Skip to main content
PMC home page
Virologica Sinica logoLink to Virologica Sinica
. 2017 Nov 16;32(6):476–484. doi: 10.1007/s12250-017-3987-6

Developing a bacteriophage cocktail for biocontrol of potato bacterial wilt

Cuihua Wei 1,2, Junli Liu 3, Alice Nyambura Maina 4,5, Francis B Mwaura 4, Junping Yu 1, Chenghui Yan 1,2, Ruofang Zhang 3,✉, Hongping Wei 1,6,✉
PMCID: PMC6598922  PMID: 29168148

Abstract

Bacterial wilt is a devastating disease of potato and can cause an 80% production loss. To control wilt using bacteriophage therapy, we isolated and characterized twelve lytic bacteriophages from different water sources in Kenya and China. Based on the lytic curves of the phages with the pathogen Ralstonia solanacearum, one optimal bacteriophage cocktail, P1, containing six phage isolations was formulated and used for studying wilt prevention and treatment efficiency in potato plants growing in pots. The preliminary tests showed that the phage cocktail was very effective in preventing potato bacterial wilt by injection of the phages into the plants or decontamination of sterilized soil spiked with R. solanacearum. Eighty percent of potato plants could be protected from the bacterial wilt (caused by R. solanacearum reference strain GIM1.74 and field isolates), and the P1 cocktail could kill 98% of live bacteria spiked in the sterilized soil at one week after spraying. However, the treatment efficiencies of P1 depended on the timing of application of the phages, the susceptibility of the plants to the bacterial wilt, as well as the virulence of the bacteria infected, suggesting that it is important to apply the phage therapy as soon as possible once there are early signs of the bacterial wilt. These results provide the basis for the development of bacteriophagebased biocontrol of potato bacterial wilt as an alternative to the use of antibiotics.

graphic file with name 12250_2017_3987_Fig1_HTML.jpg

Electronic Supplementary Material

Supplementary material is available for this article at 10.1007/s12250-017-3987-6 and is accessible for authorized users.

Keywords: Ralstonia solanacearum, bacterial wilt, potato, bacteriophage therapy

Electronic supplementary material

12250_2017_3987_MOESM1_ESM.pdf (1.3MB, pdf)

Developing a bacteriophage cocktail for biocontrol of potato bacterial wilt

Acknowledgments

This study was supported financially by the Sino-Africa Joint Research Center (SAJC201605) and the Chinese Academy of Sciences (ZDRW-ZS-2016-4). We thank Dr. Ding Gao from the core facility center, Wuhan Institute of Virology, CAS for the help on TEM observation.

Contributor Information

Ruofang Zhang, Email: ruofang_zhang@163.com.

Hongping Wei, Email: hpwei@wh.iov.cn.

