美味牛肝菌菌塘中菌根促生菌的筛选与鉴定
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摘要
为了获得黑松Pinus thunbergii-美味牛肝菌Boletus edulis(Be)菌塘中的菌根促生菌(mycorrhiza helper bacteria,MHB),为美味牛肝菌的人工培育提供理论基础,通过比较野生牛肝菌菌塘细菌胞外代谢产物对菌丝生长的促进作用以及细菌+Be真菌双接处理黑松幼苗对其苗高、地径、茎根比、干重、根侵染率的影响,筛选出菌根促生菌。结果表明,菌株B2和K2代谢产物可以显著促进Be菌丝生长,盆栽实验结果表明双接种Be+B2、Be+K2的黑松苗,苗高分别增长了17.71%和16.82%,地径增长了41.65%和35.63%,干重提高了44.71%和48.78%,根侵染率提高了23.20%和21.50%,茎根比下降了46.41%和35.40%。可见菌株B2和K2是黑松-美味牛肝菌的菌根促生菌。通过细菌的形态、生化特性、16SrDNA序列测定,菌株B2和K2均鉴定为蜡样芽孢杆菌Bacillus cereus。蜡样芽孢杆菌有望进一步开发为美味牛肝菌的菌塘生物肥料。
Mycorrhiza helper bacteria(MHB) in rhizosphere soil of Pinus thunbergii and Boletus edulis(Be), were isolated, and their positive effect on growth of ectomycorrhizal fungus Be was investigated. The effect of Be and MHB co-inoculation on pine seedling height, ground diameter of stem, stem root ratio, seeding dry weight and root infection rate was also observed. Theresult revealed that the metabolites of bacterial strains B2 and K2 significantly promoted in vitro hyphal growth of Be. Co-inoculation of Be+B2 and Be+K2 in pot experiment showed that stem root ratio decreased by 46.41% and 35.40% respectively. However, seedling height, ground diameter, dry weight, root infection rate increased by 17.71% and 16.82%, 41.65% and 35.6%, 44.71% and 48.78%, and 23.2% and 21.5%, respectively. It was concluded that B2 and K2 can be considered as MHB strains. B2 and K2 were identified as Bacillus cereus based on morphological, physiological, and biochemical analyses in combination with analysis of 16 S rDNA gene sequences. Bacillus cereus is expected to be developed as biological fertilizer in colonized soil of Boletus edulis.
Mycorrhiza helper bacteria(MHB) in rhizosphere soil of Pinus thunbergii and Boletus edulis(Be), were isolated, and their positive effect on growth of ectomycorrhizal fungus Be was investigated. The effect of Be and MHB co-inoculation on pine seedling height, ground diameter of stem, stem root ratio, seeding dry weight and root infection rate was also observed. Theresult revealed that the metabolites of bacterial strains B2 and K2 significantly promoted in vitro hyphal growth of Be. Co-inoculation of Be+B2 and Be+K2 in pot experiment showed that stem root ratio decreased by 46.41% and 35.40% respectively. However, seedling height, ground diameter, dry weight, root infection rate increased by 17.71% and 16.82%, 41.65% and 35.6%, 44.71% and 48.78%, and 23.2% and 21.5%, respectively. It was concluded that B2 and K2 can be considered as MHB strains. B2 and K2 were identified as Bacillus cereus based on morphological, physiological, and biochemical analyses in combination with analysis of 16 S rDNA gene sequences. Bacillus cereus is expected to be developed as biological fertilizer in colonized soil of Boletus edulis.
