甜瓜细菌性叶枯病病原菌鉴定及有效药剂筛选
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摘要
[目的]确定甜瓜细菌性叶枯病病原菌种类,筛选有效抑菌药剂。[方法]划线分离法获得分离物经柯氏验证菌株致病性,结合生理生化和分子生物学方法进行鉴定,抑菌圈法测定室内毒力。[结果]病原菌鉴定为丁香假单胞丁香致病变种(Pseudomonas syringae pv. syringae);在制剂推荐剂量下,0.3%四霉素水剂20 000～33 000 mg/L、30%乙蒜素水剂1429～2000 mg/L、80%乙蒜素水剂556～667 mg/L对病菌TG01的抑菌圈直径>15 mm;3%噻霉酮微乳剂2000～3333 mg/L、20%溴硝醇可湿性粉剂667～1000 mg/L、60%进口叶枯唑粉剂3333～5000 mg/L对TG01的抑菌圈直径10～15 mm;47%春雷·王铜WP 1667～2000 mg/L对TG01的抑菌圈直径8.33～7.33 mm;其他供试药剂无抑菌活性。[结论]明确了甜瓜细菌性叶枯病的病原菌为P. syringae pv. syringae,筛选出7种对该病菌有抑制作用的药剂。
[Aims]This study aims to identify the pathogen which seriously caused leaf blight disease to Cucumis melo,and screen out effective bactericides against this pathogen. [Methods]Isolates from the diseased leaves were obtained by streak plate method. Identification of the pathogenic strains was carried out according to the cultural characteristics,physiological and biochemical analysis and molecular biology identification. The effective bactericides to the pathogen were determined by using inhibitory halo method. [Results]The pathogen of bacteria leaf blight was identified as Pseudomonas syringae pv. syringae. The diameters of inhibitory haloes of tetramycin 0.3% AS with 20 000-33 000 mg/L,ethylicin 30% AS with 1429-2000 mg/L, and ethylicin 80% AS with 556-667 mg/L against the strain TG01 were more than 15 mm. The diameters of inhibitory haloes of benziothiazolinone 3% ME with 2000-3333 mg/L, bronopol 20%WP with 667-1000 mg/L, bismerthiazol 60% DP with 3333-5000 mg/L against TG01 were ranged from 10 to 15 mm.The diameters of inhibitory haloes of kasugamycin·copper oxychloride 47% WP at 1667-2000 mg/L were only8.33-7.33 mm. While other tested reagents did not show any inhibitory effects. [Conclusions]The bacterial leaf blight of Cucumis melo is caused by P. syringae pv. syringae, and seven reagents have inhibitory effects on this pathogen.
[Aims]This study aims to identify the pathogen which seriously caused leaf blight disease to Cucumis melo,and screen out effective bactericides against this pathogen. [Methods]Isolates from the diseased leaves were obtained by streak plate method. Identification of the pathogenic strains was carried out according to the cultural characteristics,physiological and biochemical analysis and molecular biology identification. The effective bactericides to the pathogen were determined by using inhibitory halo method. [Results]The pathogen of bacteria leaf blight was identified as Pseudomonas syringae pv. syringae. The diameters of inhibitory haloes of tetramycin 0.3% AS with 20 000-33 000 mg/L,ethylicin 30% AS with 1429-2000 mg/L, and ethylicin 80% AS with 556-667 mg/L against the strain TG01 were more than 15 mm. The diameters of inhibitory haloes of benziothiazolinone 3% ME with 2000-3333 mg/L, bronopol 20%WP with 667-1000 mg/L, bismerthiazol 60% DP with 3333-5000 mg/L against TG01 were ranged from 10 to 15 mm.The diameters of inhibitory haloes of kasugamycin·copper oxychloride 47% WP at 1667-2000 mg/L were only8.33-7.33 mm. While other tested reagents did not show any inhibitory effects. [Conclusions]The bacterial leaf blight of Cucumis melo is caused by P. syringae pv. syringae, and seven reagents have inhibitory effects on this pathogen.
