In Vitro Assay of Lytic Bacteriophage to Suppress the Growth of Ralstonia syzygii subsp. indonesiensis, the Causal Pathogen of Potato Wilt Disease

Authors

  • Nur Azizah Department of Agrotechnology, Faculty of Agriculture, Universitas Sumatera Utara, Medan, Indonesia
  • Irda Safni 1Department of Agrotechnology, Faculty of Agriculture, Universitas Sumatera Utara, Medan, Indonesia

DOI:

https://doi.org/10.32734/injar.v7i2.14192

Keywords:

bacteriophage, bacterial wilt disease, in vitro assay, potato, Ralstonia syzygii subsp. indonesiensis

Abstract

Bacterial wilt disease, the one of the major diseases of potatoes, caused by Ralstonia syzygii subsp. indonesiensis (Rsi). Many efforts have been made to control bacterial wilt disease, including physical control, chemicals, and the use of bacteriophages. Previous studies have shown that bacteriophage application in controlling plant diseases is a fast-expanding area and has great potential to replace chemical methods. This study aims to determine the potential of lytic bacteriophage in suppressing the growth of Rsi in vitro. This study used a Non-Factorial, Completely Randomized Design with 3 replications and 6 treatment levels: R1 (Rsi isolate 1 without bacteriophage), R2 (Rsi isolate 1 with bacteriophage), R3 (Rsi isolate 2 without bacteriophage), R4 (Rsi isolate 2 with bacteriophage), R5 (Rsi isolate 3 without bacteriophage), and R6 (Rsi isolate 3 with bacteriophage). The results showed that bacteriophage could reduce the Rsi population at 24 hours by looking at the optical density (OD) value of 600 nm wavelength and growing on NA medium using the spread-plate method. The best treatment was at R6 with a population of 1 x 108cfu/ml Rsi. This research suggests that bacteriophage has the potential to suppress the growth of Rsi, which causes potato bacterial wilt disease, in vitro.

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References

Produksi Tanaman Sayuran Menurut Kabupaten/ Kotadan Jenis Tanaman di Provinsi Sumatera Utara 2019 dan 2020. [Production of vegetables according to district/city and type of plant in North Sumatra province 2019 and 2020], Badan Pusat Statistik Provinsi Sumatera Utara, Indonesia, 2021.

J. Mansfield, S. Genin, S. Magori, V. Citovsky, M. Sriariyanum, P. Ronald, M. A. X. Dow, V. Verdier, S. V. Beer, M. A. Machado, and I. A. N. Toth. 2012. Top 10 plant pathogenic bacteria in molecular plant pathology. Molecular Plant Pathology, vol 13, no. 6, pp. 614-629.

I. Safni, I. Cleenwerk, P. De Vos, M. Fegan, L. Sly, and U. Kappler. 2014. Polyphasic taxonomic revision of the Ralstonia solanacearum spesies complex: Proposal to emend the description of Ralstonia solanacearum and Ralstonia syzygii and reclassify current R.syzygii strains as Ralstonia syzygii subsp.syzygii subsp.nov., R. sonalacearum phylotipe IV strains as Ralstonia syzygii subsp. indonesiensis subsp. nov., Banana Blood Disease Bacterium strains as Ralstonia syzygii subsp. celebesensis subsp. nov., and R. solanacearum phylotipe I and III strains as Ralstonia pseudosolanacearum sp.nov. International Journal of Systematic and Evolutionary Microbiology, vo. 64, pp. 3087-3103.

J. B. Jones, G. E. Vallad, F. B. Iriarte, A. Obradović, M. H. Wernsing, L. E. Jackson, B. Balogh, J. C. Hong, and M. T. Momol, “Considerations for using bacteriophages for plant disease control,” Bacteriophage, vol. 2, no. 4, p. e23857, 2012.

C. P. Nababan, Suswati, and S. Hasibuan, “Efektivitas penggunaan biofumigan limbah brassica terhadap penyakit layu bakteri (Ralstonia solanacearum Ras3) pada tanaman kentang di Pematang Silima Huta Kabupaten Simalungun. [Effectiveness of using brassica waste biofumigant against bacterial wilt (Ralstonia solanacearum Ras 3) on potato plants in Pematang Silima Huta, Simalungun Regency],”. Agrotekma: Jurnal Agroteknologi dan Pertanian, vol. 2, no. 1, pp. 56-64, 2017.

K. Hidayati, “PenggunaanTrichoderma konginii sebagai Pengendali Penyakit Layu Bakteri oleh Ralstonia solanacearum pada Budidaya Tanaman Kentang di Dataran Medium. [Use of Trichoderma konginii as Bacterial Wilt Control by Ralstonia solanacearum in Potato Cultivation in Medium Plains],” [Thesis]. Universitas Brawijaya, Malang, Indonesia, 2012.

