Screening and Biocontrol Efficiency of Phage Cocktail Targeted on a Group of Tobacco Bacterial Wilt

doi:10.13560/j.cnki.biotech.bull.1985.2024-1000

Abstract

Abstract:

Objective This study aims to isolate and systematically characterize specific bacteriophages targeting Ralstonia solanacearum, the causal agent of tobacco bacterial wilt, from the Fuzhou tobacco region of Jiangxi province. Strains with strong complementarity were screened and phage cocktail was modulated, providing technical support for the construction of a localized phage resource library and the biocontrol of tobacco bacterial wilt. Method We screened P. chrysogenum-specific phages from the in situ inter-root soil of the incidence area for P. chrysogenum in Fuzhou, explored their basic biological properties and bacteriostatic effects, and then further combined them into phage cocktails to investigate their preventive and control effects against tobacco green blight through indoor hole-plate and pot planting experiments. Result A total of 22 strains of P. chrysogenum-specific phages were isolated from the tobacco inter-root soil in Fuzhou city, among which phages GC3 and LC5 are mycotail phages, possessing an ortho icosahedral head and a tail of about 100 nm, with a large number of genes in the genome related to the tail tubes and sheaths, which may be related to their own virulence structure. Phage YH1 and YH3 are short-tailed phages, possessing an ortho icosahedral head and a 10 nm or so YH1 and YH3 are short-tailed phages with ortho icosahedral heads and tail filaments carrying genes such as lysozyme and chitinase, which may be related to the process of infestation or lysis of bacteria. Indoor hole plate and pot experiment proved that the Cyanobacteria-specific phage cocktail improved the inhibitory effect on Cyanobacteria and reduced the occurrence of Cyanobacteria. Further studies revealed that the control effect was gradually enhanced with increasing phage cocktail inoculation concentration up to 65.59%. Conclusion In this study, we isolated P. chrysogenum-specific phage from in situ, systematically studied its basic biological properties, and improved the prevention and control effect to 65.59% by constructing phage cocktail, which provided technical support for the establishment of P. chrysogenum-specific phage resource library and phage therapy for tobacco blight.

Key words: phage cocktail, bacterial wilt, tobacco, rhizosphere soil

FENG Xiao-hu, ZHANG Wen-mei, XU Jing, XIONG Shu-bin, WANG Li-bing, SONG Wen-jing. Screening and Biocontrol Efficiency of Phage Cocktail Targeted on a Group of Tobacco Bacterial Wilt[J]. Biotechnology Bulletin, 2025, 41(11): 311-318.

Figures/Tables 6

Table 1 Screening of Bacteriophage

噬菌体编号 Phage number	平均噬菌斑直径 Mean phage diameter （mm）	地点 Location	噬菌体编号 Phage number	平均噬菌斑直径 Mean phage diameter （mm）	地点 Location
YH1	1.313±0.149	宜黄	LC1	2.080±0.097	黎川
YH2	1.064±0.156	宜黄	LC2	2.125±0.110	黎川
YH3	0.819±0.074	宜黄	LC3	2.218±0.131	黎川
YH4	1.679±0.181	宜黄	LC4	1.265±0.137	黎川
YH5	1.655±0.142	宜黄	LC5	2.458±0.055	黎川
YH6	1.643±0.142	宜黄	GC1	2.213±0.083	广昌
YH7	2.374±0.238	宜黄	GC2	1.858±0.160	广昌
YH8	1.125±0.170	宜黄	GC3	2.013±0.061	广昌
YH9	0.857±0.117	宜黄	GC4	1.605±0.032	广昌
YH10	0.868±0.192	宜黄	GC5	1.743±0.072	广昌
YH11	1.063±0.146	宜黄	GC6	2.098±0.149	广昌

Fig. 1 Plate morphology of R. solanacearum (A) and its obligate bacteriophage (B), electron microscopic morphology of 4 strains of obligate bacteriophage of R. solanacearum (C-F)

Fig. 2 Phylogenetic distance of an obligate phage of R. solanacearum

Fig. 3 Gene map of four phages specific to R. solanacearum

Fig. 4 Inhibitory effects of phage cocktail and single phage on R. solanacearum (48 h)Different lowercase letters indicate the significance of differences P<0.05. The same below

Fig. 5 Effects of different phage cocktail inoculation concentrations on the occurrence of tobacco bacterial wiltA: Occurring process of bacterial wilt. B: Severity of disease. C: Disease index at the end of potted plant

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