Development and characterization of a fusion mutant with the truncated lacZ to screen regulatory genes for phenazine biosynthesis in Pseudomonas chlororaphis G05

Chi, Xiaoyan; Wang, Yanhua; Jiang, Miao; Feng, Zhibin; Zhang, Hongqian; Zhai, Junjie; Zhang, Haoyi; Tian, Lingqian; Wu, Xue; Yang, Tingting; Huang, Run; Hu, Xiaomei; Ge, Yihe

doi:10.1016/j.biocontrol.2017.02.012

Cited by 10 publications

(5 citation statements)

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“…are paid great attention because they can alleviate plant diseases and increase crop productivity ( Baehler et al, 2005 ; Haas and Defago, 2005 ; Haas and Keel, 2003 ). Pseudomonas chlororaphis G05 is a root-colonizing biocontrol agent that bioprotects some plants from the diseases caused by fungal phytopathogens, such as Fusarium oxysporum , Rhizoctonia solani , and F. graminearum ( Chi et al, 2017 ; Ge et al, 2008 ; Huang et al, 2018 ). It has been demonstrated that antifungal compounds, phenazine-1-carboxylic acid and pyrrolnitrin that are produced in this bacterium, mainly contribute to suppression of mycelial growth of these phytopathogenic fungi ( Chi et al, 2017 ; Huang et al, 2018 ).…”

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confidence: 99%

“…Pseudomonas chlororaphis G05 is a root-colonizing biocontrol agent that bioprotects some plants from the diseases caused by fungal phytopathogens, such as Fusarium oxysporum , Rhizoctonia solani , and F. graminearum ( Chi et al, 2017 ; Ge et al, 2008 ; Huang et al, 2018 ). It has been demonstrated that antifungal compounds, phenazine-1-carboxylic acid and pyrrolnitrin that are produced in this bacterium, mainly contribute to suppression of mycelial growth of these phytopathogenic fungi ( Chi et al, 2017 ; Huang et al, 2018 ). Up to date, besides phenazines and pyrrolnitrin, more and more antifungal compounds, including pyoleuteorin (PLT), hydrogen cyanide (HCN), 2,4-diacetylphloroglucinol (DAPG), lipopeptides, furanomycin and so on, have been identified in Pseudomonads’ strains and exhibited a remarkable biocontrol ability ( Fenton et al, 1992 ; Ge et al, 2004 ; Laville et al, 1992 ; Mavrodi et al, 1998 ; Thomashow and Weller, 1988 ; Trippe et al, 2013 ; Voisard et al, 1989 ).…”

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confidence: 99%

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vfr, A Global Regulatory Gene, is Required for Pyrrolnitrin but not for Phenazine-1-carboxylic Acid Biosynthesis in Pseudomonas chlororaphis G05

Chi

Wang

et al. 2019

Plant Pathol J

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vfr, A Global Regulatory Gene, is Required for Pyrrolnitrin but not for Phenazine-1-carboxylic Acid Biosynthesis in Pseudomonas chlororaphis G05

Chi

Wang

et al. 2019

Plant Pathol J

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“…PCN appeared to be the major compound responsible for activity against F. graminearum (Hu et al, 2014). In case of P. chlororaphis G05 (Table 4), PCA contributed to the ability to suppress F. oxysporum on cucumber (Chi et al, 2017), but pyrrolnitrin rather than PCA was essential to control Fusarium head blight on wheat caused by F. graminearum (Huang et al, 2018).…”

Section: Pseudomonas Biocontrol Strains: Pseudomonas Protegens Subgroupmentioning

confidence: 99%

The use of Pseudomonas spp. as bacterial biocontrol agents to control plant diseases