引文
Artursson V,Jansson JK,2003.Use of bromodeoxyuridine immunocapture to identify active bacteria associated with arbuscular mycorrhizal hyphae.Applied Environment Microbiology,69(10):6208-6215
Bacon CW,Yates IE,Hinton DM,Meredith F,2001.Biological control of Fusarium moniliforme in maize.Environmental Health Perspectives,109(2):325-332
Chen JZ,Qin JS,Wang HC,Tan YZ,Dai X,2013.Study on the nutritional characteristics of Boletus edulis Bull.:Fr mycelium.Hubei Agricultural Sciences,52(3):607-609(in Chinese)
Dai YC,Yang ZL,2008.A revised checklist of medicinal fungi in China.Mycosystema,27(6):801-824(in Chinese)
Dai YC,Zhou LW,Yang ZL,Wen HA,Bau T,Li TH,2010.Arevised checklist of edible fungi in China.Mycosystema,29(1):1-21(in Chinese)
Fu SC,2008.Study on synthesis of ectomycorrhiza on Boletus edulis with a range of host seedling.Master Thesis,Hunan Forestry University,Changsha.1-53(in Chinese)
KozdróJ,Piotrowska-Seget Z,Krupa P,2007.Mycorrhizal fungi and ectomycorrhiza associated bacteria isolated from an industrial desert soil protect pine seedlings(II)impact.Ecotoxicology,16:449-456
Liu JG,Pei Y,Cong W,Ouyang F,2001.Studies on fermentation conditions of Bacillus cereus strain S1.Industrial Microbiology,31(1):4-7(in Chinese)
Ma HY,2013.Isolation,culture and identification of common edible fungi in Yunnan.Master Thesis,Yunnan Univesity,Kunming.1-58(in Chinese)
Mediavilla O,Olaizola J,Santos DB,Oria-De-Rueda JA,Mart?-N-Pinto P,2016.Mycorrhization between Cistus ladanifer L.and Boletus edulis Bull is enhanced by the mycorrhiza helper bacteria Pseudomonas fluorescens Migula.Mycorrhiza,26(2):161-168
Jiao WQ,2005.Screening of synergistic factors for Bacillus cereus A-47 and its synergistic mechanism.Master Thesis,China Agricultural University,Beijing.1-43(in Chinese)
Qin Y,Li QQ,Zheng YH,Guo X,2016.Bacterial diversity of shiroes soil growing three species of wild Boletus.Microbiology China,43(10):2148-2153(in Chinese)
Schrey SD,Schellhammer M,Ecke M,2005.Mycorrhiza helper bacterium Streptomyces Ac H 505 induces differential gene expression in the ectomycorrhizal fungus Amanita muscaria.New Phytologist,168(1):205-216
Sheng JM,Wu XQ,Hou LL,Ying CX,2010.Isolation and identification of a MHB strain from the rhizosphere soil of Pinus thunbergii inoculated with Boletus edulis.Chinese Journal of Applied and Environmental Biology,16(5):701-704(in Chinese)
Vivas A,Azcon R,Biro B,Barea JM,Ruizlozano JM,2005.Influence of bacterial strains isolated from lead-polluted soil and their interactions with arbuscular mycorrhizae on the growth of Trifolium pratense L.under lead toxicity.Canadian Journal of Microbiology,49(10):577-588
Vivas A,Barea JM,Biro B,Azcon R,2006.Effectiveness of autochthonous bacterium and mycorrhizal fungus on Trifolium growth,symbiotic development and soil enzymatic activities in Zn contaminated soil.Journal of Applied Microbiology,100(3):587-598
