CAI Ling,HE Liming,XIE Xin,et al.A High-quality and Long-term Preservation Method for Ganoderma lucidum Strains[J].Northern Horticulture,2024,(10):92-98.[doi:10.11937/bfyy.20234524]
一种灵芝菌种液氮保藏方法的研究
- Title:
- A High-quality and Long-term Preservation Method for Ganoderma lucidum Strains
- 文章编号:
- 1001-0009(2024)10-0092-07
- Keywords:
- Ganoderma lucidum; liquid nitrogen preservation; ISSR
- 分类号:
- S 567.3+1
- 文献标志码:
- A
- 摘要:
- 以“药灵芝1号”为试材，采用三因素三水平的液氮保藏正交实验，对萌发率、恢复时间、菌丝生长速率、菌丝体干质量4个指标进行保藏效果综合评价，以期建立一种灵芝菌种液氮保藏方法，再以10个不同灵芝菌种使用该保藏方法进行保藏效果的验证。结果表明：最优保藏方案为采用-80 ℃液氮的降温方式，10%甘油作为保护剂，常温解冻复苏。10个不同灵芝菌种应用该保藏方法后均可萌发，恢复时间范围在1~7 d，80%菌种菌丝生长速率保藏前后无显著变化、100%菌种菌丝体干质量保藏前后无显著变化，ISSR共扩增出611条片段，大小范围在300~3 000 bp，保藏前菌种和保藏后菌种在相同引物下的扩增条带一致，未发现变异现象。因此，该研究的液氮保藏方法可保障保藏菌种的萌发率、恢复时间、菌丝生长速率、菌丝体干质量等指标，适用于多种灵芝菌种的保藏，普适性强，应用范围广，且操作简便，具有良好的应用前景。
- Abstract:
- Ganoderma lucidum No.1’ was taken as the test materials,a liquid nitrogen preservation orthogonal test with three factors and three levels was conducted.Four indicators,included germination rate,recovery time,mycelial growth rate,and dry weight of mycelium,were used to comprehensively evaluate the preservation effect,in order to establish the liquid nitrogen preservation method for Ganoderma lucidum strains,10 different strains were used to verify the preservation effect of this method.The results showed that the optimal preservation scheme involved cooling with liquid nitrogen at -80 ℃,using 10% glycerol as a protective agent,and recovering at room temperature.Ten different Ganoderma lucidum strains could germinate after using this preservation method,and the recovery time was between 1-7 days.There was no significant change in mycelium growth rate before and after preservation of 80% strains,and no significant change in mycelium dry weight of 100% strains before and after preservation.A total of 611 fragments were amplified by ISSR,ranging in size from 300-3 000 bp.The amplification bands of pre-preservation and post-preservation strains had no difference with the same primers,and no variation was found.In conclusion,the liquid nitrogen preservation method in this study could guarantee the germination rate,recovery time,mycelium growth rate,mycelium dry weight and other indicators of frozen strains.At the same time,this method was suitable for the preservation of various Ganoderma lucidum strains.It had strong universality,wide application range,simple operation and good application prospect.
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备注/Memo
第一作者简介：蔡凌（1992-），女，硕士，研究实习员，现主要从事菌类药材育种与栽培等研究工作。E-mail：775007601@qq.com.责任作者：罗霞（1974-），女，博士，研究员，现主要从事菌类药材研究与开发等研究工作。E-mail：287748567@qq.com.基金项目：四川省省级科研院所基本科研业务费资助项目（2022JDKY0013）；国家现代农业产业技术体系四川道地中药材创新团队资助项目（SCCXTD-2020-19）；四川省十四五中药材育种攻关-灵芝新品种选育及配套技术研究资助项目（2021YFZ0012）；四川省科技厅区域创新合作资助项目（2021YFQ0039）；四川省区域创新合作资助项目（2022YFQ0010）；重庆市技术创新与应用发展资助项目（cstc2021jscx-gksbX0018）。收稿日期：2023-12-26