青枯病植物疫苗工程菌FJAT-91ΔepsD培养条件优化
|
摘要
为优化青枯病植物疫苗工程菌FJAT-91ΔepsD的发酵条件,考察培养温度、培养基pH值、装液量对菌株生物量的影响,通过Box-Benhnken试验设计和响应面分析法,建立以生物量为响应值的多元二次回归模型,确定菌株发酵的最优条件。结果表明,最优发酵条件为温度29℃、pH值为7.0、装液量57 m L。根据优化的条件,获得菌株FJAT-91ΔepsD的生物量为19.8×108cfu/mL,较优化前提高了47.8%。本研究获得的菌株FJAT-91ΔepsD发酵条件可提高其生物量从而降低生产成本,有助于植物疫苗制剂的应用推广。
In order to optimize the fer mentation conditions of engineering bacterial strain FJAT-91ΔepsD for plant vaccine against bacterial wilt, the effects of culture temperature, pH value of medium and volume of liquid on the biomass of bacterial strains were investigated. Through Box-Benhnken test design and response surface analysis, a multiple quadratic regression model with biomass as response value was established to determine the optimal fermentation conditions of bacterial strains. The results showed that the optimum fermentation conditions were temperature 29℃, p H 7.0 and liquid loading 57 mL. Under the optimized conditions, the biomass of strain FJAT-91ΔepsD was 19.8×108 cfu/mL, which was 47.8% higher than that before optimization. The fermentation conditions of strain FJAT-91 epsD obtained in this study could increase its biomass and reduce production costs,which was helpful to the application and popularization of plant vaccine preparations.
In order to optimize the fer mentation conditions of engineering bacterial strain FJAT-91ΔepsD for plant vaccine against bacterial wilt, the effects of culture temperature, pH value of medium and volume of liquid on the biomass of bacterial strains were investigated. Through Box-Benhnken test design and response surface analysis, a multiple quadratic regression model with biomass as response value was established to determine the optimal fermentation conditions of bacterial strains. The results showed that the optimum fermentation conditions were temperature 29℃, p H 7.0 and liquid loading 57 mL. Under the optimized conditions, the biomass of strain FJAT-91ΔepsD was 19.8×108 cfu/mL, which was 47.8% higher than that before optimization. The fermentation conditions of strain FJAT-91 epsD obtained in this study could increase its biomass and reduce production costs,which was helpful to the application and popularization of plant vaccine preparations.
引文
Chen W.Q.,Li Y.F.,and Shen T.C.,2013,Optimization of culture conditions of a moderately halophilic bacteria by response surface methodology,Shipin Gongye Keji(Science and Technology of Food Industry),34(1):140-144(陈蔚青,李一飞,申屠超,2013,响应面法优化一株中度嗜盐菌的培养条件,食品工业科技,34(1):140-144)
Chen X.Q.,Chen D.J.,Zhu Y.J.,Chen Y.P.,Zhang H.F.,and Liu B.,2018,Construction and characterization of extracellular polysaccharide deletion mutant of Ralstonia solanacearum,Weishengwu Xuebao(Acta Microbiologica Sinica),58(5):926-938(陈小强,陈德局,朱育菁,陈燕萍,张海峰,刘波,2018,青枯雷尔氏菌胞外多糖合成缺失突变体构建及其生物学特性,微生物学报,58(5):926-938)
Ghanem K.M.,Al-Fassi F.A.,and Al-Hazmi N.M.,2016,Optimization of chloroxylenol degradation by Aspergillus niger using plackett-burman design and response surface methodology,Rom.Biotech.Lett.,11(1):15040-15048
Hayward A.C.,1991,Biology and epidemiology of bacterial wilt caused by Pseudomonas solanacearum,Annu.Rev.Phytopathol.,29:65-87
Huang Z.C.,Liu Y.,Zeng F.H.,and Lu X.Y.,2008,Advances in pathogenesis of Ralstonia solanacearum and crops resistance to bacterial wilt,Redai Yaredai Zhiwu Xuebao(Journal of Tropical and Subtropical Botany),16(5):491-496(黄真池,刘媛,曾富华,卢向阳,2008,青枯菌致病机理及作物抗青枯病研究进展,热带亚热带植物学报,16(5):491-496)
Jiang G.,Wei Z.,Xu J.,Chen H.L.,Zhang Y.,She X.M.,Macho A.P.,Ding W.,and Liao B.S.,2017,Bacterial wilt in China:history,current status,and future perspectives,Front.Plant Sci.,8:1549
