Effects of Bacillus cereus F-6 on Promoting Vanilla (Vanilla planifolia Andrews.) Plant Growth and Controlling Stem and Root Rot Disease


A lipopeptide-producing bacterium, Bacillus cereus (F-6), was isolated from the rhizosphere soil of a healthy vanilla (Vanilla planifolia) plant cultivated on a plantation under 21 years of continuous cropping with vanilla. A pot experiment was conducted to investigate the effects of the green fluorescent protein-tagged F-6 (F-6-gfp) and its bio-organic fertilizer (BIO) on vanilla plant growth and stem and root rot disease, using the same plantation soil. The application of BIO significantly increased the vanilla plant root, stem and leave dry weights; however, there was not a significant difference between the F-6-gfp-inoculated treatment and the control. Meanwhile, the BIO application also significantly reduced the severity of stem and root rot disease compared to the control. The rhizosphere soil population of Fusarium was approximately 10-fold smaller in the BIO treatment compared to the control treatment at 150 days after transplantation. The number of B. cereus F-6-gfp in the rhizosphere soil of the BIO treatment remained significantly higher than that of the F-6-gfp-inoculated treatment throughout the experiment. In conclusion, F-6-gfp successfully colonized the rhizosphere soil in the BIO treatment, promoting vanilla plant growth, reducing the disease severity index, and decreasing the Fusarium population number, helping to remove barriers to the continuous cropping of vanilla.

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Zhao, Q. , Wang, H. , Zhu, Z. , Song, Y. and Yu, H. (2015) Effects of Bacillus cereus F-6 on Promoting Vanilla (Vanilla planifolia Andrews.) Plant Growth and Controlling Stem and Root Rot Disease. Agricultural Sciences, 6, 1068-1078. doi: 10.4236/as.2015.69102.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Reismunandar and Sukma E.S. (2002) Planting Vanilla. Penebar Swadaya, Bogor.
[2] Jenna, D.B. (2005) Vanilla as a Medicinal Plant. Seminars in Integrative Medicine, 3, 129-131.
[3] Xiong, W., Zhao, Q.Y., Zhao, J., Xun, W.B., Li, R., Zhang, R.F., Wu, H.S. and Shen, Q.R. (2015) Different Continuous Cropping Spans Significantly Affect Microbial Community Membership and Structure in a Vanilla-Grown Soil as Revealed by Deep Pyrosequencing. Microbial Ecology, 70, 209-218.
[4] Mazzola, M. and Manici, L.M. (2012) Apple Replant Disease: Role of Microbial Ecology in Cause and Control. Annual Review of Phytopathology, 50, 45-65.
[5] Zhang, W., Long, X.Q., Huo, X.D, Chen, Y.F. and Lou, K. (2013) 16S rRNA-Based PCR-DGGE Analysis of Actinomycete Communities in Fields with Continuous Cotton Cropping in Xinjiang, China. Microbial Ecology, 66, 385-393.
[6] Soriano-Martin, M.L., Porras-Piedra, A. and Porras-Soriano, A. (2006) Use of Microwaves in the Prevention of Fusarium oxysporum f. sp. melonis Infection during the Commercial Production of Melon Plantlets. Crop Protection, 25, 52-57.
[7] Pinaria, A.G., Liew, E.C.Y. and Burgess, L.W. (2010) Fusarium Species Associated with Vanilla Stem Rot in Indonesia. Australasian Plant Pathology, 39, 176-183.
[8] Cao, Y., Xu, Z.H., Ling, N., Yuan, Y.J., Yang, X.M., Chen, L.H., Shen, B. and Shen, Q.R. (2013) Isolation and Identification of Lipopeptides Produced by B. subtilis SQR 9 for Suppressing Fusarium Wilt of Cucumber. Scientia Horticulturae, 135, 32-39.
[9] Zhao, Q.Y., Ran, W., Wang, H., Li, X., Shen, Q.R., Shen, S.Y. and Xu, Y.C. (2013) Biocontrol of Fusarium Wilt Disease in Muskmelon with Bacillus subtilis Y-IVI. BioControl, 58, 283-292.
[10] Yu, G.Y., Sinclair, J.B., Hartman, G.L. and Bertagnolli, B.L. (2002) Production of Iturin A by Bacillus amyloliquefaciens Suppressing Rhizoctonia solani. Soil Biology and Biochemistry, 34, 955-963.
