云南玉溪地区烟田根结线虫的SCAR鉴定、分布及其生物防治研究
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摘要
云南玉溪地区是我国的烟草主产区,根结线虫病是烟草生产中的重要地下病害,危害烟草植株生长,降低产量。本研究运用SCAR-PCR方法对云南玉溪地区根结线虫进行分子鉴定,并分析不同根结线虫种类在各县区的分布情况;同时在澄江、易门两县开展利用昆虫病原线虫对根结线虫的田间防治试验。结果表明,玉溪地区主要的优势种群为花生根结线虫Meloidogyne arenaria,在所有区县均有分布,爪哇根结线虫M. javanica为次要种群,南方根结线虫M. incognita属于零星分布;田间施加昆虫病原线虫对烟草根结线虫病具有显著的抑制作用。研究结果可为烟草根结线虫病的绿色防治提供数据支持。
Yuxi is the main tobacco producing area in China. Root-knot nematode disease is the important underground diseases in tobacco production, which has endangered tobacco plant growth and reduced tobacco yield. In this study, the SCAR-PCR method was used to identify the root-knot nematodes in Yuxi, Yunnan Province, and the distribution of different root-knot nematodes in each county was analyzed.The field experiment was performed in Chengjiang and Yimen Counties in Yuxi. The results showed that Meloidogyne arenaria was the dominant species in Yuxi region, which was distributed in all counties. M. javanica was the minor species, and M. incognita was rare in Yuxi. And the application of entomopathogenic nematodes in the field could significantly inhibit the occurrence of root-knot nematodes in tobacco. The results above could provide data support for the green control of tobacco root-knot nematode disease.
Yuxi is the main tobacco producing area in China. Root-knot nematode disease is the important underground diseases in tobacco production, which has endangered tobacco plant growth and reduced tobacco yield. In this study, the SCAR-PCR method was used to identify the root-knot nematodes in Yuxi, Yunnan Province, and the distribution of different root-knot nematodes in each county was analyzed.The field experiment was performed in Chengjiang and Yimen Counties in Yuxi. The results showed that Meloidogyne arenaria was the dominant species in Yuxi region, which was distributed in all counties. M. javanica was the minor species, and M. incognita was rare in Yuxi. And the application of entomopathogenic nematodes in the field could significantly inhibit the occurrence of root-knot nematodes in tobacco. The results above could provide data support for the green control of tobacco root-knot nematode disease.
引文
[1]Abad P,Gouzy J,Aury J M,et al.Genome sequence of the metazoan plant-parasitic nematode Meloidogyne incognita[J].Nature Biotechnology,2008,26(8):909-915.
[2]Bleve-Zacheo T,Melillo M T,Castagnone-Sereno P.The contribution of biotechnology to root-knot nematode control in tomato plants[J].Pest Technology,2007,1(1):1-16.
[3]朱斌,王军,乔永信.陕南烟草根结线虫病发病规律研究初报[J].陕西农业科学,2005(1):6-8.ZHU Bin,WANG Jun,QIAO Yong-xin.Preliminary report on the pathogenesis of tobacco root-knot nematode disease in southern Shaanxi[J].Shaanxi Agricultural Science,2005(1):6-8.
[4]杨铭,秦西云,段玉琪,等.云南烟草根结线虫病发生及防治研究[J].云南农业大学学报,1995,10(2):10-15.YANG Ming,QIN Xi-yun,DUAN Yu-qi,et al.Occurrence and control of tobacco root-knot nematode disease in Yunnan[J].Journal of Yunnan Agricultural University,1995,10(2):10-15.
[5]张广民,王兴利.烟草根结线虫病综合防治技术的研究[J].中国烟草科学,1996(4):1-6.ZHANG Guang-min,WANG Xing-li.Study on integrated control technology of tobacco root-knot nematode disease[J].Chinese Tobacco Science,1996(4):1-6.
[6]Sasser J N.Identification and host-parasite relationships of certain root-knot nematodes(Meloidogyne spp.)[M].College Park:Agricultural Experiment Station,University of Marylan,1954.
[7]Gamel S,Huchet E,Le R N A,et al.Assessment of PCR-based tools for the specific identification of some temperate Meloidogyne species including M.chitwoodi,M.fallax and M.minor[J].European Journal of Plant Pathology,2014,138(4):807-817.
