十堰辣椒根腐病病原菌分离鉴定及高氏15号的生物防治作用研究
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摘要
为明确辣椒根腐病病原,从田间采集分离、纯化、鉴定、柯赫氏法则验证病原菌,并开展高氏15号生物菌剂对辣椒根腐病的防治效果和促生作用研究.结果表明,通过形态学和分子生物学鉴定(ITS、EF-1a、IGS基因序列比对及进化树分析)发现,引起该地区辣椒根腐病的病原物为茄病镰刀菌(Fusarium solani).柯赫氏法则完成了辣椒根腐病的回接鉴定试验.田间防效和促生结果显示,多粘类芽孢杆菌在240g/667m2,高氏15号菌剂在50g/穴和75g/穴,甲霜·恶霉灵1 500倍液防治辣椒根腐病效果最好,均达到70%以上.经试验处理的株高、根长、整株鲜重、根鲜重和根干重和空白对照相比有显著差异性.
To identify the pathogen of pepper root rot,the pathogenic fungi was isolated and purified from pepper stalk root disease.The pathogenic fungus was verified via molecular determination and Koch's Rule.Field control and growth promotion effect were achieved to prevent pepper root rot by Gauze No.15.The result showed that the pathogen causing root rot of pepper was identified as Fusarium solani through identification of morphological characteristics and molecular biological analysis including ITS,EF-1 aand IGS genes and phylogenetic tree in this area.Koch's Rule was finished by infestation test.In addition,the result indicated that applying 240 g/667 m2 of Paenibacillus polymyxa,50 g/den and 75 g/den of Gause No.15 and 3% hymexazol metalaxyl AS(1 500×)generated the optimal inhibition effect on the Fusarium solani(more than 70%).After treatment,the pepper height,root length,fresh weight of the whole plant and root,and dry weight of the root had significant difference comparing with control group.
To identify the pathogen of pepper root rot,the pathogenic fungi was isolated and purified from pepper stalk root disease.The pathogenic fungus was verified via molecular determination and Koch's Rule.Field control and growth promotion effect were achieved to prevent pepper root rot by Gauze No.15.The result showed that the pathogen causing root rot of pepper was identified as Fusarium solani through identification of morphological characteristics and molecular biological analysis including ITS,EF-1 aand IGS genes and phylogenetic tree in this area.Koch's Rule was finished by infestation test.In addition,the result indicated that applying 240 g/667 m2 of Paenibacillus polymyxa,50 g/den and 75 g/den of Gause No.15 and 3% hymexazol metalaxyl AS(1 500×)generated the optimal inhibition effect on the Fusarium solani(more than 70%).After treatment,the pepper height,root length,fresh weight of the whole plant and root,and dry weight of the root had significant difference comparing with control group.
引文
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[9]李大庆,郑元利,文西明,等.几种不同药剂防治辣椒根腐病田间药效试验[J].植物医生,2007(2):44-45.LI D Q,ZHENG Y L,WEN X M,et al.Field control effects of different agentia on pepper root rot[J].Plant Doctor,2007(2):44-45.(Ch).
[10]鲁海菊,沈云玫,陶宏征,等.内生木霉P3.9菌株的多功能性及其枇杷根腐病的盆栽防效[J].西北农业学报,2017,26(11):1681-1688.LU H J,SHEN Y M,TAO H Z,et al.Multifunction of endophytic trichoderma P3.9strain and control effect on loquat root rot[J].Acta Agriculturae Boreali-Occidentalis Sinica,2017,26(11):1681-1688.(Ch).
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[12]高同国,李术娜,张冬冬,等.大豆根腐病生防细菌优势菌株的筛选、鉴定及生防效果验证[J].大豆科学,2015,34(4):661-665.GAO T G,LI S N,ZHANG D D,et al.Screening and identification of biocontrol dominant bacterium against to soybean root rot[J].Soybean Science,2015,34(4):661-665.(Ch).
[13]LUGTENBERG B,KAMILOVA F.Plant-growth-promoting rhizobacteria[J].Annual Review of Microbiology,2009,63:541-556.
