Development of a multiplex PCR for the identification of Fusarium solani and F. oxysporum in a single step
16 Pág. Departamento de Protección Vegetal (INIA)
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| Language: | English |
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Springer Nature
2021-05-11
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| Subjects: | Fusarium root rot, Fusarium wilt, ITS, Molecular detection, Soil-borne fungi, Species distribution, TEF, |
| Online Access: | http://hdl.handle.net/10261/287809 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003329 https://api.elsevier.com/content/abstract/scopus_id/85105734003 |
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dig-inia-es-10261-2878092024-05-16T20:56:58Z Development of a multiplex PCR for the identification of Fusarium solani and F. oxysporum in a single step Villarino, María De Cal Cortina, Antonieta Melgarejo, Paloma Larena, Inmaculada European Commission Ministerio de Economía y Competitividad (España) Villarino, María [0000-0002-5635-8146] De Cal Cortina, Antonieta [0000-0002-7725-7782] Melgarejo, Paloma [0000-0002-3698-8896] Larena, Inmaculada [0000-0001-8424-8916] Fusarium root rot Fusarium wilt ITS Molecular detection Soil-borne fungi Species distribution TEF 16 Pág. Departamento de Protección Vegetal (INIA) Fusarium solani and F. oxysporum are plant pathogenic fungi that cause root rot and wilt, respectively, in many economically important crops. Identification of both species by morphological characteristics is complex and time-consuming, especially when a crop is host to several species, as in the case of strawberry and other many crops. The objective of this research was to develop a molecular method for the precise and sensitive identification of Fusarium solani and F. oxysporum by multiplex PCR. Specific PCR conditions were developed for the amplification of about 170-bp PCR product from the internal transcribed spacer of F. oxysporum and a PCR product of about 650 bp from the elongation factor 1-alpha gene of F. solani. The specificity of the multiplex PCR was tested using gDNA extracted from 232 strains of F. solani and F. oxysporum, 12 strains of other Fusarium spp. and 14 strains of other genera. The sensitivity of each primer set was tested individually and simultaneously with one strain of F. oxysporum and F. solani. This protocol was then used to identify both species in natural isolates from diseased plant and soil samples from strawberry nursery. Each primer set was validated using genomic DNA from non-sterile and sterile soils artificially inoculated with both species. Under these conditions, both Fusarium species were detected and distinguished over a range of concentrations (1 × 107 to 1 × 102 conidia g−1 soil) by successful amplification of corresponding PCR products for each species. This multiplex PCR technique is a fast tool to identify natural isolates of both species, F. solani and F. oxysporum, in a single PCR reaction. This study was supported by EUPHRESCO [266505 FP-7 ERANET EUPHRESCO-SPAT “Strawberry pathogens assessment and testing”] and by Ministry of Economy and Competitiveness (Spain) [RTA2013-00062-C05-E; RTA2017-00019-C03-01 (Plan Nacional de I + D)]. Peer reviewed 2023-01-30T11:35:51Z 2023-01-30T11:35:51Z 2021-05-11 artículo http://purl.org/coar/resource_type/c_6501 Journal of Plant Diseases and Protection 128(5): 1275-1290 (2021) 1861-3837 http://hdl.handle.net/10261/287809 10.1007/s41348-021-00475-6 1861-3829 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003329 2-s2.0-85105734003 https://api.elsevier.com/content/abstract/scopus_id/85105734003 en #PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/EC/FP7/266505 info:eu-repo/grantAgreement/MINECO//RTA2013-00062-C05-E info:eu-repo/grantAgreement/MINECO//RTA2017-00019-C03-01 Journal of Plant Diseases and Protection Publisher's version https://doi.org/10.1007/s41348-021-00475-6 Sí open Springer Nature |
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España |
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ES |
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dig-inia-es |
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biblioteca |
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Biblioteca del INIA España |
| language |
English |
| topic |
Fusarium root rot Fusarium wilt ITS Molecular detection Soil-borne fungi Species distribution TEF Fusarium root rot Fusarium wilt ITS Molecular detection Soil-borne fungi Species distribution TEF |
| spellingShingle |
Fusarium root rot Fusarium wilt ITS Molecular detection Soil-borne fungi Species distribution TEF Fusarium root rot Fusarium wilt ITS Molecular detection Soil-borne fungi Species distribution TEF Villarino, María De Cal Cortina, Antonieta Melgarejo, Paloma Larena, Inmaculada Development of a multiplex PCR for the identification of Fusarium solani and F. oxysporum in a single step |
| description |
16 Pág.
Departamento de Protección Vegetal (INIA) |
| author2 |
European Commission |
| author_facet |
European Commission Villarino, María De Cal Cortina, Antonieta Melgarejo, Paloma Larena, Inmaculada |
| format |
artículo |
| topic_facet |
Fusarium root rot Fusarium wilt ITS Molecular detection Soil-borne fungi Species distribution TEF |
| author |
Villarino, María De Cal Cortina, Antonieta Melgarejo, Paloma Larena, Inmaculada |
| author_sort |
Villarino, María |
| title |
Development of a multiplex PCR for the identification of Fusarium solani and F. oxysporum in a single step |
| title_short |
Development of a multiplex PCR for the identification of Fusarium solani and F. oxysporum in a single step |
| title_full |
Development of a multiplex PCR for the identification of Fusarium solani and F. oxysporum in a single step |
| title_fullStr |
Development of a multiplex PCR for the identification of Fusarium solani and F. oxysporum in a single step |
| title_full_unstemmed |
Development of a multiplex PCR for the identification of Fusarium solani and F. oxysporum in a single step |
| title_sort |
development of a multiplex pcr for the identification of fusarium solani and f. oxysporum in a single step |
| publisher |
Springer Nature |
| publishDate |
2021-05-11 |
| url |
http://hdl.handle.net/10261/287809 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003329 https://api.elsevier.com/content/abstract/scopus_id/85105734003 |
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