Suppression of Tomato Fusarium Wilt Disease by Bacteria Strains and Their Mechanism

doi:10.16409/j.cnki.2095-039x.2015.06.012

Abstract

Abstract: The authors tested the inhibition rates of four antagonistic bacteria isolated from healthy soybean roots against as many as nine fungi by in vitro tests and potting experiments in greenhouse. Results showed that all strains exhibited great inhibitory effects on growth of the fungi, with inhibition rates between 51.00%—86.81%; Similar antagonistic abilities were also detected from the cell-free solutions and volatiles. Application of peat formulation of strains SR10 and SR22 to tomato roots prior to transplanting into potting medium infested with Fusarium oxysporum f. sp. radicis lycopersici provided great protection from wilt disease under greenhouse conditions, and the disease severity were reduced by 42.33%—51.33% and no difference in comparison with the chemical treatment. Analysis of 16S rRNA indicated that SR10 and SR11 were Paenibacillus polymyxa, SR21 Pseudomonas brassicacearum and SR22 Bacillus amyloliquefaciens. All strains could produce proteases, β-1,3-glucanases, HCN, IAA, SA; SR10, SR11 and SR22 could fix nitrogen; SR21 could solubilize phosphate; SR21 and SR22 could produce siderophores; none of them could produce chitinase. PCR analysis of antibiotic biosynthetic genes confirmed that SR10、SR11 and SR22 owned the bacA and srfAA genes, both of SR10 and SR22 owned the ituD gene, SR22 owned fend and bmyB genes; SR21 owned pltD, prnD and phlD genes.

Key words: bio-control bacteria, tomato wilt, identification, antagonistic mechanism.

CLC Number:

S476

ZHANG Liang, WANG Gailan, DUAN Jiannan, ZHAO Lanfeng, LI Huaxing. Suppression of Tomato Fusarium Wilt Disease by Bacteria Strains and Their Mechanism[J]. journal1, 2015, 31(6): 897-906.

