Altered nitrogen metabolism in biocontrol strains of Penicillium rubens

Espeso, Eduardo A.; Villarino, M.; Carreras, María Alejandra; Alonso-Guirado, Lola; Alonso, Jorge; Melgarejo, P.; Larena, Inmaculada

doi:10.1016/j.fgb.2019.103263

Cited by 8 publications

(7 citation statements)

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“…Many studies show that the nature of the carbon and nitrogen sources, carbon concentrations, and the C/N ratios of the culture media strongly affect growth and sporulation of the fungi [ 55 , 56 ]. For example, PO212 displays a nitrate-deficiency (nit-) phenotype with poor colonial growth when nitrate is used as the main source of nitrogen [ 57 ]. Here the crude extract of PO212 (CE) after growing under nitrogen stress conditions (Morton’s medium) maintained the same disease control activity as the dried PO212 conidia.…”

Section: Discussionmentioning

confidence: 99%

Exploring the Extracellular Macromolecular Composition of Crude Extracts of Penicillium rubens Strain 212 for Elucidation Its Mode of Action as a Biocontrol Agent

Carreras¹,

Espeso

Gutiérrez-Docio

et al. 2020

JoF

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Penicillium rubens strain 212 (PO212) acts as an inducer of systemic resistance in tomato plants. The effect of crude extracellular extracts of PO212 on the soil-borne pathogen Fusarium oxysporum f. sp. lycopersici has been evaluated. Evidence of the involvement of soluble, thermo-labile, and proteinase-inactivated macromolecules present in PO212 crude extracts in the control of Fusarium vascular disease in tomato plants was found. Proteomic techniques and the availability of the access to the PO212 genome database have allowed the identification of glycosyl hydrolases, oxidases, and peptidases in these extracellular extracts. Furthermore, a bioassay-guided fractionation of PO212 crude extracellular extracts using an integrated membrane/solid phase extraction process was set up. This method enabled the separation of a PO212 crude extracellular extract of seven days of growth into four fractions of different molecular sizes and polarities: high molecular mass protein fraction >5 kDa, middle molecular mass protein fraction 5–1 kDa, low molecular mass metabolite fraction, and nutrients from culture medium (mainly glucose and minerals). The high and middle molecular mass protein fractions retained disease control activity in a way similar to that of the control extracts. Proteomic techniques have allowed the identification of nine putatively secreted proteins in the high molecular mass protein fraction matching those identified in the total crude extracts. Therefore, these enzymes are considered to be potentially responsible of the crude extracellular extract-induced resistance in tomato plants against F. oxysporum f. sp. lycopersici. Further studies are required to establish which of the identified proteins participate in the PO212’s action mode as a biocontrol agent.

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Section: Discussionmentioning

confidence: 99%

Exploring the Extracellular Macromolecular Composition of Crude Extracts of Penicillium rubens Strain 212 for Elucidation Its Mode of Action as a Biocontrol Agent

Carreras¹,

Espeso

Gutiérrez-Docio

et al. 2020

JoF

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“…This nitrate assimilation-deficient phenotype was due to the presence of mutations in the NirA regulator or the nitrate transporter CrnA. However, the complementation of these mutations did not help to understand the BA of PO212 (Espeso et al, 2019).…”

Section: Introductionmentioning

confidence: 95%

“…Dried conidia of PO212 were produced in a solid-state fermentation system and dried as previously described by Larena et al (2003a). Penicillium strains were grown on potato dextrose agar (PDA) or minimal medium (MM; Espeso et al, 2019) with 5 mM ammonium tartrate and D-glucose 1% (w/v) as nitrogen and carbon source, respectively, and incubated at 25°C for 5 days. For short-term storage, strains were kept at 4°C on solid media.…”

Section: Strains and Growth Conditionsmentioning

confidence: 99%

“…Oligonucleotides were designed using Vector NTI™ Suite 8 and are listed in Table 2. The oligonucleotides for the amplification of MAT1-1 and MAT1-2 were previously described in Espeso et al (2019). PCR products were analyzed in 0.8% agarose-TAE electrophoresis and when required, they were purified using the PCR clean-Up kit (Macherey-Nagel), following the manufacturer's instructions.…”

