PCR identification of petunia male sterile cytoplasm = Identificação por PCR de citoplasma macho estéril em Petunia

Petunia is a very important ornamental plant with a broad range of flower colour and size, and most of the cultivars grown are propagated through seeds. Cytoplasmic male sterility (CMS) is a maternally inherited character determined by mitochondrial genes that results in impaired pollen development....

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Main Authors: Colombo, Noemi, Hagiwara, Juan Carlos
Format: Artículo
Language:Inglés
Published: Sociedade Brasileira de Floricultura e Plantas Ornamentais 2020
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/6705
https://ornamentalhorticulture.emnuvens.com.br/rbho/article/view/2069
https://doi.org/10.1590/2447-536X.v25i4.2069
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author Colombo, Noemi
Hagiwara, Juan Carlos
author_browse Colombo, Noemi
Hagiwara, Juan Carlos
author_facet Colombo, Noemi
Hagiwara, Juan Carlos
author_sort Colombo, Noemi
collection INTA Digital
description Petunia is a very important ornamental plant with a broad range of flower colour and size, and most of the cultivars grown are propagated through seeds. Cytoplasmic male sterility (CMS) is a maternally inherited character determined by mitochondrial genes that results in impaired pollen development. The unique and well characterized male sterile cytoplasm in Petunia is a valuable resource for hybrids production because it prevents self-fertilization of mother plants and ensures the purity of F1s. Introgression of the male sterile cytoplasm in elite lines of Petunia is achieved following a backcross scheme and can be assisted using molecular markers associated to the trait of interest. The objective of this study was to develop a molecular marker to identify the male sterile cytoplasm of Petunia. A PCR-based marker amplifying a region of the mitochondrial CMS-associated urfS only in the male sterile plants was designed. Results showed differential PCR amplification of a ≈ 600 bp product in plants carrying male sterile cytoplasm in four Petunia species and their F1s and BC1 generations. A multiplex PCR reaction was subsequently set up, adding specific primers amplifying a ≈ 800 bp product from the conserved region trnT-trn-L of the chloroplast genome as a positive control in order to unambiguously identify the cytoplasm types as normal or sterile. A rapid, simple and precise molecular marker is now available for assisting breeding of F1 hybrids in Petunia.
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spelling INTA67052020-02-10T14:14:05Z PCR identification of petunia male sterile cytoplasm = Identificação por PCR de citoplasma macho estéril em Petunia Colombo, Noemi Hagiwara, Juan Carlos Petunia Esterilidad Masculina Citoplásmica Marcadores Genéticos Plantas Ornamentales Fitomejoramiento Cytoplasmic Male Sterility Genetic Markers Ornamental Plants Plant Breeding Petunia is a very important ornamental plant with a broad range of flower colour and size, and most of the cultivars grown are propagated through seeds. Cytoplasmic male sterility (CMS) is a maternally inherited character determined by mitochondrial genes that results in impaired pollen development. The unique and well characterized male sterile cytoplasm in Petunia is a valuable resource for hybrids production because it prevents self-fertilization of mother plants and ensures the purity of F1s. Introgression of the male sterile cytoplasm in elite lines of Petunia is achieved following a backcross scheme and can be assisted using molecular markers associated to the trait of interest. The objective of this study was to develop a molecular marker to identify the male sterile cytoplasm of Petunia. A PCR-based marker amplifying a region of the mitochondrial CMS-associated urfS only in the male sterile plants was designed. Results showed differential PCR amplification of a ≈ 600 bp product in plants carrying male sterile cytoplasm in four Petunia species and their F1s and BC1 generations. A multiplex PCR reaction was subsequently set up, adding specific primers amplifying a ≈ 800 bp product