References

  1. Adams MH, Anderson ES, Gots JS, Jacob F, Wollman EL. Bacteriophages. New York-London: Interscience Publishers, Inc; 1959. pp. 96–119. [Google Scholar]
  2. Addy HS, Askora A, Kawasaki T, Fujie M, Yamada T. Util-ization of filamentous phage ϕRSM3 to control bcacterial wilt caused by Ralstonia solanacearum. Plant Dis. 2012;96:1204–1209. doi: 10.1094/PDIS-12-11-1023-RE. [DOI] [PubMed] [Google Scholar]
  3. Bae JY, Wu J, Lee HJ, Jo EJ, Murugaiyan S, Chung ES, Lee SW. Biocontrol potential of a lytic bacteriophage PE204 against bacterial wilt of tomato. J Microbio Biotechn. 2012;22:1613–1620. doi: 10.4014/jmb.1208.08072. [DOI] [PubMed] [Google Scholar]
  4. Barrell PJ, Meiyalaghan S, Jacobs JME, Conner AJ. Applications of biotechnology and genomics in potato improvement. Plant Biotechnol J. 2013;11:907–920. doi: 10.1111/pbi.12099. [DOI] [PubMed] [Google Scholar]
  5. Bhunchoth A, Phironrit N, Leksomboon C, Chatchawankanphanich O, Kotera S, Narulita E, Kawasaki T, Fujie M, Yamada T. Isolation of Ralstonia solanacearum infecting bacteriophages from tomato fields in Chiang Mai, Thailand, and their experimental use as biocontrol agents. Appl Environ Microbiol. 2015;118:1023–1033. doi: 10.1111/jam.12763. [DOI] [PubMed] [Google Scholar]
  6. Carlton RM. Phage therapy: past history and future prospects. Arch Immunol Ther Ex. 1999;47:267–274. [PubMed] [Google Scholar]
  7. Chen WY. Effects of avirulent bacteriocin-producing strains of Pseudomonas solanacearum on the control of bacterial wilt of tobacco. Plant Pathol. 1984;33:245–253. doi: 10.1111/j.1365-3059.1984.tb02646.x. [DOI] [Google Scholar]
  8. Ciampi-Panno L, Fernandez C, Bustamante P, Andrade N, Ojeda S, Contreras A. Biological control of bacterial wilt of potatoes caused by Pseudomonas solanacearum. Am J Potato Res. 1989;66:315–332. doi: 10.1007/BF02854019. [DOI] [Google Scholar]
  9. Czajkowski R, Pérombelond MCM, Van-Veen JA, Vander-Wolf JM. Control of blackleg and tuber soft rot of potato caused by Pectobacterium and Dickeyaspecies: a review. Plant Pathol. 2011;60:999–1013. doi: 10.1111/j.1365-3059.2011.02470.x. [DOI] [Google Scholar]
  10. Denny TP. Plant pathogenic Ralstonia species. In: Gnanamenisham SS, editor. Plant-associated bacteria. Dordrecht: Springer; 2007. pp. 573–644. [Google Scholar]
  11. Farag MA, Al-Mahdy DA, ET-Dine RS, Fahmy S, Yassin A, Porzel A, Brandt W. Structure-activity relationships of antimicrobial gallic acid derivatives from pomegranate and acacia fruit extracts against potato bacterial wilt pathogen. Chem and Biodivers. 2015;12:955–962. doi: 10.1002/cbdv.201400194. [DOI] [PubMed] [Google Scholar]
  12. Fujie M, Takamoto H, Kawasaki T, Fujiwara A, Yamada T. Monitoring growth and movement of Ralstonia solanacearum cells harboring plasmid pRSS12 derived from bacteriophage phiRSS1. J Biosci Bioeng. 2010;109:153–158. doi: 10.1016/j.jbiosc.2009.07.012. [DOI] [PubMed] [Google Scholar]
  13. Fujiwara A, Fujisawa M, Hamasaki R, Kawasaki T, Fujie M, Yamada T. Biocontrol of Ralstonia solanacearum by treatment with lytic bacteriophages. Appl Environ Microbiol. 2011;77:4155–4162. doi: 10.1128/AEM.02847-10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hanumanthappa KM, Palaniswamy M, Angayarkanni J. Isolation of lytic bacteriophage against Ralstonia solanacearum causing wilting symptoms in ginger (Zingiber officinale) and potato (Solanumtuberosum) plants. Int J Biol Sci. 2013;2:78–84. [Google Scholar]
  15. Hayward AC. Biology and epidemiology of bacterial wilt caused by Pseudomonas solanacearum. Annu Rev Phytopathol. 1991;29:65–87. doi: 10.1146/annurev.py.29.090191.000433. [DOI] [PubMed] [Google Scholar]
  16. Ji XL, Zhang CJ, Fang Y, Zhang Q, Lin L, Tang B, Wei Y. Isolation and characterization of glacier VMY22, a novel lytic cold-active bacteriophage of bacillus cereus. Virol Sin. 2015;30:52–58. doi: 10.1007/s12250-014-3529-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jones JB, Vallad GE, Iriarte FB, Obradovic A, Wernsing MH, Jackson LE, Balogh B, Hong JC, Momol MT. Considerations for using bacteriophages for plant disease control. Bacteriophage. 2012;2:208–214. doi: 10.4161/bact.23857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kwiatek M, Parasion S, Mizak L, Gryko R, Bartoszcze M, Kocik J. Characterization of a bacteriophage, isolated from a cow with mastitis, that is lytic against Staphylococcus aureus strains. Arch Virol. 2012;157:225–234. doi: 10.1007/s00705-011-1160-3. [DOI] [PubMed] [Google Scholar]
  19. Loc-Carrillo C, Abedon ST. Pros and cons of phage therapy. Bacteriophage. 2011;1:111–114. doi: 10.4161/bact.1.2.14590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Masum MMI, Islam SMM, Islam MS, Kabir MH. Estimation of loss due to post harvest diseases of potato in markets of different districts in Bangladesh. Afr J Biotechnol. 2011;10:11892–11902. [Google Scholar]
  21. Murugaiyan S, Bae JY, Wu J, Lee SD, Um HY, Choi HK, Chung E, Lee JH, Lee SW. Characterization of filamentous bacteriophage PE226 infecting Ralstonia solanacearum strains. J Appl Microbiol. 2011;110:296–303. doi: 10.1111/j.1365-2672.2010.04882.x. [DOI] [PubMed] [Google Scholar]
  22. Popova AV, Zhilenkov EL, Myakinina VP, Krasilnikova VM, Volozhantsev NV. Isolation and characterization of wide host range lytic bacteriophage AP22 infecting Acinetobacter baumannii. FEMS Microbiol Lett. 2012;332:40–46. doi: 10.1111/j.1574-6968.2012.02573.x. [DOI] [PubMed] [Google Scholar]
  23. Roberts DP, Denny TP, Schell MA. Cloning of the egl gene of Pseudomonas solanacearum and analysis of its role in phytopathogenicity. J Bacteriol. 1988;170:1445–1451. doi: 10.1128/JB.170.4.1445-1451.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sagar V, Jeevalatha A, Mian S, Chakrabarti SK, Gurjar MS, Arora RK, Sharma S, Bakade RR, Singh BP. Potato bacterial wilt in India caused by strains of phylotype I, II and IV of Ralstonia solanacearum. Eur J of Plant Patho. 2013;l138:51–65. [Google Scholar]
  25. Stratton CW. Dead bugs don’t mutate: susceptibility issues in the emergence of bacterial resistance. Emerg Infect Dis. 2003;9:10–16. doi: 10.3201/eid0901.020172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wicker E, Grassart L, Coranson-Beaudu R, Mian D, Guilbaud C, Fegan M, Prior P. Ralstonia solanacearum strains from Martinique (French West Indies) exhibiting a new pathogenic potential. Appl Environ Microbiol. 2007;73:6790–6801. doi: 10.1128/AEM.00841-07. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ye JX, Kostrzynska M, Dunfield K, Warriner K. Control of Salmonella on sprouting mung bean and alfalfa seeds by using a biocontrol preparation based on antagonistic bacteria and lytic bacteriophages. J Food Protect. 2010;73:9–17. doi: 10.4315/0362-028X-73.1.9. [DOI] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

12250_2017_3987_MOESM1_ESM.pdf (1.3MB, pdf)

Developing a bacteriophage cocktail for biocontrol of potato bacterial wilt

Articles from Virologica Sinica are provided here courtesy of Wuhan Institute of Virology, Chinese Academy of Sciences

RESOURCES