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Revival of saprotrophic and mycorrhizal basidiomycete cultures after 20 years in cold storage in sterile water.
Revival of saprotrophic and mycorrhizal basidiomycete cultures after 20 years in cold storage in sterile water.
Subject: Water, Distilled (Properties)
Fungi (Storage)
Cold storage (Methods)
Cultures (Biology) (Management)
Author: Richter, Dana L.
Pub Date: 08/01/2008
Publication: Name: Canadian Journal of Microbiology Publisher: NRC Research Press Audience: Academic Format: Magazine/Journal Subject: Biological sciences Copyright: COPYRIGHT 2008 NRC Research Press ISSN: 0008-4166
Issue: Date: August, 2008 Source Volume: 54 Source Issue: 8
Topic: Event Code: 200 Management dynamics Computer Subject: Company business management
Product: Product Code: 3841333 Culture-Based Tests NAICS Code: 339112 Surgical and Medical Instrument Manufacturing SIC Code: 3841 Surgical and medical instruments
Geographic: Geographic Scope: United States Geographic Code: 1USA United States
Abstract: Vegetatively colonized agar cores of 69 basidiomycete fungus isolates (48 species in 30 genera and 17 families) were stored at 5 [degrees]C in tubes of sterile distilled water without manipulation for 20 years. These were represented by 34 isolates of saprotrophic fungi (29 species in 19 genera) and 35 isolates of mycorrhizal fungi (19 species in 11 genera). Viability was evaluated based on revived growth on agar media at room temperature. Fifty-seven of the 69 isolates (82.6%) grew vigorously when revived after storage for 20 years; of the 34 saprotrophic fungus isolates, 30 revived (88.2%); of the 35 mycorrhizal fungus isolates, 27 revived (77.1%). Thirteen isolates of Laccaria were all viable after 20 years, indicating cold storage in sterile water to be a good method for maintaining this important genus of mycorrhizal fungi. In general, however, mycorrhizal fungus species demonstrated lower viability than saprotrophic fungi.
Key words: culture maintenance, culture viability, fungal preservation, long-term storage, mycorrhizal fungi, saprotrophic fungi, vegetative cultures.
Resume : Des echantillons d'agar colonises de facon vegetative comprenant 69 isolats de champignons basidiomycetes (48 especes divisees en 30 genres et 17 familles) ont ete entreposes a 5[degrees]C dans des tubes d'eau sterile distillee sans manipulation pendant 20 ans. Ceux-ci etaient representes par 34 isolats de champignons saprotrophes (29 especes divisees en 19 genres) et 35 isolats de mycorrhizes (19 especes divisees en 11 genres). La viabilite a ete evaluee par une reprise de la croissance sur agar a la temperature de la piece. Cinquante-sept des 69 isolats (82.6 %) poussaient vigoureusement lorsque ravives apres un entreposage de 20 ans; des 34 isolats de champignons saprotrophes, 30 ont ete ravives (88.2 %); des 35 isolats de mycorrhizes, 27 ont ete ravives (77.1 %). Les 13 isolats de Laccaria etaient tous vivants apres 20 ans, indiquant que l'entreposage au froid dans l'eau sterile est une bonne methode de conservation de ce genre important de mycorrhizes. En general, cependant, les especes de mycorrhizes demontraient une plus faible viabilite que les champignons saprotrophes.
Mots-cles : maintien des cultures, viabilite des cultures, preservation des champignons, entreposage a long-terme, mycorrhizes, champignons saprotrophes, cultures vegetatives.
[Traduit par la Redaction]
Introduction
Long-term storage of fungus cultures to ensure isolate stability is an important part of any mycology research laboratory. Interest in preserving microbial genomic diversity has also led to interest in maintaining cultures of fungi (Smith et al. 1994). However, maintaining genetic stability can be problematic. For example, characteristics of fungi such as pathogenicity, virulence, and growth rate are known to change over time when mycelium is continually sub-cultured on agar (Marx et al. 1984; Hung and Molina 1986; Richter et al. 2004). Advantages and disadvantages of the various methods of maintaining fungus cultures over long periods of time for research are thoroughly discussed by Smith and Onions (1983). A simple method of maintaining fungus cultures that has been successful in several laboratories in recent years is storage under sterile water in normal refrigeration (Marx and Daniel 1976; Ellis 1979; Richter and Bruhn 1989; Johnson and Martin 1992; Burdsall and Dorworth 1994; Smith et al. 1994).