[8]宋志强.江苏省蔬菜根结线虫的分布调查、种类鉴定及定量检测技术研究[D].南京:南京农业大学,2013:32-34.SONG Zhi-qiang.Distribution survey,species identification and quantitative detection of root-knot nematodes on vegetables in Jiangsu Province[D].Nanjing:Nanjing Agricultural University,2013:32-34.
[9]Georgis R,Manweiler S A.Entomopathogenic nematodes:A developing biological control technology[J].Agricultural Zoology Reviews,1994,6:63-94.
[10]宋洁,许艳丽,姚钦.昆虫病原线虫共生细菌对植物寄生线虫卵的毒性研究[J].大豆科学,2014,33(6):896-899.SONG Jie,XU Yan-li,YAO Qin.Toxicity of entomopathogenic nematode symbiotic bacteria to eggs of plant parasitic nematodes[J].Soybean Science,2014,33(6):896-899.
[11]Molina J P,Dolinski C,Souza R M,et al.Effect of entomopathogenic nematodes(Rhabditida:Steinernematidae and Heterorhabditidae)on Meloidogyne mayaguensis Rammah and Hirschmann(Tylenchida:Meloidoginidae)infection in tomato plants[J].Journal of Nematology,2007,39(4):338-342.
[12]Valle E E D,Doucet M E.Effects of Galleria mellonella cadavers infected with Heterorhabditis bacteriophora and Steinernema rarum,their macerates and dead infective juveniles on Meloidogyne javanica suppression[J].Revista De La Facultad De Ciencias Agrarias,2014,46(1):205-211.
[13]Valle E E D,Lax P,Rondán Due?as J,et al.Effects of insect cadavers infected by Heterorhabditis bacteriophora and Steinernema diaprepesi on Meloidogyne incognita parasitism in pepper and summer squash plants[J].Ciencia E Investigación Agraria,2013,40(1):109-118.
[14]王宇坤,徐冰,王小平,等.昆虫病原线虫对烟草根结线虫及蚜虫病虫害以及蚜茧蜂寄生的影响[J].农业资源与环境学报,2017,34(3):293-298.WANG Yu-kun,XU Bing,WANG Xiao-ping,et al.Effects of entomopathogenic nematodes on the development of root-knot nematode and aphid,and on the parasitism of aphid parasitoid in tobacco[J].Journal of Agricultural Resources and Environment,2017,34(3):293-298.
[15]张瑞平,曾庆宾,余伟,等.烟草根结线虫病不同防控措施的田间筛选[J].中国烟草科学,2016,37(4):54-59.ZHANG Rui-ping,ZENG Qing-bin,YU Wei,et al.Field screening of different control measures for tobacco root knot nematode[J].Chinese Tobacco Science,2016,37(4):54-59.
[16]Meng Q P,Long H,Xu J H.PCR assays for rapid and sensitive identification of three major root-knot nematodes,Meloidogyne incognita,M.javanica and M.arenaria[J].Acta Phytopathologica Sinica,2004,34(3):204-210.
[17]Zijlstra C,Donkers-Venne D,Fargette M.Identification of Meloidogyne incognita,M.javanica and M.arenaria using sequence characterised amplified region(SCAR)based PCR assays[J].Nematology,2000,2(8):847-853.
[18]卓侃,廖金铃.植物线虫分子鉴定研究进展[J].植物保护,2015(6):1-8.ZHUO Kan,LIAO Jin-ling.Advances in molecular identification of plant nematodes[J].Plant Protection,2015(6):1-8.
[19]杨艳梅,梁艳,袁绍杰,等.云南省部分烟区根结线虫形态和分子鉴定[J].南方农业学报,2017,48(2):284-291.YANG Yan-mei,LIANG Yan,YUAN Shao-jie,et al.Morphological and molecular identification of root-knot nematodes in some tobacco areas of Yunnan Province[J].Southern Agricultural Journal,2017,48(2):284-291.
[20]A?t Hamza M,Ali N,Tavoillot J,et al.Diversity of root-knot nematodes in Moroccan olive nurseries and orchards:Does Meloidogyne javanica disperse according to invasion processes?[J].BMC Ecology,2017,17(1):41.
