Pre-treatment and Suitable Reagent Enabled a Reliable and Consistent for Molecular Detection of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc4)
Fusarium wilt which is caused by the soil-borne fungus, Fusarium oxysporum f.sp. cubense (Foc), is one of important diseases in banana plant. Foc tropical race 4 (Foc4) is the most pathogenic race which could infect various banana cultivars including Cavendish cultivar which was previously considere...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Bogor Agricultural University
2019-12-01
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| Series: | Hayati Journal of Biosciences |
| Subjects: | |
| Online Access: | https://journal.ipb.ac.id/index.php/hayati/article/view/28885 |
| _version_ | 1819034341788876800 |
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| author | Listya Utami Karmawan Fenny Martha Dwivany Rizkita Rachmi Esyanti I Nyoman Pugeg Aryantha |
| author_facet | Listya Utami Karmawan Fenny Martha Dwivany Rizkita Rachmi Esyanti I Nyoman Pugeg Aryantha |
| author_sort | Listya Utami Karmawan |
| collection | DOAJ |
| description | Fusarium wilt which is caused by the soil-borne fungus, Fusarium oxysporum f.sp. cubense (Foc), is one of important diseases in banana plant. Foc tropical race 4 (Foc4) is the most pathogenic race which could infect various banana cultivars including Cavendish cultivar which was previously considered as resistant cultivar. Molecular detection of Foc using PCR analysis is indispensable to determine the race of Foc. We demonstrate a faster DNA isolation procedure described in previous method by substituting sodium acetate precipitation with ammonium acetate precipitation without affecting the result. Based on our experience, some fungal genomes were troublesome to be amplified. We suggested pre-treatment step prior to amplification procedure by incubating fungal DNA in 65°C for 10 minutes for any samples of fungal genome, including stubborn samples, before mixing into PCR mix reagent. PCR reagents should be tested for stubborn samples since some of the reagents were unable to amplify the desired DNA fragment. Pre-treatment and the choice of robust PCR reagent should be taken into consideration for a reliable and consistent Foc4 molecular detection result. |
| first_indexed | 2024-12-21T07:32:12Z |
| format | Article |
| id | doaj.art-5e2c3408edbc4ffa83989e56edcb7440 |
| institution | Directory Open Access Journal |
| issn | 1978-3019 2086-4094 |
| language | English |
| last_indexed | 2024-12-21T07:32:12Z |
| publishDate | 2019-12-01 |
| publisher | Bogor Agricultural University |
| record_format | Article |
| series | Hayati Journal of Biosciences |
| spelling | doaj.art-5e2c3408edbc4ffa83989e56edcb74402022-12-21T19:11:31ZengBogor Agricultural UniversityHayati Journal of Biosciences1978-30192086-40942019-12-0126410.4308/hjb.26.4.%xPre-treatment and Suitable Reagent Enabled a Reliable and Consistent for Molecular Detection of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc4)Listya Utami Karmawan0Fenny Martha Dwivany1Rizkita Rachmi Esyanti2I Nyoman Pugeg Aryantha3School of Life Sciences and Technology, Institut Teknologi Bandung, Bandung Faculty of Biotechnology, Atma Jaya Catholic University of Indonesia, JakartaSchool of Life Sciences and Technology, Institut Teknologi Bandung, Bandung Biosciences and Biotechnology Research Centre, Institut Teknologi Bandung, BandungSchool of Life Sciences and Technology, Institut Teknologi Bandung, BandungSchool of Life Sciences and Technology, Institut Teknologi Bandung, Bandun Biosciences and Biotechnology Research Centre, Institut Teknologi Bandung, BandungFusarium wilt which is caused by the soil-borne fungus, Fusarium oxysporum f.sp. cubense (Foc), is one of important diseases in banana plant. Foc tropical race 4 (Foc4) is the most pathogenic race which could infect various banana cultivars including Cavendish cultivar which was previously considered as resistant cultivar. Molecular detection of Foc using PCR analysis is indispensable to determine the race of Foc. We demonstrate a faster DNA isolation procedure described in previous method by substituting sodium acetate precipitation with ammonium acetate precipitation without affecting the result. Based on our experience, some fungal genomes were troublesome to be amplified. We suggested pre-treatment step prior to amplification procedure by incubating fungal DNA in 65°C for 10 minutes for any samples of fungal genome, including stubborn samples, before mixing into PCR mix reagent. PCR reagents should be tested for stubborn samples since some of the reagents were unable to amplify the desired DNA fragment. Pre-treatment and the choice of robust PCR reagent should be taken into consideration for a reliable and consistent Foc4 molecular detection result.https://journal.ipb.ac.id/index.php/hayati/article/view/28885DNA isolationgenome pre-treatmentFoc detection methodbananaFusarium wiltfood and agriculture |
| spellingShingle | Listya Utami Karmawan Fenny Martha Dwivany Rizkita Rachmi Esyanti I Nyoman Pugeg Aryantha Pre-treatment and Suitable Reagent Enabled a Reliable and Consistent for Molecular Detection of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc4) Hayati Journal of Biosciences DNA isolation genome pre-treatment Foc detection method banana Fusarium wilt food and agriculture |
| title | Pre-treatment and Suitable Reagent Enabled a Reliable and Consistent for Molecular Detection of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc4) |
| title_full | Pre-treatment and Suitable Reagent Enabled a Reliable and Consistent for Molecular Detection of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc4) |
| title_fullStr | Pre-treatment and Suitable Reagent Enabled a Reliable and Consistent for Molecular Detection of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc4) |
| title_full_unstemmed | Pre-treatment and Suitable Reagent Enabled a Reliable and Consistent for Molecular Detection of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc4) |
| title_short | Pre-treatment and Suitable Reagent Enabled a Reliable and Consistent for Molecular Detection of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc4) |
| title_sort | pre treatment and suitable reagent enabled a reliable and consistent for molecular detection of fusarium oxysporum f sp cubense tropical race 4 foc4 |
| topic | DNA isolation genome pre-treatment Foc detection method banana Fusarium wilt food and agriculture |
| url | https://journal.ipb.ac.id/index.php/hayati/article/view/28885 |
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