[14]汪华.一种防治蔬菜根部病害的多孔烟管菌高氏15号及其制剂:CN 201510564715.4[P].2015-09-08.WANG H.Gauze No.15from Blerkandera adusta to control vegetable root rot diseases:CN 201510564715.4[P].2015-09-08.(Ch).
[15]LESLIE J F,SUMMERELL B A.The Fusarium Laboratory Manual[M].Hoboken:Wiley,2007.
[16]ZHU Z,ZHENG L,PAN L,et al.Identification and characterization of Fusariumspecies associated with wilt of Eleocharis dulcis(Chinese water chestnut)in China[J].Plant Disease,2014,98(7):977-987.
[17]SHEN S S,CHOI O H,PARK S H,et al.Root colonizing and biocontrol competency of Serratia plymuthica A21-4against Phytophthora blight of pepper[J].The Plant Pathology Journal,2005,21(1):64-67.
[18]KLOEPPER J W.A review of issues related to measuring colonization of plant roots by bacteria[J].Canadian Journal of Microbiology,1992,38(6):667-672.
[19]李曹龙,秦艳,欧秀玲,等.高温产蛋白酶菌株的筛选及产酶条件的研究[J].中国酿造,2012,31(12):98-101.LI C L,QIN Y,OU X L,et al.Screening of thermostable protease-producing bacterium and its enzyme-producing condition[J].China Brewing,2012,31(12):98-101.(Ch).
[20]刘晓玲,王金晶,李永仙,等.产β-1,3-1,4-葡聚糖酶特基拉芽孢杆菌Bacillus tequilensis CGX5-1发酵培养基的优化[J].食品与生物技术学报,2013,32(6):645-650.LIU X L,WANG J J,LI Y X,et al.Medium optimization for the production ofβ-1,3-1,4-Glucanase by Bacillus tequilensis CGX5-1[J].Journal of Food Science and Biotechnology,2013,32(6):645-650.(Ch).
[21]王凯.黄曲霉生防菌的筛选鉴定及高效菌株JPP1的生防机制[D].哈尔滨:哈尔滨工业大学,2013.WANG K.Research on Screening and Identification of Bbiological Control Bacteria Against Aflatoxins and Mechanism of Strain JPP1for Biocontrol[M].Harbin:Harbin Institute of Technology,2013.(Ch).
[22]冯金龙.马铃薯病原真菌拮抗性细菌鉴定及其生物学功能研究[D].兰州:甘肃农业大学,2013.FENG J L.Potato Pathogenic Fungi Antagonistic Bacterial Identification and Its Bioligical Function Study[D].Lanzhou:Gansu Agricultural University,2013.(Ch).
[23]BAK W C.Characteristics of bed-log of shiitake damaged by Bjerkandera adusta and antagonism between these two fungi[J].The Korean Journal of Mycology,2011,39(1):44-47.
[24]张旭辉,张红楠,李勇,等.抑制西瓜蔓枯病菌的生防真菌筛选、鉴定及发酵条件优化[J].中国生物工程杂志,2017,37(5):76-86.ZHANG X H,ZHANG H N,LI Y,et al.Screening and identification of biocontrol fungi against Didymella bryoniae and optimization of fermentation conditions[J].China Biotechnology,2017,37(5):76-86.(Ch).
[25]汪文强,李勇,张旭辉,等.烟管菌M-1对柑橘炭疽病菌的抑制作用及发酵条件优化[J].中国生物防治学报,2018,34(3):423-430.WANG W Q,LI Y,ZHANG X H,et al.Antagonism of Bjerkandera adusta M-1 against colletotrichum gloeosprioides and optimization of fermentation conditions[J].Chinese Journal of Biological Control,2018,34(3):423-430.(Ch).
[26]MIRIK M,AYSAN Y,CINAR O.Biological control of bacterial spot disease of pepper with bacillus strains[J].Turkish Journal of Agruculture and Foorestry,2008,32(5):381-390.
[27]CONSTANTINO M,GMEZLVAREZ R,LVAREZSOLSJ D,et al.Effect of in oculation with rhizobacteria and arbuscular aycorrhizal fungi on growth and yield of Capsicum chinense Jacquin[J].Journal of Agriculture and Rural Development in the Tropics and Subtropics,2008,109(2):169-180.
