云南烟草黑胫病菌对甲霜灵抗药性评估及相关防治技术研究
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摘要
本文对云南烟区烟草黑胫病菌对“甲霜灵”抗性水平进行了调查,了解抗性发生状况,对烟草黑胫病菌对“甲霜灵”抗药性发生的原因及形成机理进行了探索,研究和制定抗性治理的技术措施,防止或延缓烟草黑胫病对“甲霜灵”抗药性的产生,达到合理有效使用农药,减少用药量,降低生产成本,减少烟叶农药残留及提高烟叶安全性,延长农药使用寿命。其研究结果如下:
1、在云南省烟区分离鉴定出57个烟草黑胫病菌菌株,黑胫病菌分离率为42.22%;
2、测定了57株分离纯化的烟草黑胫病菌对甲霜灵的抗性水平。不同菌株对甲霜灵的EC50值在0.0129～1.6307μg/mL之间,不同地方烟草黑胫病菌对甲霜灵抗性水平存在较大差异;
3、从各处理后分离菌株的抗药测定结果看,黑胫病菌的抗药性与施药次数和施药浓度与呈正相关。
4、分离和筛选出280个对烟草疫霉有拮抗作用的菌株。经过温室盆栽拮抗菌防治烟草黑胫病实验表明,有12株拮抗细菌对黑胫病的抑制效果。在江川、蒙自县选择了以往发黑胫病比较重的地块,用对烟草黑胫病有抑制作用的拮抗细菌进行了田间小区试验,其中GP13、HK282、HJ01912、B254等4个生防菌的防治效果分别为71.84%、58.82%、73.80%和49.96%。
5、上述生防菌在4000亩大田生产防治示范表明,其防治效果达63.2%～65.2%,抗性治理效果明显。
To prevent or delay the Tobacco Tibia in the formation of metalaxyl resistance, and to achieve rational and effective use of pesticides, reduce the dosage, lower production costs, reducing pesticide residue of tobacco leaf and improve the safety of pesticides to extend the service life of purpose. This article was about metalaxyl resistance level survey in Yunnan Province, a situation of resistance was understood, tobacco black shank resistant bacteria to metalaxyl and the causes and formation mechanism of resistance management techniques were explored, and formulate measures was studied .The results are as follows:
1, 57 strains of phytophthora nicotianae were isolated in Yunnan Province , and the isolation rate was 42.22 %.
2, The metalaxyl resistance level was detected for the 57 strains. The EC50 values different strains Metalaxyl were among 0.0129～1.6307μg/ml, there is great difference of the level of metalaxyl resistance between different Phytophthora nicotianae strains;
3, The strains was isolated from the treatment plant with metalaxyl. The metalaxyl resistance of these strains were determend , the results showed that strains resistant was positively correlated with the frequency of application, concentration and application of metalaxyl.
4, 280 strains were isolated that had antagonistic activity to phytophthora nicotianae. 12 antagonistic bacteria showed inhibit the effectiveness to tobacco black shank with the greenhouse test. The antagonism inhibit bacteria were test to contrl tobacco black shank with field plot trial in Jiangchuan County.and Mengzi County. The control effects of GP13, HK282, HJ01912, B254 were 71.84%, 58.82%, 73.80%, 49.96% respectively.
5, resistance management showed evident result with control effect of 63.2%～65.2% by 266.7htm~2 production apllication.
