金针菇菌种中短期保藏因素的正交分析

Influence factors on short-and medium-term spawn preservation of Flammulina velutipes using orthogonal experimental design

【背景】金针菇(Flammulina velutipes)是我国一种重要的栽培食用菌,年产量超过250万t,规模已跃居世界首位。菌种保藏技术是金针菇栽培和新品种研发的基础,但相关研究十分薄弱,已成为制约我国金针菇产业进一步发展的瓶颈问题。【目的】探索不同保藏因素对金针菇优良菌种中短期保藏的影响,为建立高效、低成本、易操作的保藏方法奠定基础。【方法】以温度、甘油、海藻糖、甘露醇以及保护剂体积5个因素进行正交试验。经12个月保藏,考察金针菇菌种在木屑培养基中的菌丝生长速度,通过极差分析和回归分析解析保藏因素的效应。【结果】温度、海藻糖、甘油和甘露醇对金针菇菌种的中短期保藏有极显著的影响,保护剂体积的影响不显著。温度是最重要的影响因子,与其它4个因素的互作效应均达到极显著水平。20°C是较好的短期保藏温度,-80°C为理想的中期保藏温度。渗透型与非渗透型保护剂间的互作效应对金针菇菌种的中短期保藏有极显著影响,海藻糖和甘露醇间的互作效应不显著。高浓度的海藻糖、甘油及甘露醇均不利于金针菇菌种的中短期保藏。保藏效果较佳的保护剂为10%甘油和0.3 mol/L甘露醇混合液。【结论】建立的菌种中短期保藏方法填补了金针菇产业发展的空白,研究结果可为其它大型真菌的中短期保藏提供重要参考。

[Background] Flammulina velutipes is an important edible mushroom cultivated in China with an annual output of more than 2 500 000 tons and the cultivation scale became first place in the world. Spawn preservation is the foundation of cultivation and breeding of new varieties. How ever, weak research about spawn preservation has become a bottleneck that restricts the further development of F. velutipes industry in China. [Objective] To establish an efficient, low-cost and easy method for short-and medium-term preservation of F. velutipes spawn by evaluated the preservation factors. [Methods] The orthogonal experiments were carried out with temperature, glycerol, trehalose, mannitol and the volume of protectant as the experimental factors. After 12 months of preservation, the growth rate of mycelia on sawdust medium was investigated. Range analysis and regression analysis were used to detect the effect of preservation factors. [Results] The results showed that temperature, trehalose, glycerol and mannitol had very significant inf luences on the short-and medium-term preservation of F. velutipes spawn, while the effect of protectant volume was not significant. Temperature was the most important factor, and the interaction effects of temperature and other 4 factors were all very significant. The proper short-term preservation temperature was 20 °C, and the proper medium-term preservation temperature was-80 °C. The interaction between osmotic and nonosmotic protectant had a significant effect on spawn preservation. The interaction effect between osmotic protectants（trehalose and mannitol） was not significant. High concentrations of trehalose, glycerol and mannitol were not conducive to the shortand medium-term spawn preservation of F. velutipes. The best protectant was a mixture of 10% glycerol and 0.3 mol/L mannitol. [Conclusion] The spawn preservation method established by this study filled the blank of F. velutipes industry, and the results can also provide important reference value for short-and medium-term spawn preservation of other macro fungi.