Earth and Life Institute, Applied Microbiology, Mycology, Université Catholique de Louvain, Croix du Sud 2 bte L7.05.06, 1348 Louvain-la-Neuve, Belgium.
Mycorrhiza. 2013 Aug;23(6):463-71. doi: 10.1007/s00572-013-0489-8. Epub 2013 Mar 2.
The use of ectomycorrhizal (ECM) fungi for afforestation, bioremediation, and timber production requires their maintenance over long periods under conditions that preserve their genetic, phenotypic, and physiological stability. Cryopreservation is nowadays considered as the most suitable method to maintain the phenotypic and genetic stability of a large number of filamentous fungi including the ECM fungi. Here, we compared the ability of eight ECM fungal isolates to colonize Pinus sylvestris roots and to transport inorganic phosphate (Pi) and NH4 (+) from the substrate to the plant after cryopreservation for 6 months at -130 °C or after storage at 4 °C. Overall, the mode of preservation had no significant effect on the colonization rates of P. sylvestris, the concentrations of ergosterol in the roots and substrate, and the uptake of Pi and NH4 (+). Comparing the isolates, differences were sometimes observed with one or the other method of preservation. Suillus bovinus exhibited a reduced ability to form mycorrhizas and to take up Pi following cryopreservation, while one Suillus luteus isolate exhibited a decreased ability to take up NH4 (+). Conversely, Hebeloma crustuliniforme, Laccaria bicolor, Paxillus involutus, and Pisolithus tinctorius exhibited a reduced ability to form mycorrhizas after storage at 4 °C, although this did not result in a reduced uptake of Pi and NH4 (+). Cryopreservation appeared as a reliable method to maintain important phenotypic characteristics (i.e., root colonization and nutrient acquisition) of most of the ECM fungal isolates studied. For 50 % of the ECM fungal isolates, the colonization rate was even higher with the cultures cryopreserved at -130 °C as compared to those stored at 4 °C.
外生菌根(ECM)真菌在造林、生物修复和木材生产中的应用需要在长期条件下进行维护,以保持其遗传、表型和生理稳定性。如今,冷冻保存被认为是维持大量丝状真菌(包括 ECM 真菌)表型和遗传稳定性的最适宜方法。在这里,我们比较了 8 个 ECM 真菌分离物在-130°C 冷冻保存 6 个月或在 4°C 储存后,对欧洲赤松根系的定殖能力,以及从基质向植物运输无机磷(Pi)和铵(NH4+)的能力。总体而言,保存方式对欧洲赤松的定殖率、根系和基质中麦角固醇的浓度以及 Pi 和 NH4+的吸收没有显著影响。在比较分离物时,有时会发现一种或另一种保存方法存在差异。牛肝菌(Suillus bovinus)在冷冻保存后形成菌根和吸收 Pi 的能力降低,而一种栓菌(Suillus luteus)分离物吸收 NH4+的能力降低。相反,皱盖乳菇(Hebeloma crustuliniforme)、双色蜡蘑(Laccaria bicolor)、粗皮侧耳(Paxillus involutus)和聚藻(Pisolithus tinctorius)在 4°C 储存后形成菌根的能力降低,尽管这并没有导致 Pi 和 NH4+的吸收减少。冷冻保存被认为是维持大多数研究的外生菌根真菌分离物重要表型特征(即根系定殖和养分吸收)的可靠方法。对于 50%的外生菌根真菌分离物,与在 4°C 储存的培养物相比,在-130°C 冷冻保存的培养物的定殖率更高。