14-PSIL Combining Algal and Plant Photosynthesis (CAPP2)
Lead Research Organisation:
University of Edinburgh
Department Name: Sch of Biological Sciences
Abstract
In most plants, growth rate is limited by the rate at which carbon dioxide from the atmosphere is taken up and converted to sugars in the process of photosynthesis. The enzyme responsible for the first step in this process, Rubisco, does not work at its potential maximum efficiency at the current levels of carbon dioxide present in the atmosphere. If levels were much higher, photosynthesis would be increased and plant productivity would be higher. There is an immediate requirement for increased crop productivity to provide food for the rising population of the planet. Our project addresses this problem. We are studying a mechanism present in unicellular green algae that results in high concentrations of carbon dioxide inside their photosynthesising cells (called a Carbon Concentrating Mechanism, or CCM), enabling Rubisco to work at maximum efficiency. During the initial CAPP1 programme, we discovered important new information about this mechanism, and using new and rapid methods we have identified novel algal genes and additional regulatory components which allow the CCM to operate in association with a specific micro-compartment called a pyrenoid. We have also successfully introduced some of these components into a model higher plant, Arabidopsis, and also successfully introduced a modified form of Rubisco which may facilitate aggregation into the pyrenoid. The ambitious goals of the CAPP2 extension will be to combine the expression of the CCM and pyrenoid in the Advanced Plant. Firstly, we will continue to identify genes required by the algae to achieve high concentrations of carbon dioxide inside the cells, and develop new markers and sensors to reveal the location and activity of these genes when expressed in the higher plant. Secondly, we will identify additional regulatory elements needed to form a pyrenoid, as well as exploring the impact on Rubisco enzyme efficiency and light utilisation. Thirdly, we will continue to introduce successive components into our model Advanced Plant so as to "stack" up the activities of CCM components and examine the extent of pyrenoid formation and enhanced productivity associated with the CCM. This work will provide new insights into how plants and algae acquire and use carbon dioxide from the atmosphere, of great importance in predicting and coping with the current rapid changes in the atmosphere and hence in climate. The work will also contribute to strategies to increase global food security, because it will indicate new ways in which crop productivity can be increased.
Technical Summary
The CAPP2 programme will continue to discover and characterise new components of the Chlamydomonas CCM, in addition to known bicarbonate pump(s) and carbonic anhydrases, and identify regulatory elements, protein modifications, linkers and chaperones needed to aggregate Rubisco into a pyrenoid and progressively incorporate key components into Arabidopsis. Our approaches include:
1) Identify and characterize components of the pyrenoid in Chlamydomonas, including growth phenotyping, complementation to identify the mutant locus and fluorescence tagging to identify prenoid-associated proteins. Pyrenoid defects will be screened from microscopy, Rubisco kinetics, CCM physiology and light use.
2) Pyrenoid structure, assembly and regulation will be defined using block-face scanning EM in Chlamydomonas and Advanced Plant chloroplasts. Chaperones and linkers will be identified using AUC and BN-PAGE, and other post-translational modifications including phosphorylation, ubiquitination and S-S bond forma tion by LC-MS/MS.
3) To assemble a pyrenoid in the Advanced Plant gene cassettes will be assembled and/or stacked into delivery vectors for transformation. Additional "known" CCM components will be introduced and location tested by fluorescent tags (e.g. GFP), and levels of untagged components by polyclonal antibodies. Repression of all native Rubisco in Arabidopsis double mutants, and CCM characteristics will be combined in a single plant.
4) The advanced plant lines will be characterized physiologically to determine effectiveness of introduced CCM.
5) A FRET nanosensor will be developed to allow bicarbonate pools accumulated through the CCM to be visualized.
6) Modelling of inorganic carbon accumulation and effectiveness of pyrenoid operation will be undertaken in collaboration with colleagues at UIUC Illinois.
7) A central repository of genetic tools and materials will be developed for open access to the scientific and commercial communities.
1) Identify and characterize components of the pyrenoid in Chlamydomonas, including growth phenotyping, complementation to identify the mutant locus and fluorescence tagging to identify prenoid-associated proteins. Pyrenoid defects will be screened from microscopy, Rubisco kinetics, CCM physiology and light use.
2) Pyrenoid structure, assembly and regulation will be defined using block-face scanning EM in Chlamydomonas and Advanced Plant chloroplasts. Chaperones and linkers will be identified using AUC and BN-PAGE, and other post-translational modifications including phosphorylation, ubiquitination and S-S bond forma tion by LC-MS/MS.
3) To assemble a pyrenoid in the Advanced Plant gene cassettes will be assembled and/or stacked into delivery vectors for transformation. Additional "known" CCM components will be introduced and location tested by fluorescent tags (e.g. GFP), and levels of untagged components by polyclonal antibodies. Repression of all native Rubisco in Arabidopsis double mutants, and CCM characteristics will be combined in a single plant.
4) The advanced plant lines will be characterized physiologically to determine effectiveness of introduced CCM.
5) A FRET nanosensor will be developed to allow bicarbonate pools accumulated through the CCM to be visualized.
6) Modelling of inorganic carbon accumulation and effectiveness of pyrenoid operation will be undertaken in collaboration with colleagues at UIUC Illinois.
7) A central repository of genetic tools and materials will be developed for open access to the scientific and commercial communities.
Planned Impact
1. Academics and researchers in all fields of plant research.
2. Annotators in genomics and metabolomics, database and germplasm curators.
3. UK, US and international science base.
4.Agro-industry including biotechnologists and plant breeders seeking to increase plant productivity and/or harvest index; metabolic engineers and metabolic modellers.
5. Agricultural community and advisors.
6. Postdoctoral researchers employed on the project.
7. Public
8. The next generation: school children and undergraduate students
9. Multinational and Government Agencies
How will they benefit from this research?
