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Agrobacterium-mediated gene delivery and transient expression in the red macroalga Chondrus crispus

Molecular resources and transgenic studies in red algae are lagging behind those for green algae. The Agrobacterium-mediated gene-transfer method routinely used in plant transformation has not been fully utilised in the red algae, which, as an important source of phycocolloids, warrant more studies....

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Main Authors: Ramessur, Anusha Devi, Bothwell, John H., Maggs, Christine A., Gan, Sook Yee, Phang, Siew Moi
Format: Article
Published: De Gruyter 2018
Subjects:
Q Science (General)
QH Natural history
Online Access:http://eprints.um.edu.my/20929/
https://doi.org/10.1515/bot-2018-0028
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spelling my.um.eprints.209292019-04-16T07:33:52Z http://eprints.um.edu.my/20929/ Agrobacterium-mediated gene delivery and transient expression in the red macroalga Chondrus crispus Ramessur, Anusha Devi Bothwell, John H. Maggs, Christine A. Gan, Sook Yee Phang, Siew Moi Q Science (General) QH Natural history Molecular resources and transgenic studies in red algae are lagging behind those for green algae. The Agrobacterium-mediated gene-transfer method routinely used in plant transformation has not been fully utilised in the red algae, which, as an important source of phycocolloids, warrant more studies. In this regard, a stepwise methodology was developed for Agrobacterium-mediated transformation of the carrageenophyte Chondrus crispus using pCAMBIA 1301 and a construct featuring a codon-optimized beta-glucuronidase (GUS) reporter gene driven by the endogenous Chondrus actin promoter. The effects of several factors on transformation efficiency were investigated. An intimate association of Chondrus and bacterial cells was observed using scanning electron microscopy. GUS transient expression within Chondrus cortical and medullary cells with both expression cassettes testified to the amenability of Chondrus to Agrobacterium-mediated transformation. Darker staining, indicative of higher GUS activity, was observed with the Chondrus-specific construct, suggesting its superiority over the pCAMBIA 1301. Presence of acetosyringone, the wounding method and the type of co-cultivation medium significantly affected the transformation outcome and efficiency. The Agrobacterium-mediated transient expression presented here constitutes a first step towards tailoring a transformation strategy for Chondrus, which can serve to facilitate further transgenic studies in this important red alga. De Gruyter 2018 Article PeerReviewed Ramessur, Anusha Devi and Bothwell, John H. and Maggs, Christine A. and Gan, Sook Yee and Phang, Siew Moi (2018) Agrobacterium-mediated gene delivery and transient expression in the red macroalga Chondrus crispus. Botanica Marina, 61 (5). pp. 499-510. ISSN 0006-8055 https://doi.org/10.1515/bot-2018-0028 doi:10.1515/bot-2018-0028
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
topic Q Science (General)
QH Natural history
spellingShingle Q Science (General)
QH Natural history
Ramessur, Anusha Devi
Bothwell, John H.
Maggs, Christine A.
Gan, Sook Yee
Phang, Siew Moi
Agrobacterium-mediated gene delivery and transient expression in the red macroalga Chondrus crispus
description Molecular resources and transgenic studies in red algae are lagging behind those for green algae. The Agrobacterium-mediated gene-transfer method routinely used in plant transformation has not been fully utilised in the red algae, which, as an important source of phycocolloids, warrant more studies. In this regard, a stepwise methodology was developed for Agrobacterium-mediated transformation of the carrageenophyte Chondrus crispus using pCAMBIA 1301 and a construct featuring a codon-optimized beta-glucuronidase (GUS) reporter gene driven by the endogenous Chondrus actin promoter. The effects of several factors on transformation efficiency were investigated. An intimate association of Chondrus and bacterial cells was observed using scanning electron microscopy. GUS transient expression within Chondrus cortical and medullary cells with both expression cassettes testified to the amenability of Chondrus to Agrobacterium-mediated transformation. Darker staining, indicative of higher GUS activity, was observed with the Chondrus-specific construct, suggesting its superiority over the pCAMBIA 1301. Presence of acetosyringone, the wounding method and the type of co-cultivation medium significantly affected the transformation outcome and efficiency. The Agrobacterium-mediated transient expression presented here constitutes a first step towards tailoring a transformation strategy for Chondrus, which can serve to facilitate further transgenic studies in this important red alga.
format Article
author Ramessur, Anusha Devi
Bothwell, John H.
Maggs, Christine A.
Gan, Sook Yee
Phang, Siew Moi
author_facet Ramessur, Anusha Devi
Bothwell, John H.
Maggs, Christine A.
Gan, Sook Yee
Phang, Siew Moi
author_sort Ramessur, Anusha Devi
title Agrobacterium-mediated gene delivery and transient expression in the red macroalga Chondrus crispus
title_short Agrobacterium-mediated gene delivery and transient expression in the red macroalga Chondrus crispus
title_full Agrobacterium-mediated gene delivery and transient expression in the red macroalga Chondrus crispus
title_fullStr Agrobacterium-mediated gene delivery and transient expression in the red macroalga Chondrus crispus
title_full_unstemmed Agrobacterium-mediated gene delivery and transient expression in the red macroalga Chondrus crispus
title_sort agrobacterium-mediated gene delivery and transient expression in the red macroalga chondrus crispus
publisher De Gruyter
publishDate 2018
url http://eprints.um.edu.my/20929/
https://doi.org/10.1515/bot-2018-0028
_version_ 1643691421253763072
score 13.211869

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