Richter and Bruhn (1989) stored 135 basidiomycete fungus isolates, represented by 83 species in 38 genera, in sterile cold water (5 [degrees]C); of those that were revived after 8-48 months, 35 out of a total of 37 isolates of saprotrophic fungi were viable (95%), while only 53 out of a total of 98 isolates of mycorrhizal fungi were viable (54%). The original sterile water tubes containing cores of the isolates that were revived and viable were placed back in cold storage and left unmanipulated for 20 years. Of the original 135 isolates (Richter and Bruhn 1989), 88 isolates remained. However, due to difficulties inherent in storage conditions over such an extended period, several tubes dried or became contaminated. Thus, for this study, 69 isolates (34 saprotrophic fungi, 35 mycorrhizal fungi) were taken out of storage and attempts made to revive them after 20 years of storage in sterile cold water.
Materials and methods
Fungus isolates were originally obtained from basidiome tissue. Saprotrophic fungi were isolated on potato dextrose agar (Difco) or 2% malt extract agar (Difco), while mycorrhizal fungi were isolated on Modified Melin Norkrans agar (Marx 1969). Isolates were placed into sterile cold water storage 3-10 months after the time of isolation following the methods of Marx and Daniel (1976) (see Richter and Bruhn 1989). In this process, a sterile cork borer (8 mm diameter) was used to cut colonized agar cores from the margin of actively growing cultures in Petri dishes; agar was approximately 5 mm thick. Eight to 12 cores of each fungus were placed in 20 mL of sterile distilled water in a 20 mm x 150 mm glass culture tube; screw tops were placed on tightly and sealed with several wraps of Parafilm to minimize the chance of contamination or evaporation.
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For revival, 3 cores of each isolate were taken out of the sterile water tube and placed mycelium-side down on the surface of fresh agar media in Petri plates containing the medium on which they were originally grown and stored. Plates were incubated at room temperature and monitored weekly for up to 6 weeks for evidence of growth. Growth was compared with an actively growing culture of the fungus species freshly transferred from an agar slant. After examination, isolates were rated either viable or nonviable based on resumed growth or lack of growth, respectively.
Results and discussion
Results are shown in Table 1. For comparison, isolates are grouped in families as in the earlier paper that reported these isolates (Richter and Bruhn 1989). However, family affiliations have changed for some of the fungi and the modern classification is used here (Kirk et al. 2001).
The 69 fungus isolates for which revival was attempted were represented by 48 species, in 30 genera, in 17 families; 34 of these isolates were saprotrophic fungi (29 species, in 19 genera), and 35 isolates were mycorrhizal fungi (19 species, in 11 genera).
Of the 69 isolates, 57 (82.6%) grew vigorously when revived after 20 years of storage in sterile cold water; 30 of the 34 saprotrophic fungus isolates revived (88.2%), and 27 of the 35 mycorrhizal fungus isolates revived (77.1%).
For the viable isolates, all 3 of the cores that were retrieved from sterile water for each fungus grew new mycelium, except for 2 isolates of Laccaria laccata, in which case only one of the cores was viable for each (Table 1).
Remarkably, all 13 isolates of the mycorrhizal Laccaria bicolor and Laccaria laccata isolates survived after 20 years. However, only 4 of the 7 mycorrhizal Suillus species isolates survived. A large group of principally saprotrophic fungi, the Tricholomataceae, were generally viable, with 14 of 17 isolates reviving. Within this family, only one of 3 isolates of the genus Tricholoma (mycorrhizal) was not viable. Good viability was also exhibited in the saprotrophic Marasmiaceae (all 5 isolates viable) and the Pleurotaceae (all 3 isolates viable).