1. Researchers will receive comprehensive new information about the CCM of algae, requirements for CO2 concentration in higher-plant chloroplasts, and mechanisms of assembly of supra-molecular complexes.
2. Researchers will have access at the point of publication to new genome annotation in Chlamydomonas, novel Arabidopsis material with altered primary carbon assimilation and models describing the relationship between the spatial distributions of inorganic carbon substrates and enzymes and the process of CO2 assimilation in chloroplasts.
3. The research will have a major impact on understanding of photosynthetic CO2 assimilation and its relationship to inorganic carbon concentrations in the chloroplast.
4. Agro-industry will receive information to underpin rational approaches to increase plant productivity, and relevant new genes and modelling methodologies.
5. The agricultural community will benefit in the longer term from sustainable crop improvements enabled by our research.
6. The PDRAs will receive a wide training in plant integrative biology, professional skills and wider training courses, and the opportunity to interact closely with researchers on an international scale. They will also receive training in transferable skills such as presentation and dissemination of results, and grant-writing. Enhanced career trajectory is already evident from support for the move of Dr McCormick to Edinburgh.
7. Our research findings relate to issues of public interest including sustainable crop production, global food security and atmospheric and climate change.
8. Our research has wide educational value, at all levels through schools and Universities
What will be done to ensure they benefit from this research?
1. Publish results in high-impact journals in a timely fashion, with open access where possible. Present research results at international meetings and institutions
2. Submit data and models to relevant international depositories. Notify new/corrected gene and enzyme annotations to community databases
3. Exploit extensive existing contacts of the PIs with other academics with relevant research interests as soon as any exploitable results/materials are generated.
4. Make informal contacts with biotechnologists as soon as exploitable results are generated; recognise and protect PI to ensure wise and fruitful exploitation. Collectively we have vibrant contacts with relevant industries.
6. Provide information and mentoring to ensure uptake of postdoctoral training schemes, including regular progress reviews and career development plans. Encourage participation in the dissemination of results, and understanding of the wider implications and applications of the research.
7. Use results as part of our regular engagement with non-academic audiences, e.g. local interest groups, schools, local and national shows, science showcases, media.
8. Involve school children and undergraduate students in a practical sense (visits, websites providing teaching resources, blogging and laboratory summer secondments for high school students and undergraduates).
9. Seek opportunities to inform the work of UN agencies and DFID (UK) in the developing world, and the CGIAR international network of germplasm repositories and strategic regional research.
2. Annotators in genomics and metabolomics, database and germplasm curators.
3. UK, US and international science base.
4.Agro-industry including biotechnologists and plant breeders seeking to increase plant productivity and/or harvest index; metabolic engineers and metabolic modellers.
5. Agricultural community and advisors.
6. Postdoctoral researchers employed on the project.
7. Public
8. The next generation: school children and undergraduate students
9. Multinational and Government Agencies
How will they benefit from this research?
1. Researchers will receive comprehensive new information about the CCM of algae, requirements for CO2 concentration in higher-plant chloroplasts, and mechanisms of assembly of supra-molecular complexes.
2. Researchers will have access at the point of publication to new genome annotation in Chlamydomonas, novel Arabidopsis material with altered primary carbon assimilation and models describing the relationship between the spatial distributions of inorganic carbon substrates and enzymes and the process of CO2 assimilation in chloroplasts.
3. The research will have a major impact on understanding of photosynthetic CO2 assimilation and its relationship to inorganic carbon concentrations in the chloroplast.
4. Agro-industry will receive information to underpin rational approaches to increase plant productivity, and relevant new genes and modelling methodologies.
5. The agricultural community will benefit in the longer term from sustainable crop improvements enabled by our research.
6. The PDRAs will receive a wide training in plant integrative biology, professional skills and wider training courses, and the opportunity to interact closely with researchers on an international scale. They will also receive training in transferable skills such as presentation and dissemination of results, and grant-writing. Enhanced career trajectory is already evident from support for the move of Dr McCormick to Edinburgh.
7. Our research findings relate to issues of public interest including sustainable crop production, global food security and atmospheric and climate change.
8. Our research has wide educational value, at all levels through schools and Universities
What will be done to ensure they benefit from this research?
1. Publish results in high-impact journals in a timely fashion, with open access where possible. Present research results at international meetings and institutions
2. Submit data and models to relevant international depositories. Notify new/corrected gene and enzyme annotations to community databases
3. Exploit extensive existing contacts of the PIs with other academics with relevant research interests as soon as any exploitable results/materials are generated.
4. Make informal contacts with biotechnologists as soon as exploitable results are generated; recognise and protect PI to ensure wise and fruitful exploitation. Collectively we have vibrant contacts with relevant industries.
6. Provide information and mentoring to ensure uptake of postdoctoral training schemes, including regular progress reviews and career development plans. Encourage participation in the dissemination of results, and understanding of the wider implications and applications of the research.
7. Use results as part of our regular engagement with non-academic audiences, e.g. local interest groups, schools, local and national shows, science showcases, media.
8. Involve school children and undergraduate students in a practical sense (visits, websites providing teaching resources, blogging and laboratory summer secondments for high school students and undergraduates).
9. Seek opportunities to inform the work of UN agencies and DFID (UK) in the developing world, and the CGIAR international network of germplasm repositories and strategic regional research.
Organisations
- University of Edinburgh (Lead Research Organisation)
- UNIVERSITY OF EDINBURGH (Collaboration)
- University of Illinois at Urbana-Champaign (Collaboration)
- Australian National University (ANU) (Collaboration)
- University of York (Collaboration)
- John Innes Centre (Collaboration)
- Carnegie Institution for Science (CIS) (