Aim: This study was carried out in order to evaluate the potential of bacteriophages in controlling tomato bacterial wilt disease caused by Ralstoniasolanacearum. Study design: A purposive sampling technique was used to collect samples from bacterial wilt hot spot tomato growing areas in Kenya. Place and duration of study: The research work was done at Jomo Kenyatta University of Agriculture and Technology, between June 2020 and July 2021. Methodology: Thirty diseased plants and corresponding soil were collected from five counties, Nyeri, Kajiado, Nyandarua, Kiambu and Taita Taveta. Bacteria were isolated and characterized, and then used as hosts to propagate the phages. Tests done were gram stain, oxidation test, potassium hydroxide solubility test, H2S production test catalase test, NaCl test and sugar fermentation test. Molecular analysis and phenotyping were also done in order to identify the bacteria. The bacteriophages were then isolated through a double overlay method using R.solanacearum as the host. They were characterized and assayed in a greenhouse setting to determine their effectiveness in controlling bacterial wilt. Results: Six host bacteria were isolated and all belonged to biovar II. Four phages were observed based on morphology. Upon characterization the phages were stable at 30？C, PH range between 6 - 7 and tolerance of more than an hour under UV light. In the greenhouse experiment, treatment of plants with bacteriophage prevented wilting after subsequent inoculation with the pathogen. A bacteriophage mix of SN1 and WT1 were used for efficacy tests due to their efficiency in plating and infection. Phage SN1 and WT1 exhibited high lytic activity and relatively high thermotolerance and acid tolerance, thereby showing great potential in the biocontrol of bacterial wilt infection across a variety of conditions. Conclusion: The results obtained in this research show that bacteriophages offer potential for the biocontrol of bacterial wilt.
References
[1]
Rusmania, N. (2015) 空間像再生型立体映像の研究動向. Nhk技研, 151, 10-17. https://www.nhk.or.jp/strl/publica/rd/151/3.html
[2]
Rahman, M., Islam, M., Rahman, T. and Meah, M. (2013) Biochemical Characterization of Ralstonia solanacerum Causing Bacterial Wilt of Brinjal in Bangladesh. Progressive Agriculture, 21, 9-19. https://doi.org/10.3329/pa.v21i1-2.16744
[3]
Mehta, R. (2017) History of Tomato (Poor Man’s Apple). Journal of Humanities and Social Science, 22, 31-34.
[4]
Singh, V.K., Singh, A.K., and Kumar, A. (2017) Disease Management of Tomato through PGPB: Current Trends and Future Perspective. 3 Biotech, 7, Article No. 255. https://doi.org/10.1007/s13205-017-0896-1
[5]
Din, N., Ahmad, M., Siddique, M., Ali, A., Naz, I., Ullah, N., and Ahmad, F. (2016) Phytobiocidal Management of Bacterial Wilt of Tomato Caused by Ralstonia solanacearum (Smith) Yabuuchi. Spanish Journal of Agricultural Research, 14, e1006. https://doi.org/10.5424/sjar/2016143-9012
[6]
Balemi, T., Negisho, K., Abdissa, Y., Tekalign, T., Pant, L.M., Babula, D., et al. (2005) Cultivation of Tomato: Production, Processing and Marketing. Brassica Oleracea, 12.
[7]
Yahia, E.M. and Brecht, J.K. (2012) Tomatoes. In: Rees, D., Farrell, G. and Orchard, J., Eds., Crop Post-Harvest: Science and Technology: Perishables, Blackwell Publishing Ltd, 5-23. https://doi.org/10.1002/9781444354652.ch2
[8]
Muthoni, J., Kabira, J., Shimelis, H. and Melis, R. (2014) Spread of Bacterial Wilt Disease of Potatoes in Kenya: Who is to Blame? International Journal of Horticulture, 4 ,10-15.
[9]
Nabiswa, A. (2020) Tomato Production, Distribution and Marketing in Kenya, A Baseline Survey of Kirinyaga District Report.
