Heterologous Expression of Carboxysomes in Plants

TitleHeterologous Expression of Carboxysomes in Plants
Brief TitleCarboxysomes in Plants
Collection TypeDataset
Access PrivilegesRIPE Lab
DOI - Digital Object Identifier10.25911/5b4edb4bada74
Metadata LanguageEnglish
Data LanguageEnglish
Significance StatementThis dataset reports the first successful expression of encapsulated Rubisco enzymes in simplified carboxysomes within tobacco chloroplasts. This work forms the basis of attempts to generate functional CO2 concentrating mechanisms within C3 plant chloroplasts to enhance photosynthesis and yield.
Brief DescriptionA long-term strategy to enhance global crop photosynthesis and yield involves the introduction of cyanobacterial CO2-concentrating-mechanisms (CCMs) into plant chloroplasts. Cyanobacterial CCMs enable relatively rapid CO2-fixation by elevating intracellular inorganic carbon as bicarbonate, then concentrating it as CO2 around the enzyme Rubisco in specialized protein micro-compartments called carboxysomes. To date, chloroplastic expression of carboxysomes has been elusive, requiring coordinated expression of almost a dozen proteins. Here we successfully produce simplified carboxysomes, isometric with those of the source organism Cyanobium, within tobacco chloroplasts. We replace the endogenous Rubisco large subunit gene with cyanobacterial Form-1A Rubisco large and small subunit genes, along with genes for two key α-carboxysome structural proteins. This minimal gene set produces carboxysomes which encapsulate the introduced Rubisco and enable autotrophic growth at elevated CO2. This result demonstrates the formation of α-carboxysomes from a reduced gene set, informing the step-wise construction of fully-functional α-carboxysomes in chloroplasts.
Full DescriptionThe data held within this document are the raw data collected for experiments on the heterologous expression of alpha-carboxysomes from the marine cyanobacterium Cyanobium marinum PCC7001 in chloroplasts of Nicotiana tabacum. Genes were introduced by biolistic bombardment to replace the endogenous Rubisco large subunit gene with the Rubisco large and small subunit gens from Cyanobium, or these genes in concert with the carboxysomal protein genes csoS1A and csoS2.

Data presented here include; raw data images of Southern blots confirming transgene insertion, raw images of transmission electron micrographs of transgenic plant chloroplasts and isolated carboxysomes, SDS-PAGE and immunoblot images, raw data for Nanosight N300 particle size analysis (including video data), gas exchange data for wild-type and transgenic plants, membrane inlet mass spectrometric measurement data of CO2 utilization by leaf discs of wild-type and transgenic plants, and growth data of each plant type.

Nanosight N300 data can be reanalyzed using the appropriate proprietary software. Raw numerical data are supplied as .csv files for analysis by standard spreadsheet and graphing software. Images are supplied as .tif files.
Contact Emailben.long@anu.edu.au
Contact AddressResearch School of Biology, Linnaeus Way, The Australian National University, 2601 ACT
Contact Phone Number0261259930
Principal InvestigatorBen Long
CollaboratorsRobert Sharwood
Benjamin Rae
Sarah Kaines
Yi-Leen Lim
Nghiem Nguyen
Baxter Massey
Soumi Bala
Susanne von Caemmerer
Murray Badger
Fields of Research060702 - Plant Cell and Molecular Biology
060503 - Microbial Genetics
060113 - Synthetic Biology
060199 - Biochemistry and Cell Biology not elsewhere classified
Type of Research ActivityApplied Research
Date Coverage
Date FromDate To
Date of data creation2017
Year of data publication2018
Creator(s) for Citation
Given NameSurname
Wei YihHee
Publisher for CitationThe Australian National University Data Commons
Data Size5.3 GB
Data Management PlanNo
Status: Published
Published To:
- Australian National University
Identifier: anudc:5524


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