|Table of Contents|

Optimization of Protoplast Preparation and Regeneration Conditions of Agaricus blazei

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2020年03
Page:
128-137
Research Field:
Publishing date:

Info

Title:
Optimization of Protoplast Preparation and Regeneration Conditions of Agaricus blazei
Author(s):
ZOU Zhangyi1; DENG Baiwan1; 2
(1.School of Biological Science and Engineering,Shaanxi University of Technology,Hanzhong,Shaanxi 723001;2.Shaanxi Engineering Research Center of Edible and Medicated Fungi,Hanzhong,Shaanxi 723001)
Keywords:
Agaricus blazei; protoplast yield; regeneration rate; single factor test; orthogonal experiment
PACS:
-
DOI:
10.11937/bfyy.20192866
Abstract:
In order to improve the production and regeneration rate of Agaricus blazei Murrill,this experiment used A.blazei Murrill JS01 as the material,through single factor test and orthogonal experiment,using SPSS 22.0 software to analyze,optimize the preparation and regeneration of Agaricus blazei Murrill.This experiment was the first time to increase the regeneration rate of Agaricus blazei Murrill protoplast by adding cell wall precursors and nutritional factors to the regeneration medium.The results showed that the optimal preparation conditions of protoplast were as followed,fungal age 7 days,0.6 mol·L-1 KCl as osmotic stabilizer,lysozyme enzyme concentration 1.5%,enzymatic temperature 30 ℃,enzymatic hydrolysis time 4.0 hours,and pH 6.0.Under this condition,the protoplast yield was as high as 1.97×107 ·mL-1;the fungal age was 6 days,0.6 mol·L-1 MgSO4 was used as the osmotic stabilizer,the lysozyme enzyme concentration was 2.0%,and the enzymatic temperature was 30 ℃.When the time was 3.0 hours,pH 6.5,the regeneration medium was GM,the regeneration rate of protoplast was the highest,1.12%.After adding cellobiose and vitamin B1 to the regeneration medium,the regeneration rate was 1.17%,which was 4.46% better than the optimized result.This study provided a theoretical basis for genetic research on A.blazei Murrill in purification and rejuvenation of fungal,protoplast mutagenesis,protoplast fusion breeding,and protoplast genome rearrangement.

References:

[1]顾可飞,邵毅,李晓贝,等.松茸与姬松茸营养成分差异性研究[J].农产品质量与安全,2017(1):73-78.[2]杨丹妮,汪振炯,吕胜男.超声波协同酶法提取姬松茸基质多糖及其抗肿瘤活性研究[J].食品科技,2017,42(11):212-217.[3]李晶,魏健.姬松茸多糖增强免疫功能及抗疲劳作用的研究[J].食品研究与开发,2017,38(5):201-204.[4]欧阳玉倩.巴氏蘑菇多糖降血糖机制的研究[D].太谷:山西农业大学,2014.[5]房雷雷,赵肖通,张彦青,等.姬松茸多糖诱导巨噬细胞释放NO的机制[J].食品与机械,2018,34(5):16-19.[6]刘玮,杨继国,任杰,等.姬松茸多糖ABD的结构表征及抗炎活性研究[J].现代食品科技,2017,33(5):27-32,26.[7]FIRENZUOLI F,GORI L,LOMBARDO G.The medicinal mushroom Agaricus blazei Murrill:Review of literature and pharmaco-toxicological problems[J].Evidence Based Complementary Alternative Medicine,2008,5(1):3-15.[8]刘晓庆,秦春青,孙培龙,等.姬松茸子实体多糖高速剪切提取及其抗氧化活性[J].食药用菌,2017,25(2):113-119.[9]WU S H,LI F,JIA S Y,et al.Drying effects on the antioxidant properties of polysaccharides obtained from Agaricus blazei Murrill[J].Carbohydrate Polymers,2014,103(1):414-417.[10]彭勇胜,王江之,黄程,等.响应面法优化姬松茸多糖的提取工艺[J].现代食品科技,2011,27(9):1119-1122,1104.[11]张慧,江秀红,林新坚,等.离子束选育姬松茸新菌株的鉴定[J].食用菌学报,2010,17(3):16-19.[12]刘朋虎,江枝和,雷锦桂,等.60Co与紫外复合诱变选育姬松茸新品种：福姬77[J].核农学报,2014,28(3):365-370.[13]贺建超,贺聪莹,卢美欢,等.双孢蘑菇和姬松茸灭活原生质体融合育种研究[J].中国食用菌,2014,33(1):9-10.[14]张卉,李长彪,陈明波,等.原生质体紫外诱变选育姬松茸新菌株[J].微生物学杂志,2004(6):56-57.[15]陆利霞,谷文英,丁霄霖.姬松茸原生质体诱变育种研究[J].生物技术,2002(6):12-13.[16]谭周进,杨海君,林曙,等.利用原生质体融合技术选育微生物菌种[J].核农学报,2005(1):75-79.[17]王春晖.食用菌领域原生质体技术研究应用浅析[J].中国食用菌,2011,30(3):3-5.[18]李云梦.平菇菌株提纯复壮技术应用效果研究[D].邯郸:河北工程大学,2018.[19]李小六,张运峰,张淑红,等.原生质体EMS诱变选育杏鲍菇高产菌丝多糖菌株[J].核农学报,2017,31(8):1463-1468.[20]徐凯,张伟国.原生质体融合选育耐高温柠檬酸生产菌[J].生物加工过程,2015,13(5):26-30.[21]张素敏,王国增,叶秀云,等.基因组重排技术选育纤维素酶高产菌株[J].中国食品学报,2016,16(11):105-111.[22]常水晶,张敏.“血球计数板”演示教具的制作及使用[J].中学生物学,2018,34(8):50-51.[23]赵竟男,江梅,苏晓庆.灭蚊真菌贵阳腐霉原生质体的制备和再生研究[J].贵阳医学院学报,2008(2):111-114.[24]王伟伟,肖燕,张艺夕,等.马铃薯晚疫病菌原生质体制备及再生体系的研究[J].生物技术通报,2018,34(4):77-82.[25]王金斌,李文,蒋玮,等.褐色双孢蘑菇原生质体制备、再生条件的研究[J].核农学报,2019,33(2):278-289.[26]蒋宏.酿酒酵母和水生假丝酵母原生质体融合研究[D].成都:四川大学,2006.[27]王辅德,宛新杉,叶叙丰,等.草菇和银丝草菇的原生质体分离及菌丝再生[J].上海农业学报,1988(1):17-22.[28]马欣燕.印度块菌与赤芝原生质体融合研究[D].天津:天津师范大学,2016.[29]刘祖同,罗信昌.食用蕈菌生物技术及应用[M].北京:清华大学出版社,2002.[30]张卉,刘长江.姬松茸原生质体形成和再生的研究[J].微生物学杂志,2003(3):18-20,23.

Memo

Memo:
-
Last Update: 2020-02-27