1、在云南省烟区分离鉴定出57个烟草黑胫病菌菌株,黑胫病菌分离率为42.22%;
2、测定了57株分离纯化的烟草黑胫病菌对甲霜灵的抗性水平。不同菌株对甲霜灵的EC50值在0.0129～1.6307μg/mL之间,不同地方烟草黑胫病菌对甲霜灵抗性水平存在较大差异;
3、从各处理后分离菌株的抗药测定结果看,黑胫病菌的抗药性与施药次数和施药浓度与呈正相关。
4、分离和筛选出280个对烟草疫霉有拮抗作用的菌株。经过温室盆栽拮抗菌防治烟草黑胫病实验表明,有12株拮抗细菌对黑胫病的抑制效果。在江川、蒙自县选择了以往发黑胫病比较重的地块,用对烟草黑胫病有抑制作用的拮抗细菌进行了田间小区试验,其中GP13、HK282、HJ01912、B254等4个生防菌的防治效果分别为71.84%、58.82%、73.80%和49.96%。
5、上述生防菌在4000亩大田生产防治示范表明,其防治效果达63.2%～65.2%,抗性治理效果明显。
To prevent or delay the Tobacco Tibia in the formation of metalaxyl resistance, and to achieve rational and effective use of pesticides, reduce the dosage, lower production costs, reducing pesticide residue of tobacco leaf and improve the safety of pesticides to extend the service life of purpose. This article was about metalaxyl resistance level survey in Yunnan Province, a situation of resistance was understood, tobacco black shank resistant bacteria to metalaxyl and the causes and formation mechanism of resistance management techniques were explored, and formulate measures was studied .The results are as follows:
1, 57 strains of phytophthora nicotianae were isolated in Yunnan Province , and the isolation rate was 42.22 %.
2, The metalaxyl resistance level was detected for the 57 strains. The EC50 values different strains Metalaxyl were among 0.0129～1.6307μg/ml, there is great difference of the level of metalaxyl resistance between different Phytophthora nicotianae strains;
3, The strains was isolated from the treatment plant with metalaxyl. The metalaxyl resistance of these strains were determend , the results showed that strains resistant was positively correlated with the frequency of application, concentration and application of metalaxyl.
4, 280 strains were isolated that had antagonistic activity to phytophthora nicotianae. 12 antagonistic bacteria showed inhibit the effectiveness to tobacco black shank with the greenhouse test. The antagonism inhibit bacteria were test to contrl tobacco black shank with field plot trial in Jiangchuan County.and Mengzi County. The control effects of GP13, HK282, HJ01912, B254 were 71.84%, 58.82%, 73.80%, 49.96% respectively.
5, resistance management showed evident result with control effect of 63.2%～65.2% by 266.7htm~2 production apllication.
引文
Neurospora Newslett.29:16-17
[12]Cui,W.,Ross,E.B.,Matthew,D.T.,el al.,2002.An osmosensing histidine kinase mediates dicarboximide fungicide resistance in Botryotinia fuckeliana(Botrytis cinerea)[J].Fungal Genetics and Biology.36:187-198.
[13]Takayuki,M.,Kaori,K.,Tomohiro,O.,et al.,2005.A two-component histidine kinase of the rice blast fungus is involved in osmotic stress response and fungicide action[J].Fungal Genetics and Biology.42:200-212.
[14]Koenraadt,H.,Somerville,S.C.,Jones,A.L.,1992.Characterization of mutations in the beta-tubulin gene of benomyl-resistant field strains of Venturia inaequalis and other plant pathogenic fungi[J],phytopathology.82(11):1348-1354.
[15]陆悦健,周明国,叶钟音等,抗苯并咪唑的小麦赤霉病菌β-tubulin基因序列分析与特性研究[J].植物病理学报.2000.30(1):30-34.
[16]Kawchuk,L.M.,Hutchison,L.J.,Verhaeghe,C.A.,et al.,2002.Isolation of the β-tubulin gene and characterization of thiabendazole resistance in Gibberella.pulicaris[J].Can.J.Plant Pathol.24:233-238.
[17]林孔勋主编,杀菌剂毒理学[M].北京:中国农业出版社.1995.101-140.
[18]Fabritus,A.L.,Shattock,R.C.,Judelson,H.S.,1997.Genetic analysis of metalaxyl in sensitivity loci in Phytophthora infestants using linked DNA markers[J].Phytopathology.87:1034-1040.
[19]Shaw,D.S.,Shattock,R.C.,1991.Genetics of Phytophthora infestants:The Mendelian approach[C].In Phytophthora pp.218-230 British Mycological Society,Cambridge Univ.Press,Cambridge.
[20]Chang,T.T.,Ko,W.H.,1990.Resistance to fungicide and antibiotics in Phytophatora parasitica:genetic nature and use in hybrid deterrnination[J].Phytopathology.80:1414-1421.
