The Effect of Phenazine-1-Carboxylic Acid on Mycelial Growth of Botrytis cinerea Produced by Pseudomonas aeruginosa LV Strain

Simionato, Ane Stéfano; Navarro, Miguel Octavio Pérez; Jesus, Maria Luiza Abreu de; Barazetti, André Riedi; Silva, Caroline S. da; Simões, Glenda Cavalari; Balbi-Peña, María Isabel; Mello, João Carlos Palazzo de; Panagio, Luciano Aparecido; Almeida, Ricardo Sérgio; Andrade, Galdino; Oliveira, Admilton Gonçalves de

doi:10.3389/fmicb.2017.01102

Cited by 74 publications

(43 citation statements)

References 42 publications

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“…Commercial standard lipopeptides (surfactin and iturin, Sigma-Aldrich, United States) and CLE methanol solution (10 mg/L) were run on an HPLC system (Waters, E2695, United States) with a C 18 reverse-phase column (5 µm, 4.6 × 150 mm) und er the same conditions(46). Water and acetonitrile were selected as the mobile phase at a ratio of 20:80 by volume.…”

Section: Methodsmentioning

confidence: 99%

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Inhibitory effect ofBacillus subtilisWL-2 and its IturinA lipopeptides againstPhytophthora infestans

Wang

Zhang

et al. 2019

Preprint

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15Potato late blight triggered by Phytophthora infestans ((Mont.) de Bary) represents a 16 great food security threat worldwide and is difficult to control. Currently, Bacillus spp. 17 have been considered biocontrol agents to control many fungal diseases. Here, 18 Bacillus subtilis WL-2 was selected as the antifungal strain with the most potential 19 against P. infestans mycelium growth. Additionally, the functional metabolites 20 extracted from WL-2 were identified as IturinA-family cyclic lipopeptides (CLPs) via 21 2 high-performance liquid chromatography (HPLC) and electrospray ionization mass 22 spectrometry (ESI-MS). Analyses using scanning and transmission electron 23 microscopy (SEM and TEM) revealed that IturinA caused a change in the mycelial 24 surface and damage to the internal cell structure, including cell membrane disruption 25 and irregular organelle formation. Moreover, propidium iodide staining and nucleic 26 acid and protein release were detected to clarify the cell membrane damage caused by 27 IturinA. Additionally, IturinA triggered reactive oxygen species (ROS) generation and 28 malondialdehyde (MDA) production. Mitochondrial membrane potential (MMP), 29 mitochondrial respiratory chain complexes activity (MRCCA), respiratory control rate 30 (RCR), and oxidative phosphorylation efficiency (P/O) assays indicated that P. 31 infestans mitochondria affected by IturinA were so seriously damaged that the MMP 32 and MRCCA declined remarkably and that mitochondrial ATP production ability was 33 weakened. Therefore, IturinA induces cell membrane damage, oxidative stress, and 34 dysfunction of mitochondria, resulting in P. infestans hyphal cell death. As such, the 35 results highlight that B. subtilis WL-2 and IturinA have great potential as candidates for 36 inhibiting P. infestans mycelium growth and controlling potato late blight. 37 IMPORTANCE 38Potato (Solanum tuberosum L.) is the fourth most common global food crop, and 39 its planting area and yield increase yearly. Notably, in 2015, China initiated a potato 40 staple food conversion strategy, and by 2020, approximately 50% of potatoes will 41 be consumed as a staple food. The plant pathogen fungus Phytophthora infestans 42 ((Mont.) de Bary) is the culprit of potato late blight; however, biological agents 43 3 rather than chemicals are highly necessary to control this threatening disease. In 44 this study, we discovered an antifungal substance, IturinA, a lipopeptide produced 45 by Bacillus subtilis WL-2. Moreover, our research revealed the actual mechanism 46 of IturinA against P. infestans mycelium growth and clarified the potential of B. 65Behind rice, wheat, and corn, potato (Solanum tuberosum L.) is the fourth most 66 stable food crop in the world. However, potato production is often endangered by 67 many pathogens. Worryingly, late blight triggered by Phytophthora infestans ((Mont.) 68 de Bary) could directly reduce or even eliminate potato production, and an outbreak 69 of this disease could even result in a grievous ...

