入侵云南草地贪夜蛾的分子鉴定
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摘要
草地贪夜蛾Spodoptera frugiperda(J.E. Smith)是一种原产于美洲热带和亚热带地区的重要农业入侵害虫,目前已从缅甸进入我国云南西南部地区,且呈现蔓延之势。对入侵物种快速准确鉴定是早期预警和监测预防的关键。本研究采用分子标记手段,对采自云南5个县(市)的83份样品进行鉴定,通过PCR扩增细胞色素C氧化酶第Ⅰ亚基(COⅠ)和磷酸甘油醛异构酶(Tpi)两个基因片段并进行序列测定分析。结果表明,基于COⅠ基因片段的序列比对能够对草地贪夜蛾进行准确的物种鉴定,并且能够对其近源种甜菜夜蛾等进行鉴定。基于Tpi基因序列的特异单倍型位点比较分析,确认入侵我国的草地贪夜蛾均为"玉米型"。该研究为草地贪夜蛾的快速识别提供了方法支持,同时为进一步的虫源地解析和遗传溯源积累了宝贵的前期数据。
The fall armyworm, Spodoptera frugiperda(J.E. Smith), is an important agricultural invasive pest that is native to tropical and subtropical America. At present, the insect has crossed Myanmar and entered into southwestern Yunnan of China, and has the trend to overspread in the surrounding areas. Rapid and accurate identification is the key to early warning, monitoring and prevention of this invasive species. In this study, genetic methods based on two molecular markers, mitochondrial cytochrome oxidase subunit Ⅰ gene(CO Ⅰ) and triose-phosphate isomerase(Tpi), were used to characterize 83 samples collected from five different regions in Yunnan Province. The samples could be accurately identified to be S.frugiperda or close-related species such as S.exigua by sequence alignment based on CO Ⅰ gene fragment. Subpopulation of "corn strain" was confirmed by Tpi haplotype analysis. This study provides methodological support for rapid species identification of S.frugiperda, and provides early basic data for further origin analysis and genetic traceability.
The fall armyworm, Spodoptera frugiperda(J.E. Smith), is an important agricultural invasive pest that is native to tropical and subtropical America. At present, the insect has crossed Myanmar and entered into southwestern Yunnan of China, and has the trend to overspread in the surrounding areas. Rapid and accurate identification is the key to early warning, monitoring and prevention of this invasive species. In this study, genetic methods based on two molecular markers, mitochondrial cytochrome oxidase subunit Ⅰ gene(CO Ⅰ) and triose-phosphate isomerase(Tpi), were used to characterize 83 samples collected from five different regions in Yunnan Province. The samples could be accurately identified to be S.frugiperda or close-related species such as S.exigua by sequence alignment based on CO Ⅰ gene fragment. Subpopulation of "corn strain" was confirmed by Tpi haplotype analysis. This study provides methodological support for rapid species identification of S.frugiperda, and provides early basic data for further origin analysis and genetic traceability.
引文
[1] TODD E L, POOLE R W. Keys and illustrations for the army-worm moths of the noctuid genus Spodoptera Guenée from the Western Hemisphere [J]. Annals of the Entomological Society of America, 1980, 73(6): 722-738.
[2] 吴秋琳,姜玉英,吴孔明. 草地贪夜蛾缅甸虫源迁入中国的路径分析[J/OL]. 植物保护,2019,https://doi.org/10.16688/j.zwbh.2019047.
[3] SPARKS A N. A review of the biology of the fall armyworm[J]. Florida Entomologist, 1979,62(2): 82-87.
[4] MITCHELL E R, MCNEIL J N, WESTBROOK J K, et al. Seasonal periodicity of fall armyworm, (Lepidoptera: Noctuidae) in the Caribbean basin and northward to Canada[J]. Journal of Entomological Science, 1991, 26(1): 39-50.
[5] COCK M J W, BESEH P K, BUDDIE A G, et al. Molecular methods to detect Spodoptera frugiperda in Ghana, and implications for monitoring the spread of invasive species in developing countries [J]. Scientific Reports, 2017, 7(1): 4103.
