混合寄生真菌发酵液对大豆胞囊线虫的毒力和防效
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摘要
寄生真菌是一类对大豆胞囊线虫具有潜力的生防资源,镰孢菌属(Fusarium spp.)、厚垣轮枝菌(Verticillium chlamydosporium)和淡紫拟青霉菌(Paecilomyces lilacinus)广泛存在于连作大豆田中,为了探讨这些寄生真菌对大豆胞囊线虫的抑制和防效,采用从大豆胞囊线虫抑制性土壤中分离的上述3个属12个菌株制备发酵液,按比例配置不同组合的混合菌发酵液,实验室条件下测定对大豆胞囊线虫3号生理小种胞囊、卵孵化抑制作用和对大豆胞囊线虫2龄幼虫的致死作用,筛选抑制效果好的4个发酵液组合进行温室盆栽试验。结果显示,16组混合寄生真菌发酵液均对大豆胞囊线虫胞囊和卵孵化具有强烈的抑制作用,胞囊孵化抑制率在59.1%~87.8%,卵孵化抑制率在72.1%~84.7%,大豆胞囊线虫2龄幼虫的致死作用48 h在94%以上,72 h达到了100%。F-V+P-V+V-21、F-V+P-E+V-25、F-9+P-V+V-25和F-9+P-E+V-21的4个组合可显著降低大豆胞囊线虫胞囊、卵、根表雌虫和根内J2数量,并促进大豆生长。
Parasitic fungi is a kind of biocontrol resources with potential to control soybean cyst nematode( SCN). Fusarium spp.,Verticillium chlamydosporium and Paecilomyces lilacinus have been widely found in continuous soybean field. In order to study the inhibition and control effects of these parasitic fungi on SCN,laboratory and greenhouse experiments were conducted. The fermentation filtrates were prepared from 12 strains of the 3 genera parasitic fungi isolated from SCN inhibition soil. The inhibition of the fermentation filtrates on cyst and egg hatching of SCN and the lethal effect on the 2 nd instar larvae( J2) were determined under laboratory conditions. Four fermentation filtrates combinations with higher inhibitory effect were screened in greenhouse pot experiment. The results showed that the mixed parasitic fungal fermentation filtrates of 16 groups had strong inhibitory effect on cyst and egg hatching of SCN. The cyst hatching inhibition rate ranged from 59. 1% to 87. 8%,while the egg hatching inhibition rate ranged from 72. 1 to84. 7. The lethal effect on J2 of SCN was more than 94% at 48 h,and it reached 100% at 72 h. Combination of F-V + P-V + V-21,F-V + P-E + V-25,F-9 + P-V + V-25 and F-9 + P-E + V-21 would significantly reduce the number of cyst,female,egg and J2 of SCN in root,and promote the growth of soybean.
Parasitic fungi is a kind of biocontrol resources with potential to control soybean cyst nematode( SCN). Fusarium spp.,Verticillium chlamydosporium and Paecilomyces lilacinus have been widely found in continuous soybean field. In order to study the inhibition and control effects of these parasitic fungi on SCN,laboratory and greenhouse experiments were conducted. The fermentation filtrates were prepared from 12 strains of the 3 genera parasitic fungi isolated from SCN inhibition soil. The inhibition of the fermentation filtrates on cyst and egg hatching of SCN and the lethal effect on the 2 nd instar larvae( J2) were determined under laboratory conditions. Four fermentation filtrates combinations with higher inhibitory effect were screened in greenhouse pot experiment. The results showed that the mixed parasitic fungal fermentation filtrates of 16 groups had strong inhibitory effect on cyst and egg hatching of SCN. The cyst hatching inhibition rate ranged from 59. 1% to 87. 8%,while the egg hatching inhibition rate ranged from 72. 1 to84. 7. The lethal effect on J2 of SCN was more than 94% at 48 h,and it reached 100% at 72 h. Combination of F-V + P-V + V-21,F-V + P-E + V-25,F-9 + P-V + V-25 and F-9 + P-E + V-21 would significantly reduce the number of cyst,female,egg and J2 of SCN in root,and promote the growth of soybean.
