人参主要病原菌生防真菌的筛选及鉴定
|
摘要
【目的】筛选对人参主要病原菌具高效、广谱抑菌作用的生防真菌,丰富人参生防菌菌种资源。【方法】采用稀释平板法,从吉林省2处采样地的人参健株根际土壤中分离真菌;采用平板对峙培养法,通过初筛和复筛,筛选对人参链格孢菌(Alternaria panax)、立枯丝核菌(Rhizoctonia solani)、腐皮镰孢菌(Fusarium solani)、毁灭柱孢菌(Cylindrocarpon destructans)、恶疫霉菌(Phytophthora cactorum)、人参核盘菌(Sclerotinia schinseng)和灰葡萄孢菌(Botrytis cinerea)具有高效广谱抑菌活性的生防真菌;利用传统形态学和rDNA-ITS分子分析技术对分离到的生防真菌进行鉴定。【结果】从人参健株根际土壤中共分离得到400株真菌,其中有15株生防真菌对人参7种主要病原菌具有广谱抑菌作用,特别是对立枯丝核菌有强烈的抑制作用(抑菌率均大于50%),且FSR-106的抑菌作用最强(抑菌率87.41%,下同);FSR-121对人参链格孢菌的抑菌作用最强(60.00%);FSR-74对腐皮镰孢菌的抑菌率高达67.48%;FSR-46对毁灭柱孢菌和灰葡萄孢菌的抑菌率最大(66.67%和68.97%);FSR-97对恶疫霉菌及人参核盘菌的抑制作用最强(73.33%和66.67%)。通过形态学和rDNA-ITS序列分析鉴定发现,15株生防真菌中有8株木霉属(Trichoderma)、3株青霉属(Penicillium)、1株曲霉属(Aspergillus)、1株附球菌属(Epicoccum)、1株毛壳菌属(Chaetomium)和1株轮层炭壳属(Daldinia),其中FSR-38为黑附球菌(Epicoccum nigrum),FSR-10、FSR-43和FSR-106为深绿木霉(T.atroviride),FSR-46为长枝木霉(T.longibrachiatum),FSR-55为钩状木霉(T.hamatum),FSR-72为桔绿木霉(T.citrinoviride),FSR-74为球毛壳菌(Chaetomium globosum),FSR-97为多孢木霉(Trichoderma polysporum),FSR-91为蔡氏轮层炭壳菌(Daldinia childiae),FSR-25、FSR-67和FSR-121为青霉属(Penicilliumsp.)。【结论】从人参健株根际土壤中可快速有效地筛选获得大量广谱性生防真菌;本研究所得的15株生防真菌对7种人参主要病害防治具有较高的潜在应用价值。
【Objective】This study screened effective and broad-spectrum biocontrol fungi against main pathogens of Panax ginsengto enrich biocontrol microorganism resources.【Method】Soil fungi were isolated from rhizosphere soil of healthy Panax ginsengby dilution plate method.Biocontrol strains,with highly antibacterial activity against 7ginseng pathogens including Alternaria panax,Rhizoctonia solani,Fusarium solani,Cylindrocarpon destructans,Phytophthora cactorum,Sclerotinia schinsengand Botrytis cinerea were screened using pairing culture method.Biocontrol fungi were also identified based on morphology and rDNA-ITS sequence.【Result】A total of 400 fungi were isolated fromP.gisengsoil,among which,15 had broad spectrum antimicrobial effect on 7 P.ginsengpathogens.Especially,Rhizoctonia solani were strongly inhibited by all of them with the inhibition rate of>50%,and FSR-106 was the strongest with the inhibition rate of 87.41%.FSR-121 was the strongest against Alternaria panax with the inhibition rate of60.00%,and FSR-74 was the strongest against Fusarium solani with the inhibition rate of 67.48%.Inhibition rates of FSR-46 against Cylindrocarpon destructans(66.67%)and Botrytis cinerea(68.97%)were the highest.FSR-97 was the strongest against Phytophthora cactorum and Sclerotinia schinseng with inhibition rates of 73.33% and 66.67%,respectively.Morphological and rDNA-ITS sequence analysis showed that the 15 biocontrol fungi could be divided into 6genus,i.e.8belonged to Trichoderma,3belonged to Penicillium,one belonged to Aspergillus,one belonged to Epicoccum,one belonged to Chaetomium and one belonged to Daldinia.FSR-38 was identified as Epicoccum nigrum,FSR-10,FSR-43 and FSR-106 were T.atroviride,FSR-46 was T.longibrachiatum,FSR-55 was T.hamatum,FSR-72 was T.citrinoviride,FSR-74 was Chaetomium globosum,FSR-97 was Trichoderma polysporum,FSR-91 was Daldinia childiae,while FSR-25,FSR-67 and FSR-121 were Penicilliumsp..【Conclusion】The screening method based on rhizosphere soil of healthy gineng is a rapid and efficient way to seek for strong potential biocontrol strains.The 15 biocontrol fungi isolated in this study have good potential for the control of P.ginseng pathogens.
