Barals. Unfortunately, I found only a couple of immature asci there. In those I did not see any IKI reaction, but I am not quite convinced. I am a bit hesitant to spend more apothecia.

Martin Bemmann, 08-01-2019 12:33

Re : Orange on Ulota crispa

Did you already discard the slide of your last pictures? If not, you could try there.

Edvin Johannesen, 08-01-2019 12:34

Re : Orange on Ulota crispa

Also note the spherical algal cells in a couple of the photos. I noted the same type of algal cells in photos in Zottos folder. Can there be an association?

Edvin Johannesen, 08-01-2019 12:36

Re : Orange on Ulota crispa

Martin - so Barals or Lugol after CR treatment would work?

Martin Bemmann, 08-01-2019 12:38

Re : Orange on Ulota crispa

Oops, I don't know. Just try....

Edvin Johannesen, 08-01-2019 13:00

Re : Orange on Ulota crispa

So I tried to treat with Baral's medium after CR staining. I got some dark red staining in the ascus tip - in some asci diffuse staining, in some asci a narrow pore staining. Since this staining is not there in CR, I assume it is caused by the addition of IKI (?). I leave it to you experts to interpret.

Edvin Johannesen, 08-01-2019 13:02

Re : Orange on Ulota crispa

In any case, this staining got some really nice images of the spores, incl. septation. The spores have from 5-11 septa.

Hans-Otto Baral, 08-01-2019 13:38

Re : Orange on Ulota crispa

I can only confirm Viktorie's great idea. I must apologize not to have mentioned Raul Tena as the author of the photos in the text to this fungus.

For the iodine reaction I think one ascus shows the red very well. Interesting would be to try a KOH-pretreatment. The Congo might disturb a bit the test, mut mot much. After adding 2-5% KOH for some seconds and removal of the KOH with water, add again IKI or MLZ and, apart from the blue apical ring you should see a distinctly light blue hymenium as figured by Raul.

Indeed Rehm's remark of inamyloid asci is surprising because he used IKI, not MLZ, which was not in use at that time, and he described the red (hemiamyloid) reaction as violet when he saw it.

A DNA result would be great of course.

Zotto

Edvin Johannesen, 08-01-2019 13:47

Re : Orange on Ulota crispa

Thanks for the tip, Zotto. I will re-wet the material once more to ensure there is ample material to spend one more for the iodine reaction, based on your recommendations. Luckily, mature apothecia are not crucial for DNA studies.

Björn Nordén, 08-01-2019 13:49

Re : Orange on Ulota crispa

Hi otto,

is it possible to access that google disc resource for anyone, or is it restricted?

best

Björn

Martin Bemmann, 08-01-2019 13:59

Re : Orange on Ulota crispa

By the way, the type exists in S:

https://plants.jstor.org/stable/10.5555/al.ap.specimen.s-f12182

and here is Rehm's drawing of the spores:

http://herbarium.nrm.se/img/kbo//kryptos/kbo/kryptobase/201503/max/272714.jpg

Hans-Otto Baral, 08-01-2019 17:00

Re : Orange on Ulota crispa

Hi Björn

do you get a problem when clicking on the link?

You can also try via my homepage, but maybe the same will happen.

www.in-vivo-veritas.de

Martin, the type label is interesting, I am surprised to see large globose oil drops which look like in a living spore! In Raul's photos I only see many small droplets. These large drops Rehm did not mention, instead he described the dead spores with their obvious septa.

Zottto

Edvin Johannesen, 08-01-2019 18:04

Re : Orange on Ulota crispa

Zotto, your method worked :-). Here is the result after pre-treatment with 5% KOH, then washing with water, and finally adding IKI (Baral'ische lösung). I see no staining other than in the ascus apical appratus.

Hans-Otto Baral, 08-01-2019 18:25

Re : Orange on Ulota crispa

Good! Well, I do not know these fungi personally, and I can only make suggestions. Why Raul got this reaction with the same method I do not know.