Section: Efficacy Assaysmentioning

confidence: 99%

“…Subsequent sequencing of internal transcribed spacers (ITS) regions and the isolation and identification of 5-fluoroorotic acid (5-FOA)-resistant mutants showed that PO212 was a P. rubens strain (Villarino et al, 2016). The first study to understand the genetic basis of PO212 biocontrol activity (BA) focused on the analysis of genes involved in nitrate assimilation since PO212, and other P. rubens strains showing a BA against Fusarium wilt lacked the ability to use nitrate as the main nitrogen source (Espeso et al, 2019). This nitrate assimilation-deficient phenotype was due to the presence of mutations in the NirA regulator or the nitrate transporter CrnA.…”

Section: Introductionmentioning

confidence: 99%

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Comparative analysis of Penicillium genomes reveals the absence of a specific genetic basis for biocontrol in Penicillium rubens strain 212

et al. 2023

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Penicillium rubens strain 212 (PO212) is a filamentous fungus belonging to the division Ascomycete. PO212 acts as an effective biocontrol agent against several pathogens in a variety of horticultural crops including Fusarium oxysporum f.sp. lycopersici, causing vascular wilt disease in tomato plants. We assembled draft genomes of two P. rubens strains, the biocontrol agent PO212 and the soil isolate S27, which lacks biocontrol activity. We also performed comparative analyses of the genomic sequence of PO212 with that of the other P. rubens and P. chrysogenum strains. This is the first Penicillium strain with biocontrol activity whose genome has been sequenced and compared. PO212 genome size is 2,982 Mb, which is currently organized into 65 scaffolds and a total of 10,164 predicted Open Reading Frames (ORFs). Sequencing confirmed that PO212 belongs to P. rubens clade. The comparative analysis of the PO212 genome with the genomes of other P. rubens and Penicillium chrysogenum strains available in databases showed strong conservation among genomes, but a correlation was not found between these genomic data and the biocontrol phenotype displayed by PO212. Finally, the comparative analysis between PO212 and S27 genomes showed high sequence conservation and a low number of variations mainly located in ORF regions. These differences found in coding regions between PO212 and S27 genomes can explain neither the biocontrol activity of PO212 nor the absence of such activity in S27, opening a possible avenue toward transcriptomic and epigenetic studies that may shed light on this mechanism for fighting plant diseases caused by fungal pathogens. The genome sequences described in this study provide a useful novel resource for future research into the biology, ecology, and evolution of biological control agents.

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A role for Penicillium rubens strain 212 xylanolytic system in biocontrol of Fusarium wilt disease in tomato plants

et al. 2023

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Penicillium rubens strain 212 (PO212) is an effective inducer of resistance mechanisms in tomato plants against Fusarium oxysporum f.sp. lycopersici. During the interaction of PO212 with the plant, different classes of molecules may act as elicitors such as certain secreted endo-xylanases. The aim of this work was to elucidate the possible role of the xylanolytic system of PO212 in its biocontrol activity. We identified potential genes coding for xylanases (xlnA, xlnE, xylP), β-xylosidase (xlnD) and their transcriptional regulators (xlnR and araR) in PO212, and evaluated their transcriptional patterns in response to tomato root extracts or synthetic medium containing xylan as main carbon source. For this work we compared data from biocontrol strain PO212 with those of two strains of Penicillium, P. rubens S27, and P. chrysogenum IPLA33001, lacking the biocontrol efficacy. Time-course experiments showed the effect of these carbon sources on the expression rates of xylanase genes. To achieve a strong reduction in expression of xylanolytic genes, we generated a null allele of XlnR, as the specific regulator of the xylanase pathway. Absence of XlnR function prevented growth of PO212 on media containing xylan as main carbon source and consequently, expression levels of xylanases were downregulated. The relationship of the xlnR gene regulated pathway to the efficacy of PO212 as a biological control agent was evaluated. Null xlnR strains did not reduce either disease severity or incidence as wild-type PO212 does. Thus, there is a relationship between a correct expression of xylanolytic system and the capability of these enzymes as effective elicitors to trigger of plant defense systems in tomato plants against F. oxysporum f.sp. lycopersici.

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Altered nitrogen metabolism in biocontrol strains of Penicillium rubens

Cited by 8 publications

References 43 publications

Exploring the Extracellular Macromolecular Composition of Crude Extracts of Penicillium rubens Strain 212 for Elucidation Its Mode of Action as a Biocontrol Agent

Exploring the Extracellular Macromolecular Composition of Crude Extracts of Penicillium rubens Strain 212 for Elucidation Its Mode of Action as a Biocontrol Agent

Comparative analysis of Penicillium genomes reveals the absence of a specific genetic basis for biocontrol in Penicillium rubens strain 212

A role for Penicillium rubens strain 212 xylanolytic system in biocontrol of Fusarium wilt disease in tomato plants

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