【Objective】This study screened effective and broad-spectrum biocontrol fungi against main pathogens of Panax ginsengto enrich biocontrol microorganism resources.【Method】Soil fungi were isolated from rhizosphere soil of healthy Panax ginsengby dilution plate method.Biocontrol strains,with highly antibacterial activity against 7ginseng pathogens including Alternaria panax,Rhizoctonia solani,Fusarium solani,Cylindrocarpon destructans,Phytophthora cactorum,Sclerotinia schinsengand Botrytis cinerea were screened using pairing culture method.Biocontrol fungi were also identified based on morphology and rDNA-ITS sequence.【Result】A total of 400 fungi were isolated fromP.gisengsoil,among which,15 had broad spectrum antimicrobial effect on 7 P.ginsengpathogens.Especially,Rhizoctonia solani were strongly inhibited by all of them with the inhibition rate of>50%,and FSR-106 was the strongest with the inhibition rate of 87.41%.FSR-121 was the strongest against Alternaria panax with the inhibition rate of60.00%,and FSR-74 was the strongest against Fusarium solani with the inhibition rate of 67.48%.Inhibition rates of FSR-46 against Cylindrocarpon destructans(66.67%)and Botrytis cinerea(68.97%)were the highest.FSR-97 was the strongest against Phytophthora cactorum and Sclerotinia schinseng with inhibition rates of 73.33% and 66.67%,respectively.Morphological and rDNA-ITS sequence analysis showed that the 15 biocontrol fungi could be divided into 6genus,i.e.8belonged to Trichoderma,3belonged to Penicillium,one belonged to Aspergillus,one belonged to Epicoccum,one belonged to Chaetomium and one belonged to Daldinia.FSR-38 was identified as Epicoccum nigrum,FSR-10,FSR-43 and FSR-106 were T.atroviride,FSR-46 was T.longibrachiatum,FSR-55 was T.hamatum,FSR-72 was T.citrinoviride,FSR-74 was Chaetomium globosum,FSR-97 was Trichoderma polysporum,FSR-91 was Daldinia childiae,while FSR-25,FSR-67 and FSR-121 were Penicilliumsp..【Conclusion】The screening method based on rhizosphere soil of healthy gineng is a rapid and efficient way to seek for strong potential biocontrol strains.The 15 biocontrol fungi isolated in this study have good potential for the control of P.ginseng pathogens.
引文
[1] 杨利民,陈长宝,王秀全,等.长白山区参后地生态恢复与再利用模式及其存在的问题[J].吉林农业大学学报,2004,26(5):546-549,553.Yang L M,Chen C B,Wang X Q,et al.Ecological restoration and reused modes of old ginseng land in the Changbai mountainous area and its existing problems[J].Journal of Jilin Agricultural University,2004,26(5):546-549,553.
[2] 吴连举,赵亚会,关一鸣,等.人参连作障碍原因及其防治途径研究进展[J].特产研究,2008(2):68-72.Wu L J,Zhao Y H,Guan Y M,et al.Review on studies of the reason and control methods of succession cropping obstacle of Panax ginseng C.A.Mey[J].Special Wild Economic Animal and Plant Research,2008(2):68-72.