Vivas A,Marulanda A,Ruiz-Lozano JM,Barea JM,Azcón R,2003.Influence of a Bacillus sp.on physiological activities of arbuscular mycorrhizal fungi and on plant responses to PEG-induced drought stress.Mycorrhiza,13(5):249-256
Wang PF,2012.Optimization of culture medium for Boletus edulis mycelium and its co-culture with Cyclobalanopsis glauca.Master Thesis,Yunnan University,Kunming.1-56(in Chinese)
Wu XQ,Sun MQ,2006.Mycorrhizal formation between seven ectomycorrhizal fungi and seedlings of three pine species.Acta Ecologica Sinica,26(12):4186-4191(in Chinese)
Xu M,Fu WQ,Dai CC,Jia Y,2018.Ecological function of promoting microorganisms associated with ectomycorrhizal fungi.Chinese Journal of Ecology,37(4):1246-1256(in Chinese)
陈今朝,秦家顺,王慧超,谭永忠,戴玄,2013.美味牛肝菌菌丝体营养特性分析.湖北农业科学,52(3):607-609
戴玉成,杨祝良,2008.中国药用真菌名录及部分名称的修订.菌物学报,27(6):801-824
戴玉成,周丽伟,杨祝良,文华安,图力古尔,李泰辉,2010.中国食用菌名录.菌物学报,29(1):1-21
付绍春,2008.美味牛肝菌与不同宿主植物幼苗菌根合成技术研究.湖南林业大学农业推广硕士论文,长沙.1-53
焦文沁,2005.蜡样芽孢杆菌A-47增效因子的筛选及增效机制初探.中国农业大学硕士论文,北京.1-43
刘建国,裴炎,丛威,欧阳藩,2001.蜡状芽孢杆菌S1发酵条件的研究.工业微生物,31(1):4-7
马红英,2013.云南常见食用菌分离、培养及鉴定.云南大学硕士论文,昆明.1-58
秦余,李茜茜,郑雨焕,郭霞,2016.3种野生牛肝菌的菌塘土壤细菌多样性.微生物学通报,43(10):2148-2153
盛江梅,吴小芹,侯亮亮,应晨希,2010.一株黑松-美味牛肝菌菌根辅助细菌的筛选及鉴定.应用与环境生物学报,16(5):701-704
王鹏飞,2012.一株广义美味牛肝菌菌丝体的培养基优化及与滇青冈共培养的初步研究.云南大学硕士论文,昆明.1-56
吴小芹,孙民琴,2006.七株外生菌根真菌与三种松苗菌根的形成能力.生态学报,26(1):4186-4191
徐漫,傅婉秋,戴传超,贾永,2018.外生菌根真菌促生微生物生态功能研究进展,生态学杂志,37(4):1246-1256
Bacon CW,Yates IE,Hinton DM,Meredith F,2001.Biological control of Fusarium moniliforme in maize.Environmental Health Perspectives,109(2):325-332
Chen JZ,Qin JS,Wang HC,Tan YZ,Dai X,2013.Study on the nutritional characteristics of Boletus edulis Bull.:Fr mycelium.Hubei Agricultural Sciences,52(3):607-609(in Chinese)
Dai YC,Yang ZL,2008.A revised checklist of medicinal fungi in China.Mycosystema,27(6):801-824(in Chinese)
Dai YC,Zhou LW,Yang ZL,Wen HA,Bau T,Li TH,2010.Arevised checklist of edible fungi in China.Mycosystema,29(1):1-21(in Chinese)
Fu SC,2008.Study on synthesis of ectomycorrhiza on Boletus edulis with a range of host seedling.Master Thesis,Hunan Forestry University,Changsha.1-53(in Chinese)
KozdróJ,Piotrowska-Seget Z,Krupa P,2007.Mycorrhizal fungi and ectomycorrhiza associated bacteria isolated from an industrial desert soil protect pine seedlings(II)impact.Ecotoxicology,16:449-456
Liu JG,Pei Y,Cong W,Ouyang F,2001.Studies on fermentation conditions of Bacillus cereus strain S1.Industrial Microbiology,31(1):4-7(in Chinese)
Ma HY,2013.Isolation,culture and identification of common edible fungi in Yunnan.Master Thesis,Yunnan Univesity,Kunming.1-58(in Chinese)
