草地贪夜蛾莱氏绿僵菌的分离鉴定
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摘要
草地贪夜蛾Spodoptera frugiperda(J.E.Smith)是2019年入侵我国的一种迁飞性害虫,已成为严重影响我国玉米生产的重要害虫。目前国内关于草地贪夜蛾昆虫病原真菌的研究报道很少。2019年6月,在云南省曲靖市沾益县安东村玉米田调查发现了被虫生真菌感染的草地贪夜蛾幼虫。经室内分离培养,结合形态和ITS序列相似性分析鉴定确认,感染草地贪夜蛾幼虫病原性真菌为莱氏绿僵菌Metarhizium rileyi,编号为ZYSP190701菌株。调查表明,2019年7月17日田间草地贪夜蛾的莱氏绿僵菌感染率为15.46%±2.52%,8月2日感染率达29.83%±13.71%。用莱氏绿僵菌ZYSP190701菌株1×10~8个/mL孢子浓度接种草地贪夜蛾3龄幼虫后第7天时草地贪夜蛾的感染死亡率高达100%。上述结果表明,莱氏绿僵菌ZYSP190701对草地贪夜蛾有良好的生防潜力,具有草地贪夜蛾生防菌剂开发应用的前景。
Spodoptera frugiperda(J.E.Smith) is an outbreak invasive pest, especially for maize. To date, little information about entomopathogenic fungi of Spodoptera frugiperda is available in China. In this study, fungi infected S.frugiperda larvae were found in Andong village, Zhanyi county, Qujing city, Yunnan province. The fungi were isolated from the infected cadaver of S.frugiperda, and then the species was identified based on the morphological and molecular biology, the virulence of the fungi against the larvae of S.frugiperda was tested. Results showed that this strain was identified as Metarhizium rileyi(No. ZYSP190701). The infection rate of S.frugiperda was 15.46%±2.52% on 17 th, July and 29.83%±13.71% on 2 nd, August, respectively. The cumulative mortality of the 3 rd instar larvae of S.frugiperda 7 d after immersion inoculation at 1 ×10~8 conidia/mL was 100%. Our results demonstrated that M.rileyi ZYSP190701 is a promising candidate for S.frugiperda biocontrol in field.
Spodoptera frugiperda(J.E.Smith) is an outbreak invasive pest, especially for maize. To date, little information about entomopathogenic fungi of Spodoptera frugiperda is available in China. In this study, fungi infected S.frugiperda larvae were found in Andong village, Zhanyi county, Qujing city, Yunnan province. The fungi were isolated from the infected cadaver of S.frugiperda, and then the species was identified based on the morphological and molecular biology, the virulence of the fungi against the larvae of S.frugiperda was tested. Results showed that this strain was identified as Metarhizium rileyi(No. ZYSP190701). The infection rate of S.frugiperda was 15.46%±2.52% on 17 th, July and 29.83%±13.71% on 2 nd, August, respectively. The cumulative mortality of the 3 rd instar larvae of S.frugiperda 7 d after immersion inoculation at 1 ×10~8 conidia/mL was 100%. Our results demonstrated that M.rileyi ZYSP190701 is a promising candidate for S.frugiperda biocontrol in field.
引文
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[19] 程东美,洪婉雯,孙辉,等.草地贪夜蛾幼虫僵虫发生率调查及致病菌分离鉴定[J].环境昆虫学报:1-9.(2019-07-18).http://kns.cnki.net/kcms/detail/ 44.1640.Q.20190717.1722.002.html.
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[21] 杜广祖,郑亚强,苏锦尧,等.云南莱氏野村菌田间自然发生流行动态研究[J].环境昆虫学报,2018,40(5):160-168.
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[23] SRISUKCHAYAKUL P,WIWAT C,PANTUWATANA S.Studies on the pathogenesis of the local isolates of Nomuraea rileyi against Spodoptera litura [J].Science Asia,2005,31(3):273-276.
