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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...

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Autores principales: Yang, Hsiu-Chun, Charng, Yuh-Chyang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2014
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.
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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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