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Ombrophila(?) - ascospores with great oil content
Pierre-Yves Julien, 02-08-2022 17:48
Récolte le 31/07/2022 – Ozoir-la-Ferrière (77) – France – Etude sur matériel frais vivant.
Sur débris de branchettes, sous Carpinus betulinus, en bordure de mare forestière.
Apothécies cylindriques, stipitées, à hyménophore convexe, cespiteuses sur le substrat, blanches chez les jeunes puis grisonnant.
Taille du plus grand specimen = 3mm de diamètre pour 5mm de haut.

Asques bisériées [75 ; 90] x [7.5 ; 9.0]µm à apex de type Hymenoscyphus, IKI+ bb, crochet+.
Paraphyses pluriseptées, cylindriques sans épaississement au somment, avec une grande vacuole réfringente (uniquement dans le dernier secteur ?).
Ascospores en fuseau [14.5 ; 15.5] x [4.0 ; 4.5]µm soit Qmoy = 3.6 avec un contenu lipidique 4-5 (2 grosses gouttes principales toujours présentes et de nombreuses plus petites autour).
Pas vu de poils ni de cellules marginales particulières.

Excipulum ectal composé de cellules en textura prismatica orientées parallèlement à la surface de la fructification.
Sous-hyménium composé de cellules en boudins (moy. 30x10µm) en textura intricata non gelifiée.
Medulla composée d'hyphes grêles et allongés (entre 3 et 4µm de large) en textura intricata gelifiée.

Avec les éléments donnés, je pense que j'ai affaire à un Ombrophila (pouvez-vous confirmer?) ; ensuite dans ma doc je ne trouve aucune espèce qui possède des spores aussi grandes et avec de telles guttules.
Merci d'avance pour vos commentaires. Les images en meilleure qualité sont temporairement mises dans le lien ci-dessous :

31/07/2022 – Ozoir-la-Ferrière (77) – France – Study on fresh living material.
On decaying twigs, under Carpinus betulinus, next to a forest pond.
Cylindrical stipitate apothecia, gregarious on substrate, white at first then greyish. Hymenium constantly convex.
Biggest specimen is 3mm diameter and 5mm tall.

Asci 8-spored inoperculate, biseriate, [75 ; 90] x [7.5 ; 9.0]µm with Hymenoscyphus-type apex, IKI+ bb, croziers+.
Paraphyses cylindrical, pluriseptate, with remarkable vacuolar body filling the entire last septum.
Ascospores fusoid [14.5 ; 15.5] x [4.0 ; 4.5]µm - Qmoy = 3.6. Oil droplets level 4-5 (2 big drops are always there, numerous smaller ones around)
No hair, no striking marginal cells.

Ectal excipulum of textura prismatica oriented parallel to the surface of the fruitbody.
Subhymenium of textura intricata non gelatinized (big cells - moy. 30x10µm)
Medullary excipulum of textura intricata gelatinized (narrow and elongated cells - width 3 to 4µm)

Those elements are leading me to the genus Ombrophila (please confirm that ?) ; then I find no suitable species in my doc with spores that long and with such an oil content.
Thanks in advance for advices, pics with better graphics are temporarily accessible through the link below (for free !) :


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Michel Hairaud, 02-08-2022 18:00
Michel Hairaud
Re : Ombrophila(?) - ascospores with great oil content
Bonjour Pierre Yves, 

Content de te savoir assidu aux Helotiales ! 
Comme tu es un heureux bénéficiaire des services d', tu as reçu récemment un superbe article de Zotto dan s notre revue, sur des espèces herbicoles d'Ombrophila. 
Regardes si cette récolte ne correspondrait pas par hasard à celle évoquée sous le nom provoisoire de O. aff. ambigua ?
C'est une espèce que nous avons récoltée en Bretagne avec Brigitte et tu trouveras une bonne planche dans l'article .
Je viens récemment de la retrouver sur Lycopus europaeus dan sune zone marécageuse (sous Salix ) et je prépare un dosssier pour avancer sur ce taxon.
Ta récolte nous intéresse .. 

Amitiés Michel
Hans-Otto Baral, 03-08-2022 09:02
Hans-Otto Baral
Re : Ombrophila(?) - ascospores with great oil content
Good idea Michel to compare this with O. aff. ambigua. Another option would be O. rivulorum, especially HB 3066. This variable species is quite difficult and probably not homogeneous.

The apical ring of your fing, Pierre-Yves, is Calycina-like though with an annular protrusion below. Various Ombrophila species have that, but others remind more of Hymenoscyphus.

The crystals in the gelatinized medulla are well visible in your pics 1 and 4. Whether the spores finally get 3-septate and produce conidia as in my find remains to be clarified from overmature apos.

The substrate: O. rivulorum is mainly lignicolous, but there is a cf. rivulorum, 28.VII.2020 in my folder which is on a herb, maybe comparable to Michel's species with very long spores. And there are further samples on herbs. Could you please clarify the substrate in your case? Is it woody?
Michel Hairaud, 03-08-2022 09:52
Michel Hairaud
Re : Ombrophila(?) - ascospores with great oil content
THanks Zotto, 
If the substrate (herb/wood) is actually differentiating, Pierre Yves 's find should not be aff. ambigua. as it is on small twigs.  

I just had a look at your file rivulorum , with several collections which may include this find. It seems the work you initiated for the genus could well be continued -:) 

Pierre-Yves Julien, 03-08-2022 19:05
Re : Ombrophila(?) - ascospores with great oil content
Thank you Michel and Zotto for your comments.