引文
[1]刘增柱,周玉芝,韩静淑.大豆连、轮作土壤微生物生态分布与大豆孢囊线虫群体动态的研究[J].大豆科学,1990,9(3):206-212.LIU Z Z,ZHOU Y Z,HAN J S.Study on ecological distribution of soil microorganisms and colony dynamics cyst-forming nematodes in continuous and rotation cropping systems of soybean[J].Soybean Science,1990,9(3):206-212.
[2]许艳丽,韩晓增.大豆重迎茬研究[M].哈尔滨:哈尔滨工程大学出版社,1995:136-139.XU Y L,HAN X Z.Study on continuous cropping and alternate-year of soybean[M].Harbin:Harbin Engineering University Press,1995:136-139.
[3]HARTWIG E E.Breeding productive soybean cultivars resistant to the soybean cyst nematode for the southern United States[J].Plant Disease,1981,65(4):303-307.
[4]CHEN S Y.Suppression of Heterodera glycines in soils from fields with long-term soybean monoculture[J].Biocontrol Science and Technology,2007,17(2):125-134.
[5]刘杏忠,张东升,武修英,等.定殖于大豆孢囊线虫孢囊内真菌的初步研究[J].北京农业大学学报,1991,17(3):87-91.LIU X Z,ZHANG D S,WU X Y,et al.Preliminary studies on the parasitic from the cyst of Heterodera glycines[J].Acta Agriculture Universitatis Pekinensis,1991,17(3):87-91.
[6]靳学慧,辛惠普,郑雯,等.长期轮作和连作对土壤中大豆胞囊线虫数量的影响[J].中国油料作物学报,2006,28(2):189-193.JIN X H,XIN H P,ZHENG W,et al.The influence of soil on the long-term rotation and continuous cultivation on soybean cyst nematode[J].Chinese Journal of Oil Crop Science,2006,28(2):189-193.
[7]许艳丽.土壤环境对大豆胞囊线虫卵孵化影响及线虫分子诊断研究[D].哈尔滨:东北农业大学,2004:41-47.XU Y L.The effects of soil environment on soybean cyst nematode egg hatch and nematode molecular diagnosis[D].Harbin:Northeast Agricultural University,2004:41-47.
[8]马承铸,顾真容,钱振官,等.种植制对大豆孢囊线虫群体密度和真菌区系的影响[J].上海农业学报,1994,10(2):62-66.MA C Z,GU Z R,QIAN Z G,et al.Effects of cropping system on mycoflora and population density of soybean cyst nematode Heterodera glycines[J].Acta Agriculturae Shanghai,1994,10(2):62-66.
[9]孙玉秋,许艳丽,李春杰,等.作物轮作系统对土壤中大豆胞囊线虫胞囊量和单胞囊卵量的影响[J].农业系统科学与综合研究,2011,27(2):248-252.SUN Y Q,XU Y L,LI C J,et al.Influence of cropping rotation systems on the volume of cysts and eggs in a single-cyst of soybean cyst nematode[J].System Science and Comprehensive Studies in Agriculture,2011,27(2):248-252.
[10]孙漫红,刘杏忠.连作土壤中大豆胞囊线虫种群数量减少的原因探讨[J].植物病理学报,2000,30(4):353-356,363.SUN M H,LIU X Z.Suppressive soil of soybean cysts nematode in China[J].Acta Phytopathologica Sinica,2000,30(4):353-356,363.
[11]CRUMP D H,SAYRE R M,YOUNG L D.Occurrence of nematophagous fungi in cyst nematode populations[J].Plant Disease,1983,67(1):63-64.
[12]孙漫红,刘杏忠,晋治波.淡紫拟青霉对大豆胞囊线虫卵及2龄幼虫的影响[J].植物保护学报,2002,29(1):57-61.SUN M H,LIU X Z,JIN Z B.Effect Paecilomyces lilacinus on egg hatching and juvenile mortality of Heterodera glycines[J].Journal of Plant Protection,2002,29(1):57-61.