[3] 刘亚南,赵东岳,刘 敏,等.人参病虫害发生及农药施用现状调查[J].中国农学通报,2014,30(10):294-298.Liu Y N,Zhao D Y,Liu M,et al.Investigation of pests and diseases occurrence and pesticides application in main producing areas of Panax ginseng[J].Chinese Agricultural Science Bulletin,2014,30(10):294-298.
[4] 王 悦,林红梅,侯志广,等.人参中11种农药残留振荡提取气相色谱法测定[J].农药,2013,52(7):509-511.Wang Y,Lin H M,Hou Z G,et al.Determination of 11pesticide residues in radix ginseng by solid-phase extraction and gas chromatography[J].Agrochemicals,2013,52(7):509-511.
[5] 章强华,董 国,姚晗珺.人参常用农药国外残留限量标准的分析及对策[J].人参研究,2009(1):2-10.Zhang Q H,Dong G,Yao H J.Analysis and countermeasures on residue limits standards of ginseng common pesticide abroad[J].Ginseng Research,2009(1):2-10.
[6] 冯 家.人参常见病虫害诊断与安全用药[M].长春:吉林出版集团,2010:15-20.Feng J.Diagnosis of ginseng common pests and diseases and safe medication[M].Changchun:Jilin Publishing Group,2010:15-20.
[7] 胡 霞,苑艳辉,姚卫容,等.微生物农药发展概况[J].农药,2005,44(2):31-32.Hu X,Yuan Y H,Yao W R,et al.Status of microbial pesticide development[J].Agrochemicals,2005,44(2):31-32.
[8] 杨 峻,林荣华,袁善奎,等.我国生物源农药产业现状调研及分析[J].中国生物防治,2014,30(4):441-445.Yang J,Lin R H,Yuan S K,et al.Survey and analysis of current situation of biologically derived pesticides industry in China[J].Chinese Journal of Biological Control,2014,30(4):441-445.
[9] 刘振华,罗远婵,张道敬,等.农用微生物杀菌剂剂型研究进展[J].农药学学报,2014,16(5):497-507.Liu Z H,Luo Y C,Zhang D J,et al.Research progress and prospects of microbial pesticide formulation for plant desease control[J].Chinese Journal of Pesticide Science,2014,16(5):497-507.
[10] 王 芳,图力古尔.土壤真菌多样性研究进展[J].菌物研究,2014,12(3):178-186.Wang F,Bau Tolgor.Research advance in the diversity of soil fungi[J].Journal of Fungal Research,2014,12(3):178-186.
[11] 王秉丽,李广纪,郭玉人,等.不同野生木霉菌拮抗作用的比较[J].中国生物防治,2012,28(1):147-151.Wang B L,Li G J,Guo Y R,et al.Comparison of antagonistic effects of four Trichodermastrains[J].Chinese Journal of Biological Control,2012,28(1):147-151.
[12] 李 芳,史 怀,刘 波,等.淡紫拟青霉对尖孢镰刀菌的拮抗作用与机制分析[J].植物保护学报,2005,32(4):373-378.Li F,Shi H,Liu B,et al.Antagonistic effect of biocontrol fungus,Paecilomyces lilacinus strain NH-PL-3and its mechanism against Fusarium oxysporum[J].Acta Phytophylacica Sinica,2005,32(4):373-378.
[13] 刘永亮.辣椒疫霉拮抗真菌毛壳菌和青霉菌的分离鉴定及其生防潜力的研究[D].济南:山东农业大学,2013.Liu Y L.Isolation and identification of fungal antagonist Chaetomium and Penicillum against Phytophthora capsici and studies on their potential of biocontrol[D].Ji'nan:Shandong Agricultural University,2013.
[14] 强 薇,郭 帅,张爱华,等.人参根际土壤中拮抗人参病害微生物的筛选[J].华南农业大学学报,2014,35(5):76-81.Qiang W,Guo S,Zhang A H,et al.Screening antagonistic microorganism against ginseng diseases fromPanax ginsengrhizosphere soil[J].Journal of South China Agricultural University,2014,35(5):76-81.