[11] Bie, X.M., Lu, Z.X. and Lu, F.X. (2009) Identification of Fengycin Homologues from Bacillus subtilis with ESI-MS/ CID. Journal of Microbiological Methods, 79, 272-278.
[12] Cazorla, F.M., Romero, D., Pérez-García, A., Lugtenberg, B.J.J., de Vicente, A. and Bloemberg, G. (2007) Isolation and Characterization of Antagonistic Bacillus subtilis Strains from the Avocado Rhizoplane Displaying Biocontrol Activity. Journal of Applied Microbiology, 103, 1950-1959.
[13] Ongena, M. and Jacques, P. (2008) Bacillus lipopeptides: Versatile Weapons for Plant Disease Biocontrol. Trends in Microbiology, 16, 115-125.
[14] Ramirez, C.A. and Kloepper, J.W. (2010) Plant Growth Promotion by Bacillus amyloliquefaciens FZB45 Depends on Inoculum Rate and P-Related Soil Properties. Biology and Fertility of Soils, 46, 835-844.
[15] Liu, Y.X., Shi, J.X., Feng, Y.G., Yang, X.M., Li, X. and Shen, Q.R. (2013) Tobacco Bacterial Wilt Can Be Biologically Controlled by the Application of Antagonistic Strains in Combination with Organic Fertilizer. Biology and Fertility of Soils, 49, 447-464.
[16] Tan, S.Y., Jiang, Y., Song, S., Huang, J.F., Ling, N., Xu, Y.C. and Shen, Q.R. (2013) Two Bacillus amyloliquefaciens Strains Isolated Using the Competitive Tomato Root Enrichment Method and Their Effects on Suppressing Ralstonia solanacearum and Promoting Tomato Plant Growth. Crop Protection, 43, 134-140.
[17] Zhang, N., Wu, K., He, X., Li, S.Q., Zhang, Z.H., Shen, B., Yang, X.M., Zhang, R.F., Huang, Q.W. and Shen, Q.R. (2011) A New Bioorganic Fertilizer Can Effectively Control Banana Wilt by Strong Colonization with Bacillus subtilis N11. Plant and Soil, 11, 87-97.
[18] Huang, X.F., Liu, J., Lu, L.J., Wen, Y., Xu, J.C., Yang, D.H. and Zhou, Q. (2009) Evaluation of Screening Methods for Demulsifying Bacteria and Characterization of Lipopeptide Bio-Demulsifier Produced by Alcaligenes sp. Bioresource Technology, 100, 1358-1365.
[19] Carrillo, P.G., Mardaraz, C., Pitta-Alvarez, S.I. and Giulietti, A.M. (1996) Isolation and Selection of Biosurfactant-Producing Bacteria. World Journal of Microbiology and Biotechnology, 12, 82-84.
[20] Landy, M., Warren, G.H., Rosenman, S.B. and Colio, L.G. (1948) An Antibiotic from Bacillus subtilis Active against Pathogenic Fungi. Experimental Biology and Medicine, 67, 539-541.
[21] Akpa, E., Jacques, P., Wathelet, B., Paquot, M., Fuchs, R., Budzikiewicz, H. and Thonart, P. (2001) Influence of Culture Conditions on Lipopeptide Production by Bacillus subtilis. Applied Biochemistry and Biotechnology, 91, 551-561.
[22] Pradhan, A.K., Pradhan, N. and Mall, G. (2013) Application of Lipopeptide Biosurfactant Isolated from a Halophile: Bacillus tequilensis CH for Inhibition of Biofilm. Applied Biochemistry and Biotechnology, 171, 1362-1375.
[23] Li, L., Mo, M., Qu, Q., Luo, H. and Zhang, K.Q. (2007) Compounds Inhibitory to Nematophagous Fungi Produced by Bacillus sp. Strain H6 Isolated from Fungistatic Soil. European Journal of Plant Pathology, 117, 329-340.
[24] Rong, L., Guo, X., Chen, K., Zhu, J., Li, S. and Jiang, J. (2009) Isolation of an Isocarbophos Degrading Strain of Arthrobacter sp. scl-2 and Identification of the Degradation Pathway. Journal of Microbiology and Biotechnology, 19, 1439-1446.
[25] Turgeon, N., Laclamme, C., Ho, J. and Duchaine, C. (2006) Elaboration of an Electroporation Protocol for Bacillus cereus ATTC14579. Journal of Microbiological Methods, 67, 543-548.
