香菇蛛网病病原菌树状枝葡霉生物学特性
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摘要
香菇是我国产量最高的食用菌之一,因其独特浓郁的风味而广受赞誉。然而随着香菇栽培面积的不断扩大,其各种病害的发生也日趋频繁。近期在浙江省庆元县多个菇棚内发现一种新病害。为了明确致病菌,本研究通过对罹病香菇子实体进行组织分离和单孢分离获得纯化菌株QS02,随后结合形态学特征和分子生物学技术对其进行鉴定,并通过柯赫氏法则的验证,确定该病原菌为树状枝葡霉Cladobotryum dendroides。同时,本文还对该病原菌展开了生物学特性研究,结果表明:病原菌菌丝生长最适培养基为PDA培养基,最适温度为25℃,最适碳源为可溶性淀粉,最适氮源为牛肉膏,最适pH区间为5–6;光照对该菌菌丝生长有一定的抑制作用。这是由树状枝葡霉引起的香菇蛛网病在国内的首次报道。
A new fungal disease was found in several Lentinula edodes mushroom growing sheds in Qingyuan,Zhejiang Province. Observation of morphological characteristics, phylogenetic analysis using rDNA sequences of ITS and partial 28S rDNA, and confirmation of pathogenicity using Koch's postulates revealed that the causal agent of the cobweb disease was Cladobotryum dendroides. Biological characterization showed that the optimum growth condition for the pathogen was growing temperature of 25℃, pH value of 5–6 and culture medium using soluble starch as carbon source and beef extract as nitrogen source. PDA was also suitable medium for growth. Light showed certain inhibitory effect on pathogen growth. This is the first report of the pathogen responsible for cobweb disease on Lentinula edodes in China.
A new fungal disease was found in several Lentinula edodes mushroom growing sheds in Qingyuan,Zhejiang Province. Observation of morphological characteristics, phylogenetic analysis using rDNA sequences of ITS and partial 28S rDNA, and confirmation of pathogenicity using Koch's postulates revealed that the causal agent of the cobweb disease was Cladobotryum dendroides. Biological characterization showed that the optimum growth condition for the pathogen was growing temperature of 25℃, pH value of 5–6 and culture medium using soluble starch as carbon source and beef extract as nitrogen source. PDA was also suitable medium for growth. Light showed certain inhibitory effect on pathogen growth. This is the first report of the pathogen responsible for cobweb disease on Lentinula edodes in China.
引文
Adie B,Grogan H,Archer S,Mills P,2006.Temporal and spatial dispersal of Cladobotryum conidia in the controlled environment of a mushroom growing room.Applied&Environmental Microbiology,72(11):7212-7217
Bian YB,2013.Research progress on infectious and competitive mycelial diseases of edible mushrooms.Edible Fungi of China,20(2):1-7(in Chinese)
Chang S,Miles P,2004.Mushrooms:cultivation,nutritional value,medicinal effect,and environmental impact.American Journal of Orthopsychiatry,38(4):688-692
Cao XT,Bian YB,Xiao XJ,Li JS,Wang GZ,2015.Effect of heat stress on Lentinula edodes mycelial growth recovery and resistance to Trichoderma harzianum.Edible Fungi of China,22(4):81-85(in Chinese)
Carrasco J,Navarro MJ,Gea FJ,2017a.Cobweb,a serious pathology in mushroom crops:a review.Spanish Journal of Agricultural Research,15(2):19
Carrasco J,Navarro MJ,Santos M,Gea FJ,2017b.Effect of five fungicides with different modes of action on cobweb disease(Cladobotryum mycophilum)and mushroom yield.Annals of Applied Biology,171(1):62-69