[21]喻盛甫,胡先奇,王扬.云南烟草根结线虫优势种群动态规律研究[J].云南农业大学学报,1998(1):52-58.YU Sheng-fu,HU Xian-qi,WANG Yang.Study on dynamic law of dominant population of tobacco root-knot nematode in Yunnan[J].Journal of Yunnan Agricultural University,1998(1):52-58.
[22]张喆.云南烟草根结线虫的种类鉴定[D].广州:仲恺农业工程学院,2016.ZHANG Zhe.Identification of tobacco root-knot nematodes in Yunnan[D].Guangzhou:Zhongkai University of Agriculture and Engineering,2016.
[23]Kepenekci I,Hazir S,Lewis E E.Evaluation of entomopathogenic nematodes and the supernatants of the in-vitro culture medium of their mutualistic bacteria for the control of the root-knot nematodes Meloidogyne incognita and M.arenaria[J].Pest Management Science,2016,72(2):327-334.
[24]Sun Y,Bai G Y,Wang Y X,et al.The impact of Cu,Zn and Cr salts on the relationship between insect and plant parasitic nematodes:Areduction in biocontrol efficacy[J].Applied Soil Ecology,2016,107(2):108-115.
[25]Bi Y H,Gao C Z,Yu Z G.Rhabdopeptides from Xenorhabdus budapestensis SN84 and their nematicidal activities against Meloidogyne incognita[J].Journal of Agricultural&Food Chemistry,2018,66(15):3833-3836.
[26]An R,Orellana D,Phelan L P,et al.Entomopathogenic nematodes induce systemic resistance in tomato against Spodoptera exigua,Bemisia tabaci,and Pseudomonas syringae[J].Biological Control,2016,93:24-29.
[27]Srinivasan J,Kaplan F,Ajredini R,et al.A blend of small molecules regulates both mating and development in Caenorhabditis elegans[J].Nature,2008,454(7208):1115-1118.
[28]Macosko E Z,Pokala N,Feinberg E H,et al.A hub-and-spoke circuit drives pheromone attraction and social behaviour in C.elegans[J].Nature,2009,458(7242):1171-1175.
[29]Kaplan F,Alborn H T,von Reuss S H,et al.Interspecific nematode signals regulate dispersal behavior[J].PLoS ONE,2012,7(6):1398-1408.
[30]Lacey L A,Granatstein D,Arthurs S P,et al.Use of entomopathogenic nematodes(Steinernematidae)in conjunction with mulches for control of overwintering coding moth(Lepidoptera:Tortricidae)[J].Journal of Entomological Science,2006,41(2):107-119.
[31]白光瑛,沈广爽,马海鲲,等.施用方式和土壤深度对昆虫病原线虫越冬的影响[J].农业环境科学学报,2015,34(11):2162-2168.BAI Guang-ying,SHEN Guang-shuang,MA Hai-kun,et al.Effects of application approach and soil depth on overwintering persistence of entomopathogenic nematodes[J].Journal of Agro-Environment Science,2015,34(11):2162-2168.
[32]Shapiro-Ilan D I,Hazir S,Lete L.Viability and virulence of entomopathogenic nematodes exposed to ultraviolet radiation[J].Journal of Nematology,2015,47(3):184-189.
[33]王鑫鹏,王从丽,李春杰.昆虫病原线虫共生细菌对南方根结线虫卵孵化的影响[J].土壤与作物,2017,6(4):256-262.WANG Xin-peng,WANG Cong-li,LI Chun-jie.Effect of symbiotic bacteria of entomopathogenic nematodes on egg hatching of Meloidogyne incognita[J].Soils and Crops,2017,6(4):256-262.
[2]Bleve-Zacheo T,Melillo M T,Castagnone-Sereno P.The contribution of biotechnology to root-knot nematode control in tomato plants[J].Pest Technology,2007,1(1):1-16.
[3]朱斌,王军,乔永信.陕南烟草根结线虫病发病规律研究初报[J].陕西农业科学,2005(1):6-8.ZHU Bin,WANG Jun,QIAO Yong-xin.Preliminary report on the pathogenesis of tobacco root-knot nematode disease in southern Shaanxi[J].Shaanxi Agricultural Science,2005(1):6-8.
[4]杨铭,秦西云,段玉琪,等.云南烟草根结线虫病发生及防治研究[J].云南农业大学学报,1995,10(2):10-15.YANG Ming,QIN Xi-yun,DUAN Yu-qi,et al.Occurrence and control of tobacco root-knot nematode disease in Yunnan[J].Journal of Yunnan Agricultural University,1995,10(2):10-15.