引文
[1]王志丹,赵姜,毛世平,等.中国甜瓜产业区域优势布局研究[J].中国农业资源与区划,2014,35(1):128-133.
[2]赵廷昌,孙福在,王兵万,等.哈密瓜细菌性果斑病病原菌鉴定[J].植物病理学报,2001,31(4):357-364.
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[5]李亚利,贾迎春,蒲崇建,等.甜瓜细菌性叶斑病菌(Pseudomonas syringae pv.lachrymans)寄主范围测定[J].甘肃农业大学学报,2006(6):63-66.
[6]葛林梅,李红叶,张明方,等.甜瓜细菌性软腐病病原鉴定[J].果树学报,2006(3):427-430.
[7]ELHENDAWY H H.The Biological Control of Soft Rot Disease in Melon Caused by Erwinia carotovora subsp.carotovora Using Pseudomonas fluorescens[J].Microbiological Research,1998,153(1):55-60.
[8]王培全,殷汝松,王娟,等.薄皮甜瓜细菌性枯萎病发生原因及防治措施[J].中国园艺文摘,2015,31(9):203,216.
[9]CAUDLE J R.Control of Erwinia tracheiphila in Cucumis melo[D].Theses and Dissertations-Plant and Soil Sciences,2013:36.
[10]ROJAS E S,GLEASON M L,BATZER J C,et al.Feasibility of Delaying Removal of Row Covers to Suppress Bacterial Wilt of Muskmelon(Cucumis melo)[J].Plant Disease,2011,95(6):729-734.
[11]OBRADOVIC A,ARSENIJEVIC M.First Report of a Wilt and Stem Rot of Muskmelon and Watermelon Transplant Incited by Pseudomonas chichorii in Serbia[J].Plant Disease,2007,86(4):443-443.
[12]GOUMA H,KIJIMA T.Occurrence of New Bacterial Disease of Melon(Cucumis melo)Caused by Pseudomonas chicorii[J].Annals of the Phytopathological Society of Japan,1995,61.
[13]方中达.植病研究法[M].北京:中国农业出版社,1998.
[14]ETAL E H.Bergey's Manual of Determinative Bacteriology(9th ed)[M].Lippincott Williams&Wilkins,1994.
[15]SCHAAD N W.Laboratory Guide for Identification of Plant Pathogenic Bacteria(Third Edition)[M].Beijing:China Agricultural Science and Technology Press,2011:69-81.
[16]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[17]YAMAMOTO S,HARAYAMA S.PCR Amplification and Direct Sequencing of gyr B Genes with Universal Primers and Their Application to the Detection and Taxonomic Analysis of Pseudomonas putida Strains[J].Applied&Environmental Microbiology,1995,61(3):1104-1109.
[18]SORENSEN K N,KIM K H,TAKEMOTO J Y.PCR Detection of Cyclic Lipodepsinonapeptide-Producing Pseudomonas syringae pv.syringae and Similarity of Strains[J].Applied&Environmental Microbiology,1998,64(1):226-230.
[19]FEIL H,FEIL W S,CHAIN P,et al.Comparison of the Complete Genome Sequences of Pseudomonas syringae pv.syringae B728a and pv.tomato DC3000[J].Proceedings of the National Academy of Sciences of the United States of America,2005,102(31):11064-11069.
[20]RAVINDRAN A,JALAN N,YUAN J S,et al.Comparative Genomics of Pseudomonas syringae pv.syringae Strains B301Dand HS191 and Insights into Intrapathovar Traits Associated with Plant Pathogenesis[J].Microbiologyopen,2015,4(4):553-573.
[21]李晓颖,佘小漫.广东南瓜细菌性叶枯病及其病原鉴定[J].植物病理学报,2018,48(2):159-168.
