Piattoni F, Leonardi P, Siham B, Iotti M, Zambonelli A
Department of Agricultural Sciences, Bologna University, Viale Fanin, Bologna, Italy.
Department of Life, Health and Environmental Sciences, University of L'Aquila, Via Vetoio, Coppito 1, L'Aquila, Italy.
Cryo Letters. 2017 Jan/Feb;38(1):58-64.
Truffles (Tuber spp.) are the most precious ectomycorrhizal edible mushrooms whose biodiversity is seriously endangered.
To develop a protocol for cryopreservation of Tuber spp. mycelia using T. borchii as a model species, verifying whether conservation in liquid nitrogen may affect viability, growth rate, hyphal morphology and infectivity.
Cryopreservation was performed using sorbitol, sucrose and DMSO as cryoprotectants. The morphological parameters analyzed were: hyphal diameter, septal distance and hyphal growth unit. Cryopreserved mycelium infectivity was assessed by inoculating Quercus robur seedlings.
In T. borchii cryopreserved mycelium, the lag-phase lasted for 6-42 days but no differences in growth curve evolution, growth rate and hyphal morphology were observed except for hyphal growth unit. No differences in mycorrhizal colonization were observed between the seedlings inoculated with non-cryopreserved and cryopreserved mycelium.
The established protocol is suitable for long-term conservation of Tuber mycelium and opens up the possibility of creating a Tuber spp. germplasm bank to preserve truffle diversity.
松露（块菌属）是最珍贵的外生菌根食用菌，其生物多样性正面临严重威胁。
以博氏块菌为模式物种，制定一套块菌属菌丝体的冷冻保存方案，验证液氮保存是否会影响其活力、生长速率、菌丝形态和感染性。
使用山梨醇、蔗糖和二甲基亚砜作为冷冻保护剂进行冷冻保存。分析的形态学参数包括：菌丝直径、隔膜距离和菌丝生长单位。通过接种欧洲栓皮栎幼苗评估冷冻保存菌丝体的感染性。
在博氏块菌冷冻保存的菌丝体中，延迟期持续6 - 42天，但除菌丝生长单位外，未观察到生长曲线演变、生长速率和菌丝形态的差异。接种未冷冻保存和冷冻保存菌丝体的幼苗之间，菌根定殖没有差异。
所建立的方案适用于块菌菌丝体的长期保存，并为建立块菌属种质库以保护松露多样性开辟了可能性。