[13]SONG J,LI S X,WEI W,et al.Assessment of parasitic fungi for reducing soybean cyst nematode with suppressive soil in soybean fields of northeast China[J].Acta Agriculturae Scandinavica,Section B—Soil and Plant Science,2017,67(8):730-736.
[14]宋洁,许艳丽,姚钦.大豆胞囊线虫主要寄生真菌对大豆耕作系统的响应[J].大豆科学,2016,35(3):461-467.SONG J,XU Y L,YAO Q.Response of major parasitic fungi of soybean cyst nematode on different cultivation systems[J].Soybean Science,2016,35(3):461-467.
[15]SONG J,LI S X,XU Y L,et al.Diversity of parasitic fungi from soybean cyst nematode associated with long-term continuous cropping of soybean in black soil[J].Acta Agriculturae Scandinavica,Section B—Soil and Plant Science,2016,66(5):432–442.
[16]陈立杰,刘晓杰,段玉玺,等.轮作和连作田大豆胞囊线虫胞囊上真菌定殖动态[J],大豆科学,2009,28(2):266-270.CHEN L J,LIU X J,DUAN Y X,et al.Dynamics of fungal colonization on cyst in rotation and continuous cropping of soybean[J].Soybean Science,2009,28(2):266-270.
[17]林茂松.土壤真菌防治大豆胞囊线虫的效果测定[J].中国油料,1990,12(3):63-65.LIN M S.Control effect to soybean cyst nematode by fungi from eggs of Heterodera glycines[J].Chinese Oil,1990,12(3):63-65.
[18]范圣长,段玉玺,陈立杰.大豆胞囊线虫胞囊内寄生真菌研究[J].大豆科学,2004,23(1):71-74.FAN S C,DUAN Y X,CHEN L J.The research on the cyst entoparasitic fungi of soybean cyst nematode[J].Soybean Science,2004,23(1):71-74.
[19]FITTERS P F L,DEN BELDER E.A time-lapse technique to study the effect of fungal products on embryogenesis of nematode eggs[J].Mededelingen van de Faculteit Landbouwwetenschappen-Rijksuniversiteit Gent,1993,58(2B):751-756.
[20]赵晓晖,许艳丽.镰刀菌发酵液对大豆胞囊线虫的抑制作用[J].大豆科学,2011,30(3):468-470,474.ZHAO X H,XU Y L.Inhibition of Fusarium spp.fermented filtrates on soybean cyst nematode[J].Soybean Science,2011,30(3):468-470,474.
[21]赵晓晖,许艳丽.厚垣轮枝菌发酵液对大豆胞囊线虫的抑制作用[J].大豆科技,2011,20(4):20-23.ZHAO X H,XU Y L.Inhibition of Verticillium chlamydosporium fermented filtrates on soybean cyst nematode[J].Soybean Science and Technology,2011,20(4):20-23.
[22]KHAN A,WILLIAMA K L,NEVALAINEN H K M.Effect of Paecilomyces lilacinus protease and chitinase on the eggshell structure and hatching of Meloidogyne jovanica juveniles[J].Biological Control 2004,31(3):346-352.
[23]宋洁.连作土壤寄生真菌多样性及对大豆胞囊线虫抑制作用[D].哈尔滨:东北农业大学,2016:58-64,75-92.SONG J.Diversity and suppressive effect of parasitic fungi on soyrean cyst nematode in soybean monoculture soil[D].Harbin:Northeast Agricultural University,2016:58-64,75-92.
[24]鲁建聪.寄生真菌代谢物对大豆胞囊线虫抑制及防治[D].哈尔滨:东北农业大学,2015:16-28.LU J C.Suppressness and biocontrol of parasitic fungal fermentation filtrate on soybean cyst nematode[D].Harbin:Northeast Agricultural University,2015:16-28.