[15] Ryu H,Park H,Suh D S,et al.Biological control of Colletotrichum panacicola on Panax ginseng by Bacillus subtilis HK-CSM-1[J].J Ginseng Res,2014,38:215-219.
[16] Song M,Yun H Y,Kim Y H.Antagonistic Bacillus species as a biological control of ginseng root rot caused by Fusariumcf.incarnatum[J].J Ginseng Res,2014,38:136-145.
[17] 赵阿娜,丁万隆.木霉菌剂对人参根部病害防治效果评价[J].中草药,2006(10):1552-1554.Zhao A N,Ding W L.Estimation of control effect of Trichodermafungicides on Panax ginsengroot disease[J].Chinese Traditional and Herbal Drugs,2006(10):1552-1554.
[18] 周如军,傅俊范,史会岩,等.人参锈腐病拮抗生防菌的筛选与鉴定[J].沈阳农业大学学报,2010,41(4):422-426.Zhou R J,Fu J F,Shi H Y,et al.Screening and identification of antagonistic strains against Cylindrocarpon destructans[J].Journal of Shenyang Agricultural University,2010,41(4):422-426.
[19] 杨 骁,李长田.人参内生生防真菌的筛选与鉴定[J].东北师范大学学报(自然科学版),2013,45(4):107-113.Yang X,Li C T.Screening and identification of endogeny biocontrol fungi of Panax ginseng[J].Journal of Northeast Normal University(Natural Science Edition),2013,45(4):107-113.
[20] 魏景超.真菌鉴定手册[M].上海:上海科学技术出版社,1979.Wei J C.Fungal identification manual[M].Shanghai:Shanghai Scientific Technology Press,1979.
[21] 陈凤毛.真菌ITS区序列结构及其应用[J].林业科技开发,2007,21(2):5-7.Chen F M.The sequence of ITS region and its application of fungus[J].China Forestry Science and Technology,2007,21(2):5-7.
[22] 张广志,杨合同,文成敬.木霉菌形态学分类检索与分子生物学鉴定[J].山东农业大学学报(自然科学版),2011,42(2):309-316.Zhang G Z,Yang H T,Wen C J.The key to the morphological classification and the molecular identification of Trichoderma spp.[J].Journal of Shandong Agricultural University(Natural Science Edition),2011,42(2):309-316.
[23] 齐祖同.中国真菌志:第五卷:曲霉属及其相关有性型[M].北京:科学出版社,1997.Qi Z T.Chinese fungi:the fifth volume:Aspergillus and its related sexual type[M].Beijing:Science Press,1997.
[24] 孔华忠.中国真菌志:第三十五卷:青霉属及其相关有性型[M].北京:科学出版社,2007.Kong H Z.Chinese fungi:the thirty fifth volume:Penicillium and its related sexual type[M].Beijing:Science Press,2007.
[25] 白金铠.中国真菌志:第十五卷:球壳孢目,茎点霉属,叶点霉属[M].北京:科学出版社,2003.Bai J K.Chinese fungi:the fifteenth volume:Sphaeropsidales,Phornaand Phyllosticta[M].Beijing:Science Press,2003.
[26] 吴连举,关一鸣,逄世峰,等.利用拮抗微生物防治人参、西洋参土传病害研究进展[J].安徽农业科学,2010,38(28):15630-15631,15657.Wu L J,Guan Y M,Pang S F,et al.Research progress on biological control of soil-borne diseases in ginseng and American ginssseng by anti-micoorganism[J].Journal of Anhui Agri Sci,2010,38(28):15630-15631,15657.
[27] 任守让,王瑞霞.对人参锈腐病菌的拮抗真菌及其防病效果[J].吉林农业大学学报,1998,20(S1):120.Ren S R,Wang R X.Controlling effects of antagonistic fungi to Cylindrocarpon destructans in ginseng[J].Journal of Jilin Agricultural University,1998,20(S1):120.
