The first record of <em>Densocarpa crocea</em> in Italy
10.6092/issn.2531-7342/8130
Per la prima volta Densocarpa crocea (Quél.) Healy &amp; M.E. (= Stephensia crocea Quél.) è stata trovata in Italia, ai confini della zona A del Parco Regionale dell’Abbazia di Monteveglio (SCI IT4050016). Questa specie europea è stata finora segnalata solo in Francia, Olanda e Spagna. L’ascoma è stato descritto morfologicamente e molecolarmente caratterizzato tramite sequenziamento delle regioni ITS del DNA ribosomiale. Gli ascomi di D. crocea si distinguono morfologicamente da quelli di Hydnocystis bombycina (Vittad.) Healy &amp; M.E. Sm. [= Stephensia bombycina (Vittad.) Tul. &amp; C. Tul.] solo per le dimensioni inferiori, per il colore rosso aranciato e per le dimensioni sporali (9-15 µm) che sono molto inferiori rispetto a quelle di H. bombycina.
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Mirco Iotti
Italian Journal of Mycology
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The first record of <em>Densocarpa crocea</em> in Italy
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2018-04-26
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Ital J Mycol
Mirco Iotti
2018/04/27
10.6092/issn.2531-7342/8130
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University of L'Aquila
2531-7342
ITS sequence
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The first record of <em>Densocarpa crocea</em> in Italy
https://italianmycology.unibo.it/article/download/8130/7836
47
A. Zambonelli, P. Leonardi, M. Morara, M. Iotti            Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342
DOI: https://doi.org/10.6092/issn.2531-7342/8130



 Brief note

 The first record of Densocarpa crocea in Italy
 _______________________________________________________________________________________

       Alessandra Zambonelli1, Pamela Leonardi1, Marco Morara2, Mirco Iotti3
 1
   Dipartimento di Scienze e Tecnologie Agroalimentari, University of Bologna, viale Fanin 46, 40127
   Bologna,Italy
 2
   Unione Micologica Italiana, via Fanin 46, 40127 Bologna,Italy
 3
   Department of Life, Health and Environmental Sciences, University of L’Aquila, via Vetoio, Coppito 1,
  67100 L’Aquila, Italy

 Correspondig Author e-mail: alessandr.zambonelli@unibo.it

 Abstract

     The first record of Densocarpa crocea (Quél.) Healy & M.E. Sm in Italy is reported. The specimen was
     morphologically described and the ITS sequence was deposited in GenBank.

     Keywords: Densocarpa crocea; morphological description; ITS sequence


 Riassunto

     Per la prima volta Densocarpa crocea (Quél.) Healy & M.E. (= Stephensia crocea Quél.) è stata trovata in
     Italia, ai confini della zona A del Parco Regionale dell’Abbazia di Monteveglio (SCI IT4050016). Questa
     specie europea è stata finora segnalata solo in Francia, Olanda e Spagna. L’ascoma è stato descritto
     morfologicamente e molecolarmente caratterizzato tramite sequenziamento delle regioni ITS del DNA
     ribosomiale.

     Gli ascomi di D. crocea si distinguono morfologicamente da quelli di Hydnocystis bombycina (Vittad.) Healy
     & M.E. Sm. [= Stephensia bombycina (Vittad.) Tul. & C. Tul.] solo per le dimensioni inferiori, per il colore
     rosso aranciato e per le dimensioni sporali (9-15 µm) che sono molto inferiori rispetto a quelle di H.
     bombycina.

     Parole chiave: Densocarpa crocea; descrizione morfologica; sequenza ITS

 Introduction

 The taxonomy of ascomycetes has been revolutionized by the introduction of phylogeny-based approaches in
 the last 20 years (Lumbsch, 2000). A recent study of Kumar et al. (2017) revised the classification of the
 Pyronemataceae with smooth, globose, hyaline spores, grouped in the Tarzetta-Geopyxis lineage by
 Tedersoo et al. (2006). This linkage includes the truffle genera Paurocotylis and Stephensia, and the cup
 fungus genera Geopyxis and Tarzetta (Tedersoo et al., 2006). Kumar et al. (2017) study supports the
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A. Zambonelli, P. Leonardi, M. Morara, M. Iotti             Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342
DOI: https://doi.org/10.6092/issn.2531-7342/8130



 abandonment of the genus Stephensia (Vittad.) Tul. & Tul. and suggests to transfer Stephensia bombycina
 (Vittad.) Tul. & C. Tul. to Hydnocystis, Stephensia bynumii Trappe, Bushnell & Castellano to Paurocotylis,
 and Stephensia shanori (Gilkey) Gilkey and Stephensia crocea Quél. to Densocarpa. Of these species, only
 H. bombycina and D. crocea are reported in Europe (Montecchi and Sarasini, 2000; Pegler et al., 1993; de
 Vries, 1985). While H. bombycina is quite common (Montecchi and Sarasini, 2000; Gori, 2005, Morara et
 al., 2009), D. crocea was never found in Italy.
 D. crocea was firstly found in France by Quélet (1886) who gave a synthetic description of the species.
 Then,      it    was    reported      in      several     lists   of      new      rare   fungi    of    France
 (/https://www.biodiversitylibrary.org/name/Stephensia%20crocea) and, later, in Nederland where a detailed
 description of its ascomata was made by de Vries (1985). Recently, D. crocea has been found, for the first
 time, in Spain (Rubio et al., 2006). In this brief note, we report the first record of Densocarpa crocea (Quél.)
 Healy & M.E. Sm in Italy.