Of the 6 isolates of gasteromycetes, only one was not viable, this being the mycorrhizal species Scleroderma citrinum. This is not unexpected, since in the previous study (Richter and Bruhn 1989), 22 isolates in the genus Scleroderma exhibited very low survivability even after just 1 year of storage in sterile cold water.
The fungi in this study were essentially a select group of isolates in that these were isolates that had been stored, revived, and survived up to 4 years in cold water storage from the previous study (generally 2 years storage for most isolates) (Richter and Bruhn 1989). When the percentage of survival is calculated based on the original number of isolates (135) stored (minus 19 isolates not attempted in 2006 due to dryness and (or) contamination), this results in an overall survival rate after 20 years of 49.1% for all fungi. However, this percentage is lowered by the high number of mycorrhizal fungi in the original set. For the saprotrophic fungi alone, the survival rate from the original isolates after 20 years storage was 93.8%. In contrast, for the mycorrhizal fungi the survival rate after 20 years was only 32.1%. However, the percentage of survival for the mycorrhizal fungi is skewed downward due to the high number (22) of isolates of Scleroderma, which had a very low survival rate even after just 12 months of storage (Richter and Bruhn 1989). If isolates of Scleroderma are removed from the data set, the percentage of survival of mycorrhizal fungi after 20 years in sterile cold water becomes 43.5%.
Table 2 compares survival by genus after 2-4 years (Richter and Bruhn 1989) and 20 years of storage (this study), for those genera where 2 or more isolates were originally stored in sterile cold water. Genera of both mycorrhizal and saprotrophic fungi vary in their survivability after long-term storage in sterile cold water. For example, as mentioned above, the mycorrhizal genus Laccaria appears extremely well-suited for sterile cold-water storage. In contrast, the mycorrhizal genera Boletus, Lactarius, Paxillus, Scleroderma, and Thelephora are unsuited for this type of culture storage. Other mycorrhizal genera are intermediate in their survival success after long-term storage. In general, most isolates in genera of saprotrophic fungi survived after 20 years of storage in sterile cold water, with the exception of Clitocybe, of which 3 of 8 isolates failed to revive.
Burdsall and Dorworth (1994) also demonstrated a high rate of survivability (94%) of saprotrophic basidiomycete fungus cultures in sterile cold water for up to 7 years of storage. However, Johnson and Martin (1992), who stored their cultures of saprotrophic basidiomycete fungi at room temperature in sterile water for 10 years, reported only 26% survivability.
Smith et al. (1994), who stored cultures of mycorrhizal fungi for up to 20 months, demonstrated that temperature is a factor to be considered when storing cultures; in their study 95% of cultures revived at 18 [degrees]C, while at 4 [degrees]C only 78% of cultures revived. Marx and Daniel (1976), who stored mycorrhizal fungus cultures for up to 3 years in sterile cold water, showed that survival was 100% after 1 year but reduced to 95% and 64% after 2 and 3 years of storage, respectively. Based on this, although no other studies have reported revival of cultures of mycorrhizal fungi after 20 years of storage, 32.9% appears to be an expected rate of survival for this length of time.
It is of further interest that 21 of the isolates that were revived after 20 years in sterile cold water in this study did not survive after 8-12 years of transfer annually on agar slants (see Table 1). This was true for 11 of the 13 Laccaria isolates, further indicating sterile cold water to be a superior method for maintaining isolates of this genus of fungi. The mycorrhizal genus Suillus also had a high percentage of isolates that faired better stored in sterile cold water than by transferring annually on agar slants.
In conclusion, sterile cold water storage is a simple and effective method of long-term storage of basidiomycete fungus cultures; however, functional group and family must be taken into account when considering this method for use by laboratories. Overall, based on this study, this method of long-term storage is highly suitable for isolates of saprotrophic basidiomycete fungi, but this same generalization cannot be made for mycorrhizal fungi. Although sterile cold water storage appears to be a particularly good method to store isolates of the mycorrhizal genus Laccaria, it is not suitable for storing isolates of many other genera. Families and genera of mycorrhizal fungi exhibit a highly variable response to long-term storage in sterile cold water.
