Detection of changes in mould cell wall stress-related gene expression by a novel reverse transcription real-time PCR method
The cell wall integrity (CWI) pathway is activated in response to cell wall stresses due to different food-related environments. Rho1 is one of the main regulators within such pathway. The objective of this work was to design an easy-to-use RT-qPCR technique for the evaluation of the Rho1 gene expre...
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2018
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01681605_v275_n_p17_daCruzCabral http://hdl.handle.net/20.500.12110/paper_01681605_v275_n_p17_daCruzCabral |
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paper:paper_01681605_v275_n_p17_daCruzCabral2023-06-08T15:17:23Z Detection of changes in mould cell wall stress-related gene expression by a novel reverse transcription real-time PCR method Cell wall integrity Filamentous fungi Rho1 gene Stress protein protein PgAFP unclassified drug antifungal agent fungal protein primer DNA Rho guanine nucleotide binding protein Article Aspergillus flavus computer model controlled study fungal cell wall fungal gene fungus growth gene expression gene overexpression genetic conservation mould nonhuman nucleotide sequence Penicillium Penicillium chrysogenum Penicillium polonucum quantitative analysis real time polymerase chain reaction reverse transcription polymerase chain reaction Rho1 gene sensitivity and specificity species sporogenesis wall stress analysis cell wall food contamination food control gene expression gene expression regulation genetics metabolism procedures real time polymerase chain reaction reverse transcription Antifungal Agents Aspergillus flavus Base Sequence Cell Wall DNA Primers Food Contamination Food Microbiology Fungal Proteins Gene Expression Gene Expression Regulation, Fungal Penicillium Real-Time Polymerase Chain Reaction Reverse Transcription rho GTP-Binding Proteins The cell wall integrity (CWI) pathway is activated in response to cell wall stresses due to different food-related environments. Rho1 is one of the main regulators within such pathway. The objective of this work was to design an easy-to-use RT-qPCR technique for the evaluation of the Rho1 gene expression useful to measure responses to the presence of cell wall stressors such as the antifungal protein PgAFP. Two primer pairs were designed from published conserved regions. Their specificity initially was determined by in silico analysis for several fungal species. After optimising the qPCR, the primer pair Rho1-F1/R2 was selected due to the lowest Cq values obtained and its specificity. The qPCR method showed efficiencies between 97.5% and 100.5%. Applicability of the designed qPCR method was evaluated in the presence of the stressor PgAFP. The PgAFP-resistant Penicillium polonicum and the PgAFP-sensitive Aspergillus flavus showed Rho1 gene over- and under- expression, respectively, indicating that the CWI pathway is activated in the former species but not activated in the latter one in response to the stress caused by PgAFP. This novel qPCR methodology able to detect changes in CWI-related gene expression in filamentous fungi will be useful in future studies to evaluate physiological mould responses to different food environmental challenges. © 2018 Elsevier B.V. 2018 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01681605_v275_n_p17_daCruzCabral http://hdl.handle.net/20.500.12110/paper_01681605_v275_n_p17_daCruzCabral |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Cell wall integrity Filamentous fungi Rho1 gene Stress protein protein PgAFP unclassified drug antifungal agent fungal protein primer DNA Rho guanine nucleotide binding protein Article Aspergillus flavus computer model controlled study fungal cell wall fungal gene fungus growth gene expression gene overexpression genetic conservation mould nonhuman nucleotide sequence Penicillium Penicillium chrysogenum Penicillium polonucum quantitative analysis real time polymerase chain reaction reverse transcription polymerase chain reaction Rho1 gene sensitivity and specificity species sporogenesis wall stress analysis cell wall food contamination food control gene expression gene expression regulation genetics metabolism procedures real time polymerase chain reaction reverse transcription Antifungal Agents Aspergillus flavus Base Sequence Cell Wall DNA Primers Food Contamination Food Microbiology Fungal Proteins Gene Expression Gene Expression Regulation, Fungal Penicillium Real-Time Polymerase Chain Reaction Reverse Transcription rho GTP-Binding Proteins |
| spellingShingle |