[21]Goettel,M.S.,Leger,R.J.,Bhairi,S.,1990.Pathogenicity and growth of metarhizium anisopliae stably transformed to benomyl resistance[J].Curr.Genet.17:129-1321.
[22]Yang Qian.,2000.Advanced Study on Plant Pest Biological Control[M].Harbin:Heilongjiang Science and Technology Press.
[23]Shew H D.In vitro growth response of Phytophthora parasitica var.nicotianae isolates to metalaxyl[J].Plant Disease,1984,68:764-766.
[24]Shew H D.Response of Phytophthora parasitica var.nicotianae isolates to metalaxyl exposure[J].Plant Disease,1985,69:559-562.
[25]王革,郑小波,陆家云等,云南烟草黑胫病菌对甲霜灵抗性的检测[J].南京农业大学学报,1997,20(4):105-107.
[26]袁宗胜,张广民,刘延荣等,烟草黑胫病菌对甲霜灵的敏感性测定[J].中国烟草科学,2001,4:9-12.
[27]Ferrin D M,Kabashima J N.In vitro insensitivity to metalaxyl of isolates of Phytophthora citricola and Phytophthora parasitica from ornamental hosts in South California[J].Plant Disease,1991,75:1041-1044.
[28]方中达.植病研究方法[M].北京:中国农业出版社,1998.208
[29]董汉松,曲建军,王智发.烟草病原真菌研究中的-种选择分离技术[J].中国烟草,1991,(3):1-5
[30]唐启义,冯明光.实用统计分析及其DPS数据处理系统[M].北京:科学出版社,2002.648
[31]郑小波。疫霉菌及其研究技术[M].北京:中匡农业出版社,P1-123,1997,
[34]马国胜,高智谋,陈娟.烟草黑胫病菌研究进展(Ⅲ)[J].烟草科技,2004,2:44-48.
[35]周明国.植物病原菌抗药性[J].农药市场信息,2001,(8):23-26.
[36]闫秀琴,刘慧平,韩巨才.我国植物病原菌抗药性的研究进展[J].农药,2001,40(12).4-6.
[37]刘波,刘经芬,叶钟音.灰霉病菌对苯并咪唑类杀菌剂的抗药性监测及抗性生理的研究[J].莱阳农学院学报,2003(3):217-222.
[38]罗汝南,黄健坤,戚佩坤.柑橘绿霉对苯并咪唑类杀菌剂抗药性的研究[J].植物保护学报,1985,12(4):269-273.
[39]李宝笃 李金玉.柑桔青霉病菌对多菌灵的抗药性检测[J],植物保护学报,1993,20(2):168-169.
[40]叶钟音,周明国.江淮地区小麦赤霉病对多菌灵耐药性的测定[J].植物保护学报,1985,12(3):188-190.
[41]ZHOU Ming-guo(周明国).Fungicide resistance of plant pathogens(植物病原物抗药性)[A].Zhou Ming-guo(周明国).Chemical Control of Plant Disease in China(中国植 物病害化学防治研究)[M].Beijing(北京)Chinese Agricultural Science and Technology Press(中国农业科技出版社),1998.50-61.
[42]刘英华,王开运,姜兴印等.植物病原菌抗性生物学及生化机制研究进展[J].世界农药,2004,26(3):16-18.
[43]杨谦.植物病原菌抗药性分子生物学[M].北京:科学出版社,2003.
[44]黄大肪,王侠,周淑芝.甜菜褐斑病菌对笨并咪唑类杀菌剂抗药性研究[J].植物保护学报,1982.9(2):131-135.
[45]李宝笃,沈崇尧.植物病原真菌对杀菌剂的抗性及对策[J].植物病理报.1994,24(3),193-196.
[46]张承来,欧晓明.植物病原物对杀菌剂的抗药性机制概述[J].湖南化工,2000,30(5)7-10.
[47]蔡-江.植物病原菌的抗药性与对策[J].农药,1984(1):22-25.
[48]Daggett S S,Gotz E,Therrien C D.Phenotypic changes in population of Phytophthora infestans from eastern Germany[J].Phytopathology,1993,83:319-323.
