A multiplex qPCR TaqMan-assay to detect fungal antagonism between Trichoderma atroviride (Hypocreaceae) and Botrytis cinerea (Sclerotiniaceae) in blackberry fruits using a de novo tef1-α- and an IGS-sequence based probes

Hilje-Rodríguez, Irena; Albertazzi, Federico J.; Rivera-Coto, Germán; Molina‐Bravo, Ramón

doi:10.1016/j.btre.2020.e00447

Cited by 4 publications

(5 citation statements)

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“…T. atroviride is widely used as a biocontrol agent against phytopathogens, such as Phytophthora cinnamomi [ 54 ], Botrytis cinerea [ 55 ], and Fusarium spp. [ 56 ].…”

Section: Resultsmentioning

confidence: 99%

Light-Induced Changes in Secondary Metabolite Production of Trichoderma atroviride

Missbach,

Flatschacher,

Bueschl

et al. 2023

JoF

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Many studies aim at maximizing fungal secondary metabolite production but the influence of light during cultivation has often been neglected. Here, we combined an untargeted isotope-assisted liquid chromatography–high-resolution mass spectrometry-based metabolomics approach with standardized cultivation of Trichoderma atroviride under three defined light regimes (darkness (PD), reduced light (RL) exposure, and 12/12 h light/dark cycle (LD)) to systematically determine the effect of light on secondary metabolite production. Comparative analyses revealed a similar metabolite profile upon cultivation in PD and RL, whereas LD treatment had an inhibiting effect on both the number and abundance of metabolites. Additionally, the spatial distribution of the detected metabolites for PD and RL was analyzed. From the more than 500 detected metabolites, only 25 were exclusively produced upon fungal growth in darkness and 85 were significantly more abundant in darkness. The majority were detected under both cultivation conditions and annotation revealed a cluster of substances whose production followed the pattern observed for the well-known T. atroviride metabolite 6-pentyl-alpha-pyrone. We conclude that cultivation of T. atroviride under RL can be used to maximize secondary metabolite production.

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“…T. atroviride is widely used as a biocontrol agent against phytopathogens, such as Phytophthora cinnamomi [ 54 ], Botrytis cinerea [ 55 ], and Fusarium spp. [ 56 ].…”

Section: Resultsmentioning

confidence: 99%

Light-Induced Changes in Secondary Metabolite Production of Trichoderma atroviride

Missbach,

Flatschacher,

Bueschl

et al. 2023

JoF

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“…Botrytis cinerea (BcLLCR) mycelia and conidia from infected blackberry fruits brought from the field [2] were isolated with a dissecting needle, placed over a microscope slide and observed under a light microscope for identification. Fungal tissues were cultured in Petri dishes with 20 mL PDA + 25% lactic acid.…”

Section: *Methods Detailsmentioning

confidence: 99%

“…A Trichoderma atroviride isolate, BV1CR [2] , was reactivated and cultured by following the methodology described above for B. cinerea . The fungal isolate was preserved at 4°C [3] in vials with PDA + 25% lactic acid and mineral oil ( Fig.…”

Section: *Methods Detailsmentioning

confidence: 99%

“…Inoculated fruits were monitored for 10 d after inoculations ( Fig. 3 ; [2] ). Fruits showed the presence of other fungi growing from inside even in mock-water treatments, but to a lesser extent compared to inoculated and co-inoculated fungi ( Fig.…”

Section: *Methods Detailsmentioning

confidence: 99%

“…This fungal growth did not interfere with qPCR analysis of BcLLCR and BV1CR. The interaction between these two fungi on blackberries was assessed by qPCR analysis using specific primer-probe sets that detected and quantified each organism and their growth separately in composite samples [2] .

Fig.…”

Section: *Methods Detailsmentioning

confidence: 99%

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Inoculation and co-inoculation of two monosporic fungi onto surface-sterile blackberry fruits for quantification experiments

Hilje-Rodríguez

Molina‐Bravo

2020

MethodsX

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Trichoderma is used as a biocontrol agent against different plant pathogens in different crops. In Costa Rica, Trichoderma isolates from blackberry fruits ( Rubus adenotrichos Schltdl.) have shown antagonism in laboratory and field trials against Botrytis cinerea . Quantifying fungal antagonistic activity directly on target organs or target tissues is of interest to estimate the performance of biocontrol agents. However, this is difficult due to the lack of visual manifestations of fungal structures. As part of a larger study to quantify antagonistic activity by quantitative PCR, we detail here a method to isolate and purify each fungus and then inoculate or co-inoculate them onto surface-sterilized blackberry fruits. • A procedure to co-inoculate the surfaces of blackberry fruits with monosporic fungal suspensions for molecular analyses is described. • The protocol described herein was implemented for subsequent qPCR analysis.

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A duplex droplet digital PCR assay for quantification of Alternaria spp. and Botrytis cinerea on sweet cherry at different growth stages

Larrabee

Voegel

Nelson

2021

Canadian Journal of Plant Pathology

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A multiplex qPCR TaqMan-assay to detect fungal antagonism between Trichoderma atroviride (Hypocreaceae) and Botrytis cinerea (Sclerotiniaceae) in blackberry fruits using a de novo tef1-α- and an IGS-sequence based probes

Cited by 4 publications

References 60 publications

Light-Induced Changes in Secondary Metabolite Production of Trichoderma atroviride

Light-Induced Changes in Secondary Metabolite Production of Trichoderma atroviride

Inoculation and co-inoculation of two monosporic fungi onto surface-sterile blackberry fruits for quantification experiments

A duplex droplet digital PCR assay for quantification of Alternaria spp. and Botrytis cinerea on sweet cherry at different growth stages

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