[28] 邢云章,马凤茹.绿色木霉防治人参根腐病的研究[J].特产科学实验,1983(4):16-18.Xing Y Z,Ma F R.Study of Trichoderma viride to control Fusarium solani[J].Specialty Experiment,1983(4):16-18.
[29] 赵阿娜,丁万隆,朱殿龙.拮抗人参根部病原菌木霉的筛选及其生物学特性初步研究[J].中国中药杂志,2006,31(20):1671-1674.Zhao A N,Ding W L,Zhu D L.Screening strains for Trichoderma spp.for strong antagonism against ginseng root pathogens and study on their biological characters[J].China Journal of Chinese Materia Medica,2006,31(20):1671-1674.
[30] 张 瑾,张树武,徐秉良,等.长枝木霉菌抑菌谱测定及其抑菌作用机理研究[J].中国生态农业学报,2014,22(6):661-667.Zhang J,Zhang S W,Xu B L,et al.Determining antifungal spectrum and mechanism of Trichoderma longibrachiatum in vitro[J].Chinese Journal of Eco-Agriculture,2014,22(6):661-667.
[31] 胡明江,张秀省,曹 兴,等.康宁木霉SMF2防治大白菜软腐病研究[J].北方园艺,2009(6):102-103.Hu M J,Zhang X S,Cao X,et al.Studies on the control effect of Trichoderma kningii Oud.against soft rot of Chinese cabbage[J].Northern Horticulture,2009(6):102-103.
[32] 梁巧兰,王 芳,魏列新,等.深绿木霉T2菌株对百合疫霉拮抗作用及机制[J].植物保护,2011,37(6):164-167.Liang Q L,Wang F,Wei L X,et al.Antagonistic mechanism of Trichoderma atroviride T2strain against Phytophthora nicotianae[J].Plant Protection,2011,37(6):164-167.
[33] 刘晓芸.对致病疫霉具拮抗作用的真菌XF的鉴定及抑菌活性研究[D].河北保定:河北农业大学,2007.Liu X Y.Identification and antifungal activity of XF,an antagonistic fungus against Phytophthora infestans[D].Hebei Baoding:Hebei Agricultural University,2007.
[34] 王艳红,吴晓民,朱艳萍,等.温郁金内生真菌Chaetomium globosum L18对植物病原菌的抑菌谱及拮抗机理[J].生态学报,2012,32(7):2040-2046.Wang Y H,Wu X M,Zhu Y P,et al.Inhibition effects and mechanisms of the endophytic fungus Chaetomium globosum L18from Curcuma wenyujin[J].Acata Ecologica Sinica,2012,32(7):2040-2046.
[35] 马 艳,常志州,赵江涛,等.一株疫病拮抗青霉P.st10菌株的抗菌活性及其对辣椒疫病的盆栽防效[J].中国生物防治,2006,22(3):239-243.Ma Y,Chang Z Z,Zhao J T,et al.Antifungal activity of Penicillium striatisporum strain P.st10and its inhibition against pepper blight caused by Phytophthoraspp.[J].Chinese Journal of Biological Control,2006,22(3):239-243.
[36] 窦红涛,李若瑜,万 哲,等.临床常见镰刀菌rDNA ITS序列分析[J].临床和实验医学杂志,2006,5(6):646-648.Dou H T,Li R Y,Wan Z,et al.Sequence analysis of ribosomal DNA ITS of clinical comm on Fusariumspecies[J].Journal of Clinical and Experimental Medicine,2006,5(6):646-648.
[37] 隋 心,冯富娟,娄 鑫,等.土壤微生物总DNA的提取以及土壤真菌ITS-PCR体系的建立[J].中国酿造,2011(9):166-169.Sui X,Feng F J,Lou X,et al.Total DNA from soil microorganisms and construction of ITS-PCR system for soil fungi[J].China Brewing,2011(9):166-169.