[49]孙大业,郭艳林,马力耕等.细胞信号转导[M].北京:科学出版社,2001:136-156.
[50]周嘉平等.烟草抗黑胫病突变体的细胞筛选[J].遗传学报,1990,17(3):180-188.
[51]王荔,等.运用致病毒素筛选抗烟草黑胫病的细胞突变体[J].云南农业大学学报,1999,14(1):16-21
[52]朱振元等.化学合成寡糖诱导烟草抗黑胫病的初步研究[J].植物病理学报,2004,34(3):231-236.
[53]杨献营.烟草病害防治的新途径--诱导抗病性的研究进展[J].中国烟草,1993,(4):29-31.
[54]方敦煌等.烟草黑胫病菌的拮抗根际细菌的筛选[J].云南农业大学学报,2001,16(2):93-97.
[55]北京农业大学植保系植物生态病理教研室编译.植物根际生态学与根病生物防治进展[M].北京:中国人民大学出版社,1991.122-129.
[56]朱贤朝,郭振业,刘保安.山东省烟草黑胫病生理小种的研究[J].植物病理学报,1987,17(2):90-95.
[12]Cui,W.,Ross,E.B.,Matthew,D.T.,el al.,2002.An osmosensing histidine kinase mediates dicarboximide fungicide resistance in Botryotinia fuckeliana(Botrytis cinerea)[J].Fungal Genetics and Biology.36:187-198.
[13]Takayuki,M.,Kaori,K.,Tomohiro,O.,et al.,2005.A two-component histidine kinase of the rice blast fungus is involved in osmotic stress response and fungicide action[J].Fungal Genetics and Biology.42:200-212.
[14]Koenraadt,H.,Somerville,S.C.,Jones,A.L.,1992.Characterization of mutations in the beta-tubulin gene of benomyl-resistant field strains of Venturia inaequalis and other plant pathogenic fungi[J],phytopathology.82(11):1348-1354.
[15]陆悦健,周明国,叶钟音等,抗苯并咪唑的小麦赤霉病菌β-tubulin基因序列分析与特性研究[J].植物病理学报.2000.30(1):30-34.
[16]Kawchuk,L.M.,Hutchison,L.J.,Verhaeghe,C.A.,et al.,2002.Isolation of the β-tubulin gene and characterization of thiabendazole resistance in Gibberella.pulicaris[J].Can.J.Plant Pathol.24:233-238.
[17]林孔勋主编,杀菌剂毒理学[M].北京:中国农业出版社.1995.101-140.
[18]Fabritus,A.L.,Shattock,R.C.,Judelson,H.S.,1997.Genetic analysis of metalaxyl in sensitivity loci in Phytophthora infestants using linked DNA markers[J].Phytopathology.87:1034-1040.
[19]Shaw,D.S.,Shattock,R.C.,1991.Genetics of Phytophthora infestants:The Mendelian approach[C].In Phytophthora pp.218-230 British Mycological Society,Cambridge Univ.Press,Cambridge.
[20]Chang,T.T.,Ko,W.H.,1990.Resistance to fungicide and antibiotics in Phytophatora parasitica:genetic nature and use in hybrid deterrnination[J].Phytopathology.80:1414-1421.
[21]Goettel,M.S.,Leger,R.J.,Bhairi,S.,1990.Pathogenicity and growth of metarhizium anisopliae stably transformed to benomyl resistance[J].Curr.Genet.17:129-1321.
[22]Yang Qian.,2000.Advanced Study on Plant Pest Biological Control[M].Harbin:Heilongjiang Science and Technology Press.
[23]Shew H D.In vitro growth response of Phytophthora parasitica var.nicotianae isolates to metalaxyl[J].Plant Disease,1984,68:764-766.
[24]Shew H D.Response of Phytophthora parasitica var.nicotianae isolates to metalaxyl exposure[J].Plant Disease,1985,69:559-562.
[25]王革,郑小波,陆家云等,云南烟草黑胫病菌对甲霜灵抗性的检测[J].南京农业大学学报,1997,20(4):105-107.
[26]袁宗胜,张广民,刘延荣等,烟草黑胫病菌对甲霜灵的敏感性测定[J].中国烟草科学,2001,4:9-12.