[25]鲁建聪,许艳丽,宋洁.土壤扰动对寄生真菌代谢物温室防治大豆胞囊线虫的影响[J].大豆科学,2016,35(1):106-111.LU J C,XU Y L,SONG J.Effect of soil disturbance to the biological control of fermentation filtrate of parasitic fungi on soybean cyst nematode in glasshouse[J].Soybean Science,2016,35(1):106-111.
[26]王波,李红梅,王碧,等.淡紫拟青霉与放线菌代谢物复配对南方根结线虫的防治[J].南京农业大学学报,2009,32(1):55-60.WANG B,LI H M,WANG B,et al.Biological control of Meloidogyne incognita by combination of Paecilomyces lilacinus and Actinomycetes spp.[J].Journal of Nanjing Agricultural University,2009,32(1):55-60.
[27]KLEIN E,OFEK M,KATAN J,et al.Soil suppressiveness to Fusarium disease:Shifts in root microbiome associated with reduction of pathogen root colonization[J].Biological Contral,2013,103(1):23-33.
[28]JENKINS W R.A rapid centrifugal-flotation technique for separating nematodes from soil[J].Plant Disease Reporter,1964,48(9):692.
[29]ABALLAY E,PRODAN S,MRTENSSON A,et al.Assessment of rhizobacteria from grapevine for their suppressive effect on the parasitic nematode Xiphinema index[J].Crop Protection,2012,42:36-41.
[30]林茂松,沈纪冬,文玲,等.淡紫拟青霉代谢产物生物测定和生理活性物质分析[J].江苏农业学报,1999,15(4):226-228.LIN M S,SHEN J D,WEN L,et al.Bioassay of metabolic products of Pacilomyces lilacinus and analysis of its physiologically active substance[J].Jiangsu Journal of Agricultural Science,1999,15(4):226-228.
[31]REGAIEG H,CIANCIO A,RAOUANI N H,et al.Effects of culture filtrates from the nematophagous fungus Verticillium leptobactrum on viability of the root-knot nematode Meloidogyne incognita[J].World Journal of Microbiology and Biotechnology,2010,26(12):2285–2289.
[32]CHEN S Y,DICKSON D W,MITCHELL D J.Viability of Heterodera glycines exposed to fungal filtrates[J].The Journal of Nematology,2000,32(2):190-197.
[2]许艳丽,韩晓增.大豆重迎茬研究[M].哈尔滨:哈尔滨工程大学出版社,1995:136-139.XU Y L,HAN X Z.Study on continuous cropping and alternate-year of soybean[M].Harbin:Harbin Engineering University Press,1995:136-139.
[3]HARTWIG E E.Breeding productive soybean cultivars resistant to the soybean cyst nematode for the southern United States[J].Plant Disease,1981,65(4):303-307.
[4]CHEN S Y.Suppression of Heterodera glycines in soils from fields with long-term soybean monoculture[J].Biocontrol Science and Technology,2007,17(2):125-134.
[5]刘杏忠,张东升,武修英,等.定殖于大豆孢囊线虫孢囊内真菌的初步研究[J].北京农业大学学报,1991,17(3):87-91.LIU X Z,ZHANG D S,WU X Y,et al.Preliminary studies on the parasitic from the cyst of Heterodera glycines[J].Acta Agriculture Universitatis Pekinensis,1991,17(3):87-91.
[6]靳学慧,辛惠普,郑雯,等.长期轮作和连作对土壤中大豆胞囊线虫数量的影响[J].中国油料作物学报,2006,28(2):189-193.JIN X H,XIN H P,ZHENG W,et al.The influence of soil on the long-term rotation and continuous cultivation on soybean cyst nematode[J].Chinese Journal of Oil Crop Science,2006,28(2):189-193.
[7]许艳丽.土壤环境对大豆胞囊线虫卵孵化影响及线虫分子诊断研究[D].哈尔滨:东北农业大学,2004:41-47.XU Y L.The effects of soil environment on soybean cyst nematode egg hatch and nematode molecular diagnosis[D].Harbin:Northeast Agricultural University,2004:41-47.