[5]张广民,王兴利.烟草根结线虫病综合防治技术的研究[J].中国烟草科学,1996(4):1-6.ZHANG Guang-min,WANG Xing-li.Study on integrated control technology of tobacco root-knot nematode disease[J].Chinese Tobacco Science,1996(4):1-6.
[6]Sasser J N.Identification and host-parasite relationships of certain root-knot nematodes(Meloidogyne spp.)[M].College Park:Agricultural Experiment Station,University of Marylan,1954.
[7]Gamel S,Huchet E,Le R N A,et al.Assessment of PCR-based tools for the specific identification of some temperate Meloidogyne species including M.chitwoodi,M.fallax and M.minor[J].European Journal of Plant Pathology,2014,138(4):807-817.
[8]宋志强.江苏省蔬菜根结线虫的分布调查、种类鉴定及定量检测技术研究[D].南京:南京农业大学,2013:32-34.SONG Zhi-qiang.Distribution survey,species identification and quantitative detection of root-knot nematodes on vegetables in Jiangsu Province[D].Nanjing:Nanjing Agricultural University,2013:32-34.
[9]Georgis R,Manweiler S A.Entomopathogenic nematodes:A developing biological control technology[J].Agricultural Zoology Reviews,1994,6:63-94.
[10]宋洁,许艳丽,姚钦.昆虫病原线虫共生细菌对植物寄生线虫卵的毒性研究[J].大豆科学,2014,33(6):896-899.SONG Jie,XU Yan-li,YAO Qin.Toxicity of entomopathogenic nematode symbiotic bacteria to eggs of plant parasitic nematodes[J].Soybean Science,2014,33(6):896-899.
[11]Molina J P,Dolinski C,Souza R M,et al.Effect of entomopathogenic nematodes(Rhabditida:Steinernematidae and Heterorhabditidae)on Meloidogyne mayaguensis Rammah and Hirschmann(Tylenchida:Meloidoginidae)infection in tomato plants[J].Journal of Nematology,2007,39(4):338-342.
[12]Valle E E D,Doucet M E.Effects of Galleria mellonella cadavers infected with Heterorhabditis bacteriophora and Steinernema rarum,their macerates and dead infective juveniles on Meloidogyne javanica suppression[J].Revista De La Facultad De Ciencias Agrarias,2014,46(1):205-211.
[13]Valle E E D,Lax P,Rondán Due?as J,et al.Effects of insect cadavers infected by Heterorhabditis bacteriophora and Steinernema diaprepesi on Meloidogyne incognita parasitism in pepper and summer squash plants[J].Ciencia E Investigación Agraria,2013,40(1):109-118.
[14]王宇坤,徐冰,王小平,等.昆虫病原线虫对烟草根结线虫及蚜虫病虫害以及蚜茧蜂寄生的影响[J].农业资源与环境学报,2017,34(3):293-298.WANG Yu-kun,XU Bing,WANG Xiao-ping,et al.Effects of entomopathogenic nematodes on the development of root-knot nematode and aphid,and on the parasitism of aphid parasitoid in tobacco[J].Journal of Agricultural Resources and Environment,2017,34(3):293-298.
[15]张瑞平,曾庆宾,余伟,等.烟草根结线虫病不同防控措施的田间筛选[J].中国烟草科学,2016,37(4):54-59.ZHANG Rui-ping,ZENG Qing-bin,YU Wei,et al.Field screening of different control measures for tobacco root knot nematode[J].Chinese Tobacco Science,2016,37(4):54-59.
[16]Meng Q P,Long H,Xu J H.PCR assays for rapid and sensitive identification of three major root-knot nematodes,Meloidogyne incognita,M.javanica and M.arenaria[J].Acta Phytopathologica Sinica,2004,34(3):204-210.
[17]Zijlstra C,Donkers-Venne D,Fargette M.Identification of Meloidogyne incognita,M.javanica and M.arenaria using sequence characterised amplified region(SCAR)based PCR assays[J].Nematology,2000,2(8):847-853.
[18]卓侃,廖金铃.植物线虫分子鉴定研究进展[J].植物保护,2015(6):1-8.ZHUO Kan,LIAO Jin-ling.Advances in molecular identification of plant nematodes[J].Plant Protection,2015(6):1-8.
