苍术根际橘绿木霉的生防功能及培养条件的探索  

Biocontrol Effect and Culture Condition Optimization of Trichoderma citrinoviride Strain Isolated from Rhizosphere of Atractylodes lancea

作　　者：李阔 王升[2,3] 王红阳 彭政 郭秀芝 王铁霖 王月枫[2,3] 郭兰萍 LI Kuo;WANG Sheng;WANG Hongyang;PENG Zheng;GUO Xiuzhi;WANG Tielin;WANG Yuefeng;GUO Lanping(School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,China;State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Key Laboratory of Biology and Cultivation of Herb Medicine,Ministry of Agriculture and Rural Affairs,Beijing 100700,China)

机构地区：[1]广东药科大学中药学院,广州510006 [2]中国中医科学院中药资源中心,道地药材品质保障与资源持续利用全国重点实验室,北京100700 [3]农业农村部中药材生物学与栽培重点实验室,北京100700

出　　处：《中国实验方剂学杂志》2024年第15期192-199,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家重点研发计划项目(2023YFC3503801);中国中医科学院科技创新工程项目(CI2021A03905,CI2023E002);国家中医药管理局中医药创新团队及人才支持计划-道地药材生态化与资源可持续利用项目(ZYYCXTD-D-202005);全国重点实验室中国博士后科学基金项目(2022T150730)。

摘　　要：目的:苍术根腐病是苍术种植生产上最为严重的病害之一。木霉菌是有效防治植物根腐病且对环境污染小的生防菌。该研究旨在从苍术根际中分离和研究具有根腐病防治功能的木霉菌,用于解决苍术种植生产过程面临的病害防治问题。方法:稀释涂布法和内转录间隔区(ITS)测序分离并鉴定木霉T2204;对峙培养实验和对扣培养实验验证T2204及其挥发性物质对两株病原菌的抑制活性;通过盆栽实验和气相色谱-质谱法(GC-MS)探究T2204对苍术根腐病的生防潜力及对苍术根茎4种主要挥发油成分的含量积累的影响;通过不同培养基、光照、温度、pH、碳氮源条件培养探究T2204适宜培养条件。结果:分离得到对2株强致病力苍术根腐病病原真菌具有直接抑制能力的一株木霉T2204,鉴定为橘绿木霉(Trichoderma citrinoviride);对峙培养实验显示T2204对2株镰刀菌的抑菌率分别达到77.90%和76.80%;对扣培养实验显示抑菌率分别达57.11%和81.11%。盆栽实验证明,T2204不同处理后苍术幼苗受根腐病侵染后成活率达到100%,而不接种T2204的实验组仅为50%;生长150 d后,接种T2204的苍术植株根茎干重比无处理对照(CK)组提升32%(P<0.05),同时根茎所含的苍术素浓度比CK组提升11%;适宜培养条件筛选结果:T2204适宜生长条件为培养基(PDA和PSA)、光照(24 h连续光照和光暗交替12 h/12 h)、温度(25~30℃)、pH 5~6、碳源(葡萄糖、D-果糖、可溶性淀粉和麦芽糖)、氮源(磷酸二氢铵和硫酸铵);适宜产孢条件为培养基(PSA和CMA)、光照(光暗交替12 h/12 h)、温度(20~30℃)、pH 8、碳源(蔗糖)、氮源(硝酸钠)。结论:T2204可以提升对苍术根腐病的抵御力,并促进苍术的生长及有效成分的积累,为了后续开展大规模的应用和产业化研究奠定基础。Objective:Root rot is one of the most serious diseases in the cultivation and production of Atractylodes lancea.Trichoderma spp.are effective in the biocontrol of root rot without causing environmental pollution.This study aims to isolate and study a Trichoderma strain capable preventing and controlling root rot from the rhizosphere of A.lancea and to solve the problem of disease prevention and control in the planting and production of A.lancea.Method:Trichoderma T2204 was isolated by the dilution-coating method and identified by ITS sequencing.The inhibitory activities of T2204 and its volatiles against two pathogenic fungal strains were examined by dual-culture and co-culture experiments.The biocontrol potential of T2204 on root rot of A.lancea and the effect of T2204 on the accumulation of medicinal compounds in the rhizosphere of A.lancea were investigated by pot experiments and GC-MS,respectively.In addition,the optimal medium,photoperiod,temperature,pH,and carbon and nitrogen sources for the culture of T2204 were explored.Result:The Trichoderma isolate T2204 was identified as T.citrinoviride and had direct inhibitory effects on two highly pathogenic strains causing root rot.In the dual-culture experiments with the two pathogenic strains,T2204 showcased the inhibition rates of 77.90%and 76.80%,respectively.In the co-culture experiments with the two pathogenic strains,the volatile organic compounds produced by T2204 showed the inhibition rates of 57.11%and 81.11%,respectively.The pot experiments showed that the survival rate of A.lancea seedlings infected by root rot reached 100%after inoculation with T2204 and was only 50%in the case without inoculation of T2204.After 150 days of cultivation,the dry weight and atractylodin content of the rhizome of A.lancea plants treated with T2204 increased by 32%(P<0.05)and 11%,respectively,compared with the untreated group.The optimal conditions for the growth of T2204 were PDA or PSA medium,photoperiod of 12 h dark/12 h light,25-30°C,pH 5-6,carbon sources of glucose,D-

关 键 词：苍术 橘绿木霉 根腐病 生物防治 促生 促进次生代谢 

分 类 号：R284.2[医药卫生—中药学] R285[医药卫生—中医学] R289R287R22R2-031R33R24