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Section: Methodsmentioning

confidence: 99%

“…The injection volume was 1 mL per min, and the eluate was monitored at 214 nm. According to the peak times of standard lipopeptides, the peaks of potential lipopeptides contained in CLE were collected and dried at room temperature(45, 46).…”

Section: Methodsmentioning

confidence: 99%

Inhibitory effect ofBacillus subtilisWL-2 and its IturinA lipopeptides againstPhytophthora infestans

Wang

Zhang

et al. 2019

Preprint

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“…To investigate if the level of reporter activity could be used to estimate levels of the biosynthetic product, β ‐galactosidase‐mediated indication of recombinant phenazine production in P. putida KT2440 was chosen as an example. The chorismic acid‐derived group of phenazine compounds displays antibacterial and antifungal properties (Simionato et al ., ; Zhang et al ., ; Zhu et al ., ) and P. putida KT2440, a safe and widely established representative workhorse of the Pseudomonas clade, is able to withstand production of the compound (Domröse et al ., ).…”

Section: An Example Of Ytrex Application: Generation and Lacz‐based Cmentioning

confidence: 99%

Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida

Weihmann

Domröse

Drepper

et al. 2019

Microbial Biotechnology

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SummaryBacterial gene clusters, which represent a genetic treasure trove for secondary metabolite pathways, often need to be activated in a heterologous host to access the valuable biosynthetic products. We provide here a detailed protocol for the application of the yTREX ‘gene cluster transplantation tool’: Via yeast recombinational cloning, a gene cluster of interest can be cloned in the yTREX vector, which enables the robust conjugational transfer of the gene cluster to bacteria like Pseudomonas putida, and their subsequent transposon Tn5‐based insertion into the host chromosome. Depending on the gene cluster architecture and chromosomal insertion site, the respective pathway genes can be transcribed effectively from a chromosomal promoter, thereby enabling the biosynthesis of a natural product. We describe workflows for the design of a gene cluster expression cassette, cloning of the cassette in the yTREX vector by yeast recombineering, and subsequent transfer and expression in P. putida. As an example for yTREX‐based transplantation of a natural product biosynthesis, we provide details on the cloning and activation of the phenazine‐1‐carboxylic acid biosynthetic genes from Pseudomonas aeruginosa in P. putida KT2440 as well as the use of β‐galactosidase‐encoding lacZ as a reporter of production levels.

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“…Recent research has shown strain of Pseudomonas aeruginosa secreting an organometallic compound with inhibitory activity against MDR bacteria, including carbapenemase-producing Klebsiella pneumoniae and methicillin-resistant Staphylococcus aureus [9,10]. Additionally, the same strain produced a phenazine-1-carboxylic acid (in low quantity) showing antifungal potential against Botrytis cinerea [11,12].…”

Section: Beneficial Roles Of Pseudomonas Species In Medicinementioning

confidence: 99%

The Beneficial Roles ofPseudomonasin Medicine, Industries, and Environment: A Review

Anayo¹,

Scholastica²,

Peter³

et al. 2019

Pseudomonas Aeruginosa - An Armory Within

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This chapter intends to consider Pseudomonas versatility as regards the beneficial uses of Pseudomonas for production of primary metabolites including enzymes. This chapter will consider the use of Pseudomonas for production of secondary metabolites from various pathways which are equally useful in the industries and in medicine. Pseudomonas pigments and its usefulness in bioelectricity, medicine, etc. will equally be considered in this chapter. The authors are to integrate knowledge in versatility of Pseudomonas for use as agents of microbial production of biosurfactants for environmental cleanup, restoration, and remediation.

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The Effect of Phenazine-1-Carboxylic Acid on Mycelial Growth of Botrytis cinerea Produced by Pseudomonas aeruginosa LV Strain

Cited by 74 publications

References 42 publications

Inhibitory effect ofBacillus subtilisWL-2 and its IturinA lipopeptides againstPhytophthora infestans

Inhibitory effect ofBacillus subtilisWL-2 and its IturinA lipopeptides againstPhytophthora infestans

Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida

The Beneficial Roles ofPseudomonasin Medicine, Industries, and Environment: A Review

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