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Map-based cloning of a recessive genic male sterility locus in Brassica napus L. and development of its functional marker.

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Title: Map-based cloning of a recessive genic male sterility locus in Brassica napus L. and development of its functional marker.
Authors: Li, Ji1, Hong, Dengfeng1, He, Junping, Ma, Lei, Wan, Lili1, Liu, Pingwu1, Yang, Guangsheng1 gsyang@mail.hzau.edu.cn
Source: Theoretical & Applied Genetics. Jul2012, Vol. 125 Issue 2, p223-234. 12p.
Subjects: Brassica, Genetic research, Gene mapping, Male sterility in plants, Open reading frames (Genetics), Chloroplasts, Plant breeding, Molecular cloning, Locus in plant genetics
Abstract: We previously mapped one male-sterile gene ( Bnms3) from an extensively used recessive genic male sterility line (9012AB) in Brassica napus to a 0.14-cM genomic region. In this study, two highly homologous BAC contigs possibly containing the candidate BnMs3 gene were identified using a map-based cloning strategy. A BnMs3-linked SCAR marker (DM1) capable of differentiating the subgenomes between B. rapa and the B. oleracea aided mapping of BnMs3 on the contig derived from the B. napus chromosome C9. One representative BAC clone was sequenced from each of the two contigs and resulted in a larger number of markers according to the sequence difference between the two clones. To isolate BnMs3, these markers were then analyzed in another two BC populations with different genetic backgrounds. This assay allowed for a delimitation of the mutated functional region of BnMs3 to a 9.3-kb DNA fragment. Gene prediction suggested that one complete open reading frame (ORF, ORF2) and partial CDS fragments of ORF1 and ORF3 reside in this fragment. Sequence comparison and genetic transformation eventually indicated that ORF1 (designated as BnaC9.Tic40), an analogue of the Arabidopsis gene AT5G16620 which encodes a translocon of the inner envelope of chloroplasts 40 (Tic40), is the only candidate gene of BnMs3. Furthermore, two distinct mutation types in ORF1 both causing the male-sterile phenotype were individually revealed from 9012A and the temporary maintainer line T45. The molecular mechanism of this male sterility as well as the application of BnMs3-associated functional and cosegregated markers in true breeding programs was also discussed. [ABSTRACT FROM AUTHOR]
Copyright of Theoretical & Applied Genetics is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: Map-based cloning of a recessive genic male sterility locus in Brassica napus L. and development of its functional marker.
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  Data: <searchLink fieldCode="AR" term="%22Li%2C+Ji%22">Li, Ji</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Hong%2C+Dengfeng%22">Hong, Dengfeng</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22He%2C+Junping%22">He, Junping</searchLink><br /><searchLink fieldCode="AR" term="%22Ma%2C+Lei%22">Ma, Lei</searchLink><br /><searchLink fieldCode="AR" term="%22Wan%2C+Lili%22">Wan, Lili</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Liu%2C+Pingwu%22">Liu, Pingwu</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Yang%2C+Guangsheng%22">Yang, Guangsheng</searchLink><relatesTo>1</relatesTo><i> gsyang@mail.hzau.edu.cn</i>
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  Data: <searchLink fieldCode="DE" term="%22Brassica%22">Brassica</searchLink><br /><searchLink fieldCode="DE" term="%22Genetic+research%22">Genetic research</searchLink><br /><searchLink fieldCode="DE" term="%22Gene+mapping%22">Gene mapping</searchLink><br /><searchLink fieldCode="DE" term="%22Male+sterility+in+plants%22">Male sterility in plants</searchLink><br /><searchLink fieldCode="DE" term="%22Open+reading+frames+%28Genetics%29%22">Open reading frames (Genetics)</searchLink><br /><searchLink fieldCode="DE" term="%22Chloroplasts%22">Chloroplasts</searchLink><br /><searchLink fieldCode="DE" term="%22Plant+breeding%22">Plant breeding</searchLink><br /><searchLink fieldCode="DE" term="%22Molecular+cloning%22">Molecular cloning</searchLink><br /><searchLink fieldCode="DE" term="%22Locus+in+plant+genetics%22">Locus in plant genetics</searchLink>
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  Data: We previously mapped one male-sterile gene ( Bnms3) from an extensively used recessive genic male sterility line (9012AB) in Brassica napus to a 0.14-cM genomic region. In this study, two highly homologous BAC contigs possibly containing the candidate BnMs3 gene were identified using a map-based cloning strategy. A BnMs3-linked SCAR marker (DM1) capable of differentiating the subgenomes between B. rapa and the B. oleracea aided mapping of BnMs3 on the contig derived from the B. napus chromosome C9. One representative BAC clone was sequenced from each of the two contigs and resulted in a larger number of markers according to the sequence difference between the two clones. To isolate BnMs3, these markers were then analyzed in another two BC populations with different genetic backgrounds. This assay allowed for a delimitation of the mutated functional region of BnMs3 to a 9.3-kb DNA fragment. Gene prediction suggested that one complete open reading frame (ORF, ORF2) and partial CDS fragments of ORF1 and ORF3 reside in this fragment. Sequence comparison and genetic transformation eventually indicated that ORF1 (designated as BnaC9.Tic40), an analogue of the Arabidopsis gene AT5G16620 which encodes a translocon of the inner envelope of chloroplasts 40 (Tic40), is the only candidate gene of BnMs3. Furthermore, two distinct mutation types in ORF1 both causing the male-sterile phenotype were individually revealed from 9012A and the temporary maintainer line T45. The molecular mechanism of this male sterility as well as the application of BnMs3-associated functional and cosegregated markers in true breeding programs was also discussed. [ABSTRACT FROM AUTHOR]
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  Data: <i>Copyright of Theoretical & Applied Genetics is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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      – SubjectFull: Brassica
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      – SubjectFull: Gene mapping
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      – SubjectFull: Molecular cloning
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      – SubjectFull: Locus in plant genetics
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      – TitleFull: Map-based cloning of a recessive genic male sterility locus in Brassica napus L. and development of its functional marker.
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