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Home , Coprinellus, Coprinopsis, Coprinus, Hormographiella, Psathyrella, Psathyrella candolleana

MYCELIAL CHARACTERISTICS OF SEVERAL COLLECTIONS

Susanna Badalyan, Lilit Melikyan, Tatevik Shahbazyan

Yerevan State University, Laboratory of Fungal Biology and Biotechnology, 1 Aleg Manoogian St., 0025, Yerevan, Armenia

8th International Conference on Biology and Mushroom Products, 19-22 November, 2014, New Delhi, India Introduction

Based on Index Fungorum presently the Psathyrella together with 24 other genera (, Cioprinopsis, , Lacrimaria, , etc.) from former family Coprinaceae Sing. is included in the family , phylum [1]. Modern molecular data show that the genus Psathyrella is polyphyletic and species of this genus phylogenetically are closer to species from new clades Coprinellus and . Currently Coprinellus and Psathyrella are discussing as sister clades [2]. Taxonomically significant morphological, ecological, physiological and genetic characteristics of mycelia of Basidiomycetes together with characters of fruiting bodies are playing important role in their modern phylogenetic reconstruction, as well as further biotechnological cultivation. Study of biological characteristics of mycelia of different Psathyrella collections and evaluation of their taxonomic significance is topical.

a b c d e

Fig. 1. Fruiting bodies of studied species: comatus (a), (b), (d) Coprinopsis strossmayeri (d) and (e). Table 1. Collections of tested cultures Material and Clade Species Strain Origine

Methods Pc-5 Armenia, 2011 Studies of macromorphological P. candolleana Pc-6C Armenia, 2013 characters of 12 Psathyrella Pc-620 Iran, 2008 collections, such as colony Ps/I-1S Armenia, 2002 morphology, texture and Ps/I-1C Armenia, 2002 Ps/II-2S Armenia, 2002 pigmentation, as well as growth Psathyrella Ps/II-2C Armenia, 2002 rate were carried out on malt- Psathyrella sp. Ps/II-3 Armenia, 2002 extract and potato-dextrose agar Ps/II-4 Armenia, 2002 o (MEA, PDA), at 25 C, pH 5.5 Ps/III-7S Armenia, 2002 (Table 1) [3]. Ps/III-7C Armenia, 2002 For micromorphological studies Ps/III-8S Armenia, 2002 sterile cover slips overgrown by C. disseminatus Cd-30 Armenia, 1998 aerial mycelia were used for Coprinellus C. radians L2C Armenia, 2002 microscopic observation [3]. C. flocculosus Cf-1C Armenia, 2002 Interspecies antagonism was C. atramentaria Ca-01-2C Armenia, 2013 studied between species from Coprinopsis C. strossmayeri Cs-20 Armenia, 2011 C. cinerea AmutBmut USA, 2002 clades Coprinus, Coprinellus, C108 USA, 2002 Coprinopsis and Psathyrella in Coprinus C. comatus Cc-10C Armenia, 2012 dual culture experiment [4,5]. Cc-IV Armenia, 1993 Results and Discussion

All studied Psathyrella collections formed white cottony colonies with well developed aerial mycelium on both tested agar media. The colonies were fast growing (up to 15.4 mm/d) on MEA, however on PDA they formed denser colonies growing with decreased (up to 11.0 mm/d) growth rate (Table 2, Fig. 2).

Table 2. Average growth rates (in mm/d) of Psathyrella collections on agar media

Species Strains MEA PDA P. Pc-5 15.3 11.0 candolleana Pc-620 10.3 7.2 a b c d Ps/I-1S 4.7 3.7 Ps/I-1C 12.7 6.8 Ps/I-2S 8.2 6.7 Psathyrella Ps/I-2C 10.8 5.7 a' b' c' d' sp. Ps/II-3 9.5 7.7 Fig. 2. Colony morphology of Psathyrella collections on Ps/II-4 11.1 5.3 MEA (a,b,c,d) and PDA (a’,b’,c’,d’): P. candolleana, str. Ps/III-7S 15.3 10.2 Pc-620 (a,a′) and Pc-5 (b,b′), Psathyrella sp. str. Ps/III- Ps/III-7C 15.4 10.9 7C (c,c′) and Ps/III-8S (d,d′). Ps/III-8S 13.6 8.4

Hyphae in Psathyrella collections with round shape often clamps. They are morphologically similar to hyphae of Coprinopsis species (Fig. 3). Not well developed irregular hyphal loops, previously described in Coprinopsis species were also observed in Psathyrella collections. In old cultures rare chlamydospores and type arthrospores (oidia) described in other Psathyrellaceae species were observed [1]. Crystals were abundantly present in agar media (Fig. 3 m-p).