[2] 吴连举,赵亚会,关一鸣,等.人参连作障碍原因及其防治途径研究进展[J].特产研究,2008(2):68-72.Wu L J,Zhao Y H,Guan Y M,et al.Review on studies of the reason and control methods of succession cropping obstacle of Panax ginseng C.A.Mey[J].Special Wild Economic Animal and Plant Research,2008(2):68-72.
[3] 刘亚南,赵东岳,刘 敏,等.人参病虫害发生及农药施用现状调查[J].中国农学通报,2014,30(10):294-298.Liu Y N,Zhao D Y,Liu M,et al.Investigation of pests and diseases occurrence and pesticides application in main producing areas of Panax ginseng[J].Chinese Agricultural Science Bulletin,2014,30(10):294-298.
[4] 王 悦,林红梅,侯志广,等.人参中11种农药残留振荡提取气相色谱法测定[J].农药,2013,52(7):509-511.Wang Y,Lin H M,Hou Z G,et al.Determination of 11pesticide residues in radix ginseng by solid-phase extraction and gas chromatography[J].Agrochemicals,2013,52(7):509-511.
[5] 章强华,董 国,姚晗珺.人参常用农药国外残留限量标准的分析及对策[J].人参研究,2009(1):2-10.Zhang Q H,Dong G,Yao H J.Analysis and countermeasures on residue limits standards of ginseng common pesticide abroad[J].Ginseng Research,2009(1):2-10.
[6] 冯 家.人参常见病虫害诊断与安全用药[M].长春:吉林出版集团,2010:15-20.Feng J.Diagnosis of ginseng common pests and diseases and safe medication[M].Changchun:Jilin Publishing Group,2010:15-20.
[7] 胡 霞,苑艳辉,姚卫容,等.微生物农药发展概况[J].农药,2005,44(2):31-32.Hu X,Yuan Y H,Yao W R,et al.Status of microbial pesticide development[J].Agrochemicals,2005,44(2):31-32.
[8] 杨 峻,林荣华,袁善奎,等.我国生物源农药产业现状调研及分析[J].中国生物防治,2014,30(4):441-445.Yang J,Lin R H,Yuan S K,et al.Survey and analysis of current situation of biologically derived pesticides industry in China[J].Chinese Journal of Biological Control,2014,30(4):441-445.
[9] 刘振华,罗远婵,张道敬,等.农用微生物杀菌剂剂型研究进展[J].农药学学报,2014,16(5):497-507.Liu Z H,Luo Y C,Zhang D J,et al.Research progress and prospects of microbial pesticide formulation for plant desease control[J].Chinese Journal of Pesticide Science,2014,16(5):497-507.
[10] 王 芳,图力古尔.土壤真菌多样性研究进展[J].菌物研究,2014,12(3):178-186.Wang F,Bau Tolgor.Research advance in the diversity of soil fungi[J].Journal of Fungal Research,2014,12(3):178-186.
[11] 王秉丽,李广纪,郭玉人,等.不同野生木霉菌拮抗作用的比较[J].中国生物防治,2012,28(1):147-151.Wang B L,Li G J,Guo Y R,et al.Comparison of antagonistic effects of four Trichodermastrains[J].Chinese Journal of Biological Control,2012,28(1):147-151.
[12] 李 芳,史 怀,刘 波,等.淡紫拟青霉对尖孢镰刀菌的拮抗作用与机制分析[J].植物保护学报,2005,32(4):373-378.Li F,Shi H,Liu B,et al.Antagonistic effect of biocontrol fungus,Paecilomyces lilacinus strain NH-PL-3and its mechanism against Fusarium oxysporum[J].Acta Phytophylacica Sinica,2005,32(4):373-378.
[13] 刘永亮.辣椒疫霉拮抗真菌毛壳菌和青霉菌的分离鉴定及其生防潜力的研究[D].济南:山东农业大学,2013.Liu Y L.Isolation and identification of fungal antagonist Chaetomium and Penicillum against Phytophthora capsici and studies on their potential of biocontrol[D].Ji'nan:Shandong Agricultural University,2013.