from the conserved region trnT-trn-L of the chloroplast genome as a positive control in order to unambiguously identify the cytoplasm types as normal or sterile. A rapid, simple and precise molecular marker is now available for assisting breeding of F1 hybrids in Petunia. A petúnia é uma planta ornamental muito importante, com uma ampla gama de cores e tamanhos de flores, e a maioria das cultivares são propagadas através de sementes. A esterilidade citoplasmática masculina (CMS) é um caráter hereditário da mãe, determinado por genes mitocondriais que resultam em prejuízo ao desenvolvimento do pólen. O citoplasma macho estéril único e bem caracterizado em Petúnia é um recurso valioso para a produção de híbridos, pois previne a autofecundação das plantasmãe e garante a pureza do F1. A introgressão do citoplasma macho estéril nas linhas de elite de Petúnia é alcançada seguindo um esquema de retrocruzamento e pode ser assistida usando marcadores moleculares associados à característica de interesse. O objetivo deste estudo foi desenvolver um marcador molecular para identificar o citoplasma macho estéril de Petunia. Foi desenhado um marcador baseado em PCR que amplificou uma região de urfS associada à CMS mitocondrial apenas em plantas masculinas estéreis. Os resultados mostraram amplificação por PCR diferencial de um produto de ≈ 600 bp em plantas portadoras de citoplasma estéril masculino em quatro espécies de Petúnia e suas gerações F1s e BC1. Uma reação de PCR multiplex foi subsequentemente configurada, adicionando iniciadores específicos amplificando um produto de ≈ 800 bp da região conservada trnT-trn-L do genoma do cloroplasto como um controle positivo, para identificar inequivocamente os tipos de citoplasma como normal ou estéril. Um marcador molecular rápido, simples e preciso está agora disponível para auxiliar no melhoramento genético de híbridos F1 em Petunia. Instituto de Genética Fil: Colombo, Noemi. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina Fil: Hagiwara, Juan Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Floricultura; Argentina 2020-02-10T14:05:59Z 2020-02-10T14:05:59Z 2019-12 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/6705 https://ornamentalhorticulture.emnuvens.com.br/rbho/article/view/2069 2447-536X https://doi.org/10.1590/2447-536X.v25i4.2069 eng info:eu-repograntAgreement/INTA/PNHFA/1106092/AR./Mejoramiento genético de plantas ornamentales a partir de germoplasma nativo y su introducción en los mercados. info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Sociedade Brasileira de Floricultura e Plantas Ornamentais Ornamental horticulture 25 (4) : 345-350. (Diciembre 2019)
spellingShingle Petunia
Esterilidad Masculina Citoplásmica
Marcadores Genéticos
Plantas Ornamentales
Fitomejoramiento
Cytoplasmic Male Sterility
Genetic Markers
Ornamental Plants
Plant Breeding
Colombo, Noemi
Hagiwara, Juan Carlos
PCR identification of petunia male sterile cytoplasm = Identificação por PCR de citoplasma macho estéril em Petunia
title PCR identification of petunia male sterile cytoplasm = Identificação por PCR de citoplasma macho estéril em Petunia
title_full PCR identification of petunia male sterile cytoplasm = Identificação por PCR de citoplasma macho estéril em Petunia
title_fullStr PCR identification of petunia male sterile cytoplasm = Identificação por PCR de citoplasma macho estéril em Petunia
title_full_unstemmed PCR identification of petunia male sterile cytoplasm = Identificação por PCR de citoplasma macho estéril em Petunia
title_short PCR identification of petunia male sterile cytoplasm = Identificação por PCR de citoplasma macho estéril em Petunia
title_sort pcr identification of petunia male sterile cytoplasm identificacao por pcr de citoplasma macho esteril em petunia
topic Petunia
Esterilidad Masculina Citoplásmica
Marcadores Genéticos
Plantas Ornamentales
Fitomejoramiento
Cytoplasmic Male Sterility
Genetic Markers
Ornamental Plants
Plant Breeding
url http://hdl.handle.net/20.500.12123/6705
https://ornamentalhorticulture.emnuvens.com.br/rbho/article/view/2069
https://doi.org/10.1590/2447-536X.v25i4.2069
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