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. T...
| Main Authors: | , , , , , , , , , |
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| Format: | Journal Article |
| Language: | Inglés |
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MDPI
2021
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| Online Access: | https://hdl.handle.net/10568/113757 |
| _version_ | 1855543437493796864 |
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| author | Li, Shu He, Ping Fan, Huacai Liu, Lina Yin, Kesuo Yang, Baoming Li, Yongping Huang, Su-Mei Li, Xundong Zheng, Si-Jun |
| author_browse | Fan, Huacai He, Ping Huang, Su-Mei Li, Shu Li, Xundong Li, Yongping Liu, Lina Yang, Baoming Yin, Kesuo 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 |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| 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 |
| id | CGSpace113757 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | CGSpace1137572025-11-11T23:16:56Z 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 https://hdl.handle.net/10568/113757 en Open Access application/pdf MDPI 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 |
| spellingShingle | 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 |
| 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_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_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_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 |
| topic | bacillus biological control genetic markers transcription factors fusarium wilt plant disease control control biológico marcadores genéticos factores de transcripción |
| url | https://hdl.handle.net/10568/113757 |
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