Acknowledgements
The former Institute of Wood Research at Michigan Technological University and the wood science program of Dr. Peter E. Laks allowed for long-term maintenance of these fungus cultures. Ms. Laura C. Kangas and Ms. Maureen L. Habarth are thanked for laboratory assistance and a careful reading of the manuscript.
Received 13 February 2008. Revision received 7 May 2008. Accepted 13 May 2008. Published on the NRC Research Press Web site at cjm.nrc.ca on 28 June 2008.
References
Burdsall, H.H., Jr., and Dorworth, E.B. 1994. Preserving cultures of wood-decaying Basidiomycotina using sterile distilled water in cryovials. Mycologia, 86: 275-280. doi:10.2307/3760650.
Ellis, J.J. 1979. Preserving fungus strains in sterile water. Mycologia, 71: 1072-1075. doi:10.2307/3759297.
Hung, L.L., and Molina, R. 1986. Temperature and time in storage influence the efficacy of selected isolates of fungi in commercially produced ectomycorrhizal inoculum. For. Sci. 32: 534-545.
Johnson, G.C., and Martin, A.K. 1992. Survival of wood-inhabiting fungi stored for 10 years in water and under oil. Can. J. Microbiol. 38: 861-864. doi:10.1139/m92-140.
Kirk, P.M., Cannon, P.F., David, J.C., and Stalpers, J.A. 2001. Ainsworth and Bisby's dictionary of the fungi. 9th ed. CABI, Oxfordshire, UK.
Marx, D.H. 1969. The influence of ectotrophic mycorrhizal fungi on the resistance of pine roots to pathogenic infection. I. Antagonism of mycorrhizal fungi to root pathogenic fungi and soil bacteria. Phytopathology, 59: 153-163.
Marx, D.H., and Daniel, W.J. 1976. Maintaining cultures of ectomycorrhizal and plant pathogenic fungi in sterile water cold storage. Can. J. Microbiol. 22: 338-341. PMID:1252993. doi:10. 1139/m76-051.
Marx, D.H., Cordell, C.E., Kenney, D.S., Mexal, J.G., Artman, J.D., Riffle, J.W., and Molina, R.J. 1984. Commercial vegetative inoculum of Pisolithus tinctorius and inoculation techniques for development of ectomycorrhizae on bare-root tree seedlings. For. Sci. Monogr. 25: 1-101.
Richter, D.L., and Bruhn, J.N. 1989. Revival of saprotrophic and mycorrhizal basidiomycete cultures from cold storage in sterile water. Can. J. Microbiol. 35: 1055-1060. doi:10.1139/m89-176.
Richter, D.L., Laks, P.E., Larsen, K.M., and Stephens, A.L. 2004. Comparison of isolates and strains within the brown rot fungus genus Gloeophyllum using the soil block decay method. For. Prod. J. 55: 72-75.
Smith, D., and Onions, A.H.S. 1983. The preservation and maintenance of living fungi. Commonwealth Mycology Institute, Kew.
Smith, J.E., McKay, D., and Molina, R. 1994. Survival of mycorrhizal fungal isolates stored in sterile water at tow temperatures and retrieved on solid and liquid nutrient media. Can. J. Microbiol. 40: 736-742. doi:10.1139/m94-117.
D.L. Richter. School of Forest Resources and Environmental Science, Michigan Technological University, 1400 Townsend Drive, Houghton, MI 49931, USA. (dlrichte@mtu.edu).
Table 1. Survival of basidiomycete cultures in sterile cold water
after 20 years of storage.
Fungus species by family Isolate No. *
Agaricaceae
Leucoagaricus naucinus (Fr.) Singer DR-83
Macrolepiota procera (Scop.:Fr.) Singer DR-127
Bolbitaceae
Hebeloma crustuliniforme (Bull. DR-32
ex St. Am.) Quel.