[6] CHAPMAN D, PURSE B V, ROY H E, et al. Global trade networks determine the distribution of invasive non-native species[J].Global Ecology and Biogeography,2017,26(8):907-917.
[7] GOERGEN G, KUMAR P L, SANKUNG S B, et al. First report of outbreaks of the fall armyworm Spodoptera frugiperda (J E Smith) (Lepidoptera, Noctuidae), a new alien invasive pest in West and Central Africa [J/OL]. PLoS ONE, 2016, 11(10): e0165632.
[8] 郭井菲,赵建周,何康来. 警惕危险性害虫草地贪夜蛾入侵中国[J]. 植物保护,2018,44(6):1-10.
[9] SHARANABASAPPA D, KALLESHWARASWAMY C M,ASOKAN R, et al. First report of the fall armyworm, Spodoptera frugiperda (J E Smith) (Lepidoptera: Noctuidae), an alien invasive pest on maize in India [J]. Pest Management in Horticultural Ecosystems, 2018, 24(1): 23-29.
[10] CRUZ-ESTEBAN S, ROJAS J C, SANCHEZ-GUILLEN D, et al. Geographic variation in pheromone component ratio and antennal responses, but not in attraction, to sex pheromones among fall armyworm populations infesting corn in Mexico [J]. Journal of Pest Science, 2018, 91(3): 973-983.
[11] PASHLEY D P, MARTIN J A. Reproductive incompatibility between host strains of the fall armyworm (Lepidoptera: Noctuidue)[J]. Annals of the Entomological Society of America, 1987, 80(6): 731-733.
[12] MCMICHAEL M, PROWELL D P. Differences in amplified fragment-length polymorphisms in fall armyworm (Lepidoptera: Noctuidae) host strains [J]. Annals of the Entomological Society of America, 1999, 92(2): 175-181.
[13] MEAGHER R L, CALLO-MEAGHER M. Identifying host of fall armyworm (Lepidoptera: Noctuidae) in Florida using mitochondrial markers [J]. Florida Entomologist, 2003, 86(4): 450-455.
[14] CANO-CALLE D, ARANGO-ISAZA R E, SALDAMANDOBENJUMEA C I. Molecular identification of Spodoptera frugiperda (Lepidoptera: Noctuidae) corn and rice strains in Colombia by using a PCR-RFLP of the mitochondrial gene cytochrome oxydase Ⅰ (COⅠ) and a PCR of the gene FR (for rice)[J]. Annals of the Entomological Society of America, 2015, 108(2): 172-180.
[15] NAGOSHI R N, MEAGHER R L, ADAMCZYK J J. New restriction fragment length polymorphisms in the Cytochrome oxidase I gene facilitate host strain identification of fall armyworm (Lepidoptera: Noctuidae) populations in the southeastern United States [J]. Journal of Economic Entomology, 2006, 99(3): 671-677.
[16] LU Y J, ADANG M J. Distinguishing fall armyworm (Lepidoptera: Noctuidae) strains using a diagnostic mitochondrial DNA marker [J]. Florida Entomologist, 1996, 79: 48-55.
[17] NAGOSHI R N, MURUA M G, HAY-ROE M, et al. Genetic characterization of fall armyworm (Lepidoptera: Noctuidae) host strains in Argentina [J]. Journal of Economic Entomology, 2012, 105(2): 418-428.
[18] NAGOSHI R N. The fall armyworm triosephosphate isomerase (Tpi) gene as a marker of strain identity and interstrain mating [J]. Annals of the Entomological Society of America, 2010, 103(2): 283-292.
[19] PROWELL D P, MCMICHAEL M, SILVAIN J F. Multilocus genetic analysis of host use, introgression, and speciation in host strains of fall armyworm (Lepidoptera: Noctuidae)[J]. Annals of the Entomological Society of America, 2004, 97(5): 1034-1044.
[20] LU Y J, KOCHERT G D, ISENHOUR D J. Molecular characterization of a strain-specific repeated DNA sequence in the fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)[J]. Insect Molecular Biology, 1994, 3(2): 123-130.
