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Application of an inducible transposon with anther culture in generation of di-haploid homologous mutants
BACKGROUND: Insertional mutagenesis represents one of the most effective ways to acquire information about a plant gene’s function. However, it is hindered by the autosomal genome being diploid and therefore, most mutations being recessive. The problem is addressed by inducing the transposition duri...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5432829/ https://www.ncbi.nlm.nih.gov/pubmed/28510931 http://dx.doi.org/10.1186/1999-3110-55-27 |
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author | Yang, Hsiu-Chun Charng, Yuh-Chyang |
author_facet | Yang, Hsiu-Chun Charng, Yuh-Chyang |
author_sort | Yang, Hsiu-Chun |
collection | PubMed |
description | BACKGROUND: Insertional mutagenesis represents one of the most effective ways to acquire information about a plant gene’s function. However, it is hindered by the autosomal genome being diploid and therefore, most mutations being recessive. The problem is addressed by inducing the transposition during anther culture so that selected mutations can be transmitted and then regenerated to a homozygous state. RESULTS: To this end, we treated transgenic rice floral tissues containing the inducible transposon with an inducer, salicylic acid. Excision events were detected in regenerated calli and subsequent plantlets. DNA blot and PCR assay were used to determine the homogeneity of knockout mutants. About 5% of the mutants containing transposition events were homozygous. Furthermore, the inducible transposon was active during calli regeneration. CONCLUSIONS: This strategy could be applicable to improve transposition efficiency in microspore development stages to create stable di-haploid mutants in plants. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1999-3110-55-27) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5432829 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-54328292017-05-31 Application of an inducible transposon with anther culture in generation of di-haploid homologous mutants Yang, Hsiu-Chun Charng, Yuh-Chyang Bot Stud Research BACKGROUND: Insertional mutagenesis represents one of the most effective ways to acquire information about a plant gene’s function. However, it is hindered by the autosomal genome being diploid and therefore, most mutations being recessive. The problem is addressed by inducing the transposition during anther culture so that selected mutations can be transmitted and then regenerated to a homozygous state. RESULTS: To this end, we treated transgenic rice floral tissues containing the inducible transposon with an inducer, salicylic acid. Excision events were detected in regenerated calli and subsequent plantlets. DNA blot and PCR assay were used to determine the homogeneity of knockout mutants. About 5% of the mutants containing transposition events were homozygous. Furthermore, the inducible transposon was active during calli regeneration. CONCLUSIONS: This strategy could be applicable to improve transposition efficiency in microspore development stages to create stable di-haploid mutants in plants. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1999-3110-55-27) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2014-02-24 /pmc/articles/PMC5432829/ /pubmed/28510931 http://dx.doi.org/10.1186/1999-3110-55-27 Text en © Yang and Charng; licensee Springer. 2014 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. |
spellingShingle | Research Yang, Hsiu-Chun Charng, Yuh-Chyang Application of an inducible transposon with anther culture in generation of di-haploid homologous mutants |
title | Application of an inducible transposon with anther culture in generation of di-haploid homologous mutants |
title_full | Application of an inducible transposon with anther culture in generation of di-haploid homologous mutants |
title_fullStr | Application of an inducible transposon with anther culture in generation of di-haploid homologous mutants |
title_full_unstemmed | Application of an inducible transposon with anther culture in generation of di-haploid homologous mutants |
title_short | Application of an inducible transposon with anther culture in generation of di-haploid homologous mutants |
title_sort | application of an inducible transposon with anther culture in generation of di-haploid homologous mutants |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5432829/ https://www.ncbi.nlm.nih.gov/pubmed/28510931 http://dx.doi.org/10.1186/1999-3110-55-27 |
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