Antagonistic Efficacy and Growth-Promoting Effect of Bacillus methylotrophicus Isolated from Dendrobium huoshanense

doi:10.13560/j.cnki.biotech.bull.1985.2016.08.029

Abstract

Abstract: Isolating endophytic bacteria form Dendrobium huoshanense aimed to obtain endophytic bacteria that have broad-spectrum antimicrobial activity and plant growth-promoting trait. Plate confrontation method was applied to screen the endophytic bacteria with antimicrobial activities against the black spot disease of D. huoshanense,of which the antimicrobial spectrum and plant growth-promoting（PGP）traits were determined. Total 22 endophytes were isolated from D. huoshanense,and strain RA presented strong inhibitory activities against Alternaria tenuissima and it was identified as Bacillus methylotrophicus on the basis of 16S rDNA sequence analysis and was selected for further studies. Strain RA was capable of co-producing IAA,siderophore,protease,biosurfactant,and biofilm,and significantly inhibited 8 plant pathogens and had broad-spectrum antimicrobial effects. A significant improvement in corn seedlings growth was observed when seeds of corn were bacterized prior to sowing. In conclusion,results clearly suggest that strain RA is a potential candidate for use as biocontrol agent and in biofertilizer.

Key words: Dendrobium huoshanense, endophyte, Bacillus methylotrophicus, antagonistic efficacy, growth promotion

WU Li-qin ,GU Hai-ke ,WANG Qing, SHANG Hong-zhong, LIU Gui-jun ,BAO Fang. Antagonistic Efficacy and Growth-Promoting Effect of Bacillus methylotrophicus Isolated from Dendrobium huoshanense[J]. Biotechnology Bulletin, 2016, 32(8): 200-206.

References

[1] 姚革, 张帆, 李舟. 土壤添加剂防治细菌性青枯病初报[J]. 生物防治通报, 1994, 10（3）：106-109.
[2] Hallmann J, Quadt-Hallmann A, Mahaffee WF, et al. Bacterial endophytes in agricultural crops[J]. Canadian Journal of Microbiology, 1997, 43（10）：895-914.
[3] Surette MA, Sturz AV, Lada RR, et al. Bacterial endophytes in processing carrots（Daucus carota L. var. sativus）：their localization, population density, biodiversity and their effects on plant growth[J]. Plant and Soil, 2003, 253（2）：381-390.
[4] Nejad P, Johnson PA. Endophytic bacteria induce growth promotion and wilt disease suppression in oilseed rape and tomato[J]. Biological Control, 2000, 18（3）：208-215.
[5] 暴增海, 马桂珍, 张拥华, 等. 粘帚霉菌株对大豆促生及菌核病防效的研究[J]. 中国种业, 2007, 4：42-43.
[6] 康慧颖, 王伟, 刘佳莉, 等. 两株具促生作用的苜蓿内生菌的分离纯化与鉴定[J]. 微生物学通报, 2015, 42（2）：280-288.
[7] 陈卓君, 吴毅歆, 毛自朝, 等. 萝卜根内生产酸克雷伯克氏菌的分离鉴定及其促生长作用研究[J]. 西南农业学报, 2014, 27（4）：1645-1648.
[8] 周怡, 毛亮, 张婷婷, 等. 大豆内生芽孢杆菌的分离和促生菌株的筛选及鉴定[J]. 大豆科学, 2009, 28（3）：502-506.
[9] 张敬泽, 郑小军. 铁皮石斛黑斑病病原菌的鉴定和侵染过程的细胞学研究[J]. 植物病理学报, 2004, 34（1）：92-94.
[10] 张敬泽, 方钰蓉, 张海松, 等. 铁皮石斛黑斑病菌室内药效试验[J]. 植物保护, 2005, 31（1）：44-47.
[11] Patten CL, Glick BR. Role of Pseudomonas putida indole-acetic acid in development of the host plant root system[J]. Applied and Environmental Microbiology, 2002, 68（8）：3795-3801.
[12] Pikovskaya RE. Solubilization of phosphorus in soil in connection with vital activity of some microbial species[J]. Mikrobiologiya, 1948, 17：362-370.
[13] Alexander DB, Zuberer DA. Use of chrome azurol S reagents to evaluate siderophore production by rhizosphere bacteria[J]. Biology and Fertility of Soils, 1991, 12（1）：39-45.
[14] Smibert RM, Krieg NR. Phenotypic characterization[M]//Gerha- rdt P, Murray RGE, Wood WA, Krieg NR. Methods for general and molecular bacteriology. Washington, DC：American Society of Microbiology, 1994：607-654.
[15] Eden PA, Schmidt TM, Blakemore RP, et al. Phylogenetic analysis of Aquaspirillum magnetotacticum using polymerase chain reaction-amplified 16S rRNA-specific DNA[J]. International Journal of Systematic Bacteriology, 1991, 41：324-325.
[16] Tamura K, Peterson D, Peterson N, et al. MEGA5：Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Molecular Biology and Evolution, 2011, 28：2731-2739.
[17] Bhore SJ, Preveena J, Kandasamy KI. Isolation and identification of bacterial endophytes from pharmaceutical agarwood-producing Aquilaria species[J]. Phcog Res, 2013, 5：134-137.
[18] Brader G, Stephane C, Birgit M, et al. Metabolic potential of endophytic bacteria[J]. Curr Opin Biotechnol, 2014, 27：30-37.
[19] Christina A, Christapher V, Bhore SJ. Endophytic bacteria as a source of novel antibiotics：Anoverview[J]. Pharmacog Rev, 2013, 7：11-16.
[20] Jeong H, Choi SK, Kloepper JW, Ryu CM. Genome sequence of the plant endophyte Bacillus pumilus INR7, triggering induced systemic resistance in field crops[J]. Genome Announc, 2014, 2（5）：e01093-14.
[21] 赵智灵, 刘学周, 魏晓雨, 等. 人参可利用内生菌株的筛选和鉴定[J]. 中草药, 2015, 46（14）：2143-2148.
[22] Danhorn T, Fuqua C. Biofilm formation by plant-associated bacteria[J]. Annual Review of Microbiology, 2007, 61：401-422.
[23] Blom D, Fabbri C, Connor EC, et al. Production of plant growth modulating volatiles is widespread among rhizosphere bacteria and strongly depends on culture conditions[J]. Environmental Microbiology, 2011, 13：3047-3058.
[24] Gamalero E, Glick BR. Mechanisms used by plant growth-promot-ing bacteria[M]//Maheshwari DK. Bacteria in Agrobiology：Plant Nutriente Management. Berlin, Germany：Springer, 2011：17-46.
[25] Heerklotz H. Interactions of surfactants with lipid membranes[J]. Quarterly Reviews of Biophysics, 2008, 41（3-4）：205-264.