Fine Mapping and Genetic Effect Analysis of <i>Rf21(t)</i> for the Fertility Restoration of Chinsurah-Boro-II-Type Cytoplasmic Male Sterile <i>Oryza sativa</i> (ssp. <i>japonica</i>) Lines

The combination of Chinsurah Boro II (BT)-type cytoplasmic male sterility (CMS) and <i>Rf1</i>, the main fertility restorer gene (<i>Rf</i>) for CMS-BT, has been extensively utilized for the production of three-line commercial <i>japonica</i> hybrid seeds. The ide...

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Main Authors: Yuanyue Du, Liying Fan, Yunhua Gu, Chen Wang, Kai Shi, Yebin Qin, Zhejun Li, Qiaoquan Liu, Shuzhu Tang, Honggen Zhang, Zuopeng Xu
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Language:English
Published: MDPI AG 2025-07-01
Series:Agronomy
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Online Access:https://www.mdpi.com/2073-4395/15/7/1690
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author Yuanyue Du
Liying Fan
Yunhua Gu
Chen Wang
Kai Shi
Yebin Qin
Zhejun Li
Qiaoquan Liu
Shuzhu Tang
Honggen Zhang
Zuopeng Xu
author_facet Yuanyue Du
Liying Fan
Yunhua Gu
Chen Wang
Kai Shi
Yebin Qin
Zhejun Li
Qiaoquan Liu
Shuzhu Tang
Honggen Zhang
Zuopeng Xu
author_sort Yuanyue Du
collection DOAJ
description The combination of Chinsurah Boro II (BT)-type cytoplasmic male sterility (CMS) and <i>Rf1</i>, the main fertility restorer gene (<i>Rf</i>) for CMS-BT, has been extensively utilized for the production of three-line commercial <i>japonica</i> hybrid seeds. The identification of new <i>Rf</i> genes holds significance for the breeding of BT-type restorer lines, aiming to enhance the heterosis level of BT-type <i>japonica</i> hybrids. In the present study, ‘02428’, a wide-compatibility <i>japonica</i> variety, was observed to partially restore fertility to BT-type CMS lines. Genetic analysis revealed that ‘02428’ carries a dominant <i>Rf</i> gene, <i>Rf21(t)</i>, responsible for the fertility restoration of BT-type CMS lines. Leveraging bulked segregant analysis (BSA) resequencing technology and molecular markers, the <i>Rf21(t)</i> locus was identified, and mapped within a candidate interval of 6–12.5 Mb on chromosome 2. Using the iso-cytoplasmic restorer populations, <i>Rf21(t)</i> was ultimately mapped to an interval of approximately 77 kb, encompassing 12 predicted genes, including <i>LOC_Os02g17360</i>, encoding a PPR-domain-containing protein and <i>LOC_Os02g17380 (Rf2)</i>, a cloned <i>Rf</i> for Lead-rice-type CMS. A comparative sequence analysis, gene expression profiling and gene knockout experiments confirmed that <i>LOC_Os02g17360</i> and <i>LOC_Os02g17380</i> are the most likely candidates of <i>Rf21(t)</i>. Furthermore, <i>Rf21(t)</i> showed the dosage effect on the fertility restoration of BT-type CMS lines. This newly identified <i>Rf21(t)</i> represents a valuable genetic resource for the breeding of BT-type <i>japonica</i> restorer lines. Our findings offer practical insights for breeders interested in advancing BT-type <i>japonica</i> hybrid development.
