Disease Resistant Citrus Breeding Using Newly Developed High Resolution Melting and CAPS Protocols for Alternaria Brown Spot Marker Assisted Selection
Alternaria alternata is a fungus that causes a serious disease in susceptible genotypes of citrus, particularly in mandarins. The Alternaria citri toxin (ACT) produced by the pathogen induces necrotic lesions on young leaves and fruits, defoliation and fruit drop. Here, we describe two methods of...
| Autores principales: | , , , , , , , , , , , |
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| Formato: | article |
| Lenguaje: | Inglés |
| Publicado: |
2021
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.11939/6965 https://www.mdpi.com/2073-4395/10/9/1368 |
| _version_ | 1855032522395615232 |
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| author | Arlotta, Carmen Ciacciulli, Angelo Strano, Maria Concetta Cafaro, Valeria Salonia, Fabrizio Caruso, Paola Licciardello, Concetta Russo, Giuseppe Wesley Smith, Malcolm Cuenca, José Aleza, Pablo Caruso, Marco |
| author_browse | Aleza, Pablo Arlotta, Carmen Cafaro, Valeria Caruso, Marco Caruso, Paola Ciacciulli, Angelo Cuenca, José Licciardello, Concetta Russo, Giuseppe Salonia, Fabrizio Strano, Maria Concetta Wesley Smith, Malcolm |
| author_facet | Arlotta, Carmen Ciacciulli, Angelo Strano, Maria Concetta Cafaro, Valeria Salonia, Fabrizio Caruso, Paola Licciardello, Concetta Russo, Giuseppe Wesley Smith, Malcolm Cuenca, José Aleza, Pablo Caruso, Marco |
| author_sort | Arlotta, Carmen |
| collection | ReDivia |
| description | Alternaria alternata is a fungus that causes a serious disease in susceptible genotypes
of citrus, particularly in mandarins. The Alternaria citri toxin (ACT) produced by the pathogen
induces necrotic lesions on young leaves and fruits, defoliation and fruit drop. Here, we describe
two methods of marker-assisted selection (MAS) that could be used for the early identification of
Alternaria brown spot (ABS)-resistant mandarin hybrids. The first method is based on a nested PCR
coupled to high resolution melting (HRM) analysis at the SNP08 locus, which is located at 0.4 cM
from the ABS resistance locus, and was previously indicated as the most suitable for the selection of
ABS-resistant hybrids. The method was validated on 41 mandarin hybrids of the CREA germplasm
collection, and on 862 progenies generated from five crosses involving different susceptible parents.
Four out of five populations showed Mendelian segregation at the analyzed locus, while a population
involving Murcott tangor as male parent showed distorted segregation toward the susceptible hybrids.
The second method is based on a cleaved amplified polymorphic sequences (CAPS) marker that was
developed using the same primers as the nested PCR at the SNP08 locus, coupled with BccI restriction
enzyme digestion. To verify the reliability of the two genotyping methods, in vitro leaf phenotyping
was carried out by inoculating A. alternata spores onto young leaves of 101 hybrids, randomly chosen
among the susceptible and resistant progenies. The phenotyping confirmed the SNP08 genotyping
results, so the proposed method of selection based on HRM or CAPS genotyping could be routinely
used as an alternative to KBioscience competitive allele specific polymerase chain reaction (KASPar)
single nucleotide polymorphism (SNP) genotyping system to improve citrus breeding programs.