[24] IGNOFFO C M,PUTTLER B,HOSTETTER D L,et al.Susceptibility of the cabbage looper,Trichoplusia ni and the velvetbean caterpillar,Anticarsia gemmatalis,to several isolates of the entomopathogenic fungus Nomuraea rileyi [J].Journal of Invertebrate Pathology,1976,28(21):259-262.
[25] AMBETHGAR V.Potential of entomopathogenic fungi in insecticide resistance management(IRM):a review [J].Journal of Biopesticides,2009,2(2):177-193.
[26] 蒲蛰龙,李增智.昆虫真菌学[M].合肥:安徽科学技术出版社,1996:553-554.
[27] 陈吉良,黄小龙,吴安迪,等.一种快速高效提取病原真菌DNA作为PCR模板的方法[J].菌物学报,2011,30(1):147-149.
[28] KUMAR S,TAMURA K,NEI M.MEGA3:Integrated software for molecular evolutionary genetics analysis and sequence alignment [J].Briefings in Bioinformatics,2004,5(2):150-163.
[29] BOSA C F,CHáVES D,TORRES L,et al.Evaluación de aislamientos nativos de Nomuraea rileyi para el control de Spodoptera frugiperda (Lepidoptera:Noctuidae)[J].Revista Colombiana de Entomología,2004,30(1):93-97.
[2] SPARKS A N.A review of the biology of the fall armyworm[J].Florida Entomologist,1979,62(2):82-87.
[3] TODD E L,POOLE R W.Keys and illustrations for the armyworm moths of the noctuid genus Spodoptera Guenée from the Western Hemisphere [J].Annals of the Entomological Society of America,1980,73(6):722-738.
[4] 杨学礼,刘永昌,罗茗钟,等.云南省江城县首次发现迁入我国西南地区的草地贪夜蛾[J].云南农业,2019(1):72.
[5] 卢辉,唐继洪,吕宝乾,等.草地贪夜蛾的生物防治及潜在入侵风险[J].热带作物学报,2019,40(6):1237-1244.
[6] BLANCO C A,PORTILLA M,JURAT-FUENTES J L,et al.Susceptibility of isofamilies of Spodoptera frugiperda (Lepidoptera:Noctuidae) to Cry1Ac and Cry1Fa proteins of Bacillus thuringiensis [J].Southwest Entomology,2010,35(3):409-415.
[7] LEóN-GARCíA I,RODRíGUEZ-LEYVA E,ORTEGA-ARENAS L D,et al.Susceptibilidad de Spodoptera frugiperda (J.E.Smith) (Lepidoptera:Noctuidae) an insecticidas asociada a césped en Quintana Roo,México[J].Agrociencia,2012,46:279-287.
[8] 唐艺婷,李玉艳,刘晨曦,等.蠋蝽对草地贪夜蛾的捕食能力评价和捕食行为观察[J/OL].植物保护,2019,45(4):65-68.
[9] 唐艺婷,王孟卿,陈红印,等.益蝽对草地贪夜蛾捕食能力评价和捕食行为观察[J/OL].中国生物防治学报:1-7[2019-09-03].https://doi.org/10.16409/j.cnki.2095-039x.2019.04.005.
[10] 陈壮美,赵琳超,刘航,等.斯氏侧沟茧蜂对草地贪夜蛾幼虫的寄生行为及寄生效应[J].植物保护:1-6[2019-09-03].https://doi.org/10.16688/j.zwbh.2019341.
[11] SHAH P A,PELL J K.Entomopathogenic fungi as biological control agents[J].Applied Microbiology and Biotechnology,2003,61(5/6):413-423.
[12] FENG Mingguang,POPRAWSKI T J,KHACHATOURIANS G G.Production,formulation and application of the entomopathogenic fungus Beauveria bassiana for insect control:current status [J].Biocontrol Science and Technology,1994,4(1):3-34.