[27]Ferrin D M,Kabashima J N.In vitro insensitivity to metalaxyl of isolates of Phytophthora citricola and Phytophthora parasitica from ornamental hosts in South California[J].Plant Disease,1991,75:1041-1044.
[28]方中达.植病研究方法[M].北京:中国农业出版社,1998.208
[29]董汉松,曲建军,王智发.烟草病原真菌研究中的-种选择分离技术[J].中国烟草,1991,(3):1-5
[30]唐启义,冯明光.实用统计分析及其DPS数据处理系统[M].北京:科学出版社,2002.648
[31]郑小波。疫霉菌及其研究技术[M].北京:中匡农业出版社,P1-123,1997,
[34]马国胜,高智谋,陈娟.烟草黑胫病菌研究进展(Ⅲ)[J].烟草科技,2004,2:44-48.
[35]周明国.植物病原菌抗药性[J].农药市场信息,2001,(8):23-26.
[36]闫秀琴,刘慧平,韩巨才.我国植物病原菌抗药性的研究进展[J].农药,2001,40(12).4-6.
[37]刘波,刘经芬,叶钟音.灰霉病菌对苯并咪唑类杀菌剂的抗药性监测及抗性生理的研究[J].莱阳农学院学报,2003(3):217-222.
[38]罗汝南,黄健坤,戚佩坤.柑橘绿霉对苯并咪唑类杀菌剂抗药性的研究[J].植物保护学报,1985,12(4):269-273.
[39]李宝笃 李金玉.柑桔青霉病菌对多菌灵的抗药性检测[J],植物保护学报,1993,20(2):168-169.
[40]叶钟音,周明国.江淮地区小麦赤霉病对多菌灵耐药性的测定[J].植物保护学报,1985,12(3):188-190.
[41]ZHOU Ming-guo(周明国).Fungicide resistance of plant pathogens(植物病原物抗药性)[A].Zhou Ming-guo(周明国).Chemical Control of Plant Disease in China(中国植 物病害化学防治研究)[M].Beijing(北京)Chinese Agricultural Science and Technology Press(中国农业科技出版社),1998.50-61.
[42]刘英华,王开运,姜兴印等.植物病原菌抗性生物学及生化机制研究进展[J].世界农药,2004,26(3):16-18.
[43]杨谦.植物病原菌抗药性分子生物学[M].北京:科学出版社,2003.
[44]黄大肪,王侠,周淑芝.甜菜褐斑病菌对笨并咪唑类杀菌剂抗药性研究[J].植物保护学报,1982.9(2):131-135.
[45]李宝笃,沈崇尧.植物病原真菌对杀菌剂的抗性及对策[J].植物病理报.1994,24(3),193-196.
[46]张承来,欧晓明.植物病原物对杀菌剂的抗药性机制概述[J].湖南化工,2000,30(5)7-10.
[47]蔡-江.植物病原菌的抗药性与对策[J].农药,1984(1):22-25.
[48]Daggett S S,Gotz E,Therrien C D.Phenotypic changes in population of Phytophthora infestans from eastern Germany[J].Phytopathology,1993,83:319-323.
[49]孙大业,郭艳林,马力耕等.细胞信号转导[M].北京:科学出版社,2001:136-156.
[50]周嘉平等.烟草抗黑胫病突变体的细胞筛选[J].遗传学报,1990,17(3):180-188.
[51]王荔,等.运用致病毒素筛选抗烟草黑胫病的细胞突变体[J].云南农业大学学报,1999,14(1):16-21
[52]朱振元等.化学合成寡糖诱导烟草抗黑胫病的初步研究[J].植物病理学报,2004,34(3):231-236.
[53]杨献营.烟草病害防治的新途径--诱导抗病性的研究进展[J].中国烟草,1993,(4):29-31.
[54]方敦煌等.烟草黑胫病菌的拮抗根际细菌的筛选[J].云南农业大学学报,2001,16(2):93-97.
[55]北京农业大学植保系植物生态病理教研室编译.植物根际生态学与根病生物防治进展[M].北京:中国人民大学出版社,1991.122-129.
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