聚多曲霉Snef210对南方根结线虫毒性的研究
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摘要
蔬菜根结线虫病在我国乃至世界危害日益严重,安全防控该病害成为目前急需解决的农业生产问题。本研究筛选到一株可毒杀南方根结线虫Meloidogyne incognita的生防真菌Snef210,经形态学和分子生物学鉴定为聚多曲霉Aspergillus sydowii(Bain.et Sart.) Thom et Church。该菌株发酵液对南方根结线虫2龄幼虫(J_2)具有很强的毒杀作用,原液处理24 h后校正死亡率达90.1%,处理3 d后对卵孵化的抑制率达92.8%。盆栽试验中,与对照相比,菌株发酵液10倍稀释液土壤处理对番茄根部的根结减退率为66.7%。聚多曲霉代谢产物丰富,多用于抗菌和抗肿瘤的研究,本文首次发现其对南方根结线虫有效,是一类潜在的控制植物线虫病害的新型生防因子。
Disease of root-knot nematode(RKN) is becoming more serious in China and other countries. It is an urgent problem to control this nematode in agricultural production. Snef210 was screened with high efficacy against Meloidogyne incognita and identified as Aspergillus sydowii with morphological and molecular analysis. Second stage juveniles(J_2) and egg capsules were immersed in Snef210 with in vitro methods. The results showed the corrected mortality of J_2was 90.1% at 24 h. Moreover, Snef210 exhibited high ovicidal effect with the efficacy of 92.8% at 3 d. Root-knot index treated with diluted ten-fold Snef210 fermentation broth was decreased by 66.7% in pot experiment. A. sydowii is rich in metabolic products, mostly used in antibacterial and anti-tumor studies. Snef210 had strongly toxicity to M. incognita and could be regarded as a novel potential biocontrol agent against RKNs.
Disease of root-knot nematode(RKN) is becoming more serious in China and other countries. It is an urgent problem to control this nematode in agricultural production. Snef210 was screened with high efficacy against Meloidogyne incognita and identified as Aspergillus sydowii with morphological and molecular analysis. Second stage juveniles(J_2) and egg capsules were immersed in Snef210 with in vitro methods. The results showed the corrected mortality of J_2was 90.1% at 24 h. Moreover, Snef210 exhibited high ovicidal effect with the efficacy of 92.8% at 3 d. Root-knot index treated with diluted ten-fold Snef210 fermentation broth was decreased by 66.7% in pot experiment. A. sydowii is rich in metabolic products, mostly used in antibacterial and anti-tumor studies. Snef210 had strongly toxicity to M. incognita and could be regarded as a novel potential biocontrol agent against RKNs.
引文
[1] 段玉玺.植物线虫学[M].北京:科学出版社,2011:135-144.
[2] 梁建根,郑经武.设施栽培中蔬菜根结线虫生物防治研究进展[J].中国农学通报,2010,26(19):290-293.
[3] JIN Na, XUE Hui, LI Wenjing, et al. Field evaluation of Streptomyces rubrogriseus HDZ-9-47 for biocontrol of Meloidogyne incognita on tomato [J]. Journal of Integrative Agriculture, 2017, 16(6): 1347-1357.
[4] 钱振官,张大业,马承铸,等.寡孢节丛孢菌和褶生轮枝菌对根结线虫的致病力[J].生物防治通报,1993,9(2):91-92.
[5] 马金慧,朱萍萍,茆振川,等.哈茨木霉菌株TRI2的鉴定及其对黄瓜根结线虫的防治作用[J].中国农学通报,2014,30(22):263-269.
[6] 马希斌,段玉玺,陈立杰,等.青霉菌snf44-1代谢产物不同植物线虫的影响[J].农药,2007,46(8):563-565.
[7] 李颂,段玉玺,朱晓峰,等.黑曲霉次生代谢产物对番茄抗根结线虫病效果的影响[J].中国蔬菜,2011,1(4):44-49.
[8] 齐祖同.中国真菌志[M].北京:科学出版社,1997:50-51.
[9] WANG Junfeng, LIN Xiuping, QIN Chun, et al. Antimicrobial and antiviral sesquiterpenoids from sponge-associated fungus, Aspergillus sydowii ZSDS1-F6[J]. Journal of Antibiotics, 2014, 67(8): 581-583.
[10] GASHGARI R, GHERBAWY Y, AMEEN F, et al. Molecular characterization and analysis of antimicrobial activity of endophytic fungi from medicinal plants in Saudi Arabia[J/OL]. Jundishapur Journal of Microbiology, 2016, 9(1): e26157.