[13] FARIA M R D,WRAIGHT S P.Mycoinsecticides and Mycoacaricides:A comprehensive list with worldwide coverage and international classification of formulation types [J].Biological Control,2007,43(3):237-256.
[14] SUJEETH J A R P,SAHAYARAJ K.Basic and applied aspects of biopesticides [M].New Delhi:Springer,2015:31-46.
[15] 李增智.我国利用真菌防治害虫的历史、进展及现状[J].中国生物防治学报,2015,31(5):699-711.
[16] RIVERO-BORJA M,GUZMáN-FRANCO A W,RODRíGUEZ-LEYVA E,et al.Interaction of Beauveria bassiana and Metarhizium anisopliae with chlorpyrifos ethyl and spinosad in Spodoptera frugiperda larvae[J].Pest Management Science,2018,74 (9):2047-2052.
[17] GRIJALBA E P,ESPINEL C,CUARTAS P E,et al.Metarhizium rileyi biopesticide to control Spodoptera frugiperda:Stability and insecticidal activity under glasshouse conditions[J].Fungal Biology,2018,122(11):1069-1076.
[18] AKUTSE K S,KIMEMIA J W,EKESI S,et al.Ovicidal effects of entomopathogenic fungal isolates on the invasive fall armyworm Spodoptera frugiperda (Lepidoptera:Noctuidae)[J].Journal of Applied Entomology,2019,143(6):624-634.
[19] 程东美,洪婉雯,孙辉,等.草地贪夜蛾幼虫僵虫发生率调查及致病菌分离鉴定[J].环境昆虫学报:1-9.(2019-07-18).http://kns.cnki.net/kcms/detail/ 44.1640.Q.20190717.1722.002.html.
[20] KEPLER R M,HUMBER R A,BISCHOFF J F,et al.Clarification of generic and species boundaries for Metarhizium and related fungi through multigene phylogenetics [J].Mycologia,2014,106(4):811-829.
[21] 杜广祖,郑亚强,苏锦尧,等.云南莱氏野村菌田间自然发生流行动态研究[J].环境昆虫学报,2018,40(5):160-168.
[22] PERINOTTO W M S,TERRA A L M,ANGELO I C,et al.Nomuraea rileyi as biological control agents of Rhipicephalus microplus Tick [J].Parasitology Research,2012,111(4):1743-1748.
[23] SRISUKCHAYAKUL P,WIWAT C,PANTUWATANA S.Studies on the pathogenesis of the local isolates of Nomuraea rileyi against Spodoptera litura [J].Science Asia,2005,31(3):273-276.
[24] IGNOFFO C M,PUTTLER B,HOSTETTER D L,et al.Susceptibility of the cabbage looper,Trichoplusia ni and the velvetbean caterpillar,Anticarsia gemmatalis,to several isolates of the entomopathogenic fungus Nomuraea rileyi [J].Journal of Invertebrate Pathology,1976,28(21):259-262.
[25] AMBETHGAR V.Potential of entomopathogenic fungi in insecticide resistance management(IRM):a review [J].Journal of Biopesticides,2009,2(2):177-193.
[26] 蒲蛰龙,李增智.昆虫真菌学[M].合肥:安徽科学技术出版社,1996:553-554.
[27] 陈吉良,黄小龙,吴安迪,等.一种快速高效提取病原真菌DNA作为PCR模板的方法[J].菌物学报,2011,30(1):147-149.
[28] KUMAR S,TAMURA K,NEI M.MEGA3:Integrated software for molecular evolutionary genetics analysis and sequence alignment [J].Briefings in Bioinformatics,2004,5(2):150-163.
[29] BOSA C F,CHáVES D,TORRES L,et al.Evaluación de aislamientos nativos de Nomuraea rileyi para el control de Spodoptera frugiperda (Lepidoptera:Noctuidae)[J].Revista Colombiana de Entomología,2004,30(1):93-97.