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Volvopluteus Earlei, Specie Poco Comune in Tutta Europa, Nuova Per La Toscana

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Volvopluteus Earlei, Specie Poco Comune in Tutta Europa, Nuova Per La Toscana Micologia Toscana 0, 2018 (rev. 2020): 11-25 Associazione Gruppi in attesa di ISSN - © 2018 AGMT Micologici Toscani Ricevuto / Received: 23/11/2018 Accettato / Accepted: 25/11/2018 Volvopluteus earlei, specie poco comune in tutta Europa, nuova per la Toscana Francesco Giannoni Via Giovanni Battista Marino 26, 55045 Marina di Pietrasanta (LU) - I [email protected] Umberto Pera Via Zanardelli 176, 55049 Viareggio (LU) - I [email protected] Marco Della Maggiora * Via di S. Ginese 276/i, 55062 Pieve di Compito, Capannori (LU) - I [email protected] * Autore per corrispondenza Title: Volvopluteus earlei, an uncommon species throughout Europe, new for Tuscany. Keywords: Volvopluteus, Volvopluteus earlei, taxonomy, Tuscany, Italy. Riassunto Abstract Gli autori descrivono i caratteri macroscopici The authors describe the macroscopic and e microscopici di Volvopluteus earlei, una microscopic characteristics ofVolvopluteus rara specie, raccolta per la prima volta in earlei, a rare species collected for the first Toscana, reperita in ambiente antropizzato. time in Tuscany anthropized environment. La descrizione è accompagnata da The description is provided together fotografie a colori degli sporofori scattate with colour photographs of specimens in habitat e in studio. Sono inoltre fornite in both habitat and study. Moreover, foto dei particolari microscopici, nonché photos of micro-characters as well as i risultati dell’analisi filogenetica che ne molecular analysis results confirming the confermano l’identificazione. identifications are reported. Introduzione A partire dall’agosto 2003, uno degli autori (F.G.) ha raccolto, in un giardino privato, alcuni esemplari di una specie che, da subito, ha destato un certo interesse per la presenza di una volva alla base del gambo, un cappello con cuticola sericea, brillante al disco, e lamelle rosate. I primi esemplari, analizzati solo dal punto di vista macroscopico, non hanno portato ad una determinazione precisa, ma dopo anni di nascite abbastanza 11 F. Giannoni, U. Pera & M. Della Maggiora: Volvopluteus earlei, specie poco comune in tutta Europa, nuova per la Toscana. distribuite, le raccolte del 2015 sono state oggetto di studio approfondito ed hanno finalmente permesso l’identificazione della specie come Volvopluteus earlei. Nel 2017 è stata osservata una notevole produzione di sporofori per cui le raccolte collezionate a fine 2018 sono risultate dieci, per un totale di ventotto basidiomi. La determinazione della specie è stata confermata anche a mezzo di analisi molecolare eseguita su tre diverse raccolte effettuate in anni diversi. A giudicare dalla letteratura selezionata tra una vasta quantità di fonti bibliografiche, V. earlei sembra essere un’entità fungina rara in tutta Europa, mentre il presente contributo alla sua conoscenza ci risulta essere la prima segnalazione di questa specie sul territorio regionale. Materiali e Metodi Analisi morfologica. La determinazione macroscopica dei ventotto basidiomi raccolti è stata fatta utilizzando materiale fresco, quasi tutti sono stati osservati e descritti appena raccolti, di questi solo pochi esemplari erano immaturi. Per le foto a colori si è utilizzata una macchina fotografica digitale Canon EOS 70D. Le foto al microscopio sono state ottenute con una fotocamera digitale full HD TP1080HDMI. Lo studio microscopico è stato condotto con un apparecchio binoculare Nikon Eclipse E 200 dotato di oculari 10x e obiettivi 40x, 60x e 100x (immersione in olio). L’esame microscopico è stato eseguito su materiale fresco colorato con Rosso Congo ammoniacale, in un secondo tempo per alcuni controlli sono stati utilizzati anche esemplari di erbario, in quest’ultimo caso il campione esaminato è stato precedentemente reidratato con acqua distillata e colorato con Rosso Congo ammoniacale. Le misurazioni sporali sono state effettuate su 32 spore. Le misure di lunghezza e larghezza sporale sono espresse indicando tra parentesi i valori minimo e massimo, con al centro il valore medio meno la deviazione standard e il valore medio più la deviazione standard separati da un trattino. In coda sono indicati i semplici valori medi. Sono state utilizzate le seguenti abbreviazioni:avl per lunghezza media, avw per larghezza media, Q per quoziente sporale (rapporto tra lunghezza e larghezza) e avQ per quoziente sporale medio. Estrazione, amplificazione e sequenziamento del DNA. Le operazioni di estrazione, amplificazione e sequenziamento del DNA, come anche la seguente analisi filogenetica, sono state affidate al laboratorio Alvalab (2018). Tutto il DNA è stato estratto da esemplari essiccati, impiegando un protocollo modificato basato su Murray & Thompson (1980). L’amplificazione tramite PCR è stata eseguita con i prime ITS1F e ITS4 [White et al., 1990; Gardes & Bruns, 1993] per la regione ITS rDNA. Le reazioni PCR sono state eseguite 12 Micologia Toscana 0, 2018 (rev. 2020): 11-25 F. Giannoni, U. Pera & M. Della Maggiora: Volvopluteus earlei, specie poco comune in tutta Europa, nuova per la Toscana. seguendo un programma consistente in un primo step a caldo a 95 °C per 5 minuti, seguito da 35 cicli a 94 °C, 54 °C e 72 °C con durata rispettiva di 45, 30 e 45 secondi, per finire con una fase a 72 °C per 10 minuti. I prodotti della PCR sono stati trattati in gel di agarosio all’1% e le reazioni positive sono state sequenziate con uno o entrambi i prime sopra citati. I cromatogrammi sono stati verificati per rilevare eventuali errori di lettura che poi sono stati opportunamente corretti. Analisi filogenetica. La selezione delle sequenze più strettamente correlate è stata eseguita utilizzando il tool BLAST [Altschul et al., 1997] disponibile sul database pubblico in rete INSDC (International Nucleotide Sequence Database Collaboration). Le sequenze sono state prima allineate con il software MEGA 5.0 [Tamura et al., 2011], sfruttando la sua applicazione Clustal W, e poi corrette manualmente. L’allineamento finale comprende 20 sequenze con 260 bp non coincidenti sul totale di 693 bp. Il set di sequenze allineate è stato caricato con MrBayes 3.1 [Ronquist & Huelsenbeck, 2003], con cui è stata eseguita l’analisi con l’approccio dell’inferenza bayesiana (impostazioni: modello GTR+I+G, due esecuzioni simultanee, sei catene, temperatura settata a 0.2, campionamento ogni 100 generazioni) fino alla convergenza dei parametri che è stata raggiunta dopo circa 390000 generazioni con una deviazione standard risultante minore di 0.01. Infine, la ricerca completa dell’albero con più elevato valore di massima verosimiglianza è stata eseguita in MEGA 5.0, con modello GTR e 2000 repliche di bootstrap. La soglia minima di significatività è stata impostata a 0,95 per la probabilità a posteriore (PP) risultante dall’inferenza bayesiana e a 70% per la percentuale di bootstrap (BP) a seguito dell’analisi a massima verosimiglianza. Altro. La sistematica sopragenerica indicata è quella riportata da Index Fungorum [IF]. I nomi degli autori sono stati abbreviati seguendo Kirk & Ansell (2003), mentre i codici d’erbario seguono il database in rete aggiornato da Thiers (2017). Volvopluteus earlei (Murrill) Vizzini, Contu & Justo in Justo et al., Fungal Biology 115(1): 15 (2011) ≡ Volvariopsis earlei Murrill, Mycologia 3: 282 (1911) [basionimo] ≡ Volvaria earlei (Murrill) Murrill, Mycologia 4(6): 332 (1912) ≡ Volvariella earlei (Murrill) Shaffer, Mycologia 49(4): 550 (1957) = Volvariella cookei Contu, Micologia Italiana 27(3): 38 (1998) Etimologia: da nome proprio, dedicato al micologo statunitense Franklin Summer Earle (1856- 1929). Posizione sistematica sopragenerica: regno Fungi R.T. Moore, divisione Basidiomycota R.T. Moore, sottodivisione Agaricomycotina Doweld, classe Agaricomycetes Doweld, sottoclasse Micologia Toscana 0, 2018 (rev. 2020): 11-25 13 F. Giannoni, U. Pera & M. Della Maggiora: Volvopluteus earlei, specie poco comune in tutta Europa, nuova per la Toscana. Agaricomycetidae Parmasto, ordine Agaricales Underw., famiglia Pluteaceae Kotl. & Pouzar, genere Volvopluteus Vizzini, Contu & Justo. Iconografia selezionata: Contu & Signorello (2003): 23 (come Volvariella cookei); Contu (2006): 102 (come Volvariella earlei); Lezzi (2011): 27. Descrizione Macroscopica Cappello poco carnoso, 20-48 mm, inizialmente di forma campanulata poi convessa e piana a maturità, ma sempre con largo umbone ottuso centrale. La cuticola appare liscia e lucida perché viscosa (si attacca alle labbra), ma alla lente si rivela villosa, di colore bianco, bianco sporco, spesso con una leggera sfumatura rosa e/o ocra chiaro al disco, non igrofana, con margine striato scanalato, involuto negli esemplari immaturi mentre a completa maturazione spesso la cuticola si ritrae per diversi millimetri lasciando prima intravedere poi scoperte le lamelle (Figg. 4C e 4D). Lamelle moderatamente spaziate, intercalate da numerose lamellule, larghe, libere, inizialmente bianche, rosa chiaro fino a rosa palissandro a completa maturità, con filo biancastro (Fig. 4B) e abbastanza regolare. La maturazione avviene in modo disomogeneo facendo apparire la faccia della lamella marezzata. Figura 1 - Volvopluteus earlei in habitat. Raccolta del 16-07-2015, erbario AGMT n. 71, GenBank n. MK204989. Foto: F. Giannoni 14 Micologia Toscana 0, 2018 (rev. 2020): 11-25 F. Giannoni, U. Pera & M. Della Maggiora: Volvopluteus earlei, specie poco comune in tutta Europa, nuova per la Toscana. Figura 2 - Volvopluteus earlei in habitat. Raccolta del 04-07-2017, erbario AGMT n. 72, GenBank n. MK204987. Foto: F. Giannoni Sporata rosa carico.
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