Isolation and screening of antagonistic bacteria and its active substance against tobacco bacterial wilt

doi:10.14088/j.cnki.issn0439-8114.2021.18.018

Abstract

Abstract: The antagonistic bacteria were isolated from the rhizosphere soil of healthy tobacco plants by plate inhibition circle method, and the morphological culture observation, physiological and biochemical tests and 16S rDNA sequences were compared and analyzed. The results showed that 9 strains of antagonistic bacteria were isolated, among which strain FQ-7 had the strongest antibacterial activity, with the diameter of antibacterial zone reaching 16.3 mm. The strain FQ-7 was identified as Streptomyces violaceolatus. The strain was facultative aerobic, negative in contact enzyme and methyl red test; It could decompose starch and casein while showing tolerance to NaCl with the fastest growth rate at 10 g/L; It could use maltose, mannitol, glucose and sucrose while could not use lactose, sweet alcohol, sorbitol and cellobiose; Antibacterial substances from strain FQ-7 were subjected to lipophilic compounds dissolved in ethyl acetate; The relative inhibition activity maintained 88.51%~95.40%, 86.52%~99.88% and 85.52%~91.05% under different proceeding conditions of various temperature（40~121 ℃）, pH （2.0~8.0） and UV irradiation （30~120 min UV exposure time with 8 W lamp）, respectively.

Key words: tobacco bacterial wilt, antagonistic bacteria, isolation and identification, antibacterial substances

CLC Number:

S482.2

TANG Ming-qiang, LIN Tian-ran, LI Xiao-fang, ZHOU Xiao-qin, ZENG Wen-long, LIN Xiao-lu, PENG Shui-lian. Isolation and screening of antagonistic bacteria and its active substance against tobacco bacterial wilt[J]. HUBEI AGRICULTURAL SCIENCES, 2021, 60(18): 92-96.

References 24

[1]	张欣悦,罗翠琴,陈小洁,等.两株防治烟草青枯病的烟草根际拮抗菌[J]. 中国烟草学报,2020,26(1):91-99.
[2]	PEETERS N,GUIDOT A,VAILLEAU F,et al.Ralstonia solanacearum, a wide spread bacterial plant pathogen in the post-genomic era[J]. Molecular plant pathology,2013,14(7):651-662.
[3]	刘伟,刘鹏,沈小英,等.烟草青枯病拮抗芽孢杆菌的筛选、鉴定及其抑菌活性初探[J]. 西北农林科技大学学报(自然科学版),2014,42(2):123-130.
[4]	王大海,张宁波,高凯,等.4种防治措施对烟草青枯病的防治效果[J]. 安徽农业科学,2017,45(15):152-153,156.
[5]	姜乾坤,彭阁,王瑞,等.抗青枯内生细菌的筛选及其对烟草青枯病的防治效果[J]. 中国烟草科学,2017,38(5):13-17,31.
[6]	吴翔,谢丽源,甘炳成,等.一株烟草青枯拮抗细菌的筛选、鉴定和培养特性研究[J]. 中国土壤与肥料,2019(2):208-215.
[7]	张薇. 烟草青枯菌拮抗放线菌的筛选、鉴定及发酵条件研究[D]. 山东泰安:山东农业大学,2009.
[8]	陆铮铮,彭丽娟,丁海霞,等.烟草青枯菌拮抗放线菌的筛选及鉴定[J]. 中国烟草科学,2013,34(2):54-58.
[9]	罗文建,刘雨虹,施河丽,等.青枯雷尔氏菌拮抗放线菌的筛选及其抗菌活性物质的分离[J].中国烟草科学,2017,38(2):69-74.
[10]	文才艺,郑东光,陈建光,等.一株生防圆突起链霉菌的分类鉴定[J]. 微生物学通报,2011,38(5):687-693.
[11]	中国科学院微生物研究所放线菌分类组.链霉菌鉴定手册[M].北京:科学出版社,1975.240-260.
[12]	方中达. 植病研究方法[M]. 第三版.北京:中国农业出版社,1998.243-249.
[13]	阎逊初. 放线菌的分类和鉴定[M].北京:科学出版社,1992.
[14]	叶海霞,郭春晓,丁祥,等.拮抗放线菌Y-71的鉴定及其代谢活性物质的性质[J]. 微生物学通报,2010, 37(8):1182-1187.
[15]	POOMTHONGDEE N,DUANGMAL K,PATHOM-AREE W.Acidophilic actinomycetes from rhizosphere soil:Diversity and properties beneficial to plants[J]. The journal of antibiotics,2015,68(2):106-114.
[16]	WANG C,WANG Z,QIAO X,et al.Antifungal activity of volatile organic compounds from Streptomyces alboflavus TD-1[J]. FEMS microbiology letters,2013,341(1):45.
[17]	MINGMA R,PATHOM-AREE W.Isolation of rhizospheric and roots endophytic actinomycetes from Leguminosae plant and their activities to inhibit soybean pathogen, Xanthomonas campestris pv.glycine[J]. World journal of microbiology and biotechnology,2014,30(1):271-280.
[18]	PRABAVATHY V,MATHIVANAN N,MURUGESAN K.Control of blast and sheath blight diseases of rice using antifungal metabolites produced by Streptomyces sp PM5[J].Biological control,2006,39(3):313-319.
[19]	李庆蒙. 链霉菌菌株JD211的鉴定及其次级代谢产物的初步研究[D].南昌:江西农业大学,2014.
[20]	赵辉,闫华晓,王霞,等.一株产天然蓝色素海洋链霉菌的筛选鉴定[J]. 生物学杂志,2011,28(3):63-65.
[21]	王云山,王来福,虞勇,等.耐盐碱链霉菌的聚类分析与分类鉴定[J]. 河北省科学院学报,1994(3):44-49.
[22]	杜静. 土壤放线菌的分离、鉴定及其活性化合物的研究[D].哈尔滨:东北农业大学,2010.
[23]	HAYAKAWA Y,ADACHI K,IWAKIRI T,et al.Kerriamycins A, B and C, new isotetracenone antibiotics[J].Agricultural and biological chemistry,1987,51(5):1397-1405.
[24]	BENTLEY S D,CHATER K F,CERDEÑO-TÁRRAGA A-M,et al. Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2)[J]. Nature,2002,417:141-147.