[26] Ling, N., Xue, C., Huang, Q.W., Yang, X.M., Xu, Y.C. and Shen, Q.R. (2010) Development of a Mode of Application of Bioorganic Fertilizer for Improving the Biocontrol Efficacy to Fusarium Wilt. BioControl, 55, 673-683.
[27] Ren, X.L., Zhang, N., Cao, M.H., Wu, K., Shen, Q.R. and Huang, Q.W. (2012) Biological Control of Tobacco Black Shank and Colonization of Tobacco Roots by a Paenibacillus polymyxa Strain C5. Biology and Fertility of Soils, 48, 613-620.
[28] Kinsella, K., Schulthess, C.P., Morris, T.F. and Stuart, J.D. (2009) Rapid Quantification of Bacillus subtilis Antibiotics in the Rhizosphere. Soil Biology and Biochemistry, 41, 374-379.
[29] Komada, H. (1975) Development of a Selective Medium for Quantitative Isolation of Fusarium oxysporum from Natural Soil. Review of Plant Protection Research, 8, 114-125.
[30] Turner, J.T. and Backman, P.A. (1991) Factors Relating to Peanut Yield Increases after Seed Treatment with Bacillus subtilis. Plant Disease, 75, 347-353.
[31] Zhao, Q.Y., Dong, C.X., Yang, X.M., Mei, X.L., Ran, W., Shen, Q.R. and Xu, Y.C. (2011) Biocontrol of Fusarium Wilt Disease for Cucumis melo Melon Using Bio-Organic Fertilizer. Applied Soil Ecology, 47, 67-75.
[32] Cao, Y., Zhang, Z.H., Ling, N., Yuan, Y.J., Zheng, X.Y., Shen, B. and Shen, Q.R. (2011) Bacillus subtilis SQR 9 Can Control Fusarium Wilt in Cucumber by Colonizing Plant Roots. Biology and Fertility of Soils, 47, 495-506.
[33] Chan, Y.K., Savard, M.E., Reid, L.M., Cyr, T., McCormick, W.A. and Charles, S. (2009) Identification of Lipopeptide Antibiotics of a Bacillus subtilis Isolate and Their Control of Fusarium graminearum Diseases in Maize and Wheat. BioControl, 54, 567-574.
[34] Mukherjee, A.K. and Das, K. (2005) Correlation between Diverse Cyclic Lipopeptides Production and Regulation of Growth and Substrate Utilization by Bacillus subtilis Strains in a Particular Habitat. FEMS Microbiology Ecology, 54, 479-489.
[35] Marques, A.P.G.C., Pires, C., Moreira, H., Rangel, A.O.S.S. and Castro, P.M.L. (2010) Assessment of the Plant Growth Promotion Abilities of Six Bacterial Isolates Using Zea mays as Indicator Plant. Soil Biology and Biochemistry, 42, 1229-1235.
[36] Ding, C.Y., Shen, Q.R., Zhang, R.F. and Chen, W. (2013) Evaluation of Rhizosphere Bacteria and Derived Bio-Organic Fertilizers as Potential Biocontrol Agents against Bacterial Wilt (Ralstonia solanacearum) of Potato. Plant and Soil, 366, 453-466.
[37] Zhang, F.G., Yuan, J., Yang, X.M., Cui, Y.Q., Chen, L.H., Ran, W. and Shen, Q.R. (2013) Putative Trichoderma harzianum Mutant Promotes Cucumber Growth by Enhanced Production of Indole Acetic Acid and Plant Colonization. Plant and Soil, 368, 433-444.
[38] Zhao, Q.Y., Shen, Q.R., Ran, W., Xiao, T.J., Xu, D.B. and Xu, Y.C. (2011) Inoculation of Soil by Bacillus subtilis Y-IVI Improves Plant Growth and Colonization of the Rhizosphere and Interior Tissues of Muskmelon (Cucumis melo L.). Biology and Fertility of Soils, 47, 507-514.
[39] Borrero, C., Ordovas, J., Trillas, M.I. and Aviles, M. (2006) Tomato Fusarium Wilt Suppressiveness. The Relationship between the Organic Plant Growth Media and Their Microbial Communities as Characterized by Biolog®. Soil Biology and Biochemistry, 38, 1631-1637.
[40] Zhang, S.S., Raza, W., Yang, X.M., Hu, J., Huang, Q.W., Xu, Y.C., Liu, X.H., Ran, W. and Shen, Q.R. (2008) Control of Fusarium Wilt Disease of Cucumber Plants with the Application of a Bioorganic Fertilizer. Biology and Fertility of Soils, 44, 1073-1080.

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