[8]马承铸,顾真容,钱振官,等.种植制对大豆孢囊线虫群体密度和真菌区系的影响[J].上海农业学报,1994,10(2):62-66.MA C Z,GU Z R,QIAN Z G,et al.Effects of cropping system on mycoflora and population density of soybean cyst nematode Heterodera glycines[J].Acta Agriculturae Shanghai,1994,10(2):62-66.
[9]孙玉秋,许艳丽,李春杰,等.作物轮作系统对土壤中大豆胞囊线虫胞囊量和单胞囊卵量的影响[J].农业系统科学与综合研究,2011,27(2):248-252.SUN Y Q,XU Y L,LI C J,et al.Influence of cropping rotation systems on the volume of cysts and eggs in a single-cyst of soybean cyst nematode[J].System Science and Comprehensive Studies in Agriculture,2011,27(2):248-252.
[10]孙漫红,刘杏忠.连作土壤中大豆胞囊线虫种群数量减少的原因探讨[J].植物病理学报,2000,30(4):353-356,363.SUN M H,LIU X Z.Suppressive soil of soybean cysts nematode in China[J].Acta Phytopathologica Sinica,2000,30(4):353-356,363.
[11]CRUMP D H,SAYRE R M,YOUNG L D.Occurrence of nematophagous fungi in cyst nematode populations[J].Plant Disease,1983,67(1):63-64.
[12]孙漫红,刘杏忠,晋治波.淡紫拟青霉对大豆胞囊线虫卵及2龄幼虫的影响[J].植物保护学报,2002,29(1):57-61.SUN M H,LIU X Z,JIN Z B.Effect Paecilomyces lilacinus on egg hatching and juvenile mortality of Heterodera glycines[J].Journal of Plant Protection,2002,29(1):57-61.
[13]SONG J,LI S X,WEI W,et al.Assessment of parasitic fungi for reducing soybean cyst nematode with suppressive soil in soybean fields of northeast China[J].Acta Agriculturae Scandinavica,Section B—Soil and Plant Science,2017,67(8):730-736.
[14]宋洁,许艳丽,姚钦.大豆胞囊线虫主要寄生真菌对大豆耕作系统的响应[J].大豆科学,2016,35(3):461-467.SONG J,XU Y L,YAO Q.Response of major parasitic fungi of soybean cyst nematode on different cultivation systems[J].Soybean Science,2016,35(3):461-467.
[15]SONG J,LI S X,XU Y L,et al.Diversity of parasitic fungi from soybean cyst nematode associated with long-term continuous cropping of soybean in black soil[J].Acta Agriculturae Scandinavica,Section B—Soil and Plant Science,2016,66(5):432–442.
[16]陈立杰,刘晓杰,段玉玺,等.轮作和连作田大豆胞囊线虫胞囊上真菌定殖动态[J],大豆科学,2009,28(2):266-270.CHEN L J,LIU X J,DUAN Y X,et al.Dynamics of fungal colonization on cyst in rotation and continuous cropping of soybean[J].Soybean Science,2009,28(2):266-270.
[17]林茂松.土壤真菌防治大豆胞囊线虫的效果测定[J].中国油料,1990,12(3):63-65.LIN M S.Control effect to soybean cyst nematode by fungi from eggs of Heterodera glycines[J].Chinese Oil,1990,12(3):63-65.
[18]范圣长,段玉玺,陈立杰.大豆胞囊线虫胞囊内寄生真菌研究[J].大豆科学,2004,23(1):71-74.FAN S C,DUAN Y X,CHEN L J.The research on the cyst entoparasitic fungi of soybean cyst nematode[J].Soybean Science,2004,23(1):71-74.
[19]FITTERS P F L,DEN BELDER E.A time-lapse technique to study the effect of fungal products on embryogenesis of nematode eggs[J].Mededelingen van de Faculteit Landbouwwetenschappen-Rijksuniversiteit Gent,1993,58(2B):751-756.