[2]李传宝.冬暖式大棚辣椒病害防治措施[J].辣椒杂志,2004(1):25-26.LI C B.The prevention and control measures of pepper disease in winter greenhouse[J].Pepper Magazine,2004(1):25-26.(Ch).
[3]吕佩珂,刘文珍,段半锁,等.中国蔬菜病虫原色图谱续集[M].呼和浩特:远方出版社,1996.LYU P K,LIU W Z,DUAN B S,et al.Sequel to the Primary Chromatogram of Diseases and Insects in China[M].Hohhot:Distant Press,1996.(Ch).
[4]郝蓉蓉,郭海东,杨成德,等.西藏设施辣椒根腐病病原的分离及鉴定[J].西北农业学报,2015,24(12):139-143.HAO R R,GUO H D,YANG C D,et al.Isolation and detection of the pathogen caused root rot in Tibet[J].Acta Agriculturae Boreali-Occidentalis Sinica,2015,24(12):139-143.(Ch).
[5]张亭亭,刘长远,赵奎华,等.辽宁省辣椒土传病害镰刀菌鉴定及rDNA-ITS序列分析[J].植物保护,2011,37(5):101-105.ZHANG T T,LIU C Y,ZHAO K H,et al.Identification of phytopathogenic Fusarium causing pepper soil borne diseases in Liaoning Province and sequence analysis of ribosomal DNA-ITS[J].Plant Protection,2011,37(5):101-105.(Ch).
[6]BOOTH C.The Genus Fusarium[M].Beijing:Agricultural Press,1988.
[7]郭普.植保大典[M].北京:中国三峡出版社农业科教出版中心,2006:307-308.GUO P.Encyclopedia Plant Protection[M].Beijing:Agricultural Science and Education Publishing Center of the Three Gorges Press in China,2006:307-308.(Ch).
[8]陈彦,刘长远,王晓红,等.辣椒根腐病化学防治田间试验[J].植物保护,2008,34(6):150-152.CHEN Y,LIU C Y,WANG X H,et al.Field efficacy of chemical agents against pepper root rot[J].Plant Protection,2008,34(6):150-152.(Ch).
[9]李大庆,郑元利,文西明,等.几种不同药剂防治辣椒根腐病田间药效试验[J].植物医生,2007(2):44-45.LI D Q,ZHENG Y L,WEN X M,et al.Field control effects of different agentia on pepper root rot[J].Plant Doctor,2007(2):44-45.(Ch).
[10]鲁海菊,沈云玫,陶宏征,等.内生木霉P3.9菌株的多功能性及其枇杷根腐病的盆栽防效[J].西北农业学报,2017,26(11):1681-1688.LU H J,SHEN Y M,TAO H Z,et al.Multifunction of endophytic trichoderma P3.9strain and control effect on loquat root rot[J].Acta Agriculturae Boreali-Occidentalis Sinica,2017,26(11):1681-1688.(Ch).
[11]张雷.解淀粉芽孢杆菌15-1-1发酵条件优化及对辣椒根腐病防治效果初探[D].哈尔滨:东北农业大学,2014.ZHANG L.Studies of Optimization of Fermentation Conditions and Biocontrol Effects Against Fusarium solar of Bacillus amyloliguefaciens 15-1-1[D].Harbin:Northeast Agricultural University,2014.(Ch).
[12]高同国,李术娜,张冬冬,等.大豆根腐病生防细菌优势菌株的筛选、鉴定及生防效果验证[J].大豆科学,2015,34(4):661-665.GAO T G,LI S N,ZHANG D D,et al.Screening and identification of biocontrol dominant bacterium against to soybean root rot[J].Soybean Science,2015,34(4):661-665.(Ch).
[13]LUGTENBERG B,KAMILOVA F.Plant-growth-promoting rhizobacteria[J].Annual Review of Microbiology,2009,63:541-556.
[14]汪华.一种防治蔬菜根部病害的多孔烟管菌高氏15号及其制剂:CN 201510564715.4[P].2015-09-08.WANG H.Gauze No.15from Blerkandera adusta to control vegetable root rot diseases:CN 201510564715.4[P].2015-09-08.(Ch).