Ogawa K.,Mori T.,Matsusaki H.,and Matsuzoe N.,2012,Simplified evaluation method for suppressing bacterial wilt disease by treating non-pathogenic Ralstonia solanacearum mutant selection in eggplant,Hortic.Res.,11(3):393-398
Poueymiro M.,Cazale A.C.,Francois J.M.,Parrou J.L.,Peeters N.,and Genin S.,2014,A Ralstonia solanacearum typeⅢeffector directs the production of the plant signal metabolite trehalose-6-phosphate,MBio.,5(6):e02065-14
Rajagopal K.,2016,Statistical optimization of medium components by plackett burman design and response surface methodology for enhanced antioxidant activity by Xylaria feejeensis hmjau22039,Int.Pharm.Pharm.Sci.,8(11):159-164
Tanaka H.,Negishi H.,and Maeda H.,1990,Control of tobacco bacterial wilt by an avirulent strain of Pseudomonas solanacearum m4s and its bacteriophage,Ann.Phytopath.Soc.Japan,56(2):243-246
Wicker E.,Grassart L.,Coransonbeaudu R.,Mian D.,Guilbaud C.,Fegan M.,and Prior P.,2007,Ralstonia solanacearum strains from martinique(French west indies)exhibiting a new pathogenic potential,Appl.Environ.Microbiol.,73(21):6790-6801
Xiao T.,Xiao C.G.,Zou Y.,and Yuan X.L.,2008,A preliminary study on the control of tobacco bacterial wilt by avirulent strains of Ralstonia solanacearum,Zhiwu Baohu(Plant Protection),34(2):79-82(肖田,肖崇刚,邹阳,袁希雷,2008,青枯雷尔氏菌无致病力菌株对烟草青枯病的控病作用初步研究,植物保护,34(2):79-82)
Chen X.Q.,Chen D.J.,Zhu Y.J.,Chen Y.P.,Zhang H.F.,and Liu B.,2018,Construction and characterization of extracellular polysaccharide deletion mutant of Ralstonia solanacearum,Weishengwu Xuebao(Acta Microbiologica Sinica),58(5):926-938(陈小强,陈德局,朱育菁,陈燕萍,张海峰,刘波,2018,青枯雷尔氏菌胞外多糖合成缺失突变体构建及其生物学特性,微生物学报,58(5):926-938)
Ghanem K.M.,Al-Fassi F.A.,and Al-Hazmi N.M.,2016,Optimization of chloroxylenol degradation by Aspergillus niger using plackett-burman design and response surface methodology,Rom.Biotech.Lett.,11(1):15040-15048
Hayward A.C.,1991,Biology and epidemiology of bacterial wilt caused by Pseudomonas solanacearum,Annu.Rev.Phytopathol.,29:65-87
Huang Z.C.,Liu Y.,Zeng F.H.,and Lu X.Y.,2008,Advances in pathogenesis of Ralstonia solanacearum and crops resistance to bacterial wilt,Redai Yaredai Zhiwu Xuebao(Journal of Tropical and Subtropical Botany),16(5):491-496(黄真池,刘媛,曾富华,卢向阳,2008,青枯菌致病机理及作物抗青枯病研究进展,热带亚热带植物学报,16(5):491-496)
Jiang G.,Wei Z.,Xu J.,Chen H.L.,Zhang Y.,She X.M.,Macho A.P.,Ding W.,and Liao B.S.,2017,Bacterial wilt in China:history,current status,and future perspectives,Front.Plant Sci.,8:1549
Ogawa K.,Mori T.,Matsusaki H.,and Matsuzoe N.,2012,Simplified evaluation method for suppressing bacterial wilt disease by treating non-pathogenic Ralstonia solanacearum mutant selection in eggplant,Hortic.Res.,11(3):393-398
Poueymiro M.,Cazale A.C.,Francois J.M.,Parrou J.L.,Peeters N.,and Genin S.,2014,A Ralstonia solanacearum typeⅢeffector directs the production of the plant signal metabolite trehalose-6-phosphate,MBio.,5(6):e02065-14
Rajagopal K.,2016,Statistical optimization of medium components by plackett burman design and response surface methodology for enhanced antioxidant activity by Xylaria feejeensis hmjau22039,Int.Pharm.Pharm.Sci.,8(11):159-164
Tanaka H.,Negishi H.,and Maeda H.,1990,Control of tobacco bacterial wilt by an avirulent strain of Pseudomonas solanacearum m4s and its bacteriophage,Ann.Phytopath.Soc.Japan,56(2):243-246
Wicker E.,Grassart L.,Coransonbeaudu R.,Mian D.,Guilbaud C.,Fegan M.,and Prior P.,2007,Ralstonia solanacearum strains from martinique(French west indies)exhibiting a new pathogenic potential,Appl.Environ.Microbiol.,73(21):6790-6801
Xiao T.,Xiao C.G.,Zou Y.,and Yuan X.L.,2008,A preliminary study on the control of tobacco bacterial wilt by avirulent strains of Ralstonia solanacearum,Zhiwu Baohu(Plant Protection),34(2):79-82(肖田,肖崇刚,邹阳,袁希雷,2008,青枯雷尔氏菌无致病力菌株对烟草青枯病的控病作用初步研究,植物保护,34(2):79-82)