[10]
Rostand, R.C., Joel, M., Viviene, M. and Steven, R. (2018) Isolation and Characterization of Ralstonia solanacearum Strains Causing Bacterial Wilt of Potato in Nakuru County of Kenya. African Journal of Biotechnology, 17, 1455-1465. https://doi.org/10.5897/AJB2018.16659
[11]
Burton-Freeman, B. and Reimers, K. (2011) Tomato Consumption and Health: Emerging Benefits. American Journal of Lifestyle Medicine, 5, 182-191. https://doi.org/10.1177/1559827610387488
[12]
Susianto, G., Farid, M.M., Dhany, N.R. and Addy, H.S. (2014) Host Range for Bacteriophages that Infect Bacterial Blight Pathogen on Soybean. Procedia Environmental Sciences, 20, 760-766. https://doi.org/10.1016/j.proenv.2014.03.091
[13]
Addy, H.S., Askora, A., Kawasaki, T., Fujie, M., and Yamada, T. (2012) Utilization of Filamentous Phage φRSM3 to Control Bacterial Wilt Caused by Ralstonia Solanacearum. Plant Disease, 96, 1204-1209. https://doi.org/10.1094/PDIS-12-11-1023-RE
[14]
Kalpage, M.D. and De Costa, D.M. (2015) Isolation of Bacteriophages and Determination of Their Efficiency in Controlling Ralstonia solanacearum Causing Bacterial Wilt of Tomato. Tropical Agricultural Research, 26, 140-151. https://doi.org/10.4038/tar.v26i1.8079
[15]
Chandra, M., Thakur, S., Narang, D., Saxena, H.M., Saxena, H.M. and Angad, G. (2011) Isolation of a Bacteriophage Against Salmonella Dublin and Determination of Its Physical Resistance under varied in Vitro Conditions. Article in African Journal of Microbiology Research, 5, 2044-2047. https://www.researchgate.net/publication/262480159
[16]
Aguk, J.A., Karanja, N., Schulte-Geldermann, E., Bruns, C., Kinyua, Z. and Parker, M. (2018) Control of Bacterial Wilt (Ralstonia solanacearum) in Potato (Solanum tuberosum) Using Rhizobacteria and Arbuscular Mycorrhiza Fungi. African Journal of Food, Agriculture, Nutrition and Development, 18, 13371-13387. https://doi.org/10.18697/ajfand.82.16905
[17]
Mamay, Wahyuningrum, D. and Hertadi, R. (2015) Isolation and Characterization of Levan from Moderate Halophilic Bacteria Bacillus licheniformis BK AG21. Procedia Chemistry, 16, 292-298. https://doi.org/10.1016/j.proche.2015.12.055
[18]
Al-Heeti, A.A. and Osman Ellazeik, Y.A. (2021) Biological Pest Control: Advantages and Challenges in the Arab World. https://www.researchgate.net/publication/350357648
[19]
Huerta, A.I., Milling, A. and Allen, C. (2015) Tropical Strains of Ralstonia solanacearum Outcompete Race 3 Biovar 2 Strains at Lowland Tropical Temperatures. Applied and Environmental Microbiology, 81, 3542-3551. https://doi.org/10.1128/AEM.04123-14
[20]
Wullings, B.A., Van Beuningen, A.R., Janse, J.D. and Akkermans, A.D.L. (1998) Detection of Ralstonia solanacearum, Which Causes Brown Rot of Potato, by Fluorescent in Situ Hybridization with 23S rRNA-Targeted Probes. Applied and Environmental Microbiology, 64, 4546-4554. https://doi.org/10.1128/aem.64.11.4546-4554.1998
[21]
Razia, S., Chowdhury, M.S.M., Aminuzzaman, F.M., Sultana, N. and Islam, M. (2021) Morphological, Pathological, Biochemical and Molecular Characterization of Ralstonia solanacearum Isolates in Bangladesh. American Journal of Molecular Biology, 11, 142-164. https://doi.org/10.4236/ajmb.2021.114012
[22]
Lemessa Ocho, F. (2006) Biochemical, Pathological and Genetic Characterization of Strains of Ralstonia solanacearum (Smith) from Ethiopia and Biocontrol of R. solanacearum with Bacterial Antagonists. https://repo.uni-hannover.de/items/f2da7aa6-aa96-43ca-9704-9c928e894ca9
[23]
She, X., Yu, L., Lan, G., Tang, Y. and He, Z. (2017) Identification and Genetic Characterization of Ralstonia solanacearum Species Complex Isolates from Cucurbita Maxima in China. Frontiers in Plant Science, 8, Article 1794. https://doi.org/10.3389/fpls.2017.01794
[24]