[15]LESLIE J F,SUMMERELL B A.The Fusarium Laboratory Manual[M].Hoboken:Wiley,2007.
[16]ZHU Z,ZHENG L,PAN L,et al.Identification and characterization of Fusariumspecies associated with wilt of Eleocharis dulcis(Chinese water chestnut)in China[J].Plant Disease,2014,98(7):977-987.
[17]SHEN S S,CHOI O H,PARK S H,et al.Root colonizing and biocontrol competency of Serratia plymuthica A21-4against Phytophthora blight of pepper[J].The Plant Pathology Journal,2005,21(1):64-67.
[18]KLOEPPER J W.A review of issues related to measuring colonization of plant roots by bacteria[J].Canadian Journal of Microbiology,1992,38(6):667-672.
[19]李曹龙,秦艳,欧秀玲,等.高温产蛋白酶菌株的筛选及产酶条件的研究[J].中国酿造,2012,31(12):98-101.LI C L,QIN Y,OU X L,et al.Screening of thermostable protease-producing bacterium and its enzyme-producing condition[J].China Brewing,2012,31(12):98-101.(Ch).
[20]刘晓玲,王金晶,李永仙,等.产β-1,3-1,4-葡聚糖酶特基拉芽孢杆菌Bacillus tequilensis CGX5-1发酵培养基的优化[J].食品与生物技术学报,2013,32(6):645-650.LIU X L,WANG J J,LI Y X,et al.Medium optimization for the production ofβ-1,3-1,4-Glucanase by Bacillus tequilensis CGX5-1[J].Journal of Food Science and Biotechnology,2013,32(6):645-650.(Ch).
[21]王凯.黄曲霉生防菌的筛选鉴定及高效菌株JPP1的生防机制[D].哈尔滨:哈尔滨工业大学,2013.WANG K.Research on Screening and Identification of Bbiological Control Bacteria Against Aflatoxins and Mechanism of Strain JPP1for Biocontrol[M].Harbin:Harbin Institute of Technology,2013.(Ch).
[22]冯金龙.马铃薯病原真菌拮抗性细菌鉴定及其生物学功能研究[D].兰州:甘肃农业大学,2013.FENG J L.Potato Pathogenic Fungi Antagonistic Bacterial Identification and Its Bioligical Function Study[D].Lanzhou:Gansu Agricultural University,2013.(Ch).
[23]BAK W C.Characteristics of bed-log of shiitake damaged by Bjerkandera adusta and antagonism between these two fungi[J].The Korean Journal of Mycology,2011,39(1):44-47.
[24]张旭辉,张红楠,李勇,等.抑制西瓜蔓枯病菌的生防真菌筛选、鉴定及发酵条件优化[J].中国生物工程杂志,2017,37(5):76-86.ZHANG X H,ZHANG H N,LI Y,et al.Screening and identification of biocontrol fungi against Didymella bryoniae and optimization of fermentation conditions[J].China Biotechnology,2017,37(5):76-86.(Ch).
[25]汪文强,李勇,张旭辉,等.烟管菌M-1对柑橘炭疽病菌的抑制作用及发酵条件优化[J].中国生物防治学报,2018,34(3):423-430.WANG W Q,LI Y,ZHANG X H,et al.Antagonism of Bjerkandera adusta M-1 against colletotrichum gloeosprioides and optimization of fermentation conditions[J].Chinese Journal of Biological Control,2018,34(3):423-430.(Ch).
[26]MIRIK M,AYSAN Y,CINAR O.Biological control of bacterial spot disease of pepper with bacillus strains[J].Turkish Journal of Agruculture and Foorestry,2008,32(5):381-390.
[27]CONSTANTINO M,GMEZLVAREZ R,LVAREZSOLSJ D,et al.Effect of in oculation with rhizobacteria and arbuscular aycorrhizal fungi on growth and yield of Capsicum chinense Jacquin[J].Journal of Agriculture and Rural Development in the Tropics and Subtropics,2008,109(2):169-180.
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