[2]赵廷昌,孙福在,王兵万,等.哈密瓜细菌性果斑病病原菌鉴定[J].植物病理学报,2001,31(4):357-364.
[3]赵廷昌,赵洪海,王怀松.山东省西瓜、甜瓜发生瓜类细菌性果斑病[J].植物保护,2009,35(5):170-171.
[4]缪林玉,王利民,田艳丽,等.甜瓜细菌性叶枯病菌胶体金ICA检测试纸条的研发与应用[J].农业生物技术学报,2016,24(1):134-141.
[5]李亚利,贾迎春,蒲崇建,等.甜瓜细菌性叶斑病菌(Pseudomonas syringae pv.lachrymans)寄主范围测定[J].甘肃农业大学学报,2006(6):63-66.
[6]葛林梅,李红叶,张明方,等.甜瓜细菌性软腐病病原鉴定[J].果树学报,2006(3):427-430.
[7]ELHENDAWY H H.The Biological Control of Soft Rot Disease in Melon Caused by Erwinia carotovora subsp.carotovora Using Pseudomonas fluorescens[J].Microbiological Research,1998,153(1):55-60.
[8]王培全,殷汝松,王娟,等.薄皮甜瓜细菌性枯萎病发生原因及防治措施[J].中国园艺文摘,2015,31(9):203,216.
[9]CAUDLE J R.Control of Erwinia tracheiphila in Cucumis melo[D].Theses and Dissertations-Plant and Soil Sciences,2013:36.
[10]ROJAS E S,GLEASON M L,BATZER J C,et al.Feasibility of Delaying Removal of Row Covers to Suppress Bacterial Wilt of Muskmelon(Cucumis melo)[J].Plant Disease,2011,95(6):729-734.
[11]OBRADOVIC A,ARSENIJEVIC M.First Report of a Wilt and Stem Rot of Muskmelon and Watermelon Transplant Incited by Pseudomonas chichorii in Serbia[J].Plant Disease,2007,86(4):443-443.
[12]GOUMA H,KIJIMA T.Occurrence of New Bacterial Disease of Melon(Cucumis melo)Caused by Pseudomonas chicorii[J].Annals of the Phytopathological Society of Japan,1995,61.
[13]方中达.植病研究法[M].北京:中国农业出版社,1998.
[14]ETAL E H.Bergey's Manual of Determinative Bacteriology(9th ed)[M].Lippincott Williams&Wilkins,1994.
[15]SCHAAD N W.Laboratory Guide for Identification of Plant Pathogenic Bacteria(Third Edition)[M].Beijing:China Agricultural Science and Technology Press,2011:69-81.
[16]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[17]YAMAMOTO S,HARAYAMA S.PCR Amplification and Direct Sequencing of gyr B Genes with Universal Primers and Their Application to the Detection and Taxonomic Analysis of Pseudomonas putida Strains[J].Applied&Environmental Microbiology,1995,61(3):1104-1109.
[18]SORENSEN K N,KIM K H,TAKEMOTO J Y.PCR Detection of Cyclic Lipodepsinonapeptide-Producing Pseudomonas syringae pv.syringae and Similarity of Strains[J].Applied&Environmental Microbiology,1998,64(1):226-230.
[19]FEIL H,FEIL W S,CHAIN P,et al.Comparison of the Complete Genome Sequences of Pseudomonas syringae pv.syringae B728a and pv.tomato DC3000[J].Proceedings of the National Academy of Sciences of the United States of America,2005,102(31):11064-11069.
[20]RAVINDRAN A,JALAN N,YUAN J S,et al.Comparative Genomics of Pseudomonas syringae pv.syringae Strains B301Dand HS191 and Insights into Intrapathovar Traits Associated with Plant Pathogenesis[J].Microbiologyopen,2015,4(4):553-573.
[21]李晓颖,佘小漫.广东南瓜细菌性叶枯病及其病原鉴定[J].植物病理学报,2018,48(2):159-168.