Heterologous Expression of Carboxysomes in Plants

The 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.
Type
Collection
Title
Heterologous Expression of Carboxysomes in Plants
Brief Title
Carboxysomes in Plants
Collection Type
Dataset
Access Privileges
RIPE Lab
DOI - Digital Object Identifier
10.25911/5b4edb4bada74
Metadata Language
English
Data Language
English
Significance Statement
This 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 Description
A 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 Description
The 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 Email
ben.long@anu.edu.au; dean.price@anu.edu.au; will.hee@anu.edu.au
Contact Address
Research School of Biology, Linnaeus Way, The Australian National University, 2601 ACT
Contact Phone Number
0261259930
Principal Investigator
Ben Long
Collaborators
Robert Sharwood; Benjamin Rae; Sarah Kaines; Yi-Leen Lim; Nghiem Nguyen; Baxter Massey; Soumi Bala; Susanne von Caemmerer; Murray Badger
Fields of Research
060702 - Plant Cell and Molecular Biology; 060503 - Microbial Genetics; 060113 - Synthetic Biology; 060199 - Biochemistry and Cell Biology not elsewhere classified
Type of Research Activity
Applied Research
Date Coverage
2015-06-06
2018-03-06
Date of data creation
2017
Year of data publication
2018
Creator(s) for Citation
Benedict
Long
Wei Yih
Hee
Dean
Price
Publisher for Citation
The Australian National University Data Commons
Data Size
5.3 GB
Data Management Plan
No
Status: Published
Published to:
  • Australian National University
Related items