摘要
近年来,灵芝产业高速发展,但其主要药效成分灵芝三萜的含量较低,是影响其品质的重要原因,因此探索一种提高灵芝三萜含量的生物技术手段极具应用前景。本研究以6株斜盖伞Clitopilus spp.真菌为材料分别制备多种类型的真菌诱导子,将其添加到灵芝菌丝体液体摇瓶中,研究对灵芝菌丝体生物量和灵芝三萜积累的影响。其中,Clitopilus sp.HSL-YX-7-A的高压灭菌发酵液(HSLFB)、C.hobsonii NL-19的菌丝干粉(NL-19DMP)、C.prunulus 84496的发酵液提取物(84496 FBE)和Clitopilus sp.HSL-YX-7-A的菌丝提取物(HSL ME)显著提高了灵芝菌丝体生物量和灵芝三萜含量,相对于对照组,生物量分别提高了450.09%、64.64%、46.97%和66.14%,灵芝三萜分别提高了53.01%、25.58%、25.17%和20.47%,灵芝三萜总产量则分别提高了585.15%、104.65%、86.43%和110.45%。进一步研究表明,经HSL FB、NL-19 DMP和HSL ME处理后,灵芝三萜生物合成途径关键酶基因hmgs、hmgr、mvd、fps、sqs和osc的表达量显著上调;但84496 FBE处理之后仅上调了hmgs和sqs的表达量。且6个基因的平均表达量与灵芝三萜总产量的提升率呈显著的正相关性(R^(2)=0.972,P<0.05)。本研究证实了斜盖伞诱导子能够有效提升灵芝菌丝体生物量和灵芝三萜含量,其中C.hobsonii NL-19和Clitopilus sp.HSL-YX-7-A均为栎类树种的根系共生菌。本研究进一步为开发斜盖伞真菌作为微生物菌剂提供了基础,同时为提高栎树人工林的生产力和创建“以菌养菌”的药用真菌品质提升技术体系提供了重要策略。
The industry of Ganoderma lingzhi is developing rapidly.The relative low content of ganoderma triterpenoids(GTs),the main bioactive component of G.lingzhi,however,poses a major challenge.Therefore,increasing GTs content by using a biotechnological approach holds great promises.In this work,various types of fungal elicitors from six strains of Clitopilus spp.were prepared to explore whether or how the elicitor treatments influence the mycelial biomass and GTs accumulation.Results showed that among these elicitors,the autoclaved fermentation broth of Clitopilus sp.HSL-YX-7-A(HSL FB),the dried mycelial powder of C.hobsonii NL-19(NL-19 DMP),the fermentation broth extract of C.prunulus 84496(84496 FBE)and the mycelium extract of Clitopilus sp.HSL-YX-7-A(HSL ME)significantly increased the mycelial biomass,GTs content and GTs yield compared with mycelial biomass,GTs content and GTs yield of the experimental control.The biomass increased by 450.09%,64.64%,46.97%and 66.14%,respectively;the GTs content increased by 53.01%,25.58%,25.17%and 20.47%,respectively;the GTs yield increased by 585.15%,104.65%,86.43%and 110.45%,respectively.Quantitative Real-time PCR showed that the expression levels of key enzyme genes hmgs,hmgr,mvd,fps,sqs and osc involved in the GTs biosynthesis were significantly up-regulated upon the treatments with HSL FB,NL-19 DMP and HSL ME.However,84496 FBE treatment only resulted in up-regulation of the expressions of hmgs and sqs.There was a significant positive correlation between the average expression level of the six genes and GTs production increase rate(R^(2)=0.972,P0.05).This study demonstrated that the fungal elicitors from Clitopilus spp.displayed the potentials on efficiently increasing the mycelial biomass and GTs content of G.lingzhi.Clitopilus hobsonii NL-19 and Clitopilus sp.HSL-YX-7-A are endophytic and symbiotic in Quercus root,and they are hopeful to be developed as new beneficial microbiological products.
作者
张宇薇
廖望仪
宋荣孝
彭龙
袁志林
ZHANG Yuwei;LIAO Wangyi;SONG Rongxiao;PENG Long;YUAN Zhilin(State Key Laboratory of Tree Genetics and Breeding,Chinese Academy of Forestry,Beijing 100091,China;College of Forestry,Nanjing Forestry University,Nanjing 210037,Jiangsu,China;Research Institute of Subtropical Forestry,Chinese Academy of Forestry,Hangzhou 311400,Zhejiang,China;Zhejiang Products Changle Chuangling Biological Technology Limited Company,Hangzhou 311400,Zhejiang,China)
出处
《菌物学报》
CAS
CSCD
北大核心
2023年第6期1370-1385,共16页
Mycosystema
基金
国家自然科学基金青年基金(31901290)
中央级公益性科研院所基本科研业务费专项资金(CAFYBB2020QB002)。
关键词
斜盖伞真菌
真菌诱导子
灵芝三萜
生物合成
诱导机制
Clitopilus
fungal elicitors
ganoderma triterpenoids
biosynthesis
inducing mechanisms