But now I realized in your living spore the large drops just as Rehm had drawn them!

Edvin Johannesen, 08-01-2019 18:36

Re : Orange on Ulota crispa

Yes. In some of spores it seems like the large drops press against the septa and distorts (bends) them.

Hans-Otto Baral, 08-01-2019 18:44

Re : Orange on Ulota crispa

Surely not the LBs. You perhaps mean bulging of the septa caused by differences in turgor between adjacent cells. Usually one of the two adjacent cells is dead and turgorless when such bulging (convex septum) occurs. The first spor pic shows a living spore with straight septa.

Martin Bemmann, 08-01-2019 18:59

Re : Orange on Ulota crispa

I wonder if the second drawing of a dead spore is by Rehm. Since when does millimeter paper with orange lines exist? In 1905? There is also a revision slip by Arendholz from 1982. Maybe it is drawn by him?

http://herbarium.nrm.se/img/kbo//kryptos/kbo/kryptobase/201503/max/272720.jpg

Regards

Martin

Edvin Johannesen, 08-01-2019 19:07

Re : Orange on Ulota crispa

OK. I learn every day, Zotto :-)

Zuzana Sochorová (Egertová), 08-01-2019 21:07

Re : Orange on Ulota crispa

Hi,

I have a collection of Belonium coroniforme from Slovakian mountains. It was growing in a large group, but unfortunately it was sterile. We have LSU and TEF obtained from this collection + a few more else provided by colleagues /always from Orthotrichum/. So, if someone will sequence Edvin´s collection, we can compare our data.

Best regards, Zuzana

Edvin Johannesen, 08-01-2019 21:10

Re : Orange on Ulota crispa

Great! By the way. the collector's name is Oddvar Olsen :-)

Hans-Otto Baral, 08-01-2019 21:23

Re : Orange on Ulota crispa

Hm, I am inclined to assume that millimeter paper was in use very long ago. In any case, I cannot imagine that someone else than Rehm glued this drawing between the other spore and the text. Why did he leave this space? Maybe the dead spore was drawn later by him, and the living one was first drawn as a sketch on another paper.

Belonioscyphella hypnorum is very difficult to put in a family. There is ITS, LSU and Ef1 in GenBank by Z. Egertova. Zuzana, did you compare your LSU? What is next in Blast?

Zotto

Martin Bemmann, 09-01-2019 18:59

Re : Orange on Ulota crispa

Zotto: "Indeed Rehm's remark of inamyloid asci is surprising because he used IKI, not MLZ, which was not in use at that time, and he described the red (hemiamyloid) reaction as violet when he saw it."

Rehm was certainly not aware of hemiamyloidity. He expected a blue reaction, if any. Thus it was J- for him. Where did he speak of a violet reaction?

Regards

Martin

Hans-Otto Baral, 09-01-2019 20:26

Re : Orange on Ulota crispa

Many times. I made an appendix list of them in my 1987 paper on iodine reaction. E.g., Velutarina rufoolivacea 1896: 647 (r), Heterosphaeria patella var lojkae : 203 (rb), linariae : 204 (r)

Edvin Johannesen, 09-01-2019 20:34

Re : Orange on Ulota crispa

The bryo-expertise has now confirmed that the current specimen is growing on Ulota crispa s. lat. (the Ulota crispa complex comprises U. crispa s. str., U. crispula, and U. intermedia). U. crispa was originally described as Orthotrichum crispum, and transferred to Ulota in 1818. The collector of the holotypus specimen was a bryologist and it is likely that it was growing on Orthtrichum s. str., which would imply that the fungus can grow on both Orthotrichum and Ulota spp.

Georges Greiff, 22-08-2019 15:35

Re : Orange on Ulota crispa

Dear Edwin,

Did you get anywhere with sequencing this? I have found a fungus on Ulota bruchii that seems to be in the same genus, but attacking the leaves rather than only apices. My collection has much smaller fruitbodies but otherwise looks good.