Fig. 3. Micromorphological characters of Psathyrella 0 a b c d strains on MEA at 25 C. Clamps in Ps/II-4 (a), Ps/II- 3 (b), Ps/I-2C (c), Ps/III-8S (d); hyhpal loops in Ps/III- e f g h 7S (e) and Ps/I-2C (f,g); anastomosis in Pc-5 (h); oidia in Ps/I-1S (I,j,k); i j k l chlamydospore in Ps/I-2S (l), crystals in Ps/I-2S (m), Ps/I-2C (n), Ps/II-3 (o) and Ps/III-7C (p). Size bar is 10 m n o p µm.

Study of Psathyrella and coprini (, Coprinellus disseminatus, C. radians, C. flocculosus, , C. strossmayeri, C. cinerea) collections in dual culture the highest interspecific antagonism was revealed between Psathyrella and C. comatus (family

Agaricaceae) strains (Fig. 4). Mycelial average growth rate (GRavr) indicators in C. comatus strains were decreased by 39.7 %, while in Psathyrella collections in 20.3 %. In Coprinellus species GRavr was stimulated by 60.9 % and suppressed up to 25.2 %, while in Psathyrella collections both stimulation and inhibition was around 42 %. During interaction with Coprinopsis species GRavr indicators in some Psathyrella collections were stimulated up to 90.7% and inhibited 54.0 %, while in Coprinopsis species stimulation and inhibition of GRavr indicators were 37.1 %.

a b c d Fig. 4 Interactions between Psathyrella collections and Coprinus comatus. Deadlock at the contact - type A: str. Ps/I-1S & C108 (a); primordia formation in C. comatus at the contact; str. Ps/I-1C & C108 (b); partial - type CA1: str. Ps/III-7C & Cc-IV (c) and complete - type CA2: str. Ps/III-7S & Cc-10C (d) overgrowth reactions by Psathyrella collections after deadlock. During interaction between Psathyrella and Coprinopsis collections mutual deadlock at the contact, as well as overgrowth reactions, particularly by Coprinopsis strossmayeri were described. Psathyrella collections were mainly overgrown by Coprinellus species, such as C. disseminatus and C. radians (Fig. 5).

a b c d e

f g h i j Fig. 5. Interactions between Psathyrella & Coprinellus (a,b,c,d,e) and Psathyrella & Coprinopsis (f,g,h,i,j) collections. Deadlock A type (a) between P. candolleana Pc-6 & C. disseminatus and

Psathyrella sp. Ps/I-1S & C. cinerea (f). Partial overgrowth after deadlock (type CA1) by Psathyrella sp. Ps/II-4 on C. flocculosus (b) and by Psathyrella sp. Ps/III-7S on C. atramentaria (g), by C. radians on Psathyrella sp. Ps/I-2S (d); by C. strossmayeri on P. candolleana Pc-620 (i). Complete overgrowth after deadlock (type CA2) by P. candolleana Pc-5C on C. flocculosus (c) and by P. candolleana Pc-6 on C. cinerea (h); by C. radians on Psathyrella sp. Ps/I-2S (e) and by C. strossmayeri on P. candolleana Pc-6 (j).

Table 3. Frequency of occurrence of Conclusion antagonistic reactions between Psathyrella and coprini collections Interspecific antagonism revealed between Psathyrella, Coprinellus and Coprinopsis Type of Coprinus Coprinellus Coprinopsis collections (family Psathyrellaceae) was reaction weaker and not clade specific which show Mutual deadlock their phylogenetic relatedness. A 11.1 27.8 16.7 Thus, morphological characteristics of Overgrowth by Psathyrella species mycelia and interspecific antagonism are taxonomically significant criteria. Further C 52.8 11.1 11.1 A1 studies will assist ongoing phylogenetic C 16.7 5.6 36.1 A2 reconstruction of species/clades from Total 69.5 16.7 47.2 Psathyrellaceae family. Overgrowth by coprini References C 5.5 0 2.8 C 11.1 19.4 2.8 [1] Redhead SA et al., 2001. Taxon, 50: 203-241. A1 [2] Padamsee M, 2008. Mol. Phyl. Evolution 46: 415-429. CA2 2.8 36.1 30.5 [3] Badalyan SM et al., 2011. Diversity, 3: 136-154. Total 19.4 55.5 36.1 [4]Badalyan SM et al., 2002. Phytopathol. Mediterranea. 41: 220-225. Acknowledgment: The reported study was partially supported [5] Badalyan SM, et al., 2004. Phytopathol. Mediterranea. by SCS RA, joint Armenian-Russian research project № 13RF- 43: 44-48. 110.