An Accurate, Affordable, and Precise Resazurin-Based Digital Imaging Colorimetric Assay for the Assessment of Fungicide Sensitivity Status of Fungal Populations
This study aimed at the development and validation of an accurate, more affordable, and precise digital imaging resazurin-based fungicide sensitivity colorimetric assay (COL-assay) for fungal plant pathogens from the genera <i>Mycosphaerella</i> and <i>Pyricularia</i>. This p...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-01-01
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| Series: | Agronomy |
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| Online Access: | https://www.mdpi.com/2073-4395/13/2/343 |
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| author | Tatiane Carla Silva Silvino Intra Moreira Fabio Gomes Assis Samara Nunes Campos Vicentini Abimael Gomes Silva Tamiris Yoshie Kitayama Oliveira Félix Sebastião Christiano Adriano Augusto Paiva Custódio Rui Pereira Leite Maria Cândida Godoy Gasparoto Waldir Cintra de Jesus Paulo Cezar Ceresini |
| author_facet | Tatiane Carla Silva Silvino Intra Moreira Fabio Gomes Assis Samara Nunes Campos Vicentini Abimael Gomes Silva Tamiris Yoshie Kitayama Oliveira Félix Sebastião Christiano Adriano Augusto Paiva Custódio Rui Pereira Leite Maria Cândida Godoy Gasparoto Waldir Cintra de Jesus Paulo Cezar Ceresini |
| author_sort | Tatiane Carla Silva |
| collection | DOAJ |
| description | This study aimed at the development and validation of an accurate, more affordable, and precise digital imaging resazurin-based fungicide sensitivity colorimetric assay (COL-assay) for fungal plant pathogens from the genera <i>Mycosphaerella</i> and <i>Pyricularia</i>. This proposed digital imaging assay was based on colorimetric estimates of resazurin reduction, which was used as a metabolic indicator of fungal respiration activity on microplate cultures. As fungal model systems, we used the yellow and black Sigatoka pathogens [<i>Mycosphaerella musicola</i> (Mm) and <i>M. fijiensis</i> (Mf), respectively] and the wheat blast pathogen, <i>Pyricularia oryzae</i> Triticum lineage (PoTl), which were previously characterized for QoI, DMI, and SDHI fungicide sensitivity. We then compared the classical spectrophotometry detection assay (SPEC-assay) with the proposed COL-assay based on the analyses of digital images of the microplates’ cultures captured with mobile phone cameras on a handmade trans-illuminator built for poorly equipped labs. Qualitatively, in terms of accuracy, there was full correspondence between the SPEC-assay and the COL-assay according to the fungal EC<sub>50</sub> or the relative growth classes on QoI, SDHI, and DMI fungicides for both <i>Mycosphaerella</i> and <i>Pyricularia</i> pathogens. We also observed a strong to very strong correlation coefficient between the COL-assay and the SPEC-assay fungicide sensitivity values for the QoI azoxystrobin, the SDHI fluxapyroxad, and the DMI tebuconazole. Our conclusion was that the COL-assay had a similar accuracy as the SPEC-assay (i.e., resulted in similar fungicide-sensitivity categories for both resistant or sensitive fungal isolates) and high precision. By openly sharing here the COL-assay’s full methodology, and the blueprints of the handmade trans-illuminator, we foresee its adoption by poorly equipped labs throughout the country as an affordable venue for monitoring the fungicide resistance status of populations of important fungal plant pathogens such as <i>M. fijiensis</i>, <i>M. musicola</i>, and <i>P. oryzae</i> Triticum and Oryza lineages. |
| first_indexed | 2024-03-11T09:16:39Z |
| format | Article |
| id | doaj.art-1751b44745ce4eb2b1eb6b9cff0154fc |
| institution | Directory Open Access Journal |
| issn | 2073-4395 |
| language | English |
| last_indexed | 2024-03-11T09:16:39Z |
| publishDate | 2023-01-01 |
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| series | Agronomy |
| spelling | doaj.art-1751b44745ce4eb2b1eb6b9cff0154fc2023-11-16T18:33:12ZengMDPI AGAgronomy2073-43952023-01-0113234310.3390/agronomy13020343An Accurate, Affordable, and Precise Resazurin-Based Digital Imaging Colorimetric Assay for the Assessment of Fungicide Sensitivity Status of Fungal PopulationsTatiane Carla Silva0Silvino Intra Moreira1Fabio Gomes Assis2Samara Nunes Campos Vicentini3Abimael Gomes Silva4Tamiris Yoshie Kitayama Oliveira5Félix Sebastião Christiano6Adriano Augusto Paiva Custódio7Rui Pereira Leite8Maria Cândida Godoy Gasparoto9Waldir Cintra de Jesus10Paulo Cezar Ceresini11Department of Crop Protection, Agricultural Engineering and Soil, São Paulo State University—UNESP, Ilha Solteira 15385-000, SP, BrazilDepartment of Crop Protection, Agricultural Engineering and Soil, São Paulo State University—UNESP, Ilha Solteira 15385-000, SP, BrazilDepartment of Crop Protection, Agricultural Engineering and Soil, São Paulo State University—UNESP, Ilha Solteira 15385-000, SP, BrazilDepartment of Crop Protection, Agricultural Engineering and Soil, São Paulo State University—UNESP, Ilha Solteira 