[14] 强 薇,郭 帅,张爱华,等.人参根际土壤中拮抗人参病害微生物的筛选[J].华南农业大学学报,2014,35(5):76-81.Qiang W,Guo S,Zhang A H,et al.Screening antagonistic microorganism against ginseng diseases fromPanax ginsengrhizosphere soil[J].Journal of South China Agricultural University,2014,35(5):76-81.
[15] Ryu H,Park H,Suh D S,et al.Biological control of Colletotrichum panacicola on Panax ginseng by Bacillus subtilis HK-CSM-1[J].J Ginseng Res,2014,38:215-219.
[16] Song M,Yun H Y,Kim Y H.Antagonistic Bacillus species as a biological control of ginseng root rot caused by Fusariumcf.incarnatum[J].J Ginseng Res,2014,38:136-145.
[17] 赵阿娜,丁万隆.木霉菌剂对人参根部病害防治效果评价[J].中草药,2006(10):1552-1554.Zhao A N,Ding W L.Estimation of control effect of Trichodermafungicides on Panax ginsengroot disease[J].Chinese Traditional and Herbal Drugs,2006(10):1552-1554.
[18] 周如军,傅俊范,史会岩,等.人参锈腐病拮抗生防菌的筛选与鉴定[J].沈阳农业大学学报,2010,41(4):422-426.Zhou R J,Fu J F,Shi H Y,et al.Screening and identification of antagonistic strains against Cylindrocarpon destructans[J].Journal of Shenyang Agricultural University,2010,41(4):422-426.
[19] 杨 骁,李长田.人参内生生防真菌的筛选与鉴定[J].东北师范大学学报(自然科学版),2013,45(4):107-113.Yang X,Li C T.Screening and identification of endogeny biocontrol fungi of Panax ginseng[J].Journal of Northeast Normal University(Natural Science Edition),2013,45(4):107-113.
[20] 魏景超.真菌鉴定手册[M].上海:上海科学技术出版社,1979.Wei J C.Fungal identification manual[M].Shanghai:Shanghai Scientific Technology Press,1979.
[21] 陈凤毛.真菌ITS区序列结构及其应用[J].林业科技开发,2007,21(2):5-7.Chen F M.The sequence of ITS region and its application of fungus[J].China Forestry Science and Technology,2007,21(2):5-7.
[22] 张广志,杨合同,文成敬.木霉菌形态学分类检索与分子生物学鉴定[J].山东农业大学学报(自然科学版),2011,42(2):309-316.Zhang G Z,Yang H T,Wen C J.The key to the morphological classification and the molecular identification of Trichoderma spp.[J].Journal of Shandong Agricultural University(Natural Science Edition),2011,42(2):309-316.
[23] 齐祖同.中国真菌志:第五卷:曲霉属及其相关有性型[M].北京:科学出版社,1997.Qi Z T.Chinese fungi:the fifth volume:Aspergillus and its related sexual type[M].Beijing:Science Press,1997.
[24] 孔华忠.中国真菌志:第三十五卷:青霉属及其相关有性型[M].北京:科学出版社,2007.Kong H Z.Chinese fungi:the thirty fifth volume:Penicillium and its related sexual type[M].Beijing:Science Press,2007.
[25] 白金铠.中国真菌志:第十五卷:球壳孢目,茎点霉属,叶点霉属[M].北京:科学出版社,2003.Bai J K.Chinese fungi:the fifteenth volume:Sphaeropsidales,Phornaand Phyllosticta[M].Beijing:Science Press,2003.
[26] 吴连举,关一鸣,逄世峰,等.利用拮抗微生物防治人参、西洋参土传病害研究进展[J].安徽农业科学,2010,38(28):15630-15631,15657.Wu L J,Guan Y M,Pang S F,et al.Research progress on biological control of soil-borne diseases in ginseng and American ginssseng by anti-micoorganism[J].Journal of Anhui Agri Sci,2010,38(28):15630-15631,15657.