[21] JACOBS A,VAN VUUREN A, RONG I. Characterisation of the fall armyworm (Spodoptera frugiperda J. E. Smith) (Lepidoptera: Noctuidae) from South Africa [J]. Africa Entomology, 2018, 26(1): 45-49.
[22] NAGOSHI R N, MEAGHER R L, NUSSELY G, et al. Effects of fall armyworm (Lepidoptera: Noctuidae) interstrain mating in wild populations [J]. Environmental Entomology, 2006, 35: 561-568.
[23] NAGOSHI R N, MEAGHER R L. Using intron sequence comparisons in the triose-phosphate isomerase gene to study the divergence of the fall armyworm host strains [J]. Insect Molecular Biology, 2016, 25(3): 324-337.
[24] JUA’REZ M L, SCHOFL G, VERA M T, et al. Population structure of Spodoptera frugiperda maize and rice host forms in South America: are they host strains?[J]. Entomologia Experimentalis et Applicata, 2014, 152(3): 182-199.
[25] MURU’A M G, NAGOSHI R N, DOS SANTOS D A. Demonstration using field collections that Argentina fall armyworm populations exhibit strain-specific host plant preferences[J]. Journal of Economic Entomology, 2015, 108(5): 2305-2315.
[26] NAGOSHI R N, KOFFI D, AGBOKA K, et al. Comparative molecular analyses of invasive fall armyworm in Togo reveal strong similarities to populations from the eastern United States and the Greater Antilles [J/OL]. PLoS ONE, 2017, 12(7): e0181982.
[27] NAGOSHI R N. Improvements in the identification of strains facilitate population studies of fall armyworm subgroups [J]. Annals of the Entomological Society of America,2012,105(2): 351-358.
[28] JEGER M, BRAGARD C, CAFFIER D, et al. Pest categorisation of Spodoptera frugiperda [J]. European Food Safety Authority Journal, 2017, 15(7): 4927.
[29] PASHLEY D P. Host-associated genetic differentiation in fall army worm (Lepidoptera: Noctuidae): a sibling species complex [J]. Annals of the Entomological Society of America, 1986, 79(6): 898-904.
[30] GROOT A T, MARR M, SCHOFL G, et al. Host strain specific sex pheromone variation in Spodoptera frugiperda[J]. Frontiers in Zoology, 2008, 5: 20.
[31] KOST S, HECKEL D G, YOSHIDO A, et al. A Z-linked sterility locus causes sexual abstinence in hybrid females and facilitates speciation in Spodoptera frugiperda[J].Evolution,2016,70(6):1418-1427.
[32] LIMA E R, MCNEIL J N. Female sex pheromones in the host races and hybrids of the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae)[J].Chemoecology,2009,19(1):29-36.
[33] PASHLEY D P, HAMMOND A M, HARDY T N. Reproductive isolating mechanisms in fall armyworm host strains (Lepidoptera, Noctuidae)[J]. Annals of the Entomological Society of America, 1992, 85(4): 400-405.
[34] PASHLEY D P. Quantitative genetics, development, and physiological adaptation in host strains of fall armyworm [J]. Evolution, 1988, 42(1): 93-102.
[35] PASHLEY D P, SPARKS T C, QUISENBERRY S S, et al. Two fall armyworm strains feed on corn, rice and bermudagrass [J]. Louisiana Agriculture Magazine, 1987, 30: 8-9.
[36] NAGOSHI R N, BRAMBILA J, MEAGHER R L. Use of DNA barcodes to identify invasive armyworm Spodoptera species in Florida [J]. Journal of Insect Science, 2011, 11: 154.
[37] UNBEHEND M, HANNIGER S, VASQUEZ G M, et al. Geographic variation in sexual attraction of Spodoptera frugiperda corn-and rice-strain males to pheromone lures [J]. PLoS ONE, 2014, 9: e89255.