 Materials and methods

 The ascoma of D. crocea was found in the SCI IT4050016 ”Abbazia di Monteveglio” Regional Park
 (Emilia-Romagna Region, Italy) near the zone A (coordinates 44° 17’ 56.88’’ N, 11° 05’ 9.9599” E) the 28
 November 2014, and deposited in the Herbarium CMI-Unibo (herbarium n. 4518). The habitat type is 92A0
 (Salix alba and Populus alba galleries) along the Ramato stream. The ascoma was found near plants of the
 footpath, in proximity of young plants of Quercus pubescens Willd. (Fig. 1).




 Fig. 1. Area of the SCI IT4050016 ”Abbazia di Monteveglio” Regional Park (Emilia Romagna Region, Italy) where D.
 crocea was found. On the left side there are young oaks which are not visible in the picture.
 Fig. 1. Area del parco Abbazia di Monteveglio (SCI IT4050016) dove è stata trovata D. crocea. Sulla sinistra c’è una
 sponda con giovani roverelle che non sono visibili nella foto.




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A. Zambonelli, P. Leonardi, M. Morara, M. Iotti          Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342
DOI: https://doi.org/10.6092/issn.2531-7342/8130



 Identification of the species was carried out on the basis of spore dimensions and ITS rDNA sequences.
 Spore dimensions were taken using Nis-Elements AR (v3.10) software (Zeiss) from images captured with a
 DXM1200F digital camera (Nikon). The primer pair ITS1F–ITS4 (White et al., 1990) was used to amplify
 ITS regions with the direct PCR approach (Iotti and Zambonelli, 2006). Polymerase chain reaction products
 were first purified using the NucleoSpin Extract kit (Machery-Nagel) and then sequenced in both directions
 using forward and reverse primers ITS1F and ITS4. Sequencing reactions were performed on an ABI PRISM
 3100 Genetic Analyzer (Applied Biosystems). The obtained sequence was compared against nucleotide
 database in the National Centers for Biotechnology Information GenBank (http://www.ncbi.nlm.nih.gov/) by
 using the search tool Blastn to confirm the species. The sequence of the ascoma described in this study was
 deposited in GenBank with the accession number MH087211.

 Results

 The ascoma was 3 mm in diameter, subglobose with a basal cavity, orange encrusted by a brown tomentum
 (Fig. 2a and c). The microscopic characters correspond to those previously described by de Vries (1985) and
 Kumar et al. (2017). The peridium was 400-550 µm, with the outermost layer composed by yellowish-
 brown, of interwoven granule-encrusted hyphae, large 5-10 µm which fuse with a thin layer of small-celled
 pseudoparenchyma composed by irregular arranged cells; the innermost peridium was plectenchimatous
 (Fig. 2d, e and f). The gleba was white yellowish, with a unique, large meandered channel (Fig. 2c).
 Hymenium was composed by paraphyses and asci, filling the fertile channel. Paraphyses were enlonged,
 hyaline, slightly swollen at the tips, 2.5-5 µm in diameter. Asci were cylindrical to oblong, 120-150 µm x
 17.5- 23 µm, with 6-8 uniseriate spores in a single ascus. The spores were hyaline, smooth, globose, thin-
 walled (1 µm thick), of 9-15 µm (Fig. 2b).
 The ITS sequence of the described specimen had a 99.4% of similarity (3 variable positions on 463 bp) with
 D. crocea (KT361829) deposited by Kumar et al. (2017). All the other sequences in the best blast hit list had
 a sequence similarity < 90% (date of accession 22 March 2018).

 Discussion and conclusions

 D. crocea is clearly distinguishable by H. bombycina for the smaller dimensions (3 mm and 2-3 cm
 respectively), the orange color and the smaller spores (9-15 µm and 19-22 µm) respectively. The typical
 morphological characters of this species could exclude that it was previously unnoticeable found in Italy.
 This European species was rarely found only in France, Spain and Nederland. In Nederland it is reported in
 the       red      list     as       vulnerable      species    (https://www.verspreidingsatlas.nl/0695020#;
 http://minez.nederlandsesoorten.nl/content/oranje-plooitruffel-stephensia-crocea;
 http://wikipedia.qwika.com/nl2en/Nederlandse_Rode_lijst_(paddenstoelen_K_t/m_O). This species was
 already included in the red list candidates of European Council for the Conservation of Fungi (ECCF)
 (https://www.wsl.ch/eccf/activities-en.ehtml). Considering its rarity and threaten habitat, which is commonly
 visited by truffle hunters in Italy looking for the expensive white truffle (Tuber magnatum Picco), we think
 that D. crocea should be considered priority for incorporation into the IUCN red list
 (http://iucn.ekoo.se/en/iucn/welcom).




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A. Zambonelli, P. Leonardi, M. Morara, M. Iotti                Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342
DOI: https://doi.org/10.6092/issn.2531-7342/8130




 Fig. 2. Ascoma D. crocea cut in two parts (a); gleba (c); spores (b); innermost peridium (d); outermost peridium (e);
 granule-encrusted hyphae (f). a & c bars = 1mm; b, d, e & f bars = 10 µm.
 Fig. 2. Ascoma di D. crocea tagliato in due parti (a); gleba (c); spore (b); peridio interno (d); peridio esterno (e); ife
 incrostate (f). a, c barre = 1 mm; b, d, e , f barre = 10 µm.

 Acknowledgements

 This note is dedicated to my truffle dog Geppa.

 References

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DOI: https://doi.org/10.6092/issn.2531-7342/8130


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