拮抗菌株xj063-1发酵条件的优化及室内防效测定
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摘要
为探讨菌株xj063-1的最优发酵条件,以菌体生物量和对病原菌的抑制率为指标,采用单因素方法对菌株的发酵条件进行优化,并将优化的发酵液在离体枣果上进行防效测定。结果表明,当装液量为50 m L时,拮抗菌株生物量OD600为1.987,对病原菌的抑制率为67.1%;接种量为装液量体积的4%时,生物量OD600为1.573,对病原菌的抑制率为59.7%;初始p H 7.0时,其OD600为1.905,对病原菌的抑制率为65.6%;温度在30℃、转速180 r/min时,OD600分别为1.969和1.982,对病原菌的抑制率分别为62.2%和61.2%。拮抗菌株优化后的发酵液通过预防法处理后的枣果的病斑面积为4 mm2,对照发病面积为121 mm2,抑制率为96.7%;通过治疗法处理后的枣果的病斑面积为12.25 mm2,抑制率为89.9%,表明菌株xj063-1具有较好的生防潜力。
In order to investigate best conditions for fermentation liquid of antagonistic strain xj063-1,the single factor experiments were performed with bacterial biomass and the inhibition rate of pathogenic bacteria as indicators. The results showed that the optimal fermentation conditions were composed of the following factors: liquid volume, 50 m L; inoculum, 4%( v / v); initial p H 7. 0; fermentation temperature,30℃; rotation speed,180 r / min. Bacterial biomass OD600 and the inhibition rate of pathogenic bacteria were 1. 987 /67. 1%,1. 573 /59. 7%,1. 905 /65. 6%,1. 969 /62. 2% and 1. 982 /61. 2%,respectively. When optimized filtrate was treated on jujube fruits,the lesion diameters were 4mm2 by prevention,and 12. 25 mm2 by therapy,respectively. The control effects were 96. 7% by prevention and 89. 9% by therapy,respectively. These results indicated that the antagonistic strains may provide a new biological control method in controlling Alternaria alternate.
In order to investigate best conditions for fermentation liquid of antagonistic strain xj063-1,the single factor experiments were performed with bacterial biomass and the inhibition rate of pathogenic bacteria as indicators. The results showed that the optimal fermentation conditions were composed of the following factors: liquid volume, 50 m L; inoculum, 4%( v / v); initial p H 7. 0; fermentation temperature,30℃; rotation speed,180 r / min. Bacterial biomass OD600 and the inhibition rate of pathogenic bacteria were 1. 987 /67. 1%,1. 573 /59. 7%,1. 905 /65. 6%,1. 969 /62. 2% and 1. 982 /61. 2%,respectively. When optimized filtrate was treated on jujube fruits,the lesion diameters were 4mm2 by prevention,and 12. 25 mm2 by therapy,respectively. The control effects were 96. 7% by prevention and 89. 9% by therapy,respectively. These results indicated that the antagonistic strains may provide a new biological control method in controlling Alternaria alternate.
引文
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Chen XF,Xiong RC,Xu SZ,Chen HB,Xu SL.2013.Current situation and prospect on jujube blackspot disease.Heilongjiang\sAgricultural Sciences,(10):141-144(in Chinese)[陈小飞,熊仁次,徐崇志,陈恢彪,徐胜利.2013.红枣黑斑病研究现状与展望.黑龙江农业科学,(10):141-144]
Cheng CH,Yang CA,Peng KC.2012.Antagonism of Trichoderma\sharzianum ETS 323 on Botrytis cinerea mycelium in culture conditions.Phytopathology,102(11):1054-1063
Chi ZJ,Sun J,Li J,Liu DH,Zhao SF.2014.Toxicology and field\sefficacy of 4 biological fungicides for controlling jujube black\sspot disease in Xinjang.Agrochemicals,53(7):515-517(in\sChinese)[池振江,孙洁,李进,刘多红,赵思峰.2014.4种生物农药对新疆枣果黑斑病的室内及田间药效试验.农药,53(7):515-517]
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Gao F,Li JH,Zhang NS,Wang JH,Wang ML.2014.In vitro\sscreening of antagonistic Bacillus against Alternaria alternate and\soptimization factors.Plant Protection,40(3):70-75(in Chinese)[高芬,李静虹,张娜莎,王俊红,王梦亮.2014.‘赞皇枣’黑腐病菌拮抗芽胞杆菌的筛选及发酵条件优化.植物保护,40(3):70-75]
Hong P,An GD,Hu MY,Geng P,Sang S.2013.Optimization fermentation condition for Bacillus amyloliquefaciens HF-01.Chinese Journal of Biological Control,29(4):569-578(in Chinese)[洪鹏,安国栋,胡美英,耿鹏,桑松.2013.解淀粉芽孢杆菌HF-01发酵条件优化.中国生物防治学报,29(4):569-578]
