Development and Investigation of HRM Markers to Discriminate Two Ogura Cytoplasmic Male Sterility Restorer Genes in Radish
Ogura male sterile cytoplasm is widely used for radish breeding. In this study, high-resolution melting (HRM) markers associated with <i>Rft</i> and <i>Rfo</i>, major restorer-of-fertility genes in Ogura cytoplasmic male sterility (CMS) in radish, were developed. Genetic mapp...
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| Format: | Article |
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MDPI AG
2023-12-01
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| Series: | Agronomy |
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| Online Access: | https://www.mdpi.com/2073-4395/14/1/43 |
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| author | Hong-Il Ahn Koeun Han Hee-Bum Yang Eun Su Lee Ye-Rin Lee Jinhee Kim Han Yong Park Do-Sun Kim |
| author_facet | Hong-Il Ahn Koeun Han Hee-Bum Yang Eun Su Lee Ye-Rin Lee Jinhee Kim Han Yong Park Do-Sun Kim |
| author_sort | Hong-Il Ahn |
| collection | DOAJ |
| description | Ogura male sterile cytoplasm is widely used for radish breeding. In this study, high-resolution melting (HRM) markers associated with <i>Rft</i> and <i>Rfo</i>, major restorer-of-fertility genes in Ogura cytoplasmic male sterility (CMS) in radish, were developed. Genetic mapping was carried out using F<sub>2</sub> populations derived from crosses between male-sterile Ogura CMS lines and male-fertile lines. Identification of the <i>Rft</i> and <i>Rfo</i> loci was achieved through SNP-based genotyping and linkage grouping. HRM markers were subsequently developed based on flanking sequences of SNPs linked to these loci. For the <i>Rft</i> gene, a set of 117 SNPs was selected within a candidate region on chromosome 5, and 14 HRM markers were successfully developed. Genotyping of F<sub>2</sub> showed high correlation between three markers and the phenotype. Regarding the <i>Rfo</i> gene, a set of 27 HRM markers was designed based on flanking sequences of SNPs located on chromosomes 9 and 0. Genotyping in the <i>Rfo</i> segregating population identified a single marker, RSRF27, that accurately distinguished the male sterility phenotype. Validation of the developed markers was performed in populations containing both <i>Rft</i> and <i>Rfo</i> genes, confirming their utility for genotyping and demonstrating that these two genes independently contribute to male sterility recovery. Overall, this study provides HRM markers that can be used for genotyping <i>Rft</i> and <i>Rfo</i> and contributes to a deeper understanding of male sterility restoration mechanisms in Ogura CMS. |
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| language | English |
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| publishDate | 2023-12-01 |
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| spelling | doaj.art-62b72f0211474de4b1feaa6f08be33d82024-01-26T14:21:53ZengMDPI AGAgronomy2073-43952023-12-011414310.3390/agronomy14010043Development and Investigation of HRM Markers to Discriminate Two Ogura Cytoplasmic Male Sterility Restorer Genes in RadishHong-Il Ahn0Koeun Han1Hee-Bum Yang2Eun Su Lee3Ye-Rin Lee4Jinhee Kim5Han Yong Park6Do-Sun Kim7Vegetable Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, RDA, Wanju 55365, Republic of KoreaVegetable Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, RDA, Wanju 55365, Republic of KoreaVegetable Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, RDA, Wanju 55365, Republic of KoreaVegetable Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, RDA, Wanju 55365, Republic of KoreaVegetable Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, RDA, Wanju 55365, Republic of KoreaVegetable Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, RDA, Wanju 55365, Republic of KoreaDepartment of Bioengineering, Sejong University, 209 Neungdong-ro, Gwangjin-gu, Seoul 05006, Republic of KoreaVegetable Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, RDA, Wanju 55365, Republic of KoreaOgura male sterile cytoplasm is widely used for radish breeding. In this study, high-resolution melting (HRM) markers associated with <i>Rft</i> and <i>Rfo</i>, major restorer-of-fertility genes in Ogura cytoplasmic male sterility (CMS) in radish, were developed. Genetic mapping was carried out using F<sub>2</sub> populations derived from crosses between male-sterile Ogura CMS lines and male-fertile lines. Identification of the <i>Rft</i> and <i>Rfo</i> loci was achieved through SNP-based genotyping and linkage grouping. HRM markers were subsequently developed based on flanking sequences of SNPs linked to these loci. For the <i>Rft</i> gene, a set of 117 SNPs was selected within a candidate region on chromosome 5, and 14 HRM markers were successfully developed. Genotyping of F<sub>2</sub> showed high correlation between three markers and the phenotype. Regarding the <i>Rfo</i> gene, a set of 27 HRM markers was designed based on flanking sequences of SNPs located on chromosomes 9 and 0. Genotyping in the <i>Rfo</i> segregating population identified a single marker, RSRF27, that accurately distinguished the male sterility phenotype. Validation of the developed markers was performed in populations containing both <i>Rft</i> and <i>Rfo</i> genes, confirming their utility for genotyping and demonstrating that these two genes independently contribute to male sterility recovery. Overall, this study provides HRM markers that can be used for genotyping <i>Rft</i> and <i>Rfo</i> and contributes to a deeper understanding of male sterility restoration mechanisms in Ogura CMS.https://www.mdpi.com/2073-4395/14/1/43radishrestorer-of-fertilitymolecular markerOgura CMS |
| spellingShingle | Hong-Il Ahn Koeun Han Hee-Bum Yang Eun Su Lee Ye-Rin Lee Jinhee Kim Han Yong Park Do-Sun Kim Development and Investigation of HRM Markers to Discriminate Two Ogura Cytoplasmic Male Sterility Restorer Genes in Radish Agronomy radish restorer-of-fertility molecular marker Ogura CMS |
| title | Development and Investigation of HRM Markers to Discriminate Two Ogura Cytoplasmic Male Sterility Restorer Genes in Radish |
| title_full | Development and Investigation of HRM Markers to Discriminate Two Ogura Cytoplasmic Male Sterility Restorer Genes in Radish |
| title_fullStr | Development and Investigation of HRM Markers to Discriminate Two Ogura Cytoplasmic Male Sterility Restorer Genes in Radish |
| title_full_unstemmed | Development and Investigation of HRM Markers to Discriminate Two Ogura Cytoplasmic Male Sterility Restorer Genes in Radish |
| title_short | Development and Investigation of HRM Markers to Discriminate Two Ogura Cytoplasmic Male Sterility Restorer Genes in Radish |
| title_sort | development and investigation of hrm markers to discriminate two ogura cytoplasmic male sterility restorer genes in radish |
| topic | radish restorer-of-fertility molecular marker Ogura CMS |
| url | https://www.mdpi.com/2073-4395/14/1/43 |
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