Development of a lytic Ralstonia phage cocktail and evaluation of its control efficacy against tobacco bacterial wilt

IntroductionBacterial wilt (BW) caused by Ralstonia pseudosolanacearum is a devastating soil-borne disease. Bacteriophages are important biocontrol resources that rapidly and specifically lyse host bacteria, showing good application potential in agricultural production.MethodsThis study isolated nin...

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Main Authors: Haoxin He, Ke Yi, Lei Yang, Yongfeng Jing, Lifu Kang, Zhihao Gao, Dong Xiang, Ge Tan, Yunsheng Wang, Qian Liu, Lin Xie, Shiya Jiang, Tianbo Liu, Wu Chen
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-03-01
Series:Frontiers in Plant Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fpls.2025.1554992/full
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author Haoxin He
Ke Yi
Lei Yang
Yongfeng Jing
Lifu Kang
Zhihao Gao
Dong Xiang
Ge Tan
Yunsheng Wang
Qian Liu
Lin Xie
Shiya Jiang
Tianbo Liu
Wu Chen
author_facet Haoxin He
Ke Yi
Lei Yang
Yongfeng Jing
Lifu Kang
Zhihao Gao
Dong Xiang
Ge Tan
Yunsheng Wang
Qian Liu
Lin Xie
Shiya Jiang
Tianbo Liu
Wu Chen
author_sort Haoxin He
collection DOAJ
description IntroductionBacterial wilt (BW) caused by Ralstonia pseudosolanacearum is a devastating soil-borne disease. Bacteriophages are important biocontrol resources that rapidly and specifically lyse host bacteria, showing good application potential in agricultural production.MethodsThis study isolated nine phages (YL1–YL9) and, using host range and pot experiments, identified two broader host range phages (YL1 and YL4) and two higher control efficacy phages (YL2 and YL3), which were combined to obtain five cocktails (BPC-1–BPC-5).ResultsPot experiments showed that BPC-1 (YL3 and YL4) had the highest control efficacy (99.25%). Biological characterization revealed that these four phages had substantial thermal stability and pH tolerance. Whole genome sequencing and analysis showed that YL1, YL2, YL3, and YL4 belonged to the genus Gervaisevirus. AlphaFold 3 predictions of tail fiber protein II structures showed that YL1 differed significantly from the other phages. Amino acid sequence alignment revealed that the ORF66 (YL1) “tip domain” of contained a higher proportion of aromatic and positively charged amino acids. However, the surface of the ORF69 (YL4) “tip domain” exhibited more positively charged residues than ORF66 (YL2) and ORF70 (YL3). These characteristics are hypothesized to confer a broader host range to YL1 and YL4.DiscussionThis study demonstrates that phages assembling a broad host range and high control efficacy have better biocontrol potential, providing high-quality resources for the biological control of BW.
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publisher Frontiers Media S.A.