[11] GODINHO V M, GONÇALVES V N, SANTIAGO I F, et al. Diversity and bioprospection of fungal community present in oligotrophic soil of continental Antarctica[J]. Extremophiles, 2015, 19(3): 585-596.
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[13] GLASS N L, DONALDSON G C. Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes[J]. Applied and Environmental Microbiology, 1995, 61(4): 1323-1330.
[14] CHEN S Y, DICKSON D W. A technique for determining live second-stage juveniles of Heterodera glycines [J]. Journal of Nematology,2000,32(1):117-121.
[15] 段玉玺, 靳莹莹, 王胜君, 等. 生防菌株Snef85的鉴定及其发酵液对不同种类线虫的毒力[J]. 植物保护学报,2008,35(2):132-136.
[16] 梁建根, 郑经武, 郝中娜, 等. 生防菌K-8对南方根结线虫的防治及其鉴定[J]. 中国农学通报,2011,27(21):282-286.
[17] 肖炎农, 王明祖, 付艳平, 等. 蔬菜根结线虫病情分级方法比较[J].华中农业大学学报,2000,19(4):336-338.
[18] SIDDIQUI Z A, AKHTAR M S. Effects of antagonistic fungi, plant growth-promoting rhizobacteria, and arbuscular mycorrhizal fungi alone and in combination on the reproduction of Meloidogyne incognita, and growth of tomato [J]. Journal of General Plant Pathology, 2009, 75(2):144.
[19] AFFOKPON A, COYNE D L, HTAY C C, et al. Biocontrol potential of native Trichoderma isolates against root-knot nematodes in West African vegetable production systems [J]. Soil Biology & Biochemistry, 2011, 43(3):600-608.
[20] HASHEM M, ABO-ELYOUSR K A. Management of the root-knot nematode Meloidogyne incognita, on tomato with combinations of different biocontrol organisms [J]. Crop Protection, 2011, 30(3):285-292.
[21] ZUCKERMAN B M, MATHENY M, ACOSTA N. Control of plant-parasitic nematodes by a nematicidal strain of Aspergillus niger [J]. Journal of Chemical Ecology, 1994, 20(1):33-43.
[22] JANG J Y, YONG H C, SHIN T S, et al. Biological Control of Meloidogyne incognita by Aspergillus niger F22 producing oxalic acid [J/OL]. PLoS ONE, 2016, 11(6):e0156230.
[23] KIM T Y, JANG J Y, JEON S J, et al. Nematicidal activity of kojic acid produced by Aspergillus oryzae against Meloidogyne incognita [J]. Journal of Microbiology & Biotechnology, 2016, 26(8):1383-1391.
[24] LIANG Lianming, MENG Zhaohui, YE Fengping, et al. The crystal structures of two cuticle-degrading proteases from nematophagous fungi and their contribution to infection against nematodes [J]. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology, 2010, 24(5):1391-1400.
[25] SZABó M, CSEPREGI K, GÁLBER M, et al. Control plant-parasitic nematodes with Trichoderma species and nematode-trapping fungi: The role of chi18-5, and chi18-12, genes in nematode egg-parasitism [J].Biological Control,2012,63(2):121-128.
[26] CHEN Leilei, LIU Lijun, SHI Mei, et al. Characterization and gene cloning of a novel serine protease with nematicidal activity from Trichoderma pseudokoningii SMF2 [J]. FEMS Microbiology Letters, 2009, 299(2):135-142.
[27] WANG Jieping, WANG Jiaxu, LIU Fan, et al. Enhancing the virulence of Paecilomyces lilacinus against Meloidogyne incognita eggs by overexpression of a serine protease [J]. Biotechnology Letters, 2010, 32(8):1159-1166.
[28] BILLS G F, YUE Q, CHEN L, et al. Aspergillus mulundensis sp.nov., a new species for the fungus producing the antifungal echinocandin lipopeptides, mulundocandins[J]. The Journal of Antibiotics, 2016, 69(3): 141-148.
[29] HE Fei, SUN Yulin, LIU Kaisheng, et al. Indole alkaloids from marine-derived fungus Aspergillus sydowii SCSIO 00305[J]. Journal of Antibiotics, 2012, 65(2): 109-111.
[30] KAUR A, RAJA H A, DARVEAUXB A, et al. New diketopiperazine dimer from a filamentous fungal isolate of Aspergillus sydowii [J].Magnetic Resonance in Chemistry,2015,53(8): 616-619.
[31] TRISUWAN K, RUKACHAISIRIKULV, KAEWPETM, et al. Sesquiterpene and xanthone derivatives from the sea fan-derived fungus Aspergillus sydowii PSU-F154[J]. Journal of Natural Products, 2011, 74(7): 1663-1667.