While the study confirmed that the SNP08 marker is a reliable tool for MAS of new citrus hybrids
with different genetic backgrounds, it also identified a small group of genotypes where the resistance
mechanism requires further investigation. |
| format | article |
| id | ReDivia6965 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| record_format | dspace |
| spelling | ReDivia69652025-04-25T14:48:00Z Disease Resistant Citrus Breeding Using Newly Developed High Resolution Melting and CAPS Protocols for Alternaria Brown Spot Marker Assisted Selection Arlotta, Carmen Ciacciulli, Angelo Strano, Maria Concetta Cafaro, Valeria Salonia, Fabrizio Caruso, Paola Licciardello, Concetta Russo, Giuseppe Wesley Smith, Malcolm Cuenca, José Aleza, Pablo Caruso, Marco Genotyping Single nucleotide polymorphisms H20 Plant diseases Alternaria alternata Mandarins Alternaria alternata is a fungus that causes a serious disease in susceptible genotypes of citrus, particularly in mandarins. The Alternaria citri toxin (ACT) produced by the pathogen induces necrotic lesions on young leaves and fruits, defoliation and fruit drop. Here, we describe two methods of marker-assisted selection (MAS) that could be used for the early identification of Alternaria brown spot (ABS)-resistant mandarin hybrids. The first method is based on a nested PCR coupled to high resolution melting (HRM) analysis at the SNP08 locus, which is located at 0.4 cM from the ABS resistance locus, and was previously indicated as the most suitable for the selection of ABS-resistant hybrids. The method was validated on 41 mandarin hybrids of the CREA germplasm collection, and on 862 progenies generated from five crosses involving different susceptible parents. Four out of five populations showed Mendelian segregation at the analyzed locus, while a population involving Murcott tangor as male parent showed distorted segregation toward the susceptible hybrids. The second method is based on a cleaved amplified polymorphic sequences (CAPS) marker that was developed using the same primers as the nested PCR at the SNP08 locus, coupled with BccI restriction enzyme digestion. To verify the reliability of the two genotyping methods, in vitro leaf phenotyping was carried out by inoculating A. alternata spores onto young leaves of 101 hybrids, randomly chosen among the susceptible and resistant progenies. The phenotyping confirmed the SNP08 genotyping results, so the proposed method of selection based on HRM or CAPS genotyping could be routinely used as an alternative to KBioscience competitive allele specific polymerase chain reaction (KASPar) single nucleotide polymorphism (SNP) genotyping system to improve citrus breeding programs. While the study confirmed that the SNP08 marker is a reliable tool for MAS of new citrus hybrids with different genetic backgrounds, it also identified a small group of genotypes where the resistance mechanism requires further investigation. 2021-01-13T14:37:57Z 2021-01-13T14:37:57Z 2020 article publishedVersion Arlotta, C., Ciacciulli, A., Strano, M. C., Cafaro, V., Salonia, F., Caruso, P., ... & Caruso, M. (2020). Disease Resistant Citrus Breeding Using Newly Developed High Resolution Melting and CAPS Protocols for Alternaria Brown Spot Marker Assisted Selection. Agronomy, 10(9), 1368. 2073-4395 http://hdl.handle.net/20.500.11939/6965 10.3390/agronomy10091368 https://www.mdpi.com/2073-4395/10/9/1368 en Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ openAccess electronico |
| spellingShingle | Genotyping Single nucleotide polymorphisms H20 Plant diseases Alternaria alternata Mandarins Arlotta, Carmen Ciacciulli, Angelo Strano, Maria Concetta Cafaro, Valeria Salonia, Fabrizio Caruso, Paola Licciardello, Concetta Russo, Giuseppe Wesley Smith, Malcolm Cuenca, José Aleza, Pablo Caruso, Marco Disease Resistant Citrus Breeding Using Newly Developed High Resolution Melting and CAPS Protocols for Alternaria Brown Spot Marker Assisted Selection |
| title | Disease Resistant Citrus Breeding Using Newly Developed High Resolution Melting and CAPS Protocols for Alternaria Brown Spot Marker Assisted Selection |
| title_full | Disease Resistant Citrus Breeding Using Newly Developed High Resolution Melting and CAPS Protocols for Alternaria Brown Spot Marker Assisted Selection |
| title_fullStr | Disease Resistant Citrus Breeding Using Newly Developed High Resolution Melting and CAPS Protocols for Alternaria Brown Spot Marker Assisted Selection |
| title_full_unstemmed | Disease Resistant Citrus Breeding Using Newly Developed High Resolution Melting and CAPS Protocols for Alternaria Brown Spot Marker Assisted Selection |
| title_short | Disease Resistant Citrus Breeding Using Newly Developed High Resolution Melting and CAPS Protocols for Alternaria Brown Spot Marker Assisted Selection |
| title_sort | disease resistant citrus breeding using newly developed high resolution melting and caps protocols for alternaria brown spot marker assisted selection |
| topic | Genotyping Single nucleotide polymorphisms H20 Plant diseases Alternaria alternata Mandarins |
| url | http://hdl.handle.net/20.500.11939/6965 https://www.mdpi.com/2073-4395/10/9/1368 |
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