References

[1] Ajilogba C F, Babalola O O. Integrated management strategies for tomato Fusarium wilt[J]. Biocontrol Science, 2013, 18(3): 117-127.
[2] Larkin R P, Fravel D R. Efficacy of various fungal and bacterial biocontrol organisms for control of Fusarium wilt of tomato[J]. Plant Disease, 1998, 82(9): 1022-1028.
[3] Weller D M, Raaijmakers J M, Gardener B B M, et al. Microbial populations responsible for specific soil suppressiveness to plant pathogens [J]. Annual Review of Phytopathology, 2002, 40(1): 309-348.
[4] Labuschagne N, Pretorius T, Idris A H. In plant growth and health promoting bacteria: plant growth promoting rhizobacteria as biocontrol agents against soil-borne plant diseases[M]. Berlin Heidelberg: Springer, 2011, 211-230.
[5] 徐艳辉, 李烨, 许向阳. 番茄枯萎病的研究进展[J]. 东北农业大学学报, 2008, 39(11): 128-134.
[6] Bao J R, Lazarovits G. Differential colonization of tomato roots by nonpathogenic and pathogenic Fusarium oxysporum strains may influence Fusarium wilt control[J]. Phytopathology, 2001, 91(5): 449-456.
[7] 刘邮洲, 陈志谊, 梁雪杰, 等. 番茄枯萎病和青枯病拮抗细菌的筛选、评价与鉴定[J]. 中国生物防治学报, 2012, 28(1): 101-108.
[8] Gopalakrishnan S, Pande S, Sharma M, et al. Evaluation of actinomycete isolates obtained from herbal vermicompost for the biological control of Fusarium wilt of chickpea[J]. Crop Protection, 2011, 30(8): 1070-1078.
[9] 郝晓娟, 刘波, 谢关林, 等. 短短芽孢杆菌JK-2菌株对番茄枯萎病的抑制作用及其小区防效[J]. 中国生物防治, 2007, 23(3): 223-236.
[10] Srivastava R, Khalid A, Singh U S, et al. Evaluation of arbuscular mycorrhizal fungus, fluorescent Pseudomonas and Trichoderma harzianum formulation against Fusarium oxysporum f. sp. lycopersici for the management of tomato wilt[J]. Biological Control, 2010, 53(1): 24-31.
[11] Babalola O O. Beneficial bacteria of agricultural importance[J]. Biotechnology Letters, 2010, 32(11): 1559-1570.
[12] Dutta S, Podile A R. Plant growth promoting rhizobacteria (PGPR): the bugs to debug the root zone[J]. Critical Reviews in Microbiology, 2010, 36(3): 232-244.
[13] Rosenblueth M, Martínez-Romero E. Bacterial endophytes and their interactions with hosts[J]. Molecular Plant-Microbe Interactions, 2006, 19(8): 827-837.
[14] Baysal Ö, Çalışkan M, Yeşilova Ö. An inhibitory effect of a new Bacillus subtilis strain (EU07) against Fusarium oxysporum f. sp. radicis lycopersici[J]. Physiological and Molecular Plant Pathology, 2008, 73(1): 25-32.
[15] Barahona E, Navazo A, Martínez-Granero F, et al. Pseudomonas fluorescens F113 mutant with enhanced competitive colonization ability and improved biocontrol activity against fungal root pathogens[J]. Applied and Environmental Microbiology, 2011, 77(15): 5412-5419.
[16] Bang M L, Villadsen I, Sandal T. Cloning and characterization of an endo-β-1, 3-glucanase and an aspartic protease from Phaffia rhodozyma CBS6938[J]. Applied Microbiology and Biotechnology, 1999, 51(2): 215-222.
[17] Chernin L, Ismailov Z, Haran S, et al. Chitinolytic Enterobacter agglomerans antagonistic to fungal plant pathogens[J]. Applied and Environmental Microbiology, 1995, 61(5): 1720-1726.
[18] Abo Aba S E M, Soliman E A M, Nivien A A. Enhanced production of extra cellular alkaline protease in Bacillus circulance through plasmid transfer[J]. Research Journal of Agriculture and Biological Sciences, 2006, 2(6): 526-530.
[19] Nautiyal C S. An efficient microbiological growth medium for screening phosphate solubilizing microorganisms[J]. FEMS Microbiology Letters, 1999, 170(1): 265-270.
[20] Hafeez F Y, Malik K A. Manual on bio-fertilizer technology[M]. Pakistan: NIBGE, 2000.
[21] Millar R L, Higgins V J. Association of cyanide with infection of birdsfoot trefoil by Stemphylium loti[J]. Phytopathology, 1970, 60(1): 104-110.
[22] Meyer J M, Azelvandre P, Georges C. Iron metabolism in Pseudomonas: salicylic acid, a siderophore of Pseudomonas fluorescens CHAO[J]. BioFactors (Oxford, England), 1992, 4(1): 23-27.
[23] Glickmann E, Dessaux Y. A critical examination of the specificity of the salkowski reagent for indolic compounds produced by phytopathogenic bacteria[J]. Applied and Environmental Microbiology, 1995, 61(2): 793-796.
[24] Fernando W, Ramarathnam R, Krishnamoorthy A S, et al. Identification and use of potential bacterial organic antifungal volatiles in biocontrol[J]. Soil Biology and Biochemistry, 2005, 37(5): 955-964.
[25] Khabbaz S E, Abbasi P A. Isolation, characterization, and formulation of antagonistic bacteria for the management of seedlings damping off and root rot disease of cucumber[J]. Canadian Journal of Microbiology, 2014, 60(1): 25-33.
[26] Mihuta Grimm L, Erb W A, Rowe R C. Fusarium crown and root rot of tomato in greenhouse rock wool systems: sources of inoculum and disease management with benomyl[J]. Plant Disease, 1990, 74(12): 996-1002.
[27] Galkiewicz J P, Kellogg C A. Cross-kingdom amplification using bacteria-specific primers: complications for studies of coral microbial ecology[J]. Applied and Environmental Microbiology, 2008, 74(24): 7828-7831.
[28] Schwyn B, Neilands J B. Universal chemical assay for the detection and determination of siderophores[J]. Analytical Biochemistry, 1987, 160(1): 47-56.
[29] McSpadden Gardener B B, Mavrodi D V, Thomashow L S, et al. A rapid polymerase chain reaction-based assay characterizing rhizosphere populations of 2, 4-diacetylphloroglucinol-producing bacteria[J]. Phytopathology, 2001, 91(1): 44-54.
[30] Delaney S M, Mavrodi D, Bonsall V R F. phzO, a gene for biosynthesis of 2- hydroxylated phenazine compounds in Pseudomonas aureofaciens 30-84[J]. Journal of Bacteriology, 2001, 183(1): 318-327.
[31] de Souza J T, Raaijmakers J M. Polymorphisms within the prnD and the pltC genes from pyrrolnitrin and pyoluteorin-producing Pseudomonas and Burkholderia spp.[J]. FEMS Microbiology Ecology, 2003, 43(1): 21-34.
[32] Mora I, Cabrefiga J, Montesinos E. Antimicrobial peptide genes in Bacillus strains from plant environments[J]. International Microbiology, 2012, 14(4): 213-223.
[33] Arrebola E, Jacobs R, Korsten L. Iturin A is the principal inhibitor in the biocontrol activity of Bacillus amyloliquefaciens PPCB004 against postharvest fungal pathogens[J]. Journal of Applied Microbiology, 2010, 108(2): 386-395.