[32] ALVARENGAN, BIROLLIW G, SELEGHIMM H R, et al. Biodegradation of methyl parathion by whole cells of marine-derived fungi Aspergillus sydowii and Penicillium decaturense [J]. Chemosphere, 2014, 117(1): 47-52.
[33] LOPES B P, EGEA T C, MONTEIROD A, et al. Evaluation of diuron tolerance and biotransformation by fungi from a sugarcane plantation sandy-loam soil[J]. Journal of Agricultural and Food Chemistry, 2016, 64(49): 9268-9275.
[34] BIROLLI W G, ALVARENGAN, SELEGHIMM H R, et al. Biodegradation of the pyrethroid pesticide esfenvalerate by marine-derived fungi [J]. Marine Biotechnology, 2016, 18(4): 511-520.
[35] TIAN Jiang, DONG Qiaofeng, YU Chenlei, et al. Biodegradation of the organophosphate trichlorfon and its major degradation products by a novel Aspergillus sydowii PA F-2 [J]. Journal of Agricultural and Food Chemistry, 2016,64(21):4280-4287.
[36] YU D S,SONG G,SONG L L,et al.Formaldehyde degradation by a newly isolated fungus Aspergillus sp.HUA [J].International Journal of Environmental Science and Technology,2015,12(1): 247-254.
[2] 梁建根,郑经武.设施栽培中蔬菜根结线虫生物防治研究进展[J].中国农学通报,2010,26(19):290-293.
[3] JIN Na, XUE Hui, LI Wenjing, et al. Field evaluation of Streptomyces rubrogriseus HDZ-9-47 for biocontrol of Meloidogyne incognita on tomato [J]. Journal of Integrative Agriculture, 2017, 16(6): 1347-1357.
[4] 钱振官,张大业,马承铸,等.寡孢节丛孢菌和褶生轮枝菌对根结线虫的致病力[J].生物防治通报,1993,9(2):91-92.
[5] 马金慧,朱萍萍,茆振川,等.哈茨木霉菌株TRI2的鉴定及其对黄瓜根结线虫的防治作用[J].中国农学通报,2014,30(22):263-269.
[6] 马希斌,段玉玺,陈立杰,等.青霉菌snf44-1代谢产物不同植物线虫的影响[J].农药,2007,46(8):563-565.
[7] 李颂,段玉玺,朱晓峰,等.黑曲霉次生代谢产物对番茄抗根结线虫病效果的影响[J].中国蔬菜,2011,1(4):44-49.
[8] 齐祖同.中国真菌志[M].北京:科学出版社,1997:50-51.
[9] WANG Junfeng, LIN Xiuping, QIN Chun, et al. Antimicrobial and antiviral sesquiterpenoids from sponge-associated fungus, Aspergillus sydowii ZSDS1-F6[J]. Journal of Antibiotics, 2014, 67(8): 581-583.
[10] GASHGARI R, GHERBAWY Y, AMEEN F, et al. Molecular characterization and analysis of antimicrobial activity of endophytic fungi from medicinal plants in Saudi Arabia[J/OL]. Jundishapur Journal of Microbiology, 2016, 9(1): e26157.
[11] GODINHO V M, GONÇALVES V N, SANTIAGO I F, et al. Diversity and bioprospection of fungal community present in oligotrophic soil of continental Antarctica[J]. Extremophiles, 2015, 19(3): 585-596.
[12] WHITE T J,BRUNS T,LEE S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[M]//PCR protocols: a guide to methods and applications. New York: Academic Press, 1990:315-322.
[13] GLASS N L, DONALDSON G C. Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes[J]. Applied and Environmental Microbiology, 1995, 61(4): 1323-1330.
[14] CHEN S Y, DICKSON D W. A technique for determining live second-stage juveniles of Heterodera glycines [J]. Journal of Nematology,2000,32(1):117-121.
[15] 段玉玺, 靳莹莹, 王胜君, 等. 生防菌株Snef85的鉴定及其发酵液对不同种类线虫的毒力[J]. 植物保护学报,2008,35(2):132-136.
[16] 梁建根, 郑经武, 郝中娜, 等. 生防菌K-8对南方根结线虫的防治及其鉴定[J]. 中国农学通报,2011,27(21):282-286.
[17] 肖炎农, 王明祖, 付艳平, 等. 蔬菜根结线虫病情分级方法比较[J].华中农业大学学报,2000,19(4):336-338.
[18] SIDDIQUI Z A, AKHTAR M S. Effects of antagonistic fungi, plant growth-promoting rhizobacteria, and arbuscular mycorrhizal fungi alone and in combination on the reproduction of Meloidogyne incognita, and growth of tomato [J]. Journal of General Plant Pathology, 2009, 75(2):144.
