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

Abstract:

[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. However, 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 influences 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 short- and 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.