Fang ZD,1998.Methodology of plant pathology research.3rd edition.China Agricultural Press,Beijing.37-53(in Chinese)
Fletcher JT,Gaze RH,2008.Mushroom pest and disease control:a color handbook.Manson Publishing Ltd.Academic Press,San Diego.76-80
Fu YP,Wang XX,Li D,Liu Y,Song B,Zhang CL,Wang Q,Chen MY,Zhang ZW,Li Y,2016.Identification of resistance to wet bubble disease and genetic diversity in wild and cultivated strains of Agaricus bisporus.International Journal of Molecular Sciences,17(10):1568
Grogan HM,Gaze RH,2000.Fungicide resistance among Cladobotryum spp.-causal agents of cobweb disease of the edible mushroom Agaricus bisporus.Mycological Research,104(3):357-364
Gea FJ,Carrasco J,Suz LM,Navarro MJ,2017.Characterization and pathogenicity of Cladobotryum mycophilum in Spanish Pleurotus eryngii mushroom crops and its sensitivity to fungicides.European Journal of Plant Pathology,147(1):129-139
Gea FJ,Navarro MJ,Suz LM,2018.First report of cobweb disease caused by Cladobotryum dendroides on shiitake mushroom(Lentinula edodes)in Spain.Plant Disease,102(5):1030
Hou J,2013.Study on cultivation techniques of mushroom.Horticulture&Seed,11:1-4(in Chinese)
Hu QX,Song JD,Guan DP,2008.Hazard analysis and critical control point of edible fungal disease and insect pests.Chinese Agricultural Science Bulletin,24(12):401-406(in Chinese)
Huang NL,1994.Chinese shiitake cultivation.Shanghai Scientific and Technical Literature Publishing House,Shanghai.1-57(in Chinese)
Kumar S,Stecher G,Tamura K,2016.MEGA7:molecular evolutionary genetics analysis version 7.0 for bigger datasets.Molecular Biology and Evolution,33(7):1870-1874
Lan YF,An XR,Wang QW,Tang LN,2017.Biological characteristics of Cladobotryum mycophilum causing cobweb disease on Agaricus bisporus.Edible Fungi of China,36(4):62-65(in Chinese)
Li Y,Li TH,Yang ZL,Bau T,Dai YC,2015.Atlas of Chinese macrofungal resources.Zhongyuan Nongmin Press,Zhengzhou.603-617(in Chinese)
Li Y,Shang XD,Song CY,Li ZP,Tan Q,Zhou F,2017.Study on the industrialized cultivation mode and technology of Lentinus edodes in China.Shanghai Scientific and Technical Publishers,Shanghai.3:74-75(in Chinese)
Liu ZH,Li D,Sossah FL,Okorley BA,Fu YP,Li Y,2018,Major fungal and bacterial diseases in edible fungi.Journal of Fungal Research,16(3):158-163(in Chinese)
O'Donnell K,1993.Fusarium and its near relatives.In:Reynolds DR,Taylor JW(eds.)The fungal holomorph:mitotic,meiotic and pleomorphic speciation in fungal systematics.CAB International,Wallingford,UK.225-233
P?ldmaa K,2011.Tropical species of Cladobotryum and Hypomyces producing red pigments.Studies in Mycology,68:1-34
Poto?nik I,Vukojevi?J,Staji?M,Rekanovi?E,Svetlana Milija?evi?,Stepanovi?M,Todorovi?B,2009.Toxicity of fungicides with different modes of action to Cladobotryum dendroides and Agaricus bisporus.Journal of Environmental Science and Health,Part B,44(8):5
Tamm H,P?ldmaa K,2013.Diversity,host associations,and phylogeography of temperate aurofusarin-producing Hypomyces/Cladobotryum including causal agents of cobweb disease of cultivated mushrooms.Fungal Biology,117(5):348-367