Hebeloma sp. "A" DR-11
Hebeloma sp. "B" DR-110
Boletaceae
Boletus hosenae Smith & Thiers DR-28
Leccinum scabrum (Fr.) S. F. Gray DR-22
Hydangiaceae
Laccaria bicolor (Maire) Orton DR-64
Laccaria bicolor (Maire) Orton DR-72
Laccaria bicolor (Maire) Orton DR-91
Laccaria bicolor (Maire) Orton DR-100
Laccaria bicolor (Maire) Orton DR-112
Laccaria bicolor (Maire) Orton DR-141
Laccaria laccata (Scop.:Fr.) Berk. & Br. DR-5
Laccaria laccata (Scop.:Fr.) Berk. & Br. DR-95
Laccaria laccata (Scop.:Fr.) Berk. & Br. DR-102
Laccaria laccata (Scop.:Fr.) Berk. & Br. DR-113
Laccaria laccata (Scop.:Fr.) Berk. & Br. DR-115
Laccaria laccata (Scop.:Fr.) Berk. & Br. DR-133
Laccaria laccata (Scop.:Fr.) Berk. & Br. DR-137
Hygrophoropsidaceae
Hygrophoropsis aurantiaca (Wulfen:Fr.) Maire ATCC 60968
Hygrophoropsis aurantiaca (Wulfen:Fr.) Maire DR-66
Lycoperdaceae
Calvatia gigantea (Pers.) Lloyd DR-105
Lycoperdon muscorum Morgan DR-98
Lycoperdon perlatum Pers. DR-84
Marasmiaceae
Armillaria mellea sensu lato DR-52
Armillaria mellea sensu lato DR-86
Armillaria gallica Merxm. & Romagn. DR-140
Lentinula edodes (Berk.) Pegler DR-89
Oudemansiella radicata (Re1.:Fr.) Sing. DR-88
Paxillaceae
Paxillus involutus (Batsch:Fr.) Fr. DR-117
Phallaceae
Phallus impudicus (L.) Pers. DR-90
Pleurotaceae
Pleurotus ostreatus (Jacq.:Fr.) Kum. DR-85
Pleurotus ostreatus (Jacq.:Fr.) Kum. DR-93
Pleurotus ostreatus (Jacq.:Fr.) Kum. DR-153
Plutaceae
Amanita citrina (Schaeff) S. F. Gray DR-35
Amanita flavoconia Atk. DR-94
Amanita muscaria (L.:Fr.) Hooker DR-59
Rhizopogonaceae
Rhizopogon rubescens (Tul.) Tul. DR-128
Russulaceae
Lactarius rufus (Scop.:Fr.) Fr. DR-71
Sclerodermataceae
Scleroderma citrinum Pers. DR-134
Strophariaceae
Naematoloma sp. DR-145
Pholiota flammans (Fr.) Kum. DR-78
Pholiota sp. "A" DR-50
Pholiota sp. "B" DR-146
Suillaceae
Suillus luteus (Fr.) S. F. Gray DR-37
Suillus luteus (Fr.) S. F. Gray DR-82
Suillus luteus (Fr.) S. F. Gray DR-143
Suillus neoalbidipes Palm & Stewart DR-9
Suillus neoalbidipes Palm & Stewart DR-44
Suillus pictus (Pk.) Smith & Thiers DR-21
Suillus pictus (Pk.) Smith & Thiers DR-92
Tricholomataceae
Cantharellula umbonata (Gme1.:Fr.) Singer ATCC 62011
Clitocybe clavipes (Fr.) Kum. DR-38
Clitocybe dealbata (Sow.:Fr.) Gillet DR-33
Clitocybe geotropa (Bull. ex St. Am.) Kum. DR-139
Clitocybe gibba (Pers.:Fr.) Kum. DR-16
Clitocybe hydrogramma (Bull.:Fr.) Kum. DR-67
Clitocybe odora (Bull.:Fr.) Kum. DR-3
Clitocybe sp. DR-7
Collybia sp. DR-40
Hypsizygus tessulatus (Bull.:Fr.) Singer DR-129
Lepista glaucocana (Bres.) Singer DR-138
Lepista nuda (Bull.:Fr.) Cooke DR-147
Lyophyllum decastes (Fr. ex Fr.) Singer DR-87
Panellus stypticus (Bull.:Fr.) Karst. DR-106
Tricholoma populinum Lange DR-148
Tricholoma populinum Lange DR-149
Tricholoma resplendens (Fr.) Quel. DR-79
Fungus species by family Type of fungus
Agaricaceae
Leucoagaricus naucinus (Fr.) Singer Saprotrophic
Macrolepiota procera (Scop.:Fr.) Singer Saprotrophic
Bolbitaceae
Hebeloma crustuliniforme (Bull. Mycorrhizal
ex St. Am.) Quel.