[20]赵晓晖,许艳丽.镰刀菌发酵液对大豆胞囊线虫的抑制作用[J].大豆科学,2011,30(3):468-470,474.ZHAO X H,XU Y L.Inhibition of Fusarium spp.fermented filtrates on soybean cyst nematode[J].Soybean Science,2011,30(3):468-470,474.
[21]赵晓晖,许艳丽.厚垣轮枝菌发酵液对大豆胞囊线虫的抑制作用[J].大豆科技,2011,20(4):20-23.ZHAO X H,XU Y L.Inhibition of Verticillium chlamydosporium fermented filtrates on soybean cyst nematode[J].Soybean Science and Technology,2011,20(4):20-23.
[22]KHAN A,WILLIAMA K L,NEVALAINEN H K M.Effect of Paecilomyces lilacinus protease and chitinase on the eggshell structure and hatching of Meloidogyne jovanica juveniles[J].Biological Control 2004,31(3):346-352.
[23]宋洁.连作土壤寄生真菌多样性及对大豆胞囊线虫抑制作用[D].哈尔滨:东北农业大学,2016:58-64,75-92.SONG J.Diversity and suppressive effect of parasitic fungi on soyrean cyst nematode in soybean monoculture soil[D].Harbin:Northeast Agricultural University,2016:58-64,75-92.
[24]鲁建聪.寄生真菌代谢物对大豆胞囊线虫抑制及防治[D].哈尔滨:东北农业大学,2015:16-28.LU J C.Suppressness and biocontrol of parasitic fungal fermentation filtrate on soybean cyst nematode[D].Harbin:Northeast Agricultural University,2015:16-28.
[25]鲁建聪,许艳丽,宋洁.土壤扰动对寄生真菌代谢物温室防治大豆胞囊线虫的影响[J].大豆科学,2016,35(1):106-111.LU J C,XU Y L,SONG J.Effect of soil disturbance to the biological control of fermentation filtrate of parasitic fungi on soybean cyst nematode in glasshouse[J].Soybean Science,2016,35(1):106-111.
[26]王波,李红梅,王碧,等.淡紫拟青霉与放线菌代谢物复配对南方根结线虫的防治[J].南京农业大学学报,2009,32(1):55-60.WANG B,LI H M,WANG B,et al.Biological control of Meloidogyne incognita by combination of Paecilomyces lilacinus and Actinomycetes spp.[J].Journal of Nanjing Agricultural University,2009,32(1):55-60.
[27]KLEIN E,OFEK M,KATAN J,et al.Soil suppressiveness to Fusarium disease:Shifts in root microbiome associated with reduction of pathogen root colonization[J].Biological Contral,2013,103(1):23-33.
[28]JENKINS W R.A rapid centrifugal-flotation technique for separating nematodes from soil[J].Plant Disease Reporter,1964,48(9):692.
[29]ABALLAY E,PRODAN S,MRTENSSON A,et al.Assessment of rhizobacteria from grapevine for their suppressive effect on the parasitic nematode Xiphinema index[J].Crop Protection,2012,42:36-41.
[30]林茂松,沈纪冬,文玲,等.淡紫拟青霉代谢产物生物测定和生理活性物质分析[J].江苏农业学报,1999,15(4):226-228.LIN M S,SHEN J D,WEN L,et al.Bioassay of metabolic products of Pacilomyces lilacinus and analysis of its physiologically active substance[J].Jiangsu Journal of Agricultural Science,1999,15(4):226-228.
[31]REGAIEG H,CIANCIO A,RAOUANI N H,et al.Effects of culture filtrates from the nematophagous fungus Verticillium leptobactrum on viability of the root-knot nematode Meloidogyne incognita[J].World Journal of Microbiology and Biotechnology,2010,26(12):2285–2289.
[32]CHEN S Y,DICKSON D W,MITCHELL D J.Viability of Heterodera glycines exposed to fungal filtrates[J].The Journal of Nematology,2000,32(2):190-197.