生物表面活性剂鼠李糖脂对甘蔗黑穗病菌的体外抗菌活性
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摘要
【背景】甘蔗黑穗病是一种主要的甘蔗病害,易造成甘蔗严重减产;鼠李糖脂是一种生物表面活性剂,可作为多种植物真菌病害的抑菌剂。【目的】研究生物表面活性剂鼠李糖脂对甘蔗黑穗病菌的体外抗菌活性及初步的抗菌机理。【方法】采用甘蔗黑穗病冬孢子萌发试验研究鼠李糖脂对甘蔗黑穗病冬孢子的抗菌作用。采用菌丝生长速率法和菌丝干重法对鼠李糖脂的体外抑菌试验进行检测;通过菌丝电导率的变化研究鼠李糖脂对甘蔗黑穗病菌细胞膜通透性的影响。【结果】鼠李糖脂能显著抑制甘蔗黑穗病菌孢子萌发,其中2.0 g/L鼠李糖脂对甘蔗黑穗病冬孢子萌发的抑制效果最好,抑制率达45.03%。鼠李糖脂能显著抑制甘蔗黑穗病菌双核菌丝体、单胞菌a和单胞菌b的生长。鼠李糖脂能使甘蔗黑穗病单胞菌细胞膜透性增加,与对照相比,2.0 g/L鼠李糖脂处理甘蔗黑穗病双核菌丝体0.5min后电导率升高了约9倍,处理单胞菌a30min后电导率提高了94.23%;0.1g/L鼠李糖脂处理甘蔗黑穗病单胞菌b30min后电导率升高了54.49%,随着浓度的增加,各处理电导率升高显著。【结论】鼠李糖脂对甘蔗黑穗病菌有良好的抗菌作用,有望为甘蔗黑穗病的防治提供新方法。
[Background] Sugarcane smut disease is a main disease on sugarcane that causes considerable yield losses; rhamnolipids are biosurfactants that have been studied as an antifungal agent against various of plant diseases. [Objective] To evaluate the antifungal activity of rhamnolipids against S. scitamineum in vitro, exploring the preliminary antifungal mechanism of rhamnolipids against S. scitamineum. [Methods]The teliospores of sugarcane smut germination were employed to determine the antifungal of rhamnolipids. The method of mycelial growth rate and dry weight of mycelium were used to test in in vitro test of rhamnolipid. The effect of rhamnolipids on the membrane permeability of S. scitamineum that was assayed the change of electrical conductivity of the hyphae. [Results] The results showed that rhamnolipids could significantly inhibit the spore germination of S. scitamineum. 2.0 g/L rhamnolipids reduced the germination rate of teliospores by 45.03%. Rhamnolipids significantly inhibited the mycelial growth of the diploid, haploid-a and haploid-b strain of S. scitamineum. Rhamnolipids could increase the membrane permeability of cell membrane of S. scitamineum, compared with the control, the conductivity of diploid strain raised 9-fold at 2.0 g/L rhamnolipids after 5 min,the conductivity of haploid-a strain of S.scitamineum increased by 94.23% after 30 min at 2.0 g/L rhamnolipids. The conductivity of mycelium increased by 54.49% while haploid-b strain of S. scitamineum had been handled with 0.1 g/L rhamnolipids for 30 min. Mycelial conductivity raised with the increasing of the concentration of rhamnolipids.[Conclusion] Rhamnolipids showed a high antifungal activity against S. scitamineum, which could offer a new method for controlling sugarcane smut disease.
[Background] Sugarcane smut disease is a main disease on sugarcane that causes considerable yield losses; rhamnolipids are biosurfactants that have been studied as an antifungal agent against various of plant diseases. [Objective] To evaluate the antifungal activity of rhamnolipids against S. scitamineum in vitro, exploring the preliminary antifungal mechanism of rhamnolipids against S. scitamineum. [Methods]The teliospores of sugarcane smut germination were employed to determine the antifungal of rhamnolipids. The method of mycelial growth rate and dry weight of mycelium were used to test in in vitro test of rhamnolipid. The effect of rhamnolipids on the membrane permeability of S. scitamineum that was assayed the change of electrical conductivity of the hyphae. [Results] The results showed that rhamnolipids could significantly inhibit the spore germination of S. scitamineum. 2.0 g/L rhamnolipids reduced the germination rate of teliospores by 45.03%. Rhamnolipids significantly inhibited the mycelial growth of the diploid, haploid-a and haploid-b strain of S. scitamineum. Rhamnolipids could increase the membrane permeability of cell membrane of S. scitamineum, compared with the control, the conductivity of diploid strain raised 9-fold at 2.0 g/L rhamnolipids after 5 min,the conductivity of haploid-a strain of S.scitamineum increased by 94.23% after 30 min at 2.0 g/L rhamnolipids. The conductivity of mycelium increased by 54.49% while haploid-b strain of S. scitamineum had been handled with 0.1 g/L rhamnolipids for 30 min. Mycelial conductivity raised with the increasing of the concentration of rhamnolipids.[Conclusion] Rhamnolipids showed a high antifungal activity against S. scitamineum, which could offer a new method for controlling sugarcane smut disease.
