An Agrobacterium-mediated transformation via organogenesis regeneration of a facultative CAM plant, the common ice plant Mesembryanthemum crystallinum L

The common ice plant, Mesembryanthemum crystallinum L. provides a useful model for the study of environmentally induced photosynthetic conversion and abiotic stresses tolerance. However, a procedure for the production of transgenic ice plant, which is essential for functional genomics, has not been...

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Main Authors: Sakae Agarie, Makiko Umemoto, Haruki Sunagawa, Toyoaki Anai, John C. Cushman
Format: Article
Language:English
Published: Taylor & Francis Group 2020-07-01
Series:Plant Production Science
Subjects:
Online Access:http://dx.doi.org/10.1080/1343943X.2020.1730700
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author Sakae Agarie
Makiko Umemoto
Haruki Sunagawa
Toyoaki Anai
John C. Cushman
author_facet Sakae Agarie
Makiko Umemoto
Haruki Sunagawa
Toyoaki Anai
John C. Cushman
author_sort Sakae Agarie
collection DOAJ
description The common ice plant, Mesembryanthemum crystallinum L. provides a useful model for the study of environmentally induced photosynthetic conversion and abiotic stresses tolerance. However, a procedure for the production of transgenic ice plant, which is essential for functional genomics, has not been fully established. Here we tested the factors on the transformation of cotyledonary nodes excised from the ice plant seedlings such as thidiazuron (TDZ), NaCl and phytosulfokine (PSK), a peptidyl plant growth factor using Agrobacterium tumefaciens strains EHA101 and EHA105 harboring binary vector plasmids pBI7EGFP and pCAMBIA1302, respectively. The established procedure is as follows: the explants (cotyledonary nodes) were co-cultivated with Agrobacterium for 3 days, and the explants were cultured in the medium with 0.5 mg l−1 kinetin and 100 mg l−1 carbenicillin for 72 h, and they were cultured in the medium with 0.5 mg l−1 kinetin and 100 nM PSKfor 4 weeks. Thidiazuron and NaCl enhanced the production of multiple adventitious shoot formation during regeneration but reduced the transformation efficiency due to the vitrification of adventitious shoots. PSK was effective in the production of healthy adventitious shoots. The transformation frequency at the stage of whole plants was 0.6% and 4.6% per inoculated cotyledonary nodes using the Agrobacterium strain EHA101 (pBI7EGFP) and EHA105 (pCAMBIA1302), respectively.
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spelling doaj.art-e8afd90053204afcaa33f500627b193a2022-12-22T00:21:05ZengTaylor & Francis GroupPlant Production Science1343-943X1349-10082020-07-0123334334910.1080/1343943X.2020.17307001730700An Agrobacterium-mediated transformation via organogenesis regeneration of a facultative CAM plant, the common ice plant Mesembryanthemum crystallinum LSakae Agarie0Makiko Umemoto1Haruki Sunagawa2Toyoaki Anai3John C. Cushman4Kyushu UniversitySaga UniversitySaga UniversitySaga UniversityUniversity of NevadaThe common ice plant, Mesembryanthemum crystallinum L. provides a useful model for the study of environmentally induced photosynthetic conversion and abiotic stresses tolerance. However, a procedure for the production of transgenic ice plant, which is essential for functional genomics, has not been fully established. Here we tested the factors on the transformation of cotyledonary nodes excised from the ice plant seedlings such as thidiazuron (TDZ), NaCl and phytosulfokine (PSK), a peptidyl plant growth factor using Agrobacterium tumefaciens strains EHA101 and EHA105 harboring binary vector plasmids pBI7EGFP and pCAMBIA1302, respectively. The established procedure is as follows: the explants (cotyledonary nodes) were co-cultivated with Agrobacterium for 3 days, and the explants were cultured in the medium with 0.5 mg l−1 kinetin and 100 mg l−1 carbenicillin for 72 h, and they were cultured in the medium with 0.5 mg l−1 kinetin and 100 nM PSKfor 4 weeks. Thidiazuron and NaCl enhanced the production of multiple adventitious shoot formation during regeneration but reduced the transformation efficiency due to the vitrification of adventitious shoots. PSK was effective in the production of healthy adventitious shoots. The transformation frequency at the stage of whole plants was 0.6% and 4.6% per inoculated cotyledonary nodes using the Agrobacterium strain EHA101 (pBI7EGFP) and EHA105 (pCAMBIA1302), respectively.http://dx.doi.org/10.1080/1343943X.2020.1730700agrobacteriumcamcommon ice plantorganogenesistransformation
spellingShingle Sakae Agarie
Makiko Umemoto
Haruki Sunagawa
Toyoaki Anai
John C. Cushman
An Agrobacterium-mediated transformation via organogenesis regeneration of a facultative CAM plant, the common ice plant Mesembryanthemum crystallinum L
Plant Production Science
agrobacterium
cam
common ice plant
organogenesis
transformation
title An Agrobacterium-mediated transformation via organogenesis regeneration of a facultative CAM plant, the common ice plant Mesembryanthemum crystallinum L
title_full An Agrobacterium-mediated transformation via organogenesis regeneration of a facultative CAM plant, the common ice plant Mesembryanthemum crystallinum L
title_fullStr An Agrobacterium-mediated transformation via organogenesis regeneration of a facultative CAM plant, the common ice plant Mesembryanthemum crystallinum L
title_full_unstemmed An Agrobacterium-mediated transformation via organogenesis regeneration of a facultative CAM plant, the common ice plant Mesembryanthemum crystallinum L
title_short An Agrobacterium-mediated transformation via organogenesis regeneration of a facultative CAM plant, the common ice plant Mesembryanthemum crystallinum L
title_sort agrobacterium mediated transformation via organogenesis regeneration of a facultative cam plant the common ice plant mesembryanthemum crystallinum l
topic agrobacterium
cam
common ice plant
organogenesis
transformation
url http://dx.doi.org/10.1080/1343943X.2020.1730700
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