Höfte¹

2021

Burleigh Dodds Series in Agricultural Science

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Most Pseudomonas biocontrol strains are associated with the rhizosphere of plants, where they control soil pathogens by antibiosis or competition, and leaf pathogens via induced systemic resistance. Genome mining and the division of the vastly heterogeneous genus Pseudomonas in phylogenomic (sub)groups has clarified the relation between biocontrol characteristics and phylogeny. Based on their activity, Pseudomonas biocontrol strains come in three types. A first type, represented by P. chlororaphis, P. protegens, P. corrugata and P. aeruginosa (sub)group strains, produces an arsenal of secondary metabolites with broad antimicrobial activity. The second type is found in the P. putida, P. fluorescens, P. koreensis, P. mandelii, and P. gessardii (sub)group. The spectrum of biocontrol properties of these strains is less diverse and involves siderophores and cyclic lipopeptides. The third type colonizes above-ground plant parts. Strains from this type mainly belong to the P. syringae group and are used to control postharvest pathogens. This chapter starts with recent advances in Pseudomonas taxonomy and a summary of its most important biocontrol traits. It then provides an overview of the most important Pseudomonas groups and subgroups harboring biocontrol strains. Examples of well-characterized and representative biocontrol strains show the links between phylogeny, ecology and biocontrol traits. The chapter concludes by reviewing commercially-available biocontrol strains.

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“…Digested with XbaI and BamHI, they were then cloned into the XbaI-BglII site of the shuttle vector pME6010 (Heeb et al, 2000), to generate pME10N. After sequencing, pME10N and the previously-constructed pME10Z (Chi et al, 2017) were respectively transformed into the competent G05DphzDprn::lacZ cells. The positive transformants on LB agar plates supplemented with tetracycline were confirmed by plasmid isolation and restriction enzyme digestion.…”

Section: The Phz and Prn Operon Complementation Assaymentioning

confidence: 99%

Construction of a β-galactosidase-gene-based fusion is convenient for screening candidate genes involved in regulation of pyrrolnitrin biosynthesis in <i>Pseudomonas chlororaphis</i> G05

Luo

Jiang

Feng

et al. 2018

J. Gen. Appl. Microbiol.

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In our recent work, we found that pyrrolnitrin, and not phenazines, pyrrolnitrin contributed to the suppression of the mycelia growth of Fusarium graminearum that causes heavy Fusarium head blight (FHB) disease in cereal crops. However, pyrrolnitrin production of Pseudomonas chlororaphis G05 in King's B medium was very low. Although a few regulatory genes mediating the prnABCD (the prn operon, pyrrolnitrin biosynthetic locus) expression have been identified, it is not enough for us to enhance pyrrolnitrin production by systematically constructing a genetically-engineered strain. To obtain new candidate genes involved in regulation of the prn operon expression, we successfully constructed a fusion mutant G05ΔphzΔprn::lacZ, in which most of the coding regions of the prn operon and the phzABCDEFG (the phz operon, phenazine biosynthetic locus) were deleted, and the promoter region plus the first thirty condons of the prnA was in-frame fused with the truncated lacZ gene on its chromosome. The expression of the fused lacZ reporter gene driven by the promoter of the prn operon made it easy for us to detect the level of the prn expression in terms of the color variation of colonies on LB agar plates supplemented with 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-Gal). With this fusion mutant as a recipient strain, mini-Tn5-based random insertional mutagenesis was then conducted. By picking up colonies with color change, it is possible for us to screen and identify new candidate genes involved in regulation of the prn expression. Identification of additional regulatory genes in further work could reasonably be expected to increase pyrrolnitrin production in G05 and to improve its biological control function.

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Development and characterization of a fusion mutant with the truncated lacZ to screen regulatory genes for phenazine biosynthesis in Pseudomonas chlororaphis G05

Cited by 10 publications

References 39 publications

vfr, A Global Regulatory Gene, is Required for Pyrrolnitrin but not for Phenazine-1-carboxylic Acid Biosynthesis in Pseudomonas chlororaphis G05

vfr, A Global Regulatory Gene, is Required for Pyrrolnitrin but not for Phenazine-1-carboxylic Acid Biosynthesis in Pseudomonas chlororaphis G05

The use of Pseudomonas spp. as bacterial biocontrol agents to control plant diseases

Construction of a β-galactosidase-gene-based fusion is convenient for screening candidate genes involved in regulation of pyrrolnitrin biosynthesis in <i>Pseudomonas chlororaphis</i> G05

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