N. Prihatingingsih, A. Triwidodo, H. Bambang, and W. Jaka, “Mekanisme antibiosis Bacillus substilis B315 untuk pengendalian penyakit layu bakteri kentang. [Antibiosis mechanism of Bacillus substilis B315 for controlling potato bacterial wilt],” Journal of Tropical Plant Pests and Diseases, vol. 15, no. 1, pp. 64-71, 2015.

F. A. Choliq, M. Martosudiro, Istiqomah, and M. F. Nijami, “Isolasi dan uji kemampuan bakteriofag sebagai agens pengendali penyakit layu bakteri (Ralstonia solanacearum) pada tanaman tomat. [Isolation and Testing of the Ability of Bacteriophages as Controlling Agents for Bacterial Wilt Disease (Ralstonia solanacearum) in Tomato Plants],” Viabel Jurnal Ilmiah Ilmu-Ilmu Pertanian, vol. 14, no. 1, pp. 8-20, 2020.

N. T. Vu and C. S. Oh, “Bacteriophage usage for bacterial disease management and diagnosis in plants,” The Plant Pathology Journal, vol. 36, no. 3, p. 204, 2020.

W. L. Mallmann, and C. A.R. L. Hemstreet, “Isolation of an inhibitory substance from plants,” Agricultural Research, vol. 28, pp. 599-602, 1924.

G. H. Coons and J. E. Kotila, “The transmissible lytic principle (bacteriophage) in relation to plant pathogens,” Phytopathology, vol. 15, pp. 357-370, 1925.

R. C. Thomas, “A bacteriophage in relation to Stewart’s disease of corn,” Phytopathology, vol. 25, no. 3, pp. 371-372, 1935.

R. J. Boyd, A. C. Hildebrandt, and O. N. Allen, “Retardation of crown gall enlargement after bacteriophage treatment,” Plant Disease Reporter, vol. 55, pp. 145-158, 1971.

T. T. Kuo, L. C. Chang, C. M. Yang, and S.E Yang, “Bacterial leaf blight of rice plant. IV. Effect of bacteriophage on the infectivity of Xanthomonas oryzae,” Acad Sinica Inst Bot Bot Bull, vol. 12, pp. 1-9, 1971.

H. Tanaka, H. Negishi, and H. Maeda, “Control of tobacco bacterial wilt by an avirulent strain of Pseudomonas solanacearum M4S and its bacteriophage,” Japanese Journal of Phytopathology, vol. 56, no. 2, pp. 243-246, 1990.

H. S. Addy, A. Askora, T. Kawasaki, M. Fujie, and T. Yamada, “Utilization of filamentous phage ɸRSM3 to control bacterial wilt caused by Ralstonia solanacearum,” Plant Disease, vol. 96, no. 8, pp.1204-1209, 2012.

R. S. Murthi, I. Safni, and Lisnawita, “Isolation and selection specific bacteriphage from banana in North Sumatera to biologically control Ralstonia syzygii subsp. celebesensis in vitro,” International Conference on Agriculture, Environment and Food Security, vol. 782, no 4, p. 042018, 2021.

D. R, Amaliah, “Isolasi Keragaman Bakteri Endofitdan Filosfer yang Berpotensi sebagai Agens Pengendali Hayati Penyakit Layu Bakteri (Ralstonia solanacearum) pada Tanaman Kentang (Solanum tuberosum L.). [Isolation of Diversity of Endophyte Bacteria and Phylosphere that are Potential as Biological Control of Bacterial Wilt Disease (Ralstonia solanacearum) on Potato (Solanum tuberosum L.)],” thesis, Universitas Hasanuddin, Makassar, Indonesia, 2017.

C. Allen, A. Kelman, and E. R. French, “Brown rot. In W. R. Stevenson, R. Loria, G. D. Franc, & D. P. Weingartner (Eds.),” Compendium of Potato Diseases (2 ed., pp. 11-13). St. Paul: APS Press, 2001.

E. Ryu, “A simple method of differentiation between Gram-positive and Gram-negative organisms without staining,” Kitasato Archive of Experimental Medicine, 1940, vol. 17, pp. 58-63.

R. A. Lelliot and D. E. Stead, “Methods for Diagnostics of Bacterial Diseases of Plants,” London: Blackwell Scientific Publications, 1987.

A. C. Hayward, “Phenotypic methods for the differentiation of Pseudomonas solanacearum: biovars and supplementary observations. In Techniques for Diagnosis of Pseudomonas solanacearum and for Resistance Screening against Groundnut Bacterial Wilt,” Edited by V. K. Mehan & D. McDonald. India: International Crops Research Institute for the Semi-Arid Tropics, 1995.

R. S. Murthi, “Studi Potensi Bakteriofage untuk Mengendalikan Penyakit Darah pada Tanaman Pisang (Ralstonia syzygii subsp.celebesensis). [Study of Bacteriophage Potential to Control Blood Disease in Banana (Ralstonia syzygii subsp. celebesensis)],” thesis, Universitas Sumatera Utara, Medan, Indonesia, 2021.

