A real-time fluorescent reverse transcription quantitative PCR assay for rapid detection of genetic markers’ expression associated with Fusarium Wilt of banana biocontrol activities in bacillus
Fusarium wilt of banana, caused by Fusarium oxysporum f. sp. cubense (Foc), especially Tropical Race 4 (TR4), seriously threatens banana production worldwide. There is no single effective control measure, although certain Bacillus strains secrete antibiotics as promising disease-biocontrol agents. This study identified five Bacillus strains displaying strong antibiotic activity against TR4, using a systemic assessment for presence/absence of genetic markers at genome level, and expres sion profiles at transcriptome level. A conventional PCR with 13 specific primer pairs detected biocontrol-related genes. An accurate, quantitative real-time PCR protocol with novel designed specific primers was developed to characterise strain-specific gene expression, that optimises strain culturing and RNA-isolation methodologies. Six genes responsible for synthesising non-ribosomal peptide synthetase biocontrol metabolites were detected in all five strains. Three genes were involved in synthesising three Polyketide synthetase metabolites in all five strains, but the macrolactin synthase gene mln was only detected in WBN06 and YN1282-2. All five Bacillus strains have the genes dhb and bioA, essential for synthesising bacillibactin and biotin. However, the gene sboA, involved in subtilisin synthesis, is absent in all five strains. These genes’ expression patterns were significantly different among these strains, suggesting different mechanisms involved in TR4 biocontrol. Results will help elucidate functional genes’ biocontrol mechanisms.
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| Formato: | Journal Article biblioteca |
| Idioma: | English |
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MDPI
2021-04-30
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| Materias: | bacillus, biological control, genetic markers, transcription factors, fusarium wilt, plant disease control, control biológico, marcadores genéticos, factores de transcripción, |
| Acceso en línea: | https://hdl.handle.net/10568/113757 https://doi.org/10.3390/jof7050353 |
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dig-cgspace-10568-1137572023-12-08T19:36:04Z A real-time fluorescent reverse transcription quantitative PCR assay for rapid detection of genetic markers’ expression associated with Fusarium Wilt of banana biocontrol activities in bacillus Li, Shu He, Ping Fan, Huacai Liu, Lina Yin, Kesuo Yang, Baoming Li, Yongping Huang, Su-Mei Li, Xundong Zheng, Si-Jun bacillus biological control genetic markers transcription factors fusarium wilt plant disease control control biológico marcadores genéticos factores de transcripción Fusarium wilt of banana, caused by Fusarium oxysporum f. sp. cubense (Foc), especially Tropical Race 4 (TR4), seriously threatens banana production worldwide. There is no single effective control measure, although certain Bacillus strains secrete antibiotics as promising disease-biocontrol agents. This study identified five Bacillus strains displaying strong antibiotic activity against TR4, using a systemic assessment for presence/absence of genetic markers at genome level, and expres sion profiles at transcriptome level. A conventional PCR with 13 specific primer pairs detected biocontrol-related genes. An accurate, quantitative real-time PCR protocol with novel designed specific primers was developed to characterise strain-specific gene expression, that optimises strain culturing and RNA-isolation methodologies. Six genes responsible for synthesising non-ribosomal peptide synthetase biocontrol metabolites were detected in all five strains. Three genes were involved in synthesising three Polyketide synthetase metabolites in all five strains, but the macrolactin synthase gene mln was only detected in WBN06 and YN1282-2. All five Bacillus strains have the genes dhb and bioA, essential for synthesising bacillibactin and biotin. However, the gene sboA, involved in subtilisin synthesis, is absent in all five strains. These genes’ expression patterns were significantly different among these strains, suggesting different mechanisms involved in TR4 biocontrol. Results will help elucidate functional genes’ biocontrol mechanisms. 2021-04-30 2021-05-21T10:04:41Z 2021-05-21T10:04:41Z Journal Article Li, S.; He, P.; Fan, H.; Liu, L.; Yin, K.; Yang, B.; Li, Y.; Huang, S.-M.; Li, X.; Zheng, S.-J. (2021) A real-time fluorescent reverse transcription quantitative PCR assay for rapid detection of genetic markers’ expression associated with Fusarium Wilt of banana biocontrol activities in bacillus. Journal of Fungi 7(5): 353. ISSN: 2309-608X 2309-608X https://hdl.handle.net/10568/113757 https://doi.org/10.3390/jof7050353 en CC-BY-4.0 Open Access application/pdf MDPI Journal of Fungi |