Georges

Edvin Johannesen, 22-08-2019 15:46

Re : Orange on Ulota crispa

Dear Georges,

I was told a good while ago that the sequencing (first attempt) was not successful. I will check whether it has been tried once more.

BR Edvin

Georges Greiff, 22-08-2019 17:20

Re : Orange on Ulota crispa

Thanks.

I will put up my collection now for comparison. Perhaps it is completely un-related as it has an ostiole unlike most Helotiales.

Best wishes,

Georges

Arnold Büschlen, 11-12-2019 18:18

Re : Orange on Ulota crispa

Hallo,
ich bin neu hier im Forum. Im speziellen intressiert mich die Diskussion um B. coroniforme und um seine Trägermoose. Darf ich fragen: Auf Grund welcher relevanten Merkmale wurde diese Probe als Ulota crispa bestimmt? Lagen gut ausgebildete Sporophyten vor?

Besten Dank!

Edvin Johannesen, 11-12-2019 18:32

Re : Orange on Ulota crispa

Please see my comment from Jan 9th. Here are some photos.

Arnold Büschlen, 11-12-2019 19:29

Re : Orange on Ulota crispa

Besten Dank. Ja, die hier gezeigten Sporophyten sprechen klar für die Gattung Ulota. - U. Crispa oder eventuell U. crispula? Hast du eventuell ein Bild von Endostom?

Edvin Johannesen, 11-12-2019 19:41

Re : Orange on Ulota crispa

Will this suffice? Or do you mean in the microscope?

Edvin Johannesen, 11-12-2019 19:48

Re : Orange on Ulota crispa

Tooke these now.

Arnold Büschlen, 11-12-2019 20:24

Re : Orange on Ulota crispa

Super! Ja das spricht klar für U. crispa.

Wenn es erwünscht ist, kann ein Bild mit dem inneren Peristom der 4 Ulota-Arten die auf den ersten Blick verwechselt werden können, zeigen.

B. coroniforme auf U. crispa ist sehr interessant. Die Funde von B. coroniforme in der Schweiz sind bis jetz alle auf Orthotrichum pallens gefunden worden.

Edvin Johannesen, 11-12-2019 20:29

Re : Orange on Ulota crispa

Well, I trust you when you say that it is easy to see the difference :-)

Arnold Büschlen, 11-12-2019 20:56

Re : Orange on Ulota crispa

Hier 2 weiterführende Publikationen:

- https://www.researchgate.net/publication/299083677_Integrative_taxonomy_sheds_light_on_an_old_problem_The_Ulota_crispa_complex_Orthotrichaceae_Musci
-https://rbg-web2.rbge.org.uk/bbs/activities/field%20bryology/FB117/FB117_Ulota_Tom.pdf

Edvin Johannesen, 11-12-2019 21:39

Re : Orange on Ulota crispa

Thanks.

Gilles Corriol, 09-09-2020 17:34

Re : Orange on Ulota crispa

Dear Edvin,
A just found the same fungus as you in the Pyrenees, also on Ulota apex. Have you finaly concluded with the help of colleagues and/or DNA analysis ?
If so, I would be interested in the conclusion.Thank you very much.
Gilles

Edvin Johannesen, 24-09-2020 13:08

Re : Orange on Ulota crispa

Dear Gilles,

Apologies for the late reply. The cultivation from spores failed and the sequencing produced dubious results, probably due to contamination. May I suggest you contact Zuzana Sochorová? She wrote several months ago:

I have a collection of Belonium coroniforme from Slovakian mountains. It was growing in a large group, but unfortunately it was sterile. We have LSU and TEF obtained from this collection + a few more else provided by colleagues /always from Orthotrichum/.

Unfortunately there are currently no resources to do further sequencing studies on the Norwegian collection. Please let me know if you aresuccessful in contacting Zuzana.

Regards,

Edvin

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