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spelling doaj-art-00fc2113be8149b281e8695d103fef6f2025-08-20T02:45:42ZengMDPI AGAgronomy2073-43952025-07-01157169010.3390/agronomy15071690Fine Mapping and Genetic Effect Analysis of <i>Rf21(t)</i> for the Fertility Restoration of Chinsurah-Boro-II-Type Cytoplasmic Male Sterile <i>Oryza sativa</i> (ssp. <i>japonica</i>) LinesYuanyue Du0Liying Fan1Yunhua Gu2Chen Wang3Kai Shi4Yebin Qin5Zhejun Li6Qiaoquan Liu7Shuzhu Tang8Honggen Zhang9Zuopeng Xu10Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaJiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaJiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaJiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaJiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaJiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaJiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaJiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaJiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaJiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaJiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, ChinaThe combination of Chinsurah Boro II (BT)-type cytoplasmic male sterility (CMS) and <i>Rf1</i>, the main fertility restorer gene (<i>Rf</i>) for CMS-BT, has been extensively utilized for the production of three-line commercial <i>japonica</i> hybrid seeds. The identification of new <i>Rf</i> genes holds significance for the breeding of BT-type restorer lines, aiming to enhance the heterosis level of BT-type <i>japonica</i> hybrids. In the present study, ‘02428’, a wide-compatibility <i>japonica</i> variety, was observed to partially restore fertility to BT-type CMS lines. Genetic analysis revealed that ‘02428’ carries a dominant <i>Rf</i> gene, <i>Rf21(t)</i>, responsible for the fertility restoration of BT-type CMS lines. Leveraging bulked segregant analysis (BSA) resequencing technology and molecular markers, the <i>Rf21(t)</i> locus was identified, and mapped within a candidate interval of 6–12.5 Mb on chromosome 2. Using the iso-cytoplasmic restorer populations, <i>Rf21(t)</i> was ultimately mapped to an interval of approximately 77 kb, encompassing 12 predicted genes, including <i>LOC_Os02g17360</i>, encoding a PPR-domain-containing protein and <i>LOC_Os02g17380 (Rf2)</i>, a cloned <i>Rf</i> for Lead-rice-type CMS. A comparative sequence analysis, gene expression profiling and gene knockout experiments confirmed that <i>LOC_Os02g17360</i> and <i>LOC_Os02g17380</i> are the most likely candidates of <i>Rf21(t)</i>. Furthermore, <i>Rf21(t)</i> showed the dosage effect on the fertility restoration of BT-type CMS lines. This newly identified <i>Rf21(t)</i> represents a valuable genetic resource for the breeding of BT-type <i>japonica</i> restorer lines. Our findings offer practical insights for breeders interested in advancing BT-type <i>japonica</i> hybrid development.https://www.mdpi.com/2073-4395/15/7/1690<i>japonica</i>BT-type CMSfertility restorer gene (<i>Rf</i>)gene mapping
spellingShingle Yuanyue Du
Liying Fan
Yunhua Gu
Chen Wang
Kai Shi
Yebin Qin
Zhejun Li
Qiaoquan Liu
Shuzhu Tang
Honggen Zhang
Zuopeng Xu
Fine Mapping and Genetic Effect Analysis of <i>Rf21(t)</i> for the Fertility Restoration of Chinsurah-Boro-II-Type Cytoplasmic Male Sterile <i>Oryza sativa</i> (ssp. <i>japonica</i>) Lines
Agronomy
<i>japonica</i>
BT-type CMS
fertility restorer gene (<i>Rf</i>)
gene mapping
title Fine Mapping and Genetic Effect Analysis of <i>Rf21(t)</i> for the Fertility Restoration of Chinsurah-Boro-II-Type Cytoplasmic Male Sterile <i>Oryza sativa</i> (ssp. <i>japonica</i>) Lines
title_full Fine Mapping and Genetic Effect Analysis of <i>Rf21(t)</i> for the Fertility Restoration of Chinsurah-Boro-II-Type Cytoplasmic Male Sterile <i>Oryza sativa</i> (ssp. <i>japonica</i>) Lines
title_fullStr Fine Mapping and Genetic Effect Analysis of <i>Rf21(t)</i> for the Fertility Restoration of Chinsurah-Boro-II-Type Cytoplasmic Male Sterile <i>Oryza sativa</i> (ssp. <i>japonica</i>) Lines
title_full_unstemmed Fine Mapping and Genetic Effect Analysis of <i>Rf21(t)</i> for the Fertility Restoration of Chinsurah-Boro-II-Type Cytoplasmic Male Sterile <i>Oryza sativa</i> (ssp. <i>japonica</i>) Lines
title_short Fine Mapping and Genetic Effect Analysis of <i>Rf21(t)</i> for the Fertility Restoration of Chinsurah-Boro-II-Type Cytoplasmic Male Sterile <i>Oryza sativa</i> (ssp. <i>japonica</i>) Lines
title_sort fine mapping and genetic effect analysis of i rf21 t i for the fertility restoration of chinsurah boro ii type cytoplasmic male sterile i oryza sativa i ssp i japonica i lines
topic <i>japonica</i>
BT-type CMS
fertility restorer gene (<i>Rf</i>)
gene mapping
url https://www.mdpi.com/2073-4395/15/7/1690
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