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| countrycode |
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| component |
Bibliográfico |
| access |
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| databasecode |
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| tag |
biblioteca |
| region |
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| libraryname |
Biblioteca del CGIAR |
| language |
English |
| topic |
bacillus biological control genetic markers transcription factors fusarium wilt plant disease control control biológico marcadores genéticos factores de transcripción bacillus biological control genetic markers transcription factors fusarium wilt plant disease control control biológico marcadores genéticos factores de transcripción |
| spellingShingle |
bacillus biological control genetic markers transcription factors fusarium wilt plant disease control control biológico marcadores genéticos factores de transcripción bacillus biological control genetic markers transcription factors fusarium wilt plant disease control control biológico marcadores genéticos factores de transcripción Li, Shu He, Ping Fan, Huacai Liu, Lina Yin, Kesuo Yang, Baoming Li, Yongping Huang, Su-Mei Li, Xundong Zheng, Si-Jun A real-time fluorescent reverse transcription quantitative PCR assay for rapid detection of genetic markers’ expression associated with Fusarium Wilt of banana biocontrol activities in bacillus |
| description |
Fusarium wilt of banana, caused by Fusarium oxysporum f. sp. cubense (Foc), especially Tropical Race 4 (TR4), seriously threatens banana production worldwide. There is no single effective control measure, although certain Bacillus strains secrete antibiotics as promising disease-biocontrol agents. This study identified five Bacillus strains displaying strong antibiotic activity against TR4, using a systemic assessment for presence/absence of genetic markers at genome level, and expres sion profiles at transcriptome level. A conventional PCR with 13 specific primer pairs detected biocontrol-related genes. An accurate, quantitative real-time PCR protocol with novel designed specific primers was developed to characterise strain-specific gene expression, that optimises strain culturing and RNA-isolation methodologies. Six genes responsible for synthesising non-ribosomal peptide synthetase biocontrol metabolites were detected in all five strains. Three genes were involved in synthesising three Polyketide synthetase metabolites in all five strains, but the macrolactin synthase gene mln was only detected in WBN06 and YN1282-2. All five Bacillus strains have the genes dhb and bioA, essential for synthesising bacillibactin and biotin. However, the gene sboA, involved in subtilisin synthesis, is absent in all five strains. These genes’ expression patterns were significantly different among these strains, suggesting different mechanisms involved in TR4 biocontrol. Results will help elucidate functional genes’ biocontrol mechanisms. |
| format |
Journal Article |
| topic_facet |
bacillus biological control genetic markers transcription factors fusarium wilt plant disease control control biológico marcadores genéticos factores de transcripción |
| author |
Li, Shu He, Ping Fan, Huacai Liu, Lina Yin, Kesuo Yang, Baoming Li, Yongping Huang, Su-Mei Li, Xundong Zheng, Si-Jun |
| author_facet |
Li, Shu He, Ping Fan, Huacai Liu, Lina Yin, Kesuo Yang, Baoming Li, Yongping Huang, Su-Mei Li, Xundong Zheng, Si-Jun |
| author_sort |
Li, Shu |
| title |
A real-time fluorescent reverse transcription quantitative PCR assay for rapid detection of genetic markers’ expression associated with Fusarium Wilt of banana biocontrol activities in bacillus |
| title_short |
A real-time fluorescent reverse transcription quantitative PCR assay for rapid detection of genetic markers’ expression associated with Fusarium Wilt of banana biocontrol activities in bacillus |
| title_full |
A real-time fluorescent reverse transcription quantitative PCR assay for rapid detection of genetic markers’ expression associated with Fusarium Wilt of banana biocontrol activities in bacillus |
| title_fullStr |
A real-time fluorescent reverse transcription quantitative PCR assay for rapid detection of genetic markers’ expression associated with Fusarium Wilt of banana biocontrol activities in bacillus |
| title_full_unstemmed |
A real-time fluorescent reverse transcription quantitative PCR assay for rapid detection of genetic markers’ expression associated with Fusarium Wilt of banana biocontrol activities in bacillus |
| title_sort |
real-time fluorescent reverse transcription quantitative pcr assay for rapid detection of genetic markers’ expression associated with fusarium wilt of banana biocontrol activities in bacillus |
| publisher |
MDPI |
| publishDate |
2021-04-30 |
| url |
https://hdl.handle.net/10568/113757 https://doi.org/10.3390/jof7050353 |
| work_keys_str_mv |
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