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Monitoring tritrophic biocontrol interactions between Bacillus spp., Fusarium oxysporum f. sp. cubense, tropical race 4 and banana plants in vivo based on fluorescent transformation system

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Title Monitoring tritrophic biocontrol interactions between Bacillus spp., Fusarium oxysporum f. sp. cubense, tropical race 4 and banana plants in vivo based on fluorescent transformation system
 
Creator He, Ping
Li, Shu
Xu, Shengtao
Fan, Huacai
Wang, Yongfen
Zhou, Wei
Fu, Gang
Han, Guangyu
Wang, Yunyue
Zheng, Si-Jun
 
Subject biological control
disease management
monitoring techniques
control biológico
gestión de la enfermedad
técnicas de vigilancia
 
Description Bacillus spp. are effective biocontrol agents for Fusarium wilt of banana, tropical race 4 (TR4). This study explores the colonization by B. subtilis, B. velezensis and B. amyloliquefaciens of host banana plants and elucidates the mechanism of antagonistic TR4 biocontrol. The authors selected one B. subtilis strain, three B. velezensis strains and three B. amyloliquefaciens strains that are proven to significantly inhibit TR4 in vitro, optimized the genetic transformation conditions and explored their colonization process in banana plants. The results showed that we successfully constructed an optimized fluorescent electro-transformation system (OD600 of bacteria concentration = 0.7, plasmid concentration = 50 ng/μL, plasmid volume = 2 μL, transformation voltage = 1.8kV, transformation capacitance = 400Ω) of TR4-inhibitory Bacillus spp. strains. The red fluorescent protein (RFP)-labelled strains were shown to have high stability with a plasmid-retention frequency above 98%, where bacterial growth rates and TR4 inhibition are unaffected by fluorescent plasmid insertion. In vivo colonizing observation by Laser Scanning Confocal Microscopy (LSCM) and Scanning Electron Microscopy (SEM) showed that Bacillus spp. can colonize the internal cells of banana plantlets roots. Further fluorescent observation by (LSCM) showed these RFP-labelled bacteria exhibit chemotaxis (chemotaxis ratio was 1.85±0.04) towards green fluorescent protein (GFP)-labelled TR4 hyphae in banana plants. We conclude that B. subtilis, B. velezensis and B. amyloliquefaciens can successfully colonize banana plants and interact with TR4. Monitoring its dynamic interaction with TR4 and its biocontrol mechanism is under further study.
 
Date 2021-10-13
2021-12-10T07:42:49Z
2021-12-10T07:42:49Z
 
Type Journal Article
 
Identifier He, P.; Li, S.; Xu, S.; Fan, H.; Wang, Y.; Zhou, W.; Fu, G.; Han, G.; Wang, Y.; Zheng, S.-J. (2021) Monitoring tritrophic biocontrol interactions between Bacillus spp., Fusarium oxysporum f. sp. cubense, tropical race 4 and banana plants in vivo based on fluorescent transformation system. Frontiers in Microbiology 12: 754918 13 p. ISSN: 1664-302X
1664-302X
https://hdl.handle.net/10568/116646
https://doi.org/10.3389/fmicb.2021.754918
 
Language en
 
Rights CC-BY-4.0
Open Access
 
Format 13 p.
application/pdf
 
Publisher Frontiers Media SA
 
Source Frontiers in Microbiology