Wang Q,2010.Identification of Trichoderma and screening of its biological agent.Guizhou Agriculture Sciences,38(7):117-118(in Chinese)
White TJ,Bruns T,Lee S,Taylor JW,1990.In:Innis MA,Gelfand DH,Sninsky JJ,White TJ(eds.)Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics.PCR Protocols:a guide to methods and applications.Academic Press,New York.18(1):315-322
Zhang QH,Wang W,Li CH,Wen ZQ,2015.Biological characteristics of Hypomyces aurantius parasitic on Hypsizygus marmoreus.Mycosystema,34(3):350-356(in Chinese)
边银丙,2013.食用菌菌丝体侵染性病害与竞争性病害研究进展.食用菌学报,20(2):1-7
曹现涛,边银丙,肖新军,李进山,王刚正,2015.高温胁迫对香菇菌丝生长及其抗哈茨木霉能力的影响.食用菌学报,22(4):81-85
方中达,1998.植病研究方法.第3版.北京:中国农业出版社.37-53
黄年来,1994.中国香菇栽培学.上海:上海科技文献出版社.1-57
胡清秀,宋金娣,管道平,2008.食用菌病虫害危害分析与防治关键控制点.中国农学通报,24(12):401-406
侯俊,2013.香菇栽培技术研究.园艺与种苗,11:1-4
李玉,李泰辉,杨祝良,图力古尔,戴玉成,2015.中国大型菌物资源图鉴.郑州:中原农民出版社.603-617
兰玉菲,安秀荣,王庆武,唐丽娜,2017.双孢蘑菇蛛网病病菌生物学特性研究.中国食用菌,36(4):62-65
李玉,尚晓冬,宋春艳,李正鹏,谭琦,周峰,2017.我国香菇工厂化栽培模式及技术初探.上海农业科技,3:74-75
刘正慧,李丹,Frederick Leo Sossah,Benjamin Azu Okorley,付永平,李玉,2018.食用菌主要真菌和细菌病害.菌物研究,16(3):158-163
王琪,2010.香菇木霉病菌的鉴定及室内生物制剂筛选.贵州农业科学,38(7):117-118
张琪辉,王威,李成欢,温志强,2015.斑玉蕈蛛网病的病原菌及其生物学特性.菌物学报,34(3):350-356
Bian YB,2013.Research progress on infectious and competitive mycelial diseases of edible mushrooms.Edible Fungi of China,20(2):1-7(in Chinese)
Chang S,Miles P,2004.Mushrooms:cultivation,nutritional value,medicinal effect,and environmental impact.American Journal of Orthopsychiatry,38(4):688-692
Cao XT,Bian YB,Xiao XJ,Li JS,Wang GZ,2015.Effect of heat stress on Lentinula edodes mycelial growth recovery and resistance to Trichoderma harzianum.Edible Fungi of China,22(4):81-85(in Chinese)
Carrasco J,Navarro MJ,Gea FJ,2017a.Cobweb,a serious pathology in mushroom crops:a review.Spanish Journal of Agricultural Research,15(2):19
Carrasco J,Navarro MJ,Santos M,Gea FJ,2017b.Effect of five fungicides with different modes of action on cobweb disease(Cladobotryum mycophilum)and mushroom yield.Annals of Applied Biology,171(1):62-69
Fang ZD,1998.Methodology of plant pathology research.3rd edition.China Agricultural Press,Beijing.37-53(in Chinese)
Fletcher JT,Gaze RH,2008.Mushroom pest and disease control:a color handbook.Manson Publishing Ltd.Academic Press,San Diego.76-80
Fu YP,Wang XX,Li D,Liu Y,Song B,Zhang CL,Wang Q,Chen MY,Zhang ZW,Li Y,2016.Identification of resistance to wet bubble disease and genetic diversity in wild and cultivated strains of Agaricus bisporus.International Journal of Molecular Sciences,17(10):1568
Grogan HM,Gaze RH,2000.Fungicide resistance among Cladobotryum spp.-causal agents of cobweb disease of the edible mushroom Agaricus bisporus.Mycological Research,104(3):357-364
Gea FJ,Carrasco J,Suz LM,Navarro MJ,2017.Characterization and pathogenicity of Cladobotryum mycophilum in Spanish Pleurotus eryngii mushroom crops and its sensitivity to fungicides.European Journal of Plant Pathology,147(1):129-139
Gea FJ,Navarro MJ,Suz LM,2018.First report of cobweb disease caused by Cladobotryum dendroides on shiitake mushroom(Lentinula edodes)in Spain.Plant Disease,102(5):1030
Hou J,2013.Study on cultivation techniques of mushroom.Horticulture&Seed,11:1-4(in Chinese)
