Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome Amplification

[EN] Flow cytometry is widely used to determine genome size and ploidy level in plants. This technique, when coupled with fluorescence-activated cell sorting (FACS), whole genome amplification and genotyping (WGA), opens up new opportunities for genetic studies of individualized nuclei. This strateg...

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Autores: Garavello, Miguel, Cuenca, José, Dreissig, Steven, Fuchs, Jörg, Houben, Andreas, Aleza, Pablo
Tipo de recurso: artículo
Fecha de publicación:2019
País:España
Institución:Universitat Politècnica de València (UPV)
Repositorio:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
Idioma:inglés
OAI Identifier:oai:riunet.upv.es:10251/204373
Acceso en línea:https://riunet.upv.es/handle/10251/204373
Access Level:acceso abierto
Palabra clave:SSR and SNP markers
Flow cytometry
Fluorescence-activated cell sorting
Tetraploid
Triploid
Unreduced gametes
Whole genome amplification
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spelling Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome AmplificationGaravello, MiguelCuenca, JoséDreissig, StevenFuchs, JörgHouben, AndreasAleza, PabloSSR and SNP markersFlow cytometryFluorescence-activated cell sortingTetraploidTriploidUnreduced gametesWhole genome amplification[EN] Flow cytometry is widely used to determine genome size and ploidy level in plants. This technique, when coupled with fluorescence-activated cell sorting (FACS), whole genome amplification and genotyping (WGA), opens up new opportunities for genetic studies of individualized nuclei. This strategy was used to analyze the genetic composition of single pollen nuclei of different citrus species. The flow cytometry and microscope observations allowed us to differentiate the populations of pollen nuclei present in the diploid and euploid genotypes analyzed, showing that citrus has binuclear pollen. We have identified in the ¿CSO¿ tangor an additional nuclei population composed by the vegetative plus generative nuclei. Genotyping of this nuclei population revealed that vegetative and generative nuclei show the same genetic configuration. In addition, we have demonstrated the presence of unreduced gametes in the diploid genotype ¿Mexican lime.¿ Genomic amplification is a robust method for haploid nuclei genotyping with several molecular markers, whereas in diploid nuclei using heterozygous markers showed a bias towards one of the two alleles, limiting the use of this tool in this type of nuclei. We further discuss the importance and applications of single pollen genotyping in citrus genetic studies.This work was supported by the project RTA2015-00069-00-00 from the Ministry of Economía y Competividad and Instituto Nacional de Investigación y Tecnología Agraria y Agroalimentaria and the grant Programa de Perfeccionamiento resolution 1177/14 from Consejo Directivo of Instituto Nacional de Tecnología Agropecuaria (Argentina).Frontiers Media SAInstituto Nacional de Tecnología AgropecuariaInstituto Nacional de Investigación y Tecnología Agraria y AlimentariaMinisterio de Economía y Competitividad e Instituto Nacional de Investigación y Tecnología Agraria y AlimentariaRepositorio Institucional de la Universitat Politècnica de València Riunet20192019-09-24journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://riunet.upv.es/handle/10251/204373reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valénciainstname:Universitat Politècnica de València (UPV)InglésengMinisterio de Economía y Competitividad http://dx.doi.org/10.13039/501100003329 RTA2015-00069-00-00 Mejora genética y sanitaria de cítricos mediante métodos biotecnológicosInstituto Nacional de Tecnologia Agropecuaria https://doi.org/10.13039/501100010677 1177%2F14open accesshttp://purl.org/coar/access_right/c_abf2Reconocimiento (by)http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:riunet.upv.es:10251/2043732026-06-13T07:49:27Z
dc.title.none.fl_str_mv Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome Amplification
title Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome Amplification
spellingShingle Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome Amplification
Garavello, Miguel
SSR and SNP markers
Flow cytometry
Fluorescence-activated cell sorting
Tetraploid
Triploid
Unreduced gametes
Whole genome amplification
title_short Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome Amplification
title_full Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome Amplification
title_fullStr Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome Amplification
title_full_unstemmed Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome Amplification
title_sort Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome Amplification
dc.creator.none.fl_str_mv Garavello, Miguel
Cuenca, José
Dreissig, Steven
Fuchs, Jörg
Houben, Andreas
Aleza, Pablo
author Garavello, Miguel
author_facet Garavello, Miguel
Cuenca, José
Dreissig, Steven
Fuchs, Jörg
Houben, Andreas
Aleza, Pablo
author_role author
author2 Cuenca, José
Dreissig, Steven
Fuchs, Jörg
Houben, Andreas
Aleza, Pablo
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto Nacional de Tecnología Agropecuaria
Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria
Ministerio de Economía y Competitividad e Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria
Repositorio Institucional de la Universitat Politècnica de València Riunet
dc.subject.none.fl_str_mv SSR and SNP markers
Flow cytometry
Fluorescence-activated cell sorting
Tetraploid
Triploid
Unreduced gametes
Whole genome amplification
topic SSR and SNP markers
Flow cytometry
Fluorescence-activated cell sorting
Tetraploid
Triploid
Unreduced gametes
Whole genome amplification
description [EN] Flow cytometry is widely used to determine genome size and ploidy level in plants. This technique, when coupled with fluorescence-activated cell sorting (FACS), whole genome amplification and genotyping (WGA), opens up new opportunities for genetic studies of individualized nuclei. This strategy was used to analyze the genetic composition of single pollen nuclei of different citrus species. The flow cytometry and microscope observations allowed us to differentiate the populations of pollen nuclei present in the diploid and euploid genotypes analyzed, showing that citrus has binuclear pollen. We have identified in the ¿CSO¿ tangor an additional nuclei population composed by the vegetative plus generative nuclei. Genotyping of this nuclei population revealed that vegetative and generative nuclei show the same genetic configuration. In addition, we have demonstrated the presence of unreduced gametes in the diploid genotype ¿Mexican lime.¿ Genomic amplification is a robust method for haploid nuclei genotyping with several molecular markers, whereas in diploid nuclei using heterozygous markers showed a bias towards one of the two alleles, limiting the use of this tool in this type of nuclei. We further discuss the importance and applications of single pollen genotyping in citrus genetic studies.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019-09-24
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://riunet.upv.es/handle/10251/204373
url https://riunet.upv.es/handle/10251/204373
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Ministerio de Economía y Competitividad http://dx.doi.org/10.13039/501100003329 RTA2015-00069-00-00 Mejora genética y sanitaria de cítricos mediante métodos biotecnológicos
Instituto Nacional de Tecnologia Agropecuaria https://doi.org/10.13039/501100010677 1177%2F14
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Reconocimiento (by)
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Reconocimiento (by)
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Frontiers Media SA
publisher.none.fl_str_mv Frontiers Media SA
dc.source.none.fl_str_mv reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
instname:Universitat Politècnica de València (UPV)
instname_str Universitat Politècnica de València (UPV)
reponame_str RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
collection RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,301603

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