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Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application

Sorghum is the fifth most widely planted cereal crop in the world and is commonly cultivated in arid and semi‐arid regions such as Africa. Despite its importance as a food source, sorghum genetic improvement through transgenic approaches has been limited because of an inefficient transformation syst...

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Autores principales: Che, Ping, Anand, Ajith, Wu, Emily, Sander, Jeffry D., Simon, Marissa K., Zhu, Weiwei, Sigmund, Amy L., Zastrow‐Hayes, Gina, Miller, Michael, Liu, Donglong, Lawit, Shai J., Zhao, Zuo‐Yu, Albertsen, Marc C., Jones, Todd J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5999184/
https://www.ncbi.nlm.nih.gov/pubmed/29327444
http://dx.doi.org/10.1111/pbi.12879
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author Che, Ping
Anand, Ajith
Wu, Emily
Sander, Jeffry D.
Simon, Marissa K.
Zhu, Weiwei
Sigmund, Amy L.
Zastrow‐Hayes, Gina
Miller, Michael
Liu, Donglong
Lawit, Shai J.
Zhao, Zuo‐Yu
Albertsen, Marc C.
Jones, Todd J.
author_facet Che, Ping
Anand, Ajith
Wu, Emily
Sander, Jeffry D.
Simon, Marissa K.
Zhu, Weiwei
Sigmund, Amy L.
Zastrow‐Hayes, Gina
Miller, Michael
Liu, Donglong
Lawit, Shai J.
Zhao, Zuo‐Yu
Albertsen, Marc C.
Jones, Todd J.
author_sort Che, Ping
collection PubMed
description Sorghum is the fifth most widely planted cereal crop in the world and is commonly cultivated in arid and semi‐arid regions such as Africa. Despite its importance as a food source, sorghum genetic improvement through transgenic approaches has been limited because of an inefficient transformation system. Here, we report a ternary vector (also known as cohabitating vector) system using a recently described pVIR accessory plasmid that facilitates efficient Agrobacterium‐mediated transformation of sorghum. We report regeneration frequencies ranging from 6% to 29% in Tx430 using different selectable markers and single copy, backbone free ‘quality events’ ranging from 45% to 66% of the total events produced. Furthermore, we successfully applied this ternary system to develop transformation protocols for popular but recalcitrant African varieties including Macia, Malisor 84‐7 and Tegemeo. In addition, we report the use of this technology to develop the first stable CRISPR/Cas9‐mediated gene knockouts in Tx430.
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spelling pubmed-59991842018-07-12 Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application Che, Ping Anand, Ajith Wu, Emily Sander, Jeffry D. Simon, Marissa K. Zhu, Weiwei Sigmund, Amy L. Zastrow‐Hayes, Gina Miller, Michael Liu, Donglong Lawit, Shai J. Zhao, Zuo‐Yu Albertsen, Marc C. Jones, Todd J. Plant Biotechnol J Research Articles Sorghum is the fifth most widely planted cereal crop in the world and is commonly cultivated in arid and semi‐arid regions such as Africa. Despite its importance as a food source, sorghum genetic improvement through transgenic approaches has been limited because of an inefficient transformation system. Here, we report a ternary vector (also known as cohabitating vector) system using a recently described pVIR accessory plasmid that facilitates efficient Agrobacterium‐mediated transformation of sorghum. We report regeneration frequencies ranging from 6% to 29% in Tx430 using different selectable markers and single copy, backbone free ‘quality events’ ranging from 45% to 66% of the total events produced. Furthermore, we successfully applied this ternary system to develop transformation protocols for popular but recalcitrant African varieties including Macia, Malisor 84‐7 and Tegemeo. In addition, we report the use of this technology to develop the first stable CRISPR/Cas9‐mediated gene knockouts in Tx430. John Wiley and Sons Inc. 2018-02-06 2018-07 /pmc/articles/PMC5999184/ /pubmed/29327444 http://dx.doi.org/10.1111/pbi.12879 Text en © 2018 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Che, Ping
Anand, Ajith
Wu, Emily
Sander, Jeffry D.
Simon, Marissa K.
Zhu, Weiwei
Sigmund, Amy L.
Zastrow‐Hayes, Gina
Miller, Michael
Liu, Donglong
Lawit, Shai J.
Zhao, Zuo‐Yu
Albertsen, Marc C.
Jones, Todd J.
Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application
title Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application
title_full Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application
title_fullStr Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application
title_full_unstemmed Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application
title_short Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application
title_sort developing a flexible, high‐efficiency agrobacterium‐mediated sorghum transformation system with broad application
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5999184/
https://www.ncbi.nlm.nih.gov/pubmed/29327444
http://dx.doi.org/10.1111/pbi.12879
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