Hu QX,Song JD,Guan DP,2008.Hazard analysis and critical control point of edible fungal disease and insect pests.Chinese Agricultural Science Bulletin,24(12):401-406(in Chinese)
Huang NL,1994.Chinese shiitake cultivation.Shanghai Scientific and Technical Literature Publishing House,Shanghai.1-57(in Chinese)
Kumar S,Stecher G,Tamura K,2016.MEGA7:molecular evolutionary genetics analysis version 7.0 for bigger datasets.Molecular Biology and Evolution,33(7):1870-1874
Lan YF,An XR,Wang QW,Tang LN,2017.Biological characteristics of Cladobotryum mycophilum causing cobweb disease on Agaricus bisporus.Edible Fungi of China,36(4):62-65(in Chinese)
Li Y,Li TH,Yang ZL,Bau T,Dai YC,2015.Atlas of Chinese macrofungal resources.Zhongyuan Nongmin Press,Zhengzhou.603-617(in Chinese)
Li Y,Shang XD,Song CY,Li ZP,Tan Q,Zhou F,2017.Study on the industrialized cultivation mode and technology of Lentinus edodes in China.Shanghai Scientific and Technical Publishers,Shanghai.3:74-75(in Chinese)
Liu ZH,Li D,Sossah FL,Okorley BA,Fu YP,Li Y,2018,Major fungal and bacterial diseases in edible fungi.Journal of Fungal Research,16(3):158-163(in Chinese)
O'Donnell K,1993.Fusarium and its near relatives.In:Reynolds DR,Taylor JW(eds.)The fungal holomorph:mitotic,meiotic and pleomorphic speciation in fungal systematics.CAB International,Wallingford,UK.225-233
P?ldmaa K,2011.Tropical species of Cladobotryum and Hypomyces producing red pigments.Studies in Mycology,68:1-34
Poto?nik I,Vukojevi?J,Staji?M,Rekanovi?E,Svetlana Milija?evi?,Stepanovi?M,Todorovi?B,2009.Toxicity of fungicides with different modes of action to Cladobotryum dendroides and Agaricus bisporus.Journal of Environmental Science and Health,Part B,44(8):5
Tamm H,P?ldmaa K,2013.Diversity,host associations,and phylogeography of temperate aurofusarin-producing Hypomyces/Cladobotryum including causal agents of cobweb disease of cultivated mushrooms.Fungal Biology,117(5):348-367
Wang Q,2010.Identification of Trichoderma and screening of its biological agent.Guizhou Agriculture Sciences,38(7):117-118(in Chinese)
White TJ,Bruns T,Lee S,Taylor JW,1990.In:Innis MA,Gelfand DH,Sninsky JJ,White TJ(eds.)Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics.PCR Protocols:a guide to methods and applications.Academic Press,New York.18(1):315-322
Zhang QH,Wang W,Li CH,Wen ZQ,2015.Biological characteristics of Hypomyces aurantius parasitic on Hypsizygus marmoreus.Mycosystema,34(3):350-356(in Chinese)
边银丙,2013.食用菌菌丝体侵染性病害与竞争性病害研究进展.食用菌学报,20(2):1-7
曹现涛,边银丙,肖新军,李进山,王刚正,2015.高温胁迫对香菇菌丝生长及其抗哈茨木霉能力的影响.食用菌学报,22(4):81-85
方中达,1998.植病研究方法.第3版.北京:中国农业出版社.37-53
黄年来,1994.中国香菇栽培学.上海:上海科技文献出版社.1-57
胡清秀,宋金娣,管道平,2008.食用菌病虫害危害分析与防治关键控制点.中国农学通报,24(12):401-406
侯俊,2013.香菇栽培技术研究.园艺与种苗,11:1-4
李玉,李泰辉,杨祝良,图力古尔,戴玉成,2015.中国大型菌物资源图鉴.郑州:中原农民出版社.603-617
兰玉菲,安秀荣,王庆武,唐丽娜,2017.双孢蘑菇蛛网病病菌生物学特性研究.中国食用菌,36(4):62-65
李玉,尚晓冬,宋春艳,李正鹏,谭琦,周峰,2017.我国香菇工厂化栽培模式及技术初探.上海农业科技,3:74-75
刘正慧,李丹,Frederick Leo Sossah,Benjamin Azu Okorley,付永平,李玉,2018.食用菌主要真菌和细菌病害.菌物研究,16(3):158-163
王琪,2010.香菇木霉病菌的鉴定及室内生物制剂筛选.贵州农业科学,38(7):117-118
张琪辉,王威,李成欢,温志强,2015.斑玉蕈蛛网病的病原菌及其生物学特性.菌物学报,34(3):350-356