Hebeloma sp. "A" Mycorrhizal
Hebeloma sp. "B" Mycorrhizal
Boletaceae
Boletus hosenae Smith & Thiers Mycorrhizal
Leccinum scabrum (Fr.) S. F. Gray Mycorrhizal
Hydangiaceae
Laccaria bicolor (Maire) Orton Mycorrhizal
Laccaria bicolor (Maire) Orton Mycorrhizal
Laccaria bicolor (Maire) Orton Mycorrhizal
Laccaria bicolor (Maire) Orton Mycorrhizal
Laccaria bicolor (Maire) Orton Mycorrhizal
Laccaria bicolor (Maire) Orton Mycorrhizal
Laccaria laccata (Scop.:Fr.) Berk. & Br. Mycorrhizal
Laccaria laccata (Scop.:Fr.) Berk. & Br. Mycorrhizal
Laccaria laccata (Scop.:Fr.) Berk. & Br. Mycorrhizal
Laccaria laccata (Scop.:Fr.) Berk. & Br. Mycorrhizal
Laccaria laccata (Scop.:Fr.) Berk. & Br. Mycorrhizal
Laccaria laccata (Scop.:Fr.) Berk. & Br. Mycorrhizal
Laccaria laccata (Scop.:Fr.) Berk. & Br. Mycorrhizal
Hygrophoropsidaceae
Hygrophoropsis aurantiaca (Wulfen:Fr.) Maire Saprotrophic
Hygrophoropsis aurantiaca (Wulfen:Fr.) Maire Saprotrophic
Lycoperdaceae
Calvatia gigantea (Pers.) Lloyd Saprotrophic
Lycoperdon muscorum Morgan Saprotrophic
Lycoperdon perlatum Pers. Saprotrophic
Marasmiaceae
Armillaria mellea sensu lato Saprotrophic
Armillaria mellea sensu lato Saprotrophic
Armillaria gallica Merxm. & Romagn. Saprotrophic
Lentinula edodes (Berk.) Pegler Saprotrophic
Oudemansiella radicata (Re1.:Fr.) Sing. Saprotrophic
Paxillaceae
Paxillus involutus (Batsch:Fr.) Fr. Mycorrhizal
Phallaceae
Phallus impudicus (L.) Pers. Saprotrophic
Pleurotaceae
Pleurotus ostreatus (Jacq.:Fr.) Kum. Saprotrophic
Pleurotus ostreatus (Jacq.:Fr.) Kum. Saprotrophic
Pleurotus ostreatus (Jacq.:Fr.) Kum. Saprotrophic
Plutaceae
Amanita citrina (Schaeff) S. F. Gray Mycorrhizal
Amanita flavoconia Atk. Mycorrhizal
Amanita muscaria (L.:Fr.) Hooker Mycorrhizal
Rhizopogonaceae
Rhizopogon rubescens (Tul.) Tul. Mycorrhizal
Russulaceae
Lactarius rufus (Scop.:Fr.) Fr. Mycorrhizal
Sclerodermataceae
Scleroderma citrinum Pers. Mycorrhizal
Strophariaceae
Naematoloma sp. Saprotrophic
Pholiota flammans (Fr.) Kum. Saprotrophic
Pholiota sp. "A" Saprotrophic
Pholiota sp. "B" Saprotrophic
Suillaceae
Suillus luteus (Fr.) S. F. Gray Mycorrhizal
Suillus luteus (Fr.) S. F. Gray Mycorrhizal
Suillus luteus (Fr.) S. F. Gray Mycorrhizal
Suillus neoalbidipes Palm & Stewart Mycorrhizal
Suillus neoalbidipes Palm & Stewart Mycorrhizal
Suillus pictus (Pk.) Smith & Thiers Mycorrhizal
Suillus pictus (Pk.) Smith & Thiers Mycorrhizal
Tricholomataceae
Cantharellula umbonata (Gme1.:Fr.) Singer Saprotrophic