[27] 任守让,王瑞霞.对人参锈腐病菌的拮抗真菌及其防病效果[J].吉林农业大学学报,1998,20(S1):120.Ren S R,Wang R X.Controlling effects of antagonistic fungi to Cylindrocarpon destructans in ginseng[J].Journal of Jilin Agricultural University,1998,20(S1):120.
[28] 邢云章,马凤茹.绿色木霉防治人参根腐病的研究[J].特产科学实验,1983(4):16-18.Xing Y Z,Ma F R.Study of Trichoderma viride to control Fusarium solani[J].Specialty Experiment,1983(4):16-18.
[29] 赵阿娜,丁万隆,朱殿龙.拮抗人参根部病原菌木霉的筛选及其生物学特性初步研究[J].中国中药杂志,2006,31(20):1671-1674.Zhao A N,Ding W L,Zhu D L.Screening strains for Trichoderma spp.for strong antagonism against ginseng root pathogens and study on their biological characters[J].China Journal of Chinese Materia Medica,2006,31(20):1671-1674.
[30] 张 瑾,张树武,徐秉良,等.长枝木霉菌抑菌谱测定及其抑菌作用机理研究[J].中国生态农业学报,2014,22(6):661-667.Zhang J,Zhang S W,Xu B L,et al.Determining antifungal spectrum and mechanism of Trichoderma longibrachiatum in vitro[J].Chinese Journal of Eco-Agriculture,2014,22(6):661-667.
[31] 胡明江,张秀省,曹 兴,等.康宁木霉SMF2防治大白菜软腐病研究[J].北方园艺,2009(6):102-103.Hu M J,Zhang X S,Cao X,et al.Studies on the control effect of Trichoderma kningii Oud.against soft rot of Chinese cabbage[J].Northern Horticulture,2009(6):102-103.
[32] 梁巧兰,王 芳,魏列新,等.深绿木霉T2菌株对百合疫霉拮抗作用及机制[J].植物保护,2011,37(6):164-167.Liang Q L,Wang F,Wei L X,et al.Antagonistic mechanism of Trichoderma atroviride T2strain against Phytophthora nicotianae[J].Plant Protection,2011,37(6):164-167.
[33] 刘晓芸.对致病疫霉具拮抗作用的真菌XF的鉴定及抑菌活性研究[D].河北保定:河北农业大学,2007.Liu X Y.Identification and antifungal activity of XF,an antagonistic fungus against Phytophthora infestans[D].Hebei Baoding:Hebei Agricultural University,2007.
[34] 王艳红,吴晓民,朱艳萍,等.温郁金内生真菌Chaetomium globosum L18对植物病原菌的抑菌谱及拮抗机理[J].生态学报,2012,32(7):2040-2046.Wang Y H,Wu X M,Zhu Y P,et al.Inhibition effects and mechanisms of the endophytic fungus Chaetomium globosum L18from Curcuma wenyujin[J].Acata Ecologica Sinica,2012,32(7):2040-2046.
[35] 马 艳,常志州,赵江涛,等.一株疫病拮抗青霉P.st10菌株的抗菌活性及其对辣椒疫病的盆栽防效[J].中国生物防治,2006,22(3):239-243.Ma Y,Chang Z Z,Zhao J T,et al.Antifungal activity of Penicillium striatisporum strain P.st10and its inhibition against pepper blight caused by Phytophthoraspp.[J].Chinese Journal of Biological Control,2006,22(3):239-243.
[36] 窦红涛,李若瑜,万 哲,等.临床常见镰刀菌rDNA ITS序列分析[J].临床和实验医学杂志,2006,5(6):646-648.Dou H T,Li R Y,Wan Z,et al.Sequence analysis of ribosomal DNA ITS of clinical comm on Fusariumspecies[J].Journal of Clinical and Experimental Medicine,2006,5(6):646-648.
[37] 隋 心,冯富娟,娄 鑫,等.土壤微生物总DNA的提取以及土壤真菌ITS-PCR体系的建立[J].中国酿造,2011(9):166-169.Sui X,Feng F J,Lou X,et al.Total DNA from soil microorganisms and construction of ITS-PCR system for soil fungi[J].China Brewing,2011(9):166-169.