[19] AFFOKPON A, COYNE D L, HTAY C C, et al. Biocontrol potential of native Trichoderma isolates against root-knot nematodes in West African vegetable production systems [J]. Soil Biology & Biochemistry, 2011, 43(3):600-608.
[20] HASHEM M, ABO-ELYOUSR K A. Management of the root-knot nematode Meloidogyne incognita, on tomato with combinations of different biocontrol organisms [J]. Crop Protection, 2011, 30(3):285-292.
[21] ZUCKERMAN B M, MATHENY M, ACOSTA N. Control of plant-parasitic nematodes by a nematicidal strain of Aspergillus niger [J]. Journal of Chemical Ecology, 1994, 20(1):33-43.
[22] JANG J Y, YONG H C, SHIN T S, et al. Biological Control of Meloidogyne incognita by Aspergillus niger F22 producing oxalic acid [J/OL]. PLoS ONE, 2016, 11(6):e0156230.
[23] KIM T Y, JANG J Y, JEON S J, et al. Nematicidal activity of kojic acid produced by Aspergillus oryzae against Meloidogyne incognita [J]. Journal of Microbiology & Biotechnology, 2016, 26(8):1383-1391.
[24] LIANG Lianming, MENG Zhaohui, YE Fengping, et al. The crystal structures of two cuticle-degrading proteases from nematophagous fungi and their contribution to infection against nematodes [J]. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology, 2010, 24(5):1391-1400.
[25] SZABó M, CSEPREGI K, GÁLBER M, et al. Control plant-parasitic nematodes with Trichoderma species and nematode-trapping fungi: The role of chi18-5, and chi18-12, genes in nematode egg-parasitism [J].Biological Control,2012,63(2):121-128.
[26] CHEN Leilei, LIU Lijun, SHI Mei, et al. Characterization and gene cloning of a novel serine protease with nematicidal activity from Trichoderma pseudokoningii SMF2 [J]. FEMS Microbiology Letters, 2009, 299(2):135-142.
[27] WANG Jieping, WANG Jiaxu, LIU Fan, et al. Enhancing the virulence of Paecilomyces lilacinus against Meloidogyne incognita eggs by overexpression of a serine protease [J]. Biotechnology Letters, 2010, 32(8):1159-1166.
[28] BILLS G F, YUE Q, CHEN L, et al. Aspergillus mulundensis sp.nov., a new species for the fungus producing the antifungal echinocandin lipopeptides, mulundocandins[J]. The Journal of Antibiotics, 2016, 69(3): 141-148.
[29] HE Fei, SUN Yulin, LIU Kaisheng, et al. Indole alkaloids from marine-derived fungus Aspergillus sydowii SCSIO 00305[J]. Journal of Antibiotics, 2012, 65(2): 109-111.
[30] KAUR A, RAJA H A, DARVEAUXB A, et al. New diketopiperazine dimer from a filamentous fungal isolate of Aspergillus sydowii [J].Magnetic Resonance in Chemistry,2015,53(8): 616-619.
[31] TRISUWAN K, RUKACHAISIRIKULV, KAEWPETM, et al. Sesquiterpene and xanthone derivatives from the sea fan-derived fungus Aspergillus sydowii PSU-F154[J]. Journal of Natural Products, 2011, 74(7): 1663-1667.
[32] ALVARENGAN, BIROLLIW G, SELEGHIMM H R, et al. Biodegradation of methyl parathion by whole cells of marine-derived fungi Aspergillus sydowii and Penicillium decaturense [J]. Chemosphere, 2014, 117(1): 47-52.
[33] LOPES B P, EGEA T C, MONTEIROD A, et al. Evaluation of diuron tolerance and biotransformation by fungi from a sugarcane plantation sandy-loam soil[J]. Journal of Agricultural and Food Chemistry, 2016, 64(49): 9268-9275.
[34] BIROLLI W G, ALVARENGAN, SELEGHIMM H R, et al. Biodegradation of the pyrethroid pesticide esfenvalerate by marine-derived fungi [J]. Marine Biotechnology, 2016, 18(4): 511-520.
[35] TIAN Jiang, DONG Qiaofeng, YU Chenlei, et al. Biodegradation of the organophosphate trichlorfon and its major degradation products by a novel Aspergillus sydowii PA F-2 [J]. Journal of Agricultural and Food Chemistry, 2016,64(21):4280-4287.
[36] YU D S,SONG G,SONG L L,et al.Formaldehyde degradation by a newly isolated fungus Aspergillus sp.HUA [J].International Journal of Environmental Science and Technology,2015,12(1): 247-254.