Clitocybe clavipes (Fr.) Kum. Saprotrophic
Clitocybe dealbata (Sow.:Fr.) Gillet Saprotrophic
Clitocybe geotropa (Bull. ex St. Am.) Kum. Saprotrophic
Clitocybe gibba (Pers.:Fr.) Kum. Saprotrophic
Clitocybe hydrogramma (Bull.:Fr.) Kum. Saprotrophic
Clitocybe odora (Bull.:Fr.) Kum. Saprotrophic
Clitocybe sp. Saprotrophic
Collybia sp. Saprotrophic
Hypsizygus tessulatus (Bull.:Fr.) Singer Saprotrophic
Lepista glaucocana (Bres.) Singer Saprotrophic
Lepista nuda (Bull.:Fr.) Cooke Saprotrophic
Lyophyllum decastes (Fr. ex Fr.) Singer Saprotrophic
Panellus stypticus (Bull.:Fr.) Karst. Saprotrophic
Tricholoma populinum Lange Mycorrhizal
Tricholoma populinum Lange Mycorrhizal
Tricholoma resplendens (Fr.) Quel. Mycorrhizal
Fungus species by family Viability
([dagger])
Agaricaceae
Leucoagaricus naucinus (Fr.) Singer +
Macrolepiota procera (Scop.:Fr.) Singer -
Bolbitaceae
Hebeloma crustuliniforme (Bull. +
ex St. Am.) Quel.
Hebeloma sp. "A" +
Hebeloma sp. "B" -
Boletaceae
Boletus hosenae Smith & Thiers +
Leccinum scabrum (Fr.) S. F. Gray -
Hydangiaceae
Laccaria bicolor (Maire) Orton (+)
Laccaria bicolor (Maire) Orton (+)
Laccaria bicolor (Maire) Orton +
Laccaria bicolor (Maire) Orton (+)
Laccaria bicolor (Maire) Orton (+)
Laccaria bicolor (Maire) Orton (+)
Laccaria laccata (Scop.:Fr.) Berk. & Br. (+)
Laccaria laccata (Scop.:Fr.) Berk. & Br. (+)
Laccaria laccata (Scop.:Fr.) Berk. & Br. (+)
Laccaria laccata (Scop.:Fr.) Berk. & Br. (+1)
Laccaria laccata (Scop.:Fr.) Berk. & Br. (+)
Laccaria laccata (Scop.:Fr.) Berk. & Br. (+)
Laccaria laccata (Scop.:Fr.) Berk. & Br. +
Hygrophoropsidaceae
Hygrophoropsis aurantiaca (Wulfen:Fr.) Maire +
Hygrophoropsis aurantiaca (Wulfen:Fr.) Maire +
Lycoperdaceae
Calvatia gigantea (Pers.) Lloyd +
Lycoperdon muscorum Morgan +
Lycoperdon perlatum Pers. +
Marasmiaceae
Armillaria mellea sensu lato +
Armillaria mellea sensu lato +
Armillaria gallica Merxm. & Romagn. +
Lentinula edodes (Berk.) Pegler +
Oudemansiella radicata (Re1.:Fr.) Sing. +
Paxillaceae
Paxillus involutus (Batsch:Fr.) Fr. -
Phallaceae
Phallus impudicus (L.) Pers. +
Pleurotaceae
Pleurotus ostreatus (Jacq.:Fr.) Kum. +
Pleurotus ostreatus (Jacq.:Fr.) Kum. +
Pleurotus ostreatus (Jacq.:Fr.) Kum. +
Plutaceae
Amanita citrina (Schaeff) S. F. Gray +
Amanita flavoconia Atk. +
Amanita muscaria (L.:Fr.) Hooker (+)
Rhizopogonaceae
Rhizopogon rubescens (Tul.) Tul. +
Russulaceae
Lactarius rufus (Scop.:Fr.) Fr. +
Sclerodermataceae
Scleroderma citrinum Pers. -
Strophariaceae