Mediavilla O,Olaizola J,Santos DB,Oria-De-Rueda JA,Mart?-N-Pinto P,2016.Mycorrhization between Cistus ladanifer L.and Boletus edulis Bull is enhanced by the mycorrhiza helper bacteria Pseudomonas fluorescens Migula.Mycorrhiza,26(2):161-168
Jiao WQ,2005.Screening of synergistic factors for Bacillus cereus A-47 and its synergistic mechanism.Master Thesis,China Agricultural University,Beijing.1-43(in Chinese)
Qin Y,Li QQ,Zheng YH,Guo X,2016.Bacterial diversity of shiroes soil growing three species of wild Boletus.Microbiology China,43(10):2148-2153(in Chinese)
Schrey SD,Schellhammer M,Ecke M,2005.Mycorrhiza helper bacterium Streptomyces Ac H 505 induces differential gene expression in the ectomycorrhizal fungus Amanita muscaria.New Phytologist,168(1):205-216
Sheng JM,Wu XQ,Hou LL,Ying CX,2010.Isolation and identification of a MHB strain from the rhizosphere soil of Pinus thunbergii inoculated with Boletus edulis.Chinese Journal of Applied and Environmental Biology,16(5):701-704(in Chinese)
Vivas A,Azcon R,Biro B,Barea JM,Ruizlozano JM,2005.Influence of bacterial strains isolated from lead-polluted soil and their interactions with arbuscular mycorrhizae on the growth of Trifolium pratense L.under lead toxicity.Canadian Journal of Microbiology,49(10):577-588
Vivas A,Barea JM,Biro B,Azcon R,2006.Effectiveness of autochthonous bacterium and mycorrhizal fungus on Trifolium growth,symbiotic development and soil enzymatic activities in Zn contaminated soil.Journal of Applied Microbiology,100(3):587-598
Vivas A,Marulanda A,Ruiz-Lozano JM,Barea JM,Azcón R,2003.Influence of a Bacillus sp.on physiological activities of arbuscular mycorrhizal fungi and on plant responses to PEG-induced drought stress.Mycorrhiza,13(5):249-256
Wang PF,2012.Optimization of culture medium for Boletus edulis mycelium and its co-culture with Cyclobalanopsis glauca.Master Thesis,Yunnan University,Kunming.1-56(in Chinese)
Wu XQ,Sun MQ,2006.Mycorrhizal formation between seven ectomycorrhizal fungi and seedlings of three pine species.Acta Ecologica Sinica,26(12):4186-4191(in Chinese)
Xu M,Fu WQ,Dai CC,Jia Y,2018.Ecological function of promoting microorganisms associated with ectomycorrhizal fungi.Chinese Journal of Ecology,37(4):1246-1256(in Chinese)
陈今朝,秦家顺,王慧超,谭永忠,戴玄,2013.美味牛肝菌菌丝体营养特性分析.湖北农业科学,52(3):607-609
戴玉成,杨祝良,2008.中国药用真菌名录及部分名称的修订.菌物学报,27(6):801-824
戴玉成,周丽伟,杨祝良,文华安,图力古尔,李泰辉,2010.中国食用菌名录.菌物学报,29(1):1-21
付绍春,2008.美味牛肝菌与不同宿主植物幼苗菌根合成技术研究.湖南林业大学农业推广硕士论文,长沙.1-53
焦文沁,2005.蜡样芽孢杆菌A-47增效因子的筛选及增效机制初探.中国农业大学硕士论文,北京.1-43
刘建国,裴炎,丛威,欧阳藩,2001.蜡状芽孢杆菌S1发酵条件的研究.工业微生物,31(1):4-7
马红英,2013.云南常见食用菌分离、培养及鉴定.云南大学硕士论文,昆明.1-58
秦余,李茜茜,郑雨焕,郭霞,2016.3种野生牛肝菌的菌塘土壤细菌多样性.微生物学通报,43(10):2148-2153
盛江梅,吴小芹,侯亮亮,应晨希,2010.一株黑松-美味牛肝菌菌根辅助细菌的筛选及鉴定.应用与环境生物学报,16(5):701-704
王鹏飞,2012.一株广义美味牛肝菌菌丝体的培养基优化及与滇青冈共培养的初步研究.云南大学硕士论文,昆明.1-56
吴小芹,孙民琴,2006.七株外生菌根真菌与三种松苗菌根的形成能力.生态学报,26(1):4186-4191
徐漫,傅婉秋,戴传超,贾永,2018.外生菌根真菌促生微生物生态功能研究进展,生态学杂志,37(4):1246-1256