15385-000, SP, BrazilDepartment of Crop Protection, Agricultural Engineering and Soil, São Paulo State University—UNESP, Ilha Solteira 15385-000, SP, BrazilDepartment of Crop Protection, Agricultural Engineering and Soil, São Paulo State University—UNESP, Ilha Solteira 15385-000, SP, BrazilDepartment of Crop Protection, Agricultural Engineering and Soil, São Paulo State University—UNESP, Ilha Solteira 15385-000, SP, BrazilParaná Rural Development Institute, IDR—Paraná/IAPAR, Londrina 86047-902, PR, BrazilParaná Rural Development Institute, IDR—Paraná/IAPAR, Londrina 86047-902, PR, BrazilCollege of Agricultural Sciences from Ribeira Valley, São Paulo State University—UNESP, Registro 11900-000, SP, BrazilNatural Science Center (CCN), Campus Lagoa do Sino, Federal University of São Carlos (UFSCar), Buri 18290-000, SP, BrazilDepartment of Crop Protection, Agricultural Engineering and Soil, São Paulo State University—UNESP, Ilha Solteira 15385-000, SP, BrazilThis study aimed at the development and validation of an accurate, more affordable, and precise digital imaging resazurin-based fungicide sensitivity colorimetric assay (COL-assay) for fungal plant pathogens from the genera <i>Mycosphaerella</i> and <i>Pyricularia</i>. This proposed digital imaging assay was based on colorimetric estimates of resazurin reduction, which was used as a metabolic indicator of fungal respiration activity on microplate cultures. As fungal model systems, we used the yellow and black Sigatoka pathogens [<i>Mycosphaerella musicola</i> (Mm) and <i>M. fijiensis</i> (Mf), respectively] and the wheat blast pathogen, <i>Pyricularia oryzae</i> Triticum lineage (PoTl), which were previously characterized for QoI, DMI, and SDHI fungicide sensitivity. We then compared the classical spectrophotometry detection assay (SPEC-assay) with the proposed COL-assay based on the analyses of digital images of the microplates’ cultures captured with mobile phone cameras on a handmade trans-illuminator built for poorly equipped labs. Qualitatively, in terms of accuracy, there was full correspondence between the SPEC-assay and the COL-assay according to the fungal EC<sub>50</sub> or the relative growth classes on QoI, SDHI, and DMI fungicides for both <i>Mycosphaerella</i> and <i>Pyricularia</i> pathogens. We also observed a strong to very strong correlation coefficient between the COL-assay and the SPEC-assay fungicide sensitivity values for the QoI azoxystrobin, the SDHI fluxapyroxad, and the DMI tebuconazole. Our conclusion was that the COL-assay had a similar accuracy as the SPEC-assay (i.e., resulted in similar fungicide-sensitivity categories for both resistant or sensitive fungal isolates) and high precision. By openly sharing here the COL-assay’s full methodology, and the blueprints of the handmade trans-illuminator, we foresee its adoption by poorly equipped labs throughout the country as an affordable venue for monitoring the fungicide resistance status of populations of important fungal plant pathogens such as <i>M. fijiensis</i>, <i>M. musicola</i>, and <i>P. oryzae</i> Triticum and Oryza lineages.https://www.mdpi.com/2073-4395/13/2/343fungal foliar pathogensfungicide resistancehigh-risk fungicidesQoIDMISDHI |
| spellingShingle | Tatiane Carla Silva Silvino Intra Moreira Fabio Gomes Assis Samara Nunes Campos Vicentini Abimael Gomes Silva Tamiris Yoshie Kitayama Oliveira Félix Sebastião Christiano Adriano Augusto Paiva Custódio Rui Pereira Leite Maria Cândida Godoy Gasparoto Waldir Cintra de Jesus Paulo Cezar Ceresini An Accurate, Affordable, and Precise Resazurin-Based Digital Imaging Colorimetric Assay for the Assessment of Fungicide Sensitivity Status of Fungal Populations Agronomy fungal foliar pathogens fungicide resistance high-risk fungicides QoI DMI SDHI |
| title | An Accurate, Affordable, and Precise Resazurin-Based Digital Imaging Colorimetric Assay for the Assessment of Fungicide Sensitivity Status of Fungal Populations |
| title_full | An Accurate, Affordable, and Precise Resazurin-Based Digital Imaging Colorimetric Assay for the Assessment of Fungicide Sensitivity Status of Fungal Populations |
| title_fullStr | An Accurate, Affordable, and Precise Resazurin-Based Digital Imaging Colorimetric Assay for the Assessment of Fungicide Sensitivity Status of Fungal Populations |
| title_full_unstemmed | An Accurate, Affordable, and Precise Resazurin-Based Digital Imaging Colorimetric Assay for the Assessment of Fungicide Sensitivity Status of Fungal Populations |
| title_short | An Accurate, Affordable, and Precise Resazurin-Based Digital Imaging Colorimetric Assay for the Assessment of Fungicide Sensitivity Status of Fungal Populations |
| title_sort | accurate affordable and precise resazurin based digital imaging colorimetric assay for the assessment of fungicide sensitivity status of fungal populations |
| topic | fungal foliar pathogens fungicide resistance high-risk fungicides QoI DMI SDHI |
| url | https://www.mdpi.com/2073-4395/13/2/343 |
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