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spelling doaj-art-68cafbfdd5dd4bda83d80a59f01c15002025-08-20T02:22:43ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2025-03-011610.3389/fpls.2025.15549921554992Development of a lytic Ralstonia phage cocktail and evaluation of its control efficacy against tobacco bacterial wiltHaoxin He0Ke Yi1Lei Yang2Yongfeng Jing3Lifu Kang4Zhihao Gao5Dong Xiang6Ge Tan7Yunsheng Wang8Qian Liu9Lin Xie10Shiya Jiang11Tianbo Liu12Wu Chen13College of Plant Protection, Hunan Agricultural University, Changsha, ChinaTobacco Leaf Raw Material Procurement Center, China Tobacco Hunan Industrial Co., Ltd, Changsha, ChinaTobacco Leaf Raw Material Procurement Center, China Tobacco Hunan Industrial Co., Ltd, Changsha, ChinaTobacco Leaf Raw Material Procurement Center, China Tobacco Hunan Industrial Co., Ltd, Changsha, ChinaTobacco Leaf Raw Material Procurement Center, China Tobacco Hunan Industrial Co., Ltd, Changsha, ChinaTobacco Leaf Raw Material Procurement Center, China Tobacco Hunan Industrial Co., Ltd, Changsha, ChinaTobacco Leaf Raw Material Procurement Center, China Tobacco Hunan Industrial Co., Ltd, Changsha, ChinaTobacco Leaf Raw Material Procurement Center, China Tobacco Hunan Industrial Co., Ltd, Changsha, ChinaCollege of Plant Protection, Hunan Agricultural University, Changsha, ChinaCollege of Plant Protection, Hunan Agricultural University, Changsha, ChinaCollege of Plant Protection, Hunan Agricultural University, Changsha, ChinaCollege of Plant Protection, Hunan Agricultural University, Changsha, ChinaPlant Protection Research Center, Hunan Tobacco Science Research Institute, Changsha, ChinaCollege of Plant Protection, Hunan Agricultural University, Changsha, ChinaIntroductionBacterial wilt (BW) caused by Ralstonia pseudosolanacearum is a devastating soil-borne disease. Bacteriophages are important biocontrol resources that rapidly and specifically lyse host bacteria, showing good application potential in agricultural production.MethodsThis study isolated nine phages (YL1–YL9) and, using host range and pot experiments, identified two broader host range phages (YL1 and YL4) and two higher control efficacy phages (YL2 and YL3), which were combined to obtain five cocktails (BPC-1–BPC-5).ResultsPot experiments showed that BPC-1 (YL3 and YL4) had the highest control efficacy (99.25%). Biological characterization revealed that these four phages had substantial thermal stability and pH tolerance. Whole genome sequencing and analysis showed that YL1, YL2, YL3, and YL4 belonged to the genus Gervaisevirus. AlphaFold 3 predictions of tail fiber protein II structures showed that YL1 differed significantly from the other phages. Amino acid sequence alignment revealed that the ORF66 (YL1) “tip domain” of contained a higher proportion of aromatic and positively charged amino acids. However, the surface of the ORF69 (YL4) “tip domain” exhibited more positively charged residues than ORF66 (YL2) and ORF70 (YL3). These characteristics are hypothesized to confer a broader host range to YL1 and YL4.DiscussionThis study demonstrates that phages assembling a broad host range and high control efficacy have better biocontrol potential, providing high-quality resources for the biological control of BW.https://www.frontiersin.org/articles/10.3389/fpls.2025.1554992/fullbacteria wilt (BW)Ralstonia solanacearumRalstonia phagephage cocktailcontrol efficacytail fiber
spellingShingle Haoxin He
Ke Yi
Lei Yang
Yongfeng Jing
Lifu Kang
Zhihao Gao
Dong Xiang
Ge Tan
Yunsheng Wang
Qian Liu
Lin Xie
Shiya Jiang
Tianbo Liu
Wu Chen
Development of a lytic Ralstonia phage cocktail and evaluation of its control efficacy against tobacco bacterial wilt
Frontiers in Plant Science
bacteria wilt (BW)
Ralstonia solanacearum
Ralstonia phage
phage cocktail
control efficacy
tail fiber
title Development of a lytic Ralstonia phage cocktail and evaluation of its control efficacy against tobacco bacterial wilt
title_full Development of a lytic Ralstonia phage cocktail and evaluation of its control efficacy against tobacco bacterial wilt
title_fullStr Development of a lytic Ralstonia phage cocktail and evaluation of its control efficacy against tobacco bacterial wilt
title_full_unstemmed Development of a lytic Ralstonia phage cocktail and evaluation of its control efficacy against tobacco bacterial wilt
title_short Development of a lytic Ralstonia phage cocktail and evaluation of its control efficacy against tobacco bacterial wilt
title_sort development of a lytic ralstonia phage cocktail and evaluation of its control efficacy against tobacco bacterial wilt
topic bacteria wilt (BW)
Ralstonia solanacearum
Ralstonia phage
phage cocktail
control efficacy
tail fiber
url https://www.frontiersin.org/articles/10.3389/fpls.2025.1554992/full
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