Pantoea ananatis is a plant pathogenic bacterium that severely impacts rice. In spite of its worldwide prevalence, limited studies have been conducted so far on the control of P. ananatis. Bacitracin A is a non-ribosomal peptide antibiotic with strong antibacterial activity produced by Bacillus licheniformis strain HN-5. We investigated the mechanisms of action underlying the biocontrol and bactericidal efficacy of bacitracin A against P. ananatis. Fluorescence microscopy and bacterial cell viability analyses revealed that the median effective concentration of bacitracin A against P. ananatis was 9.10 μg ml−1. Scanning and transmission electron microscopy showed that bacitracin A damaged the cell wall and membrane of P. ananatis. Quantitative real-time PCR indicated that the transcriptional expression of ftsZ, glmS, and gumD, which are involved in cell division, cell-wall biosynthesis, and extracellular polymeric substance biosynthesis, respectively, was upregulated at 12 h and significantly downregulated at 24 h after bacitracin A treatment in P. ananatis. Bacitracin A caused cell leakage and changes to membrane permeability in P. ananatis, supporting its use as a natural biocontrol agent for P. ananatis.
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Borshchevskaya L, Kalinina A, Sineokii S (2013) Design of a PCR test based on the gyrA gene sequence for the identification of closely related species of the Bacillus subtilis group. Appl Biochem Microbiol 49(7):646–655
Carrillo C, Teruel JA, Aranda FJ, Ortiz A (2003) Molecular mechanism of membrane permeabilization by the peptide antibiotic surfactin. Biochim Biophys Acta 1611(1–2):91–97
Choi O, Kim H, Lee Y, Kim J, Moon JS, Hwang I (2012) First report of sheath rot of rice caused by Pantoea ananatis in Korea. Plant Pathol J 28(3):331–332
Choi Y, Cho S, Simkhada J, Rahman M, Choi Y, Kim C, Yoo J (2017) A novel multifunctional peptide oligomer of bacitracin with possible bioindustrial and therapeutic applications from a Korean food-source Bacillus strain. PLoS ONE 12(5):e0176971
Ciesiołka J, Jeżowska-Bojczuk M, Wrzesiński J, Stokowa-Sołtys J, Nagaj J, Kasprowicz A, Błaszczyk L, Szczepanik W (2014) Antibiotic bacitracin induces hydrolytic degradation of nucleic acids. BBA-Gen Subj 1840(6):1782–1789
Cother E, Reinke R, Mckenzie C, Lanoiselet V, Noble D (2004) An unusual stem necrosis of rice caused by Pantoea ananas and the first record of this pathogen on rice in Australia. Plant Path 33(4):495–503
Coutinho T, Venter S (2009) Pantoea ananatis: an unconventional plant pathogen. Mol Plant Pathol 10(3):325–335
Drablos F, Nicholson D, Ronning M (1999) EXAFS study of zinc coordination in bacitracin A. BBA-Protern. Struct M 1431(2):433–442
Fravel D (2005) Commercialization and implementation of biocontrol. Annu Rev Phytopathol 43:337–359
Hancock R (2005) Mechanisms of action of newer antibiotics for Grampositive pathogens. Lancet Infect Dis 5(4):209–218
Jin P, Wang H, Liu W, Zhang S, Lin C, Zheng F, Guo M (2017) Bactericidal metabolites from Phellinus noxius HN-1 against Microcystis aeruginosa. Sci Rep 7(1):3132–3141
Jin P, Wang H, Tan Z, Xuan Z, Dahar G, Li Q, Miao W, Liu W (2020) Antifungal mechanism of bacillomycin D from Bacillus velezensis HN-2 against Colletotrichum gloeosporioides Penz. Pestic Biochem Phys 163:102–107
Kido K, Hasegawa M, Hioyuki M, Kobayashi M, Yuichi T (2010) Pantoea ananatis strains are differentiated into three groups based on reactions of tobacco and welsh onion and on genetic characteristics. J Gen Plant Pathol 76(3):208–218