[38] NAGOSHI R N, SILVIE P, MEAGHER R L. Comparison of haplotype frequencies differentiate fall armyworm (Lepidoptera: Noctuidae) corn-strain populations from Florida and Brazil [J]. Journal of Economic Entomology,2007,100(3): 954-961.
[39] NAGOSHI R N, MEAGHER R L, FLANDERS K, et al. Using haplotypes to monitor the migration of fall armyworm (Lepidoptera: Noctuidae) corn-strain populations from Texas and Florida [J].Journal of Economic Entomology,2008,101(3):742-749.
[40] NAGOSHI R N, ROSAS-GARCIA N M, MEAGHER R L, et al. Haplotype profile comparisons between Spodoptera frugiperda (Lepidoptera: Noctuidae) populations from Mexico with those from Puerto Rico, South America, and the United States and their implications to migratory behavior [J]. Journal of Economic Entomology, 2015, 108(1): 135-144.
[41] NAGOSHI R N, MEAGHER R L, HAY-ROE M. Inferring the annual migration patterns of fall armyworm (Lepidoptera: Noctuidae) in the United States from mitochondrial haplotypes[J]. Ecology and Evolution, 2012, 2(7): 1458-1467.
[42] GOUIN A, BRETAUDEAU A, NAM K. Two genomes of highly polyphagous lepidopteran pests (Spodoptera frugiperda, Noctuidae) with different host-plant ranges [J]. Scientific Reports, 2017, 7: 11816.
[43] LIU Q N, CHAI X Y, BIAN D D. The complete mitochondrial genome of fall armyworm Spodoptera frugiperda (Lepidoptera:Noctuidae)[J].Genes & Genomics,2016,38:205-216.
[2] 吴秋琳,姜玉英,吴孔明. 草地贪夜蛾缅甸虫源迁入中国的路径分析[J/OL]. 植物保护,2019,https://doi.org/10.16688/j.zwbh.2019047.
[3] SPARKS A N. A review of the biology of the fall armyworm[J]. Florida Entomologist, 1979,62(2): 82-87.
[4] MITCHELL E R, MCNEIL J N, WESTBROOK J K, et al. Seasonal periodicity of fall armyworm, (Lepidoptera: Noctuidae) in the Caribbean basin and northward to Canada[J]. Journal of Entomological Science, 1991, 26(1): 39-50.
[5] COCK M J W, BESEH P K, BUDDIE A G, et al. Molecular methods to detect Spodoptera frugiperda in Ghana, and implications for monitoring the spread of invasive species in developing countries [J]. Scientific Reports, 2017, 7(1): 4103.
[6] CHAPMAN D, PURSE B V, ROY H E, et al. Global trade networks determine the distribution of invasive non-native species[J].Global Ecology and Biogeography,2017,26(8):907-917.
[7] GOERGEN G, KUMAR P L, SANKUNG S B, et al. First report of outbreaks of the fall armyworm Spodoptera frugiperda (J E Smith) (Lepidoptera, Noctuidae), a new alien invasive pest in West and Central Africa [J/OL]. PLoS ONE, 2016, 11(10): e0165632.
[8] 郭井菲,赵建周,何康来. 警惕危险性害虫草地贪夜蛾入侵中国[J]. 植物保护,2018,44(6):1-10.
[9] SHARANABASAPPA D, KALLESHWARASWAMY C M,ASOKAN R, et al. First report of the fall armyworm, Spodoptera frugiperda (J E Smith) (Lepidoptera: Noctuidae), an alien invasive pest on maize in India [J]. Pest Management in Horticultural Ecosystems, 2018, 24(1): 23-29.
[10] CRUZ-ESTEBAN S, ROJAS J C, SANCHEZ-GUILLEN D, et al. Geographic variation in pheromone component ratio and antennal responses, but not in attraction, to sex pheromones among fall armyworm populations infesting corn in Mexico [J]. Journal of Pest Science, 2018, 91(3): 973-983.
[11] PASHLEY D P, MARTIN J A. Reproductive incompatibility between host strains of the fall armyworm (Lepidoptera: Noctuidue)[J]. Annals of the Entomological Society of America, 1987, 80(6): 731-733.