Hou M,Zhan FQ,Zhang HT,Hou XQ,Yang R,Long XQ.2014.Optimization of spore production medium and conditions of antagonistic strain S13 of Bacillus amyloliauefaciens against Fusarium wilt tomato.Xinjing Agriculture Sciences,51(7):1269-1276(in Chinese)[侯敏,詹发强,张慧涛,侯新强,杨蓉,龙宣杞.2014.番茄枯萎病拮抗细菌S13产芽孢发酵培养基及发酵条件优化.新疆农业科学,51(7):1269-1276]
Hu LL,Xu HH,Liao MD.2011.Optimization of culture condition\sfor a strain of PS04 producing antifungal antibiotics.Journal of\sHuazhong Agricultural University,30(3):276-279(in Chinese)[胡亮亮,徐汉虹,廖美德.2011.胶冻样类芽孢杆菌PS04产抗真菌物质培养条件的优化.华中农业大学学报,30(3):276-279]
Ju RC,Wang P,Gong CY,Zhao HT,Liu L,Li L,Yan HX,Zhang\sPP,Wang HP,Li YY.2014.Isolation,identification and fermentation optimization of an antibiotic Bacillus subtilis.Chinese\sAgricultural Science Bulletin,30(28):295-300(in Chinese)[鞠瑞成,王鹏,公春艳,赵宏涛,刘露,李丽,闫洪雪,张鹏鹏,王海朋,李园园.2014.1株枯草芽孢杆菌的分离鉴定及其发酵条件优化.中国农学通报,30(28):295-300]
Karim MIA,Lucas RJ,Osborne KJ,Rogers PL.1993.The effect of\soxygen on the sporulation and toxicity of Bacillus sphericus2362.Biotechnology Letters,15(1):47-50
Lin YB,Lu JX,Yan X,Liu BY,Fang S,Fan YQ.2010.Screening,identification and optimized fermentation condition of antagonistic actinomycetes from Rehminnae glutinoso rhizosphere.Journal of Plant Protection,37(3):234-240(in Chinese)[林雁冰,陆家贤,颜霞,刘博野,方松,范元青.2010.地黄根圈土壤拮抗放线菌筛选、鉴定及发酵条件优化.植物保护学报,37(3):234-240]
Liu JL,Xue YR,Liu CH.2014.Optimization of shake flask-fermentation conditions for Iturin A production by endophytic Bacillus amyloliquefaciens CC09.Journal of Microbiology China,41(1):75-82(in Chinese)[刘京兰,薛雅蓉,刘常宏.2014.内生解淀粉芽孢杆菌CC09产Iturin A摇瓶发酵条件优化.微生物学通报,41(1):75-82]
Liu PF,He FL,Qu CH,Ji MS,Zhao CS.2012.Fermentation conditions of antagonistic bacterium D221 against Fusarium solani.Journal of Northeast Agricultural University,43(1):110-115(in Chinese)[刘培福,何付丽,曲春鹤,纪明山,赵长山.2012.辣椒根腐病拮抗细菌D221发酵条件的研究.东北农业大学学报,43(1):110-115]
Liu XL,Liu Y,Ma R,Liang YM,Chen BJ,Yan J.2015.Identification and biological characteristics of the pathogen causing jujube black spot in Xinjiang.Journal of Northwest Forestry University,30(3):132-138(in Chinese)[刘晓琳,刘玉,马荣,梁英梅,陈宝军,闫军.2015.新疆枣果黑斑病病原鉴定及生物学特性.西北林学院学报,30(3):132-138]
Ma R,Liu XL,Sun YY,Chen M,Yan J,Chen BJ.2014.Screening and identification of the antagonistic bacteria of jujube black\sspot disease in Xinjiang.Journal of Xinjiang Agricultural University,37(6):460-464(in Chinese)[马荣,刘晓琳,孙园园,陈梦,闫军,陈宝军.2014.新疆枣果黑斑病拮抗细菌的筛选及鉴定.新疆农业大学学报,37(6):460-464]
Ma R,Zhang CY,Liu Y,Liang YM,Fan GL,Tian CM.2012.Laboratory fungicide screening for Alternaria sp.black spot pathogen on jujube in Xinjiang.Agrochemicals,51(10):767-770(in Chinese)[马荣,张传燕,刘玉,梁英梅,樊贵利,田呈明.2012.新疆红枣黑斑病病菌的室内杀菌剂筛选.农药,51(10):767-770]
Sun Z,Yang LM,Ma XJ,Zhang YM.2014.Screening and indentification of antagonistic bacteria on Alternaria panax Whetz.Journal of Jilin Agricultural University,36(3):276-281(in\sChinese)[孙卓,杨利民,马秀杰,张一鸣.2014.人参黑斑病拮抗细菌的分离筛选和鉴定.吉林农业大学学报,36(3):276-281]
Tu QH.2013.Screening of antagonistic bacteria YS-1 of postharvest\sdisease of citrus fruits and study on antimicrobial mechanism\sand properties of the antibacterial crude extract of the strain\sYS-1.Ph.D Thesis.Nanchang:Nanchang University(in Chinese)[涂起红.2013.柑橘采后病害生防菌YS-1的筛选和YS-1粗提物的抑菌机理及性质研究.博士学位论文.南昌:南昌大学]
Wang CL,Zhang LS,Gao WW,Zhang W,Qi YY,Yang QJ.2008.Antagonism and control effect produced by Bacillus strain\sTS-01 against Alternaria mali.Journal of Plant Protection,35(2):183-184(in Chinese)[王程亮,张潞生,高微微,张文,戚元勇,杨庆军.2008.芽孢杆菌TS-01对苹果斑点落叶病菌的拮抗作用及防病效果.植物保护学报,35(2):183-184]
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