Schönfeld, J., Heuer, H., Van Elsas, J.D. and Smalla, K. (2003) Specific and Sensitive Detection of Ralstonia solanacearum in Soil on the Basis of PCR Amplification of fliC Fragments. Applied and Environmental Microbiology, 69, 7248-7256. https://doi.org/10.1128/AEM.69.12.7248-7256.2003
[25]
Bhunchoth, A., Phironrit, N., Leksomboon, C., Chatchawankanphanich, O., Kotera, S., Narulita, E., Kawasaki, T., Fujie, M. and Yamada, T. (2015) Isolation of Ralstonia solanacearum-Infecting Bacteriophages from Tomato Fields in Chiang Mai, Thailand, and Their Experimental Use as Biocontrol Agents. Journal of Applied Microbiology, 118, 1023-1033. https://doi.org/10.1111/jam.12763
[26]
Rombouts, S., Volckaert, A., Venneman, S., Declercq, B., Vandenheuvel, D., Allonsius, C.N., Van Malderghem, C., Jang, H.B., Briers, Y., Noben, J.P., Klumpp, J., Van Vaerenbergh, J., Maes, M. and Lavigne, R. (2016) Characterization of Novel Bacteriophages for Biocontrol of Bacterial Blight in Leek Caused by Pseudomonas syringae pv. porri. Frontiers in Microbiology, 7, Article 279. https://doi.org/10.3389/fmicb.2016.00279
[27]
Lee, J. and Park, T.-H. (2016) Isolation and Characterization of Bacteriophages Infecting Ralstonia solanacearum from Potato Fields. Research in Plant Disease, 22, 236-242. https://doi.org/10.5423/RPD.2016.22.4.236
[28]
Fujiwara, A., Fujisawa, M., Hamasaki, R., Kawasaki, T., Fujie, M. and Yamada, T. (2011) Biocontrol of Ralstonia solanacearum by Treatment with Lytic Bacteriophages. Applied and Environmental Microbiology, 77, 4155-4162. https://doi.org/10.1128/AEM.02847-10
[29]
Mountseng, C.P., Mahot, H., Membang, G., Mboussi, S.B., Begoude, D. and Ambang, Z. (2024) Morphological and Biochemical Characterization of Strains of Ralstonia solanacearum, Causal Agent of Bacterial Wilt in Tomatoes in Cameroon and Screening of Virulent Strains. Annual Research & Review in Biology, 39, 27-42. https://doi.org/10.9734/arrb/2024/v39i52079
[30]
Wang, Z., Luo, W., Cheng, S., Zhang, H., Zong, J. and Zhang, Z. (2023) Ralstonia solanacearum—A Soil Borne Hidden Enemy of Plants: Research Development in Management Strategies, Their Action Mechanism and Challenges. Frontiers in Plant Science, 14, Article 1141902. https://doi.org/10.3389/fpls.2023.1141902
[31]
Seleim, M.A.A., Abo-Elyousr, K.A.M., Abd-El-Moneem, K.M. and Saead, F.A. (2014) First Report of Bacterial Wilt Caused by Ralstonia solanacearum Biovar 2 Race 1 on Tomato in Egypt. Plant Pathology Journal, 30, 299-303. https://doi.org/10.5423/PPJ.NT.10.2013.0101
[32]
Álvarez, B. and Biosca, E.G. (2017) Bacteriophage-Based Bacterial Wilt Biocontrol for an Environmentally Sustainable Agriculture. Frontiers in Plant Science, 8, Article 1218. https://doi.org/10.3389/fpls.2017.01218
[33]
Wdowiak, M., Paczesny, J. and Raza, S. (2022) Enhancing the Stability of Bacteriophages Using Physical, Chemical, and Nano-Based Approaches: A Review. Pharmaceutics, 14, Article 1936. https://doi.org/10.3390/pharmaceutics14091936
[34]
Ranveer, S.A., Dasriya, V., Ahmad, M.F., Dhillon, H.S., Samtiya, M., Shama, E., Anand, T., Dhewa, T., Chaudhary, V., Chaudhary, P., Behare, P., Ram, C., Puniya, D. V., Khedkar, G.D., Raposo, A., Han, H. and Puniya, A.K. (2024) Positive and Negative Aspects of Bacteriophages and Their Immense Role in the Food Chain. NPJ Science of Food, 8, Article No. 1. https://doi.org/10.1038/s41538-023-00245-8
[35]
Pawaskar, J., Joshi, M.S., Navathe, S. and Agale, R.C. (2014) Physiological and Biochemical Characters of Ralstonia solanacearum. International Journal of Research in Agricultural Sciences, 1, 2348-3997.
[36]
Jończyk, E., Kłak, M., Międzybrodzki, R. and Górski, A. (2011) The Influence of External Factors on Bacteriophages-Review. Folia Microbiologica, 56, 191-200. https://doi.org/10.1007/s12223-011-0039-8
[37]
Düzgüneş, N., Sessevmez, M. and Yildirim, M. (2021) Bacteriophage Therapy of Bacterial Infections: The Rediscovered Frontier. Pharmaceuticals, 14, 1-16. https://doi.org/10.3390/ph14010034