[12] MCMICHAEL M, PROWELL D P. Differences in amplified fragment-length polymorphisms in fall armyworm (Lepidoptera: Noctuidae) host strains [J]. Annals of the Entomological Society of America, 1999, 92(2): 175-181.
[13] MEAGHER R L, CALLO-MEAGHER M. Identifying host of fall armyworm (Lepidoptera: Noctuidae) in Florida using mitochondrial markers [J]. Florida Entomologist, 2003, 86(4): 450-455.
[14] CANO-CALLE D, ARANGO-ISAZA R E, SALDAMANDOBENJUMEA C I. Molecular identification of Spodoptera frugiperda (Lepidoptera: Noctuidae) corn and rice strains in Colombia by using a PCR-RFLP of the mitochondrial gene cytochrome oxydase Ⅰ (COⅠ) and a PCR of the gene FR (for rice)[J]. Annals of the Entomological Society of America, 2015, 108(2): 172-180.
[15] NAGOSHI R N, MEAGHER R L, ADAMCZYK J J. New restriction fragment length polymorphisms in the Cytochrome oxidase I gene facilitate host strain identification of fall armyworm (Lepidoptera: Noctuidae) populations in the southeastern United States [J]. Journal of Economic Entomology, 2006, 99(3): 671-677.
[16] LU Y J, ADANG M J. Distinguishing fall armyworm (Lepidoptera: Noctuidae) strains using a diagnostic mitochondrial DNA marker [J]. Florida Entomologist, 1996, 79: 48-55.
[17] NAGOSHI R N, MURUA M G, HAY-ROE M, et al. Genetic characterization of fall armyworm (Lepidoptera: Noctuidae) host strains in Argentina [J]. Journal of Economic Entomology, 2012, 105(2): 418-428.
[18] NAGOSHI R N. The fall armyworm triosephosphate isomerase (Tpi) gene as a marker of strain identity and interstrain mating [J]. Annals of the Entomological Society of America, 2010, 103(2): 283-292.
[19] PROWELL D P, MCMICHAEL M, SILVAIN J F. Multilocus genetic analysis of host use, introgression, and speciation in host strains of fall armyworm (Lepidoptera: Noctuidae)[J]. Annals of the Entomological Society of America, 2004, 97(5): 1034-1044.
[20] LU Y J, KOCHERT G D, ISENHOUR D J. Molecular characterization of a strain-specific repeated DNA sequence in the fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)[J]. Insect Molecular Biology, 1994, 3(2): 123-130.
[21] JACOBS A,VAN VUUREN A, RONG I. Characterisation of the fall armyworm (Spodoptera frugiperda J. E. Smith) (Lepidoptera: Noctuidae) from South Africa [J]. Africa Entomology, 2018, 26(1): 45-49.
[22] NAGOSHI R N, MEAGHER R L, NUSSELY G, et al. Effects of fall armyworm (Lepidoptera: Noctuidae) interstrain mating in wild populations [J]. Environmental Entomology, 2006, 35: 561-568.
[23] NAGOSHI R N, MEAGHER R L. Using intron sequence comparisons in the triose-phosphate isomerase gene to study the divergence of the fall armyworm host strains [J]. Insect Molecular Biology, 2016, 25(3): 324-337.
[24] JUA’REZ M L, SCHOFL G, VERA M T, et al. Population structure of Spodoptera frugiperda maize and rice host forms in South America: are they host strains?[J]. Entomologia Experimentalis et Applicata, 2014, 152(3): 182-199.
[25] MURU’A M G, NAGOSHI R N, DOS SANTOS D A. Demonstration using field collections that Argentina fall armyworm populations exhibit strain-specific host plant preferences[J]. Journal of Economic Entomology, 2015, 108(5): 2305-2315.
[26] NAGOSHI R N, KOFFI D, AGBOKA K, et al. Comparative molecular analyses of invasive fall armyworm in Togo reveal strong similarities to populations from the eastern United States and the Greater Antilles [J/OL]. PLoS ONE, 2017, 12(7): e0181982.