D. Sartika, “Efektifitas dan keamanan in vivo fage litik FR38 dari limbah domestik dalam menurunkan cemaran salmonella P38 indigenous pada sosis, susu, dan air. [Efectivity and safety in vivo lytic FR38 from domestic waste to reduce contamination of indigenous Salmonella P38 on sosage, milk and water],” thesis, Institut Pertanian Bogor, Bogor, Indonesia, 2012.

L. Goodridge, A. Gallaccio, and W. M. Griffiths, “Morphological, hostrange, and genetic characterization of two colifages,” Applied and Environmental Microbiology, vol. 69, pp. 5364-5371, 2021.

F. A. Choliq, M. Martosudiro, Istiqomah, and M. F. Nijami, “Isolasi dan uji kemampuan bakteriofag sebagai agens pengendali penyakit layu bakteri (Ralstonia solanacearum) pada tanaman tomat. [Isolation and capability essay of bacteriophage as a biocontrol agent of bacterial wilt disease (Ralstonia solanacearum) on tomato]”. J. Viabel Pertanian, vol. 14, no. 1, pp. 8-20, 2020.

S. T. Abedon and J. Yin, “Bacteriophage plaques: theory and analysis. Bacteriophages: methods and protocols,” Volume 1: Isolation, Characterization, and Interactions, Humana Press. pp. 161-174, 2009.

H. Prescott, “Laboratory Exercises in Microbiology (Exercise 48: Isolation of Escherichia coli Bacteriophages from Sewage and Determining Bacteriophage Titers),” Fifth Edition. USA:The McGraw Hill Companies, 2002.

M. T. Madigan, J. M. Martinko, P. V. Dunlap, and D. P. Clark, “Brock Biology of Microorganisms 12th edn,” International Microbiology, vol. 11, pp. 65-73, 2008.

T. P. Denny, “Plant pathogenic Ralstonia species. In Plant-associated Bacteria, pp. 573-644,” Edited by S. Gnanamanickam. Dordrecht, the Netherlands: Springer, 2006.

H. Toyoda, K. Kakutani, S. Ikeda, S. Goto, H. Tanaka, and S. Ouchi, “Characterization of deoxyribonucleic acid of virulent bacteriophage and its infectivity to host bacteria, Pseudomonas solanacearum,” Journal of Phytopathology, vol. 13, no. 1, pp.11-21, 1991.

R. Young, “Phage lysis: three steps, three choices, one outcome,” Journal of Microbiology, vol. 52, pp. 243-258, 2014.

V. A. Fischetti, “Bacteriophage lysins as effective antibacterials,” Current Opinion in Microbiology, vol.11, no. 5, pp. 393-400, 2008.

I. N. Wang, Smith, DL and Young, R. 2000. Holins: the protein clocks of bacteriophage infections,” Annual Reviews in Microbiology, vol. 54, no. 1, pp. 799-825, 2000.

I. N. Wang, J. Deaton, and R. Young, “Sizing the holin lesion with an endolysin-β-galactosidase fusion,” Journal of Bacteriology, vol. 185, no. 3, pp. 779-787, 2003.

R. Y. Young, N. Wang, and W. D. Roof, “Phages will out: strategies of host cell lysis,” Trends in Microbiology, vol.8, no. 3, pp. 120-128, 2000.

T. G. Bernhardt, I. N. Wang, D. K. Struck, and R. Young, “A protein antibiotic in the phage Qβ virion: diversity in lysis targets,” Science, vol. 292, no. 5525, pp. 2326-2329, 2001.

A. G. Cobián-Güemes, M. Youle, V. A. Cantú, B. Felts, J. Nulton, and F. Rohwer, “Viruses as winners in the game of life,” Annual Review of Virology, vol. 3, pp. 197-214, 2014.

X. Wittebole, S. De Roock, and S. M. Opal, “A historical overview of bacteriophage therapy as an alternative to antibiotics for the treatment of bacterial pathogens,” Virulence, vol. 5, no. 1, pp. 226-235, 2014.

P. D. Umrao, V. Kumar, and S. D. Kaistha, “Biocontrol potential of bacteriophage ɸsp1 against bacterial wilt-causing Ralstoniasolanacearum in Solanaceae crops,” Egyptian Journal of Biological Pest Control, vol. 31, no. 1, pp. 1-12, 2021.

X. Wang, Z. Wei, K. Yang, J. Wang, A. Jousset, Y. Xu, Q. Shen, and V. P. Friman, “Phage combination therapies for bacterial wilt disease in tomato,” Nature Biotechnology, vol. 37, no. 12, pp. 1513-1520, 2019.

Published

2024-07-07

How to Cite

Azizah, N., & Irda Safni. (2024). In Vitro Assay of Lytic Bacteriophage to Suppress the Growth of Ralstonia syzygii subsp. indonesiensis, the Causal Pathogen of Potato Wilt Disease. Indonesian Journal of Agricultural Research, 7(2), 79 - 90. https://doi.org/10.32734/injar.v7i2.14192