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Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callus
BACKGROUND: Large T-DNA fragment transfer has long been a problem for Agrobacterium-mediated transformation. Although vector systems, such as the BIBAC series, were successfully developed for the purpose, low transformation efficiencies were consistently observed. RESULTS: To gain insights of this p...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6085696/ https://www.ncbi.nlm.nih.gov/pubmed/30092808 http://dx.doi.org/10.1186/s12896-018-0459-5 |
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author | Xi, Jing Patel, Minesh Dong, Shujie Que, Qiudeng Qu, Rongda |
author_facet | Xi, Jing Patel, Minesh Dong, Shujie Que, Qiudeng Qu, Rongda |
author_sort | Xi, Jing |
collection | PubMed |
description | BACKGROUND: Large T-DNA fragment transfer has long been a problem for Agrobacterium-mediated transformation. Although vector systems, such as the BIBAC series, were successfully developed for the purpose, low transformation efficiencies were consistently observed. RESULTS: To gain insights of this problem in monocot transformation, we investigated the T-strand accumulation of various size of T-DNA in two kinds of binary vectors (one copy vs. multi-copy) upon acetosyringone (AS) induction and explored ways to improve the efficiency of the large T-DNA fragment transfer in Agrobacterium-mediated rice transformation. By performing immuno-precipitation of VirD2-T-strands and quantitative real-time PCR assays, we monitored the accumulation of the T-strands in Agrobacterium tumeficiens after AS induction. We further demonstrated that extension of AS induction time highly significantly improved large-size T-DNA transfer to rice cells. CONCLUSIONS: Our data provide valuable information of the T-strand dynamics and its impact on large T-DNA transfer in monocots, and likely dicots as well. |
format | Online Article Text |
id | pubmed-6085696 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-60856962018-08-16 Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callus Xi, Jing Patel, Minesh Dong, Shujie Que, Qiudeng Qu, Rongda BMC Biotechnol Research BACKGROUND: Large T-DNA fragment transfer has long been a problem for Agrobacterium-mediated transformation. Although vector systems, such as the BIBAC series, were successfully developed for the purpose, low transformation efficiencies were consistently observed. RESULTS: To gain insights of this problem in monocot transformation, we investigated the T-strand accumulation of various size of T-DNA in two kinds of binary vectors (one copy vs. multi-copy) upon acetosyringone (AS) induction and explored ways to improve the efficiency of the large T-DNA fragment transfer in Agrobacterium-mediated rice transformation. By performing immuno-precipitation of VirD2-T-strands and quantitative real-time PCR assays, we monitored the accumulation of the T-strands in Agrobacterium tumeficiens after AS induction. We further demonstrated that extension of AS induction time highly significantly improved large-size T-DNA transfer to rice cells. CONCLUSIONS: Our data provide valuable information of the T-strand dynamics and its impact on large T-DNA transfer in monocots, and likely dicots as well. BioMed Central 2018-08-09 /pmc/articles/PMC6085696/ /pubmed/30092808 http://dx.doi.org/10.1186/s12896-018-0459-5 Text en © The Author(s). 2018 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. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Xi, Jing Patel, Minesh Dong, Shujie Que, Qiudeng Qu, Rongda Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callus |
title | Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callus |
title_full | Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callus |
title_fullStr | Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callus |
title_full_unstemmed | Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callus |
title_short | Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callus |
title_sort | acetosyringone treatment duration affects large t-dna molecule transfer to rice callus |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6085696/ https://www.ncbi.nlm.nih.gov/pubmed/30092808 http://dx.doi.org/10.1186/s12896-018-0459-5 |
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