Kim J, Kim B, Lee C (2007) Alga-lytic of Pseudomonas fluorescens against the red tide causing marine alga Heterosigma akashiwo (Raphidophyceae). Biol Control 41(3):296–303
Konz D, Klens A, Schorgendorfer K, Marahiel M (1997) The bacitracin biosynthesis operon of Bacillus licheniformis ATCC 10716: molecular characterization of three multi-modular peptide synthetases. Chem Biol 4(12):927–937
Li J, Wang N (2011) The wxacO gene of Xanthomonas citri ssp. citri encodes a protein with a role in lipopolysaccharide biosynthesis, biofilm formation, stress tolerance and virulence. Mol Plant Pathol 12(4):381–396
Livak K, Schmittgen T (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-DDC (T) method. Methods 25(4):402–408
Maayer P, Chan W, Rubagotti E, Venter S, Toth I, Birch P, Coutinho T (2014) Analysis of the Pantoea ananatis pan-genome reveals factors underlying its ability to colonize and interact with plant, insect and vertebrate hosts. BMC Genomics 15(1):404–417
Magnet-Dana R, Peypoux F (1994) Iturins, a special class of poreforming lipopeptides: biological and physiological properties. Toxicology 87(1–3):151–174
Makovitzki A, Avrahami D, Shai Y (2006) Ultrashort antibacterial and antifungal lipopeptides. Proc Natl Acad Sci USA 103(43):15997–16002
Mondal K, Mani C, Singh J, Kim J, Mudgett M (2011) A new leaf blight of rice caused by Pantoea ananatis in India. Plant Dis 95(12):1582–1583
Morris M (1994) Primary structural confirmation of components of the bacitracin complex. Biol Mass Spectrom 23(2):61–70
Moyne A, Shelby R, Cleveland T, Tuzun S (2001) Bacillomycin D: an iturin with antifungal activity against Aspergillus flavus. J Appl Microbiol 90(4):622–629
Nasir M, Besson F (2012) Conformational analyses of bacillomycin D, a natural antimicrobial lipopeptide, alone or in interaction with lipid monolayers at the air-water interface. J Colloid Interf Sci 387(1):187–193
Oh E, Bae J, Kumar A, Choi H, Jeon B (2018) Antioxidant-based synergistic eradication of methicillin-resistant Staphylococcus aureus (MRSA) biofilms with bacitracin. Int J Antimicrob Ag. 52(1):96–99
Ongena M, Jacques P (2008) Bacillus lipopeptides: versatile weapons for plant disease biocontrol. Trends Microbiol 16(3):115–125
Ongena M, Jacques P, Touré Y, Destain J, Jabrane A, Thonart P (2005) Involvement of fengycin-type lipopeptides in the multifaceted biocontrol potential of Bacillus subtilis. Appl Microbiol Biot 69(1):29–38
Pavli V, Kmetec V (2001) Optimization of HPLC method for stability testing of bacitracin. J Pharmaceut Biomed 24(5–6):977–982
Pollock T, Thorne L, Yamazaki M, Mikolajczak M, Armentrout R (1994) Mechanism of bacitracin resistance in gram-negative bacteria that synthesize exopolysaccharides. J Bacteriol 176(20):6229–6237
Qian G, Zhou Y, Zhao Y, Song Z, Wang S, Fan J, Hu B, Venturi V, Liu F (2013) Proteomic analysis reveals novel extracellular virulence-associated proteins and functions regulated by the diffusible signal factor (DSF) in Xanthomonas oryzae pv. oryzicola. J Proteome Res 12(7):3327–3341
Romero D, de Vicente A, Olmos J, Dávila J, Pérez-García A (2007) Effect of lipopeptides of antagonistic strains of Bacillus subtilis on the morphology and ultrastructure of the cucurbit fungal pathogen Podosphaera fusca. J Appl Microbiol 103(4):969–976
Schallmey M, Singh A, Ward O (2004) Developments in the use of Bacillus species for industrial production. Can J Microbiol 50(1):1–17
Shyntum D, Theron J, Venter S, Moleleki L, Toth I, Coutinho T (2015) Pantoea ananatis utilizes a type VI secretion system for pathogenesis and bacterial competition. Mol Plant-Microbe Interact 28(4):420–431