Naematoloma sp. +
Pholiota flammans (Fr.) Kum. -
Pholiota sp. "A" +
Pholiota sp. "B" +
Suillaceae
Suillus luteus (Fr.) S. F. Gray (+)
Suillus luteus (Fr.) S. F. Gray (+)
Suillus luteus (Fr.) S. F. Gray -
Suillus neoalbidipes Palm & Stewart (+)
Suillus neoalbidipes Palm & Stewart +
Suillus pictus (Pk.) Smith & Thiers -
Suillus pictus (Pk.) Smith & Thiers (-)
Tricholomataceae
Cantharellula umbonata (Gme1.:Fr.) Singer +
Clitocybe clavipes (Fr.) Kum. (+)
Clitocybe dealbata (Sow.:Fr.) Gillet +
Clitocybe geotropa (Bull. ex St. Am.) Kum. -
Clitocybe gibba (Pers.:Fr.) Kum. (+)
Clitocybe hydrogramma (Bull.:Fr.) Kum. +
Clitocybe odora (Bull.:Fr.) Kum. -
Clitocybe sp. +
Collybia sp. +
Hypsizygus tessulatus (Bull.:Fr.) Singer (+)
Lepista glaucocana (Bres.) Singer +
Lepista nuda (Bull.:Fr.) Cooke +
Lyophyllum decastes (Fr. ex Fr.) Singer (+)
Panellus stypticus (Bull.:Fr.) Karst. +
Tricholoma populinum Lange -
Tricholoma populinum Lange +
Tricholoma resplendens (Fr.) Quel. (+)
* DR, Collection of Dana L. Richter; ATCC, American Type Culture
Collection, Beltsville, Maryland.
([dagger]) Three agar cores were plated for each fungus. +, indicates
3 cores were viable; +1, indicates only one of 3 cores was
viable; -, indicates 3 cores were not viable. Data in parentheses
indicate that the culture did not survive after 8-12 years on
agar slants transferred annually.
Table 2. Summary of survival by genus of basidiomycete cultures in
sterile cold water after 2-4 and 20 years of storage in sterile cold
water (only genera with 2 or more original isolates stored are
shown).
Total no.
Fungus genus Type of fungus of isolates
Amanita Mycorrhizal 9
Armillaria Saprotrophic 3
Boletus Mycorrhizal 9
Clitocybe Saprotrophic 8
Hebeloma Mycorrhizal 4
Hygrophoropsis Saprotrophic 2
Laccaria Mycorrhizal 15
Lactarius Mycorrhizal 9
Leccinum Mycorrhizal 2
Lepista Saprotrophic 2
Lycoperdon Saprotrophic 2
Paxillus Mycorrhizal 3
Pholiota Saprotrophic 3
Pleurotus Saprotrophic 3
Scleroderma Mycorrhizal 22
Suillus Mycorrhizal 13
Thelephora Mycorrhizal 3
Tricholoma Mycorrhizal 4
No. of isolates surviving
after:
Fungus genus 2-4 years * 20 years
Amanita 3 3
Armillaria 3 3
Boletus 1 1
Clitocybe 7 5
Hebeloma 3 2
Hygrophoropsis 2 2
Laccaria 13 13
Lactarius 1 1
Leccinum 1 1
Lepista 2 2
Lycoperdon 2 2
Paxillus 1 0
Pholiota 3 2
Pleurotus 3 3
Scleroderma 1 0
Suillus 7 4
Thelephora 0 -
Tricholoma 3 2
* Data are from Richter and Bruhn (1989).
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