[19]杨艳梅,梁艳,袁绍杰,等.云南省部分烟区根结线虫形态和分子鉴定[J].南方农业学报,2017,48(2):284-291.YANG Yan-mei,LIANG Yan,YUAN Shao-jie,et al.Morphological and molecular identification of root-knot nematodes in some tobacco areas of Yunnan Province[J].Southern Agricultural Journal,2017,48(2):284-291.
[20]A?t Hamza M,Ali N,Tavoillot J,et al.Diversity of root-knot nematodes in Moroccan olive nurseries and orchards:Does Meloidogyne javanica disperse according to invasion processes?[J].BMC Ecology,2017,17(1):41.
[21]喻盛甫,胡先奇,王扬.云南烟草根结线虫优势种群动态规律研究[J].云南农业大学学报,1998(1):52-58.YU Sheng-fu,HU Xian-qi,WANG Yang.Study on dynamic law of dominant population of tobacco root-knot nematode in Yunnan[J].Journal of Yunnan Agricultural University,1998(1):52-58.
[22]张喆.云南烟草根结线虫的种类鉴定[D].广州:仲恺农业工程学院,2016.ZHANG Zhe.Identification of tobacco root-knot nematodes in Yunnan[D].Guangzhou:Zhongkai University of Agriculture and Engineering,2016.
[23]Kepenekci I,Hazir S,Lewis E E.Evaluation of entomopathogenic nematodes and the supernatants of the in-vitro culture medium of their mutualistic bacteria for the control of the root-knot nematodes Meloidogyne incognita and M.arenaria[J].Pest Management Science,2016,72(2):327-334.
[24]Sun Y,Bai G Y,Wang Y X,et al.The impact of Cu,Zn and Cr salts on the relationship between insect and plant parasitic nematodes:Areduction in biocontrol efficacy[J].Applied Soil Ecology,2016,107(2):108-115.
[25]Bi Y H,Gao C Z,Yu Z G.Rhabdopeptides from Xenorhabdus budapestensis SN84 and their nematicidal activities against Meloidogyne incognita[J].Journal of Agricultural&Food Chemistry,2018,66(15):3833-3836.
[26]An R,Orellana D,Phelan L P,et al.Entomopathogenic nematodes induce systemic resistance in tomato against Spodoptera exigua,Bemisia tabaci,and Pseudomonas syringae[J].Biological Control,2016,93:24-29.
[27]Srinivasan J,Kaplan F,Ajredini R,et al.A blend of small molecules regulates both mating and development in Caenorhabditis elegans[J].Nature,2008,454(7208):1115-1118.
[28]Macosko E Z,Pokala N,Feinberg E H,et al.A hub-and-spoke circuit drives pheromone attraction and social behaviour in C.elegans[J].Nature,2009,458(7242):1171-1175.
[29]Kaplan F,Alborn H T,von Reuss S H,et al.Interspecific nematode signals regulate dispersal behavior[J].PLoS ONE,2012,7(6):1398-1408.
[30]Lacey L A,Granatstein D,Arthurs S P,et al.Use of entomopathogenic nematodes(Steinernematidae)in conjunction with mulches for control of overwintering coding moth(Lepidoptera:Tortricidae)[J].Journal of Entomological Science,2006,41(2):107-119.
[31]白光瑛,沈广爽,马海鲲,等.施用方式和土壤深度对昆虫病原线虫越冬的影响[J].农业环境科学学报,2015,34(11):2162-2168.BAI Guang-ying,SHEN Guang-shuang,MA Hai-kun,et al.Effects of application approach and soil depth on overwintering persistence of entomopathogenic nematodes[J].Journal of Agro-Environment Science,2015,34(11):2162-2168.
[32]Shapiro-Ilan D I,Hazir S,Lete L.Viability and virulence of entomopathogenic nematodes exposed to ultraviolet radiation[J].Journal of Nematology,2015,47(3):184-189.
[33]王鑫鹏,王从丽,李春杰.昆虫病原线虫共生细菌对南方根结线虫卵孵化的影响[J].土壤与作物,2017,6(4):256-262.WANG Xin-peng,WANG Cong-li,LI Chun-jie.Effect of symbiotic bacteria of entomopathogenic nematodes on egg hatching of Meloidogyne incognita[J].Soils and Crops,2017,6(4):256-262.
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