Cell wall integrity Filamentous fungi Rho1 gene Stress protein protein PgAFP unclassified drug antifungal agent fungal protein primer DNA Rho guanine nucleotide binding protein Article Aspergillus flavus computer model controlled study fungal cell wall fungal gene fungus growth gene expression gene overexpression genetic conservation mould nonhuman nucleotide sequence Penicillium Penicillium chrysogenum Penicillium polonucum quantitative analysis real time polymerase chain reaction reverse transcription polymerase chain reaction Rho1 gene sensitivity and specificity species sporogenesis wall stress analysis cell wall food contamination food control gene expression gene expression regulation genetics metabolism procedures real time polymerase chain reaction reverse transcription Antifungal Agents Aspergillus flavus Base Sequence Cell Wall DNA Primers Food Contamination Food Microbiology Fungal Proteins Gene Expression Gene Expression Regulation, Fungal Penicillium Real-Time Polymerase Chain Reaction Reverse Transcription rho GTP-Binding Proteins Detection of changes in mould cell wall stress-related gene expression by a novel reverse transcription real-time PCR method |
| topic_facet |
Cell wall integrity Filamentous fungi Rho1 gene Stress protein protein PgAFP unclassified drug antifungal agent fungal protein primer DNA Rho guanine nucleotide binding protein Article Aspergillus flavus computer model controlled study fungal cell wall fungal gene fungus growth gene expression gene overexpression genetic conservation mould nonhuman nucleotide sequence Penicillium Penicillium chrysogenum Penicillium polonucum quantitative analysis real time polymerase chain reaction reverse transcription polymerase chain reaction Rho1 gene sensitivity and specificity species sporogenesis wall stress analysis cell wall food contamination food control gene expression gene expression regulation genetics metabolism procedures real time polymerase chain reaction reverse transcription Antifungal Agents Aspergillus flavus Base Sequence Cell Wall DNA Primers Food Contamination Food Microbiology Fungal Proteins Gene Expression Gene Expression Regulation, Fungal Penicillium Real-Time Polymerase Chain Reaction Reverse Transcription rho GTP-Binding Proteins |
| description |
The cell wall integrity (CWI) pathway is activated in response to cell wall stresses due to different food-related environments. Rho1 is one of the main regulators within such pathway. The objective of this work was to design an easy-to-use RT-qPCR technique for the evaluation of the Rho1 gene expression useful to measure responses to the presence of cell wall stressors such as the antifungal protein PgAFP. Two primer pairs were designed from published conserved regions. Their specificity initially was determined by in silico analysis for several fungal species. After optimising the qPCR, the primer pair Rho1-F1/R2 was selected due to the lowest Cq values obtained and its specificity. The qPCR method showed efficiencies between 97.5% and 100.5%. Applicability of the designed qPCR method was evaluated in the presence of the stressor PgAFP. The PgAFP-resistant Penicillium polonicum and the PgAFP-sensitive Aspergillus flavus showed Rho1 gene over- and under- expression, respectively, indicating that the CWI pathway is activated in the former species but not activated in the latter one in response to the stress caused by PgAFP. This novel qPCR methodology able to detect changes in CWI-related gene expression in filamentous fungi will be useful in future studies to evaluate physiological mould responses to different food environmental challenges. © 2018 Elsevier B.V. |
| title |
Detection of changes in mould cell wall stress-related gene expression by a novel reverse transcription real-time PCR method |
| title_short |
Detection of changes in mould cell wall stress-related gene expression by a novel reverse transcription real-time PCR method |
| title_full |
Detection of changes in mould cell wall stress-related gene expression by a novel reverse transcription real-time PCR method |
| title_fullStr |
Detection of changes in mould cell wall stress-related gene expression by a novel reverse transcription real-time PCR method |
| title_full_unstemmed |
Detection of changes in mould cell wall stress-related gene expression by a novel reverse transcription real-time PCR method |
| title_sort |
detection of changes in mould cell wall stress-related gene expression by a novel reverse transcription real-time pcr method |
| publishDate |
2018 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01681605_v275_n_p17_daCruzCabral http://hdl.handle.net/20.500.12110/paper_01681605_v275_n_p17_daCruzCabral |
| _version_ |
1768544906967515136 |