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- Title
基于响应面法对香菇原生质体制备条件的优化.
- Authors
孙佳星; 杨红; 孙晓东; 郝然; 张伟东; 吕国忠
- Abstract
Lentinula edodes is the highest-yielding edible mushroom grown in China. Optimizing he protoplast preparation conditions to generate high quality and vitality of the fungal protoplasts is critical for breeding and genetic diversity analysis of L. edodes. The optimal action conditions and correlations of four factors, concentration of osmolymer, enzymatic type, enzymolysis duration and enzymolysis temperature, were tested and screened by single factor test. According to the results of single factor test, the duration and temperature of enzymolysis and enzyme solution were investigated by three factors and three levels of response surface method. The single factor test shows that using mixed enzyme preparation of 2% lysozyme + snail enzyme + cellulase with 0.6 mol/L mannitol as osmotic stabilizer for synergistically multiple enzyme enzymolysis was significantly effective as compared with other treatments (P<0.05). Through Box-Behnken design, the optimal preparation conditions of L. edodes protoplasts were as follows: enzymolysis duration of 3.25 h, enzymolysis concentration at 1.7%, and enzymolysis temperature at 30.5 °C. Under these conditions the yield of protoplasts was 14.02×106 CFU/mL, consistent with the theoretical yield (13.862×106 CFU/mL). The results provide a key technological guidance for further protoplast fused breeding and genetic diversity analysis of L. edodes.
- Subjects
CHINA; MANNITOL; EDIBLE mushrooms; GENETIC variation; PROTOPLASTS; LYSOZYMES; CELLULASE
- Publication
Mycosystema, 2022, Vol 41, Issue 5, p837
- ISSN
1672-6472
- Publication type
Article
- DOI
10.13346/j.mycosystema.210295