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Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants

Cowpea (Vigna unguiculata (L.) Walp.) is one of the most important legume crops planted worldwide, but despite decades of effort, cowpea transformation is still challenging due to inefficient Agrobacterium‐mediated transfer DNA delivery, transgenic selection and in vitro shoot regeneration. Here, we...

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Autores principales: Che, Ping, Chang, Shujun, Simon, Marissa K., Zhang, Zhifen, Shaharyar, Ahmed, Ourada, Jesse, O’Neill, Dennis, Torres‐Mendoza, Mijael, Guo, Yinping, Marasigan, Kathleen M., Vielle‐Calzada, Jean‐Philippe, Ozias‐Akins, Peggy, Albertsen, Marc C., Jones, Todd J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8252785/
https://www.ncbi.nlm.nih.gov/pubmed/33595147
http://dx.doi.org/10.1111/tpj.15202
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author Che, Ping
Chang, Shujun
Simon, Marissa K.
Zhang, Zhifen
Shaharyar, Ahmed
Ourada, Jesse
O’Neill, Dennis
Torres‐Mendoza, Mijael
Guo, Yinping
Marasigan, Kathleen M.
Vielle‐Calzada, Jean‐Philippe
Ozias‐Akins, Peggy
Albertsen, Marc C.
Jones, Todd J.
author_facet Che, Ping
Chang, Shujun
Simon, Marissa K.
Zhang, Zhifen
Shaharyar, Ahmed
Ourada, Jesse
O’Neill, Dennis
Torres‐Mendoza, Mijael
Guo, Yinping
Marasigan, Kathleen M.
Vielle‐Calzada, Jean‐Philippe
Ozias‐Akins, Peggy
Albertsen, Marc C.
Jones, Todd J.
author_sort Che, Ping
collection PubMed
description Cowpea (Vigna unguiculata (L.) Walp.) is one of the most important legume crops planted worldwide, but despite decades of effort, cowpea transformation is still challenging due to inefficient Agrobacterium‐mediated transfer DNA delivery, transgenic selection and in vitro shoot regeneration. Here, we report a highly efficient transformation system using embryonic axis explants isolated from imbibed mature seeds. We found that removal of the shoot apical meristem from the explants stimulated direct multiple shoot organogenesis from the cotyledonary node tissue. The application of a previously reported ternary transformation vector system provided efficient Agrobacterium‐mediated gene delivery, while the utilization of spcN as selectable marker enabled more robust transgenic selection, plant recovery and transgenic plant generation without escapes and chimera formation. Transgenic cowpea plantlets developed exclusively from the cotyledonary nodes at frequencies of 4% to 37% across a wide range of cowpea genotypes. CRISPR/Cas‐mediated gene editing was successfully demonstrated. The transformation principles established here could also be applied to other legumes to increase transformation efficiencies.
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spelling pubmed-82527852021-07-12 Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants Che, Ping Chang, Shujun Simon, Marissa K. Zhang, Zhifen Shaharyar, Ahmed Ourada, Jesse O’Neill, Dennis Torres‐Mendoza, Mijael Guo, Yinping Marasigan, Kathleen M. Vielle‐Calzada, Jean‐Philippe Ozias‐Akins, Peggy Albertsen, Marc C. Jones, Todd J. Plant J Original Articles Cowpea (Vigna unguiculata (L.) Walp.) is one of the most important legume crops planted worldwide, but despite decades of effort, cowpea transformation is still challenging due to inefficient Agrobacterium‐mediated transfer DNA delivery, transgenic selection and in vitro shoot regeneration. Here, we report a highly efficient transformation system using embryonic axis explants isolated from imbibed mature seeds. We found that removal of the shoot apical meristem from the explants stimulated direct multiple shoot organogenesis from the cotyledonary node tissue. The application of a previously reported ternary transformation vector system provided efficient Agrobacterium‐mediated gene delivery, while the utilization of spcN as selectable marker enabled more robust transgenic selection, plant recovery and transgenic plant generation without escapes and chimera formation. Transgenic cowpea plantlets developed exclusively from the cotyledonary nodes at frequencies of 4% to 37% across a wide range of cowpea genotypes. CRISPR/Cas‐mediated gene editing was successfully demonstrated. The transformation principles established here could also be applied to other legumes to increase transformation efficiencies. John Wiley and Sons Inc. 2021-03-16 2021-05 /pmc/articles/PMC8252785/ /pubmed/33595147 http://dx.doi.org/10.1111/tpj.15202 Text en © 2021 Corteva Agriscience. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Che, Ping
Chang, Shujun
Simon, Marissa K.
Zhang, Zhifen
Shaharyar, Ahmed
Ourada, Jesse
O’Neill, Dennis
Torres‐Mendoza, Mijael
Guo, Yinping
Marasigan, Kathleen M.
Vielle‐Calzada, Jean‐Philippe
Ozias‐Akins, Peggy
Albertsen, Marc C.
Jones, Todd J.
Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants
title Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants
title_full Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants
title_fullStr Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants
title_full_unstemmed Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants
title_short Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants
title_sort developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8252785/
https://www.ncbi.nlm.nih.gov/pubmed/33595147
http://dx.doi.org/10.1111/tpj.15202
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