摘要
采用原生质体紫外诱变育种方法,进行高产γ-氨基丁酸的秀珍菇菌株选育工作,对原生质体制备条件、再生条件和诱变株γ-氨基丁酸含量进行了筛选分析。结果表明:MM缓冲液中,20 mg/mL的溶壁酶能够有效去除秀珍菇细胞壁,获得2×10~6个/mL的原生质体;在含有0.5 mol/L甘露醇的YMG培养基上,原生质体再生率最高,可达到0.65%±0.13%。35 W紫外灯照射1～2 min时,致死率可达到76%～86%;菌丝体和子实体中,3株突变株γ-氨基丁酸含量较出发菌株分别提高了20.7%、196.3%、126.8%和21.3%、237.8%、97.3%。诱变菌株的获得,可为高值化富含γ-氨基丁酸秀珍菇生产菌株的培育提供优异的育种材料。
An ultraviolet ray-mediated protoplast mutagenesis method was applied to breed γ-aminobutyric acid (GABA)-rich Pleurotus geesteranus strains, screening conditions for protoplast preparation, protoplast regeneration and GABA concentrations in mutants. The results show that, concentration of protoplast of Pleurotus geesterans could reach 2×10^6 cloes/mL when 20 mg/mL(wt) lywallzyme being dissolved in MM buffer. The protoplast regeneration rate could top 0.65%±0.13% on a YMG plus 0.5 mol/L mannitol medium. The lethal ratio could range from 76% to 86% when protoplasts being irradiated under a 35 W ultraviolet lamp for 1~2 minutes. GABA concentration in mycelia and fruiting bodies from 3 mutants has increased by 20.7% and 196.3%, 126.8% and 21.3%, 237.8% and 97.3% respectively, compared to those from the original strains. The acquisition of GABA-rich mutant strains can provide excellent breeding materials for cultivation of high-value GABA-rich Pleurotus geesteranus products.
作者
张亚青
张羡
付佳伟
楼坚
刘士旺
ZHANG Yaqing;ZHANG Xian;FU Jiawei;LOU Jian;LIU Shiwang(School of Biological and Chemical Engineering,Zhejiang University of Science and Technology,Hangzhou 310023,Zhejiang,China;Zhejiang Provincial Key Laboratory for Chemical and Biological Processing Technology of Farm Produce,Hangzhou 310023,Zhejiang,China;Zhejiang Province Collaborative Innovation Center of Agricultural Biological Resources Biochemical Manufacturing,Hangzhou 310023,Zhejiang,China)
出处
《浙江科技学院学报》
CAS
2019年第3期198-205,共8页
Journal of Zhejiang University of Science and Technology
基金
浙江省重大科技专项计划项目(2015C02053)
关键词
秀珍菇
原生质体
紫外诱变育种
Γ-氨基丁酸
Pleurotus geesteranus
protoplast
ultraviolet-mediated mutagenesis breeding
GABA