Silva I, Regasini L, Petrônio M, Silva D, Bolzani V, Belasque J, Sacramento L, Ferreira H (2012) Antibacterial activity of alkyl gallates against Xanthomonas citri subsp citri. J Bacteriol 195(1):85–94
Skamnioti P, Gurr S (2009) Against the grain: safeguarding rice from rice blast disease. Trends Biotechnol 27(3):141–150
Stein T (2005) Bacillus subtilis antibiotics: Structures, syntheses and specific functions. Mol Microbiol 56(4):845–857
Stone K, Strominger J (1971) Mechanism of action of bacitracin: complexation with metalion and C55-isoprenyl pyrophosphate. P Natl Acad Sci 68(12):3223–3227
Storm D, Strominger J (1973) Complex formation between bacitracin peptides and isoprenyl pyrophosphates. The specificity of lipid-peptide interactions. J Biol Chem 248(11):3940–3945
Straus S, Hancock R (2006) Mode of action of the new antibiotic for Gram-positive pathogens daptomycin: comparison with cationic antimicrobial peptides and lipopeptides. Biochim Biophys Acta Biomembr 1758(9):1215–1223
Suleiman S, Song F, Su M, Hang T, Song M (2017) Analysis of bacitracin and its related substances by liquid chromatography tandem mass spectrometry. J Pharm Anal 7(1):48–55
Torres M, Jones J, Dangl J (2006) Reactive oxygen species signaling in response to pathogens. Plant Physiol 141(2):373–378
Wang L, Lee F, Tai C, Kasai H (2007) Comparison of gyrB gene sequences, 16SrRNA gene sequences and DNA-DNA hybridization in the Bacillus subtilis group. Int J Syst Evol Microbiol 57(8):1846–1850
Wang Q, Zheng H, Wan X, Huang H, Li J, Christopher T, Wang C, Chen S (2017) Optimization of inexpensive agricultural by-products as raw materials for bacitracin production in Bacillus licheniformis DW2. Appl Biochem Biotechnol 183(4):1146–1157
Wu L, Wu H, Chen L, Yu X, Borriss R, Gao X (2015) Difficidin and bacilysin from Bacillus amyloliquefaciens FZB42 have antibacterial activity against Xanthomonas oryzae rice pathogens. Sci Rep 5:12975–12983
Yun M, Torres P, Oirdi M, Rigano L, Gonzalez-Lamothe R, Marano M, Castagnaro A, Dankert M, Bouarab K, Vojnov A (2006) Xanthan induces plant susceptibility by suppressing callose deposition. Plant Physiol 141(1):178–187
Zeriouh H, Romero D, García-Gutiérrez L, Cazorla F, de Vicente A, Perez-Garcia A (2011) The iturin-like lipopeptides are essential components in the biological control arsenal of Bacillus subtilis against bacterial diseases of cucurbits. Mol Plant-Microbe Interact 24(12):1540–1552
Zuo W, Jin P, Dong W, Dai H, Mei W (2014) Metabolites from the endophytic fungus HP-1 of Chinese eaglewood. Chin J Nat Med 12(2):151–153
We thank Xuewen Gao (College of Plant Protection, Nanjing Agricultural University, Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing, China) for a kind support. The authors declare no competing financial interests.
The funding was provided by the National Natural Science Foundation of China (Grant No. 31960552), the Scientific Research Foundation for Advanced Talents (Grant No. KYQD(ZR)1842) and Hainan Provincial Science and Technology Foundation Youth Talent Innovation Program (Grant No. QCXM201903).
Conflict of interest
All authors declare that there is no conflict of interest in this work.
This article does not contain any studies with human participants performed by any of the authors.
Handling Editor: Jane Debode.
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Jin, P., Tan, Z., Wang, H. et al. Antimicrobial effect of Bacillus licheniformis HN-5 bacitracin A on rice pathogen Pantoea ananatis. BioControl 66, 249–257 (2021). https://doi.org/10.1007/s10526-020-10052-9
- Antimicrobial activity
- Bacitracin A
- Pantoea ananatis