Related Experiment Video
Updated: Aug 17, 2026
Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants
Published on: May 5, 2023
Long-term cryopreservation of ectomycorrhizal basidiomycetes: A comparative evaluation of the vermiculite and
Masanori Sato1, Shigeki Inaba1, Akira Nakagiri2
1NITE International Patent Organism Depositary (IPOD), 2-5-8, Kazusakamatari, Kisarazu-shi, Chiba, 292-0818, Japan.
Abstract:
This study evaluated the long-term cryopreservation efficacy of two previously developed protocols-the Modified Perlite Protocol (MPP) and the Vermiculite Protocol (VP)-using 65 ectomycorrhizal basidiomycete culture strains stored for five years in the vapor phase of a liquid nitrogen tank. Survival rates were assessed based on recovery ratios, with both MPP and VP showing high viability: 64 of 65 strains showed complete recovery under each protocol. Colony morphology remained stable across all methods, showing no visible phenotypic changes due to cryopreservation. Regarding post-thaw mycelial growth potential measured by colony diameter, VP was superior or equivalent to MPP in 49 out of 51 analyzed strains immediately post-thaw, yielding significantly greater growth in 19 of these strains. Further statistical analysis among three protocols (HPP, MPP, and VP) in a subset of 14 strains showed that VP generally produced the largest mean colony diameters immediately post-thaw in most strains and was significantly larger than MPP in six strains, although strain-dependent exceptions were observed. These findings suggest that both MPP and VP are suitable for the long-term preservation of cryosensitive basidiomycete cultures, with VP offering enhanced immediate post-thaw mycelial growth potential.
Insights
Long-term cryopreservation using the Modified Perlite Protocol (MPP) and Vermiculite Protocol (VP) proved highly effective for basidiomycete cultures. Both methods ensured high survival and stable morphology, with VP showing superior post-thaw mycelial growth.
Area of Science:
- Mycology
- Cryobiology
- Fungal Biotechnology
Background:
- Cryopreservation is crucial for maintaining fungal germplasm.
- Long-term storage requires robust and effective protocols.
- Ectomycorrhizal fungi are vital for forest ecosystems.
Purpose of the Study:
- To evaluate the long-term efficacy of the Modified Perlite Protocol (MPP) and Vermiculite Protocol (VP) for cryopreserving basidiomycete cultures.
- To assess fungal viability, morphological stability, and post-thaw growth after five years of cryopreservation.
Main Methods:
- Cryopreservation of 65 ectomycorrhizal basidiomycete strains using MPP and VP.
- Storage in the vapor phase of a liquid nitrogen tank for five years.
- Assessment of survival rates via recovery ratios and colony morphology, and measurement of post-thaw mycelial growth by colony diameter.
Main Results:
- Both MPP and VP demonstrated high viability, with 64 of 65 strains showing complete recovery.
- No visible phenotypic changes in colony morphology were observed post-thaw.
- The Vermiculite Protocol (VP) showed superior or equivalent post-thaw mycelial growth compared to MPP in most strains, significantly enhancing growth in 19 strains.
Conclusions:
- Both MPP and VP are suitable for long-term cryopreservation of sensitive basidiomycete cultures.
- VP offers enhanced immediate post-thaw mycelial growth potential, making it a potentially advantageous method.
- Strain-dependent variations in growth response highlight the need for protocol optimization based on fungal species.
