Cargando…
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...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8147159/ https://www.ncbi.nlm.nih.gov/pubmed/33946404 http://dx.doi.org/10.3390/jof7050353 |
_version_ | 1783697565079830528 |
---|---|
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 |
collection | PubMed |
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 expression 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 | Online Article Text |
id | pubmed-8147159 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-81471592021-05-26 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 J Fungi (Basel) Article 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 expression 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. MDPI 2021-04-30 /pmc/articles/PMC8147159/ /pubmed/33946404 http://dx.doi.org/10.3390/jof7050353 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article 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 | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8147159/ https://www.ncbi.nlm.nih.gov/pubmed/33946404 http://dx.doi.org/10.3390/jof7050353 |
work_keys_str_mv | AT lishu arealtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT heping arealtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT fanhuacai arealtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT liulina arealtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT yinkesuo arealtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT yangbaoming arealtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT liyongping arealtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT huangsumei arealtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT lixundong arealtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT zhengsijun arealtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT lishu realtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT heping realtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT fanhuacai realtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT liulina realtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT yinkesuo realtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT yangbaoming realtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT liyongping realtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT huangsumei realtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT lixundong realtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus AT zhengsijun realtimefluorescentreversetranscriptionquantitativepcrassayforrapiddetectionofgeneticmarkersexpressionassociatedwithfusariumwiltofbananabiocontrolactivitiesinbacillus |