引文
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[12]Guo S,Zhang ZG,Tian YQ,et al.Allelopathic effects of the root exudates in the watermelon and rootstocks on Fusarium oxysporum f.sp.niveum[J].Acta Agriculturae Boreali-Sinica,2010,25(6):160-163(in Chinese)郭尚,张作刚,田永强,等.西瓜及砧木根系分泌物对西瓜枯萎病菌的化感效应[J].华北农学报,2010,25(6):160-163
[13]Fang ZD.Plant Disease Research Methods[M].3rd ed.Beijing:China Agriculture Press,1998(in Chinese)方中达.植病研究方法[M].3版.北京:中国农业出版社,1998
[14]Liu JD,Jiang XF.Toxicity test of Jinggangmycin A,diniconazole and their mixtures to Ustilaginoidea virens[J].Modern Agrochemicals,2005,4(1):31-32,34(in Chinese)刘嘉德,蒋秀芳.井冈霉素、烯唑醇及其混剂对稻曲病菌毒力测定[J].现代农药,2005,4(1):31-32,34
[15]Xia XM,Wang KY,Fan K,et al.Changes of relative leakage and osmolarity sensitivity to tebuconazole-resistant strains of Rhizoctonia cerealis[J].Chinese Journal of Pesticide Science,2005,7(2):126-130(in Chinese)夏晓明,王开运,范昆,等.抗戊唑醇禾谷丝核菌的渗透压敏感性及相对渗率变化研究[J].农药学学报,2005,7(2):126-130
[16]Sanchez L,Courteaux B,Hubert J,et al.Rhamnolipids elicit defense responses and induce disease resistance against biotrophic,hemibiotrophic,and necrotrophic pathogens that require different signaling pathways in arabidopsis and highlight a central role for salicylic acid[J].Plant Physiology,2012,160(3):1630-1641
[17]Vatsa P,Sanchez L,Clement C,et al.Rhamnolipid biosurfactants as new players in animal and plant defense against microbes[J].International Journal of Molecular Sciences,2010,11(12):5095-5108
[18]Goswami D,Borah SN,Lahkar J,et al.Antifungal properties of rhamnolipid produced by Pseudomonas aeruginosa DS9 against Colletotrichum falcatum[J].Journal of Basic Microbiology,2015,55(11):1265-1274
[19]de Jonghe K,de Dobbelaere I,Sarrazyn R,et al.Control of Phytophthora cryptogea in the hydroponic forcing of witloof chicory with the rhamnolipid-based biosurfactant formulation PRO1[J].Plant Pathology,2005,54(2):219-226
[20]Kim BS,Lee JY,Hwang BK.In vivo control and in vitro antifungal activity of rhamnolipid B,a glycolipid antibiotic,against Phytophthora capsici and Colletotrichum orbiculare[J].Pest Management Science,2000,56(12):1029-1035
[21]Zhu GN,Lin SY,Xian XY,et al.Bioactivities of eight fungicides against Sporisorium scitaminea[J].Journal of Southern Agriculture,2014,45(6):989-994(in Chinese)朱桂宁,林珊宇,贤小勇,等.8种杀菌剂对甘蔗黑穗病菌的室内毒力测定[J].南方农业学报,2014,45(6):989-994
[22]Wu WH,Xie ZJ,He CP,et al.Biological characteristics of teliospores of Ustilago scitaminea and virulence of 2fungicides against them indoor[J].Chinese Journal of Tropical Crops,2009,30(11):1674-1678(in Chinese)吴伟怀,谢祖健,贺春萍,等.甘蔗黑穗病菌冬孢子生物学特性及杀菌剂对其萌发的影响[J].热带作物学报,2009,30(11):1674-1678
[23]Perneel M,D’Hondt L,de Maeyer K,et al.Phenazines and biosurfactants interact in the biological control of soil-borne diseases caused by Pythium spp.[J].Environmental Microbiology,2008,10(3):778-788
[24]Chapman JS.Biocide resistance mechanisms[J].International Biodeterioration&Biodegradation,2003,51(2):133-138
[25]Chen ZL,Zhu BY,Liu F,et al.Antifungal activity of two quaternary ammonium cationic surfactants against Botrytis cinerea[J].Acta Phytophylacica Sinica,2007,34(5):539-544(in Chinese)陈召亮,朱炳煜,刘峰,等.两种季铵盐阳离子表面活性剂对番茄灰霉病菌的生物活性[J].植物保护学报,2007,34(5):539-544
