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Gain-of-function in Arabidopsis (GAINA) for identifying functional genes in Hevea brasiliensis

BACKGROUND: Forward genetics approaches are not popularly applied in non-model plants due to their complex genomes, long life cycles, backward genetic studies etc. Researchers have to adopt reverse genetic methods to characterize gene functions in non-model plants individually, the efficiency of whi...

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Autores principales: Cheng, Han, Gao, Jing, Cai, Haibin, Zhu, Jianshun, Huang, Huasun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5075328/
https://www.ncbi.nlm.nih.gov/pubmed/27818891
http://dx.doi.org/10.1186/s40064-016-3523-4
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author Cheng, Han
Gao, Jing
Cai, Haibin
Zhu, Jianshun
Huang, Huasun
author_facet Cheng, Han
Gao, Jing
Cai, Haibin
Zhu, Jianshun
Huang, Huasun
author_sort Cheng, Han
collection PubMed
description BACKGROUND: Forward genetics approaches are not popularly applied in non-model plants due to their complex genomes, long life cycles, backward genetic studies etc. Researchers have to adopt reverse genetic methods to characterize gene functions in non-model plants individually, the efficiency of which is usually low. RESULTS: In this study, we report a gain-of-function in Arabidopsis (GAINA) strategy which can be used for batch identification of functional genes in a plant species. This strategy aims to obtain the gain-of-function of rubber tree genes through overexpressing transformation ready full-length cDNA libraries in Arabidopsis. An initial transformation test produced about two thousand independent transgenic Arabidopsis lines, in which multiple obvious aberrant phenotypes were observed, suggesting the gain-of-function of rubber tree genes. The transferred genes were further isolated and identified. One gene identified to be metallothionein-like protein type 3 gene was further transferred into Arabidopsis and reproduced a similar aberrant phenotype. CONCLUSION: The GAINA system proves to be an efficient tool for batch identification of functional genes in Hevea brasiliensis, and also applicable in other non-model plants. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40064-016-3523-4) contains supplementary material, which is available to authorized users.
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spelling pubmed-50753282016-11-04 Gain-of-function in Arabidopsis (GAINA) for identifying functional genes in Hevea brasiliensis Cheng, Han Gao, Jing Cai, Haibin Zhu, Jianshun Huang, Huasun Springerplus Methodology BACKGROUND: Forward genetics approaches are not popularly applied in non-model plants due to their complex genomes, long life cycles, backward genetic studies etc. Researchers have to adopt reverse genetic methods to characterize gene functions in non-model plants individually, the efficiency of which is usually low. RESULTS: In this study, we report a gain-of-function in Arabidopsis (GAINA) strategy which can be used for batch identification of functional genes in a plant species. This strategy aims to obtain the gain-of-function of rubber tree genes through overexpressing transformation ready full-length cDNA libraries in Arabidopsis. An initial transformation test produced about two thousand independent transgenic Arabidopsis lines, in which multiple obvious aberrant phenotypes were observed, suggesting the gain-of-function of rubber tree genes. The transferred genes were further isolated and identified. One gene identified to be metallothionein-like protein type 3 gene was further transferred into Arabidopsis and reproduced a similar aberrant phenotype. CONCLUSION: The GAINA system proves to be an efficient tool for batch identification of functional genes in Hevea brasiliensis, and also applicable in other non-model plants. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40064-016-3523-4) contains supplementary material, which is available to authorized users. Springer International Publishing 2016-10-22 /pmc/articles/PMC5075328/ /pubmed/27818891 http://dx.doi.org/10.1186/s40064-016-3523-4 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Methodology
Cheng, Han
Gao, Jing
Cai, Haibin
Zhu, Jianshun
Huang, Huasun
Gain-of-function in Arabidopsis (GAINA) for identifying functional genes in Hevea brasiliensis
title Gain-of-function in Arabidopsis (GAINA) for identifying functional genes in Hevea brasiliensis
title_full Gain-of-function in Arabidopsis (GAINA) for identifying functional genes in Hevea brasiliensis
title_fullStr Gain-of-function in Arabidopsis (GAINA) for identifying functional genes in Hevea brasiliensis
title_full_unstemmed Gain-of-function in Arabidopsis (GAINA) for identifying functional genes in Hevea brasiliensis
title_short Gain-of-function in Arabidopsis (GAINA) for identifying functional genes in Hevea brasiliensis
title_sort gain-of-function in arabidopsis (gaina) for identifying functional genes in hevea brasiliensis
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5075328/
https://www.ncbi.nlm.nih.gov/pubmed/27818891
http://dx.doi.org/10.1186/s40064-016-3523-4
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