摘要
以PEG为融合剂原生质体融合技术为育种技术,研究茯苓与灵芝原生质体的融合育种条件,将灵芝的漆酶基因转入茯苓中,构建茯苓育种的漆酶转化体系。通过正交实验方法,对茯苓和灵芝菌种的原生质体制备、融合再生条件进行优化,筛选融合菌株并对其进行种属鉴定和漆酶活力测定,结果表明:茯苓原生质体制备和再生的最佳条件为2.5%溶壁酶、2 h和32℃,原生质体融合的最佳条件为25%PEG4000、pH8.5、35℃和20 min;经筛选获得了可使RB-PDA平板稳定变色的融合菌株,分子鉴定表明其主要具有茯苓的遗传性状,与灵芝的亲缘关系较远,但具备部分灵芝的遗传性状;漆酶活力测定表明亲本灵芝的产漆酶性状转移到了融合菌株中。
In this paper,the fusion breeding conditions of Poria cocos and Ganoderma lucidum protoplasts were studied,used PEG as the fusion agent,protoplast fusion as the breeding technique,and laccase transformation system for Poria cocos breeding was constructed by transferred the laccase gene of Ganoderma lucidum into Poria cocos.The preparation,regeneration,strain screening,identification and activity determination of protoplasts were carried out for Poria cocos and Ganoderma lucidum.Experimental results show that the optimum conditions for preparation and regeneration of protoplasm were:2.5%lywallzyme,2 h,32℃.The optimal conditions for the fusion of Poria cocos and Ganoderma lucidum protoplast were:25%PEG4000,pH 8.5,35℃,20 min.The fusion strain with stabilized the color change of RB-PDA plate was screened and identified,the results showed that the fusion strain has the genetic character of tuckahoe,and the genetic relationship with Ganoderma lucidum is far,but has part of the genetic character of Ganoderma lucidum.It also showed that laccase production of parent Ganoderma lucidum was transferred to the fusion strain by determined the laccase activity of fusion strain.
作者
陈小红
林标声
傅明骏
张燎原
CHEN Xiaohong;LIN Biaosheng;FU Mingjun;ZHANG Liaoyuan(College of Life Science,Longyan University,Longyan 364000,China;College of Life Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
出处
《食品科技》
CAS
北大核心
2020年第9期1-7,共7页
Food Science and Technology
基金
福建省中青年教师教育科研项目(JAT60479、JAT60498)。
关键词
茯苓
灵芝
漆酶
原生质体
PEG
融合
Wolfiporia cocos
Ganoderma lucidum
laccase
protoplasts
PEG
fusion