I guess the substrate is woody; here is a photo of the section of the substrate witch is 6mm thick. I don't believe there was any Salix in the area but this is something I can check next time.

In addition to my description, I'd say that the fruit bodies are turning brown or brown-red when aging instead of greyish, see my pics from today, 3 days later.

Michel, I need some more time to read the article in its entirety but well the shape and content of your spores aff. ambigua photo 2f are actually close indeed.

Zotto, I quickly checked your HB3066, spores are close to mine as well, I need some more time to review your notes. In my doc, it is written for O. rivulorum : "Apothecia very pale flesh-rose, medulla not to faintly gelatinous", this is why I rejected this possibility.

Anyway I dried the specimen before they decay too much, I can send samples if needed.
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Hans-Otto Baral, 04-08-2022 10:16
Hans-Otto Baral
Re : Ombrophila(?) - ascospores with great oil content
o.k., this reddening is due to the VBs in the paraphyses.

As you wrote "Medulla composée d'hyphes grêles et allongés (entre 3 et 4µm de large) en textura intricata gelifiée", I concluded your fungus must be gelatinous. Gel is not easily demonstrable, the easiest is to check if the fungus behaves gelatinous when touched with a needle or when pressed under the coverglass.

I have no time for a study. Now when you have it dried, it is not easy to recognize an old apo for checking spore germination.

For the wood anatomy you need to photograph a clean cross-cut vertically, either by surface or transmitted light, best under around 30x or 100x.
Pierre-Yves Julien, 07-08-2022 14:07
Re : Ombrophila(?) - ascospores with great oil content
Hello Zotto & Michel,

Maybe I have a wrong definition of « gelifiée / gelatinized ». I stated «textura intricata gelifiée » because of the microscopic view ;
In pic "7" (what I consider the medulla is), cells are loose in the textura intricata and we can't distinguish other cells in different focal planes in the gaps => gélifié.
In pic "4" (what I consider the subhymenium is), cells are not loose in the textura intricata and we can distinguish other cells in different focal planes in the gaps => non gelifié.

The fruit body was not so much macroscopically gelatinous and I could radially cut it straight easily.
Pierre-Yves Julien, 07-08-2022 14:09
Re : Ombrophila(?) - ascospores with great oil content
I finally had time to go further in the study of this species – I managed to study an overmature fruit body (fresh one that I spared from drying) and I found a lot of spores producing conidia (see pic attached – water then congo red by capillarity) ; sometimes the overmature spore produce only 1 conidia, sometimes 2 conidia ; sometimes it is 1-septate, sometimes 0-septate.

In presence of congo red, I found some spores that seem to be wrapped in a sheath ( ?) ; It is difficult for me to say if it is an artefact, a true sheath or just the remnant of ascus, I'd be interested in your opinion. I didn't see such a thing in all my preps in water.

Still in this overmature apothecia, most of the spores where already dead but I found some living ones, which were a bit smaller and with a lesser oil content than the aforementioned description.

I add here as well a pic of an ascus base for record.
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Pierre-Yves Julien, 07-08-2022 14:11
Re : Ombrophila(?) - ascospores with great oil content
Concerning the substrate, here are some views with the steromicroscope at x7.5, x20 and x50 magnification. One end of the twig (maybe a root ?) was left raw as I found it in the forest (lower part of attached pic) ; the other end is after I cut it vertically with a sharp blade (upper part of pic). I hope it helps in the identification.

Thank you Zotto for driving me in this study and sharing your documents for comparison.

I uploaded the new pics with good graphics in another link here :
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Hans-Otto Baral, 07-08-2022 21:46
Hans-Otto Baral
Re : Ombrophila(?) - ascospores with great oil content
Splendid indeed that you found the conidia! So only 0-1-septate when overmature.

A sheath I would not say for sure. This is better to clarufy in freshly ejected spores, while this could indeed be only ascoplasma.

The anatomy: There is almost no marrow or empty space in the centre which I would expect for herbs, this speaks for a woody plant. But I cannot recognize it.
Pierre-Yves Julien, 16-08-2022 14:59
Re : Ombrophila(?) - ascospores with great oil content
In order to close the topic, I believe there is sufficient evidence to call this collection Ombrophila cf. rivulorum. 

Zotto, I've got a related question after reading your article on Ombrophila ambigua (cf. attached photo) : could there be, in Helotiales, a correlation between maturity of spores and their amount of oil content ? Thanks for your time.
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Hans-Otto Baral, 16-08-2022 17:56
Hans-Otto Baral
Re : Ombrophila(?) - ascospores with great oil content
No, this is not so easy. I surely wrote in the paper that both spore types are found inside mature asci and are forcibly ejected. It is a kind of heterospory.

Apart from such rather rare heterospory, in Helotiales and other groups exist principally two cases: (1) young asci are eguttulate and the spores when just formed are also, i.e. the LBs appear during spore development from young to mature; (2) young asci are multiguttulate and yung spores are also, only spore size and size of LBs increases during maturation.
Pierre-Yves Julien, 18-08-2022 15:10
Re : Ombrophila(?) - ascospores with great oil content
Okay, it is clearer for me now :)
Pierre-Yves Julien, 05-09-2022 08:56
Re : Ombrophila(?) - ascospores with great oil content
Hello, I went back to the spot last week and found out there were Salix trees around, I just didn't pay attention last time. Macroview of wood sample taken from a standing tree confirms as well the nature of substrate (photo#1).

More over, end of August there were even more fruitbodies close to the spot, clearly connected to roots or decaying branches of Salix (photo#2).
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