摘要
以赤芝和无柄灵芝菌株为试验材料,通过单因素及正交试验,对比分析菌龄、酶解时间、酶解温度、离心转速对灵芝原生质体制备的影响,优化制备条件,并初步探讨不同稳渗剂类型、浓度及培养方法对灵芝原生质体再生的影响。结果表明:赤芝原生质体制备最佳条件为菌龄10 d,酶解温度35℃,酶解时间3 h,4000 r/min离心,原生质体产量9.75×10^(7)/mL;无柄灵芝原生质体制备条件为菌龄7 d,酶解温度31℃,酶解时间3.5 h,3000 r/min离心,原生质体产量7.48×10^(7)/mL。灵芝菌株以0.5 mol/L甘露醇为稳渗剂,双层平板培养法(上层培养基琼脂体积分数为1.1%)进行再生,可以获得高再生率。
Using Ganoderma lingzhi and Ganoderma sessile strains as experimental materials,the effects of mycelial age,enzymolysis time,enzymolysis temperature and centrifugal speed on the preparation of Ganoderma lucidum protoplasts were compared and analyzed through single factor test and orthogonal test,the preparation conditions were optimized,and the effects of different types and concentrations of osmotic stabilizers and different culture methods on the regeneration of Ganoderma lucidum protoplasts were preliminarily discussed.The results showed that the optimal conditions for the preparation of Ganoderma lingzhi protoplasts were mycelial age of 10 days,enzymolysis temperature of 35℃,enzymolysis for 3 hours,centrifugation at 4000 r/min,and protoplast yield of 9.75×10^(7)/mL;Ganoderma sessile protoplasts were prepared at a mycelial age of 7 days,enzymolysis temperature of 31℃,enzymolysis for 3.5 hours,centrifugation at 3000 r/min,and a protoplast yield of 7.48×10^(7)/mL.Ganoderma lucidum strains were regenerated with 0.5 mol/L mannitol as osmotic stabilizer,and the double-layer plate culture method(with an upper agar concentration of 1.1%)could obtain a high regeneration rate.
作者
许琳
刘洋
张雪超
胡清秀
闫梅霞
XU Lin;LIU Yang;ZHANG Xuechao;HU Qingxiu;YAN Meixia(Institute of Special Animal and Plant Sciences,Chinese Academy of Agricultural Sciences,Changchun 130112,China;Jilin Province Horticultural Specialty Management Station,Changchun 130000,China;Institute of Agricultural Resources and Regional Planning,Chinese Academy of Agricultural Sciences,Beijing 100081,China;Tianjin Institute of Industrial Biotechnology,Chinese Academy of Sciences,Tianjin 300308,China)
出处
《吉林农业大学学报》
CSCD
北大核心
2024年第6期979-987,共9页
Journal of Jilin Agricultural University
基金
吉林省发展和改革委员会项目(20220038-5)
天津市合成生物技术创新能力提升行动项目(TSBICIP-CXRC-006)。
关键词
灵芝
原生质体
正交试验
再生率
Ganoderma lucidum
protoplast
orthogonal test
regeneration rate
作者简介
许琳,女,在读硕士,主要从事食药用菌育种;通信作者:闫梅霞,E-mail:yanmeixiac@126.com。
