摘要
建立了一种原生质体制备新方法,通过研究接种块数量和酶解时间对香菇(Lentinula edodes)和金针菇(Flammulina filiformis)原生质体制备的影响,对该方法进行了优化并利用该方法制备了糙皮侧耳(Pleurotus ostreatus)、双孢蘑菇(Agaricus bisporus)和中国美味蘑菇(A.sinodeliciosus)原生质体。结果表明:在铺有玻璃纸的PDA平板(直径90 mm)上分别接种7个香菇、金针菇菌丝块(直径4 mm),培养3 d后将带有菌丝体的玻璃纸转移到另一个无菌培养皿中,直接在菌丝上滴加3 mL1.5%(W∶V)溶壁酶溶液,酶解60 min,单个平板制备的原生质体数量为10^6~10^7个。使用该方法成功地制备了糙皮侧耳、双孢蘑菇和中国美味蘑菇原生质体。该方法与常用的原生质体制备方法相比具有制备时间短、操作简单和不易污染的优点。
Currently,the most commonly used protocol for preparing protoplasts in mushroom is complicated and time consuming.Using Lentinula edodes and Flammulina filiformis as testing subjects,we proposed a new method:seven mycelium blocks(4 mm diameter)were first inoculated onto a 90 mm PDA plate covered with cellophane film on the medium,then after incubation for 3 d,the cellophane with mycelia was transferred to a new sterile dish,added with 3 mL 1.5%(W∶V)lyticase solution on the surface of the colonies and then incubated for 60 min for cell wall degradation.Using this new method,about 10^6-10^7 protoplasts were obtained in total per plate.This method was then further validated on Pleurotus ostreatus,Agaricus bisporus and A.sinodeliciosus,respectively.Compared with the traditional methods,this new method obviated the filtration step,and thus making the protoplast preparation much more accelerated,easier and less likely to be contaminated.
作者
李良敏
章炉军
尚晓冬
宋春艳
于海龙
张美彦
谭琦
LI Liangmin;ZHANG Lujun;SHANG Xiaodong;SONG Chunyan;YU Hailong;ZHANG Meiyan;TAN Qi(College of Food Science&Technology,Shanghai Ocean University,Shanghai,201200,China;Institute of Edible Fungi,Shanghai Academy of Agricultural Sciences,Key Laboratory of Edible Fungi Resources and Utilization(South),Ministry of Agriculture and Rural Affairs,P.R.China,National Engineering Research Center of Edible Fungi,National R&D Center for Edible Fungi Processing,Shanghai Key Laboratory of Agricultural Genetics and Breeding of Shanghai,Shanghai 201403,China)
出处
《食用菌学报》
CSCD
北大核心
2020年第4期108-114,共7页
Acta Edulis Fungi
基金
上海市科技兴农项目(2019-02-08-00-08-F01114)
上海市科委国内合作项目(19395800400)
国家现代农业产业技术体系(CARS-20)。
关键词
食用菌
原生质体
玻璃纸
酶解
Edible fungi
protoplast
cellophane
enzymatic digestion