[26]Bender CL,Scholz-Schroeder BK.New insights into the biosynthesis,mode of action,and regulation of syringomycin,syringopeptin,and coronatine[A]//Ramos JL.Virulence and Gene Regulation.Berlin Heidelberg:Springer,2004:125-158
[27]Yu T,Shang YK,Shi HX,et al.Antifungal activity of cationic surfactants quarternary ammonium salt on Sclerotinia sclerotiorum[J].Chinese Journal of Pesticide Science,2008,10(1):99-104(in Chinese)于婷,尚玉珂,石怀兴,等.几种季铵盐阳离子表面活性剂对核盘菌的抗菌活性[J].农药学学报,2008,10(1):99-104
[2]Shen WK.Sugarcane disease in Guangdong sugarcane regions in recent years and integrated controlling methods[J].Sugarcane&Canesugar,2004(1):1-5(in Chinese)沈万宽.广东蔗区甘蔗病害现状与综合防治措施[J].甘蔗糖业,2004(1):1-5
[3]Albert HH,Schenck S.PCR amplification from a homolog of the bE mating-type gene as a sensitive assay for the presence of Ustilago scitaminea DNA[J].Plant Disease,1996,80(10):1189-1192
[4]Kamal A,Shaik AB,Kumar CG,et al.Metabolic profiling and biological activities of bioactive compounds produced by Pseudomonas sp.strain ICTB-745 isolated from Ladakh,India[J].Journal of Microbiology and Biotechnology,2012,22(1):69-79
[5]Stanghellini ME,Kim DH,Rasmussen SL,et al.Control of root rot of peppers caused by Phytophthora capsid with a nonionic surfactant[J].Plant Disease,1996,80(10):1113-1116
[6]Stanghellini ME,Miller RM.Biosurfactants:their identity and potential efficacy in the biological control of zoosporic plant pathogens[J].Plant Disease,1997,81(1):4-12
[7]Sha RY,Jiang LF,Meng Q,et al.Producing cell-free culture broth of rhamnolipids as a cost-effective fungicide against plant pathogens[J].Journal of Basic Microbiology,2012,52(4):458-466
[8]Lahkar J,Borah SN,Deka S,et al.Biosurfactant of Pseudomonas aeruginosa JS29 against Alternaria solani:the causal organism of early blight of tomato[J].BioControl,2015,60(3):401-411
[9]D’Aes J,De Maeyer K,Pauwelyn E,et al.Biosurfactants in plant-Pseudomonas interactions and their importance to biocontrol[J].Environmental Microbiology Reports,2010,2(3):359-372
[10]Wang L,Mao YL,Xu HL,et al.Research progress on rhamnolipid in the control of plant fungal disease[J].Guangdong Agricultural Sciences,2017,44(8):90-95(in Chinese)王蕾,毛玉玲,许汉亮,等.鼠李糖脂防治植物真菌病害研究进展[J].广东农业科学,2017,44(8):90-95
[11]Shen WK,Jiang ZD,Yang ZD,et al.New resistance identification method and resistance evaluation of sugarcane varieties to smut disease[J].Journal of Huazhong Agricultural University,2014,33(2):51-56(in Chinese)沈万宽,姜子德,杨湛端,等.甘蔗抗黑穗病的鉴定新方法及其品种抗性评价[J].华中农业大学学报,2014,33(2):51-56
[12]Guo S,Zhang ZG,Tian YQ,et al.Allelopathic effects of the root exudates in the watermelon and rootstocks on Fusarium oxysporum f.sp.niveum[J].Acta Agriculturae Boreali-Sinica,2010,25(6):160-163(in Chinese)郭尚,张作刚,田永强,等.西瓜及砧木根系分泌物对西瓜枯萎病菌的化感效应[J].华北农学报,2010,25(6):160-163
[13]Fang ZD.Plant Disease Research Methods[M].3rd ed.Beijing:China Agriculture Press,1998(in Chinese)方中达.植病研究方法[M].3版.北京:中国农业出版社,1998
[14]Liu JD,Jiang XF.Toxicity test of Jinggangmycin A,diniconazole and their mixtures to Ustilaginoidea virens[J].Modern Agrochemicals,2005,4(1):31-32,34(in Chinese)刘嘉德,蒋秀芳.井冈霉素、烯唑醇及其混剂对稻曲病菌毒力测定[J].现代农药,2005,4(1):31-32,34