[27] NAGOSHI R N. Improvements in the identification of strains facilitate population studies of fall armyworm subgroups [J]. Annals of the Entomological Society of America,2012,105(2): 351-358.
[28] JEGER M, BRAGARD C, CAFFIER D, et al. Pest categorisation of Spodoptera frugiperda [J]. European Food Safety Authority Journal, 2017, 15(7): 4927.
[29] PASHLEY D P. Host-associated genetic differentiation in fall army worm (Lepidoptera: Noctuidae): a sibling species complex [J]. Annals of the Entomological Society of America, 1986, 79(6): 898-904.
[30] GROOT A T, MARR M, SCHOFL G, et al. Host strain specific sex pheromone variation in Spodoptera frugiperda[J]. Frontiers in Zoology, 2008, 5: 20.
[31] KOST S, HECKEL D G, YOSHIDO A, et al. A Z-linked sterility locus causes sexual abstinence in hybrid females and facilitates speciation in Spodoptera frugiperda[J].Evolution,2016,70(6):1418-1427.
[32] LIMA E R, MCNEIL J N. Female sex pheromones in the host races and hybrids of the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae)[J].Chemoecology,2009,19(1):29-36.
[33] PASHLEY D P, HAMMOND A M, HARDY T N. Reproductive isolating mechanisms in fall armyworm host strains (Lepidoptera, Noctuidae)[J]. Annals of the Entomological Society of America, 1992, 85(4): 400-405.
[34] PASHLEY D P. Quantitative genetics, development, and physiological adaptation in host strains of fall armyworm [J]. Evolution, 1988, 42(1): 93-102.
[35] PASHLEY D P, SPARKS T C, QUISENBERRY S S, et al. Two fall armyworm strains feed on corn, rice and bermudagrass [J]. Louisiana Agriculture Magazine, 1987, 30: 8-9.
[36] NAGOSHI R N, BRAMBILA J, MEAGHER R L. Use of DNA barcodes to identify invasive armyworm Spodoptera species in Florida [J]. Journal of Insect Science, 2011, 11: 154.
[37] UNBEHEND M, HANNIGER S, VASQUEZ G M, et al. Geographic variation in sexual attraction of Spodoptera frugiperda corn-and rice-strain males to pheromone lures [J]. PLoS ONE, 2014, 9: e89255.
[38] NAGOSHI R N, SILVIE P, MEAGHER R L. Comparison of haplotype frequencies differentiate fall armyworm (Lepidoptera: Noctuidae) corn-strain populations from Florida and Brazil [J]. Journal of Economic Entomology,2007,100(3): 954-961.
[39] NAGOSHI R N, MEAGHER R L, FLANDERS K, et al. Using haplotypes to monitor the migration of fall armyworm (Lepidoptera: Noctuidae) corn-strain populations from Texas and Florida [J].Journal of Economic Entomology,2008,101(3):742-749.
[40] NAGOSHI R N, ROSAS-GARCIA N M, MEAGHER R L, et al. Haplotype profile comparisons between Spodoptera frugiperda (Lepidoptera: Noctuidae) populations from Mexico with those from Puerto Rico, South America, and the United States and their implications to migratory behavior [J]. Journal of Economic Entomology, 2015, 108(1): 135-144.
[41] NAGOSHI R N, MEAGHER R L, HAY-ROE M. Inferring the annual migration patterns of fall armyworm (Lepidoptera: Noctuidae) in the United States from mitochondrial haplotypes[J]. Ecology and Evolution, 2012, 2(7): 1458-1467.
[42] GOUIN A, BRETAUDEAU A, NAM K. Two genomes of highly polyphagous lepidopteran pests (Spodoptera frugiperda, Noctuidae) with different host-plant ranges [J]. Scientific Reports, 2017, 7: 11816.
[43] LIU Q N, CHAI X Y, BIAN D D. The complete mitochondrial genome of fall armyworm Spodoptera frugiperda (Lepidoptera:Noctuidae)[J].Genes & Genomics,2016,38:205-216.