[15]Xia XM,Wang KY,Fan K,et al.Changes of relative leakage and osmolarity sensitivity to tebuconazole-resistant strains of Rhizoctonia cerealis[J].Chinese Journal of Pesticide Science,2005,7(2):126-130(in Chinese)夏晓明,王开运,范昆,等.抗戊唑醇禾谷丝核菌的渗透压敏感性及相对渗率变化研究[J].农药学学报,2005,7(2):126-130
[16]Sanchez L,Courteaux B,Hubert J,et al.Rhamnolipids elicit defense responses and induce disease resistance against biotrophic,hemibiotrophic,and necrotrophic pathogens that require different signaling pathways in arabidopsis and highlight a central role for salicylic acid[J].Plant Physiology,2012,160(3):1630-1641
[17]Vatsa P,Sanchez L,Clement C,et al.Rhamnolipid biosurfactants as new players in animal and plant defense against microbes[J].International Journal of Molecular Sciences,2010,11(12):5095-5108
[18]Goswami D,Borah SN,Lahkar J,et al.Antifungal properties of rhamnolipid produced by Pseudomonas aeruginosa DS9 against Colletotrichum falcatum[J].Journal of Basic Microbiology,2015,55(11):1265-1274
[19]de Jonghe K,de Dobbelaere I,Sarrazyn R,et al.Control of Phytophthora cryptogea in the hydroponic forcing of witloof chicory with the rhamnolipid-based biosurfactant formulation PRO1[J].Plant Pathology,2005,54(2):219-226
[20]Kim BS,Lee JY,Hwang BK.In vivo control and in vitro antifungal activity of rhamnolipid B,a glycolipid antibiotic,against Phytophthora capsici and Colletotrichum orbiculare[J].Pest Management Science,2000,56(12):1029-1035
[21]Zhu GN,Lin SY,Xian XY,et al.Bioactivities of eight fungicides against Sporisorium scitaminea[J].Journal of Southern Agriculture,2014,45(6):989-994(in Chinese)朱桂宁,林珊宇,贤小勇,等.8种杀菌剂对甘蔗黑穗病菌的室内毒力测定[J].南方农业学报,2014,45(6):989-994
[22]Wu WH,Xie ZJ,He CP,et al.Biological characteristics of teliospores of Ustilago scitaminea and virulence of 2fungicides against them indoor[J].Chinese Journal of Tropical Crops,2009,30(11):1674-1678(in Chinese)吴伟怀,谢祖健,贺春萍,等.甘蔗黑穗病菌冬孢子生物学特性及杀菌剂对其萌发的影响[J].热带作物学报,2009,30(11):1674-1678
[23]Perneel M,D’Hondt L,de Maeyer K,et al.Phenazines and biosurfactants interact in the biological control of soil-borne diseases caused by Pythium spp.[J].Environmental Microbiology,2008,10(3):778-788
[24]Chapman JS.Biocide resistance mechanisms[J].International Biodeterioration&Biodegradation,2003,51(2):133-138
[25]Chen ZL,Zhu BY,Liu F,et al.Antifungal activity of two quaternary ammonium cationic surfactants against Botrytis cinerea[J].Acta Phytophylacica Sinica,2007,34(5):539-544(in Chinese)陈召亮,朱炳煜,刘峰,等.两种季铵盐阳离子表面活性剂对番茄灰霉病菌的生物活性[J].植物保护学报,2007,34(5):539-544
[26]Bender CL,Scholz-Schroeder BK.New insights into the biosynthesis,mode of action,and regulation of syringomycin,syringopeptin,and coronatine[A]//Ramos JL.Virulence and Gene Regulation.Berlin Heidelberg:Springer,2004:125-158
[27]Yu T,Shang YK,Shi HX,et al.Antifungal activity of cationic surfactants quarternary ammonium salt on Sclerotinia sclerotiorum[J].Chinese Journal of Pesticide Science,2008,10(1):99-104(in Chinese)于婷,尚玉珂,石怀兴,等.几种季铵盐阳离子表面活性剂对核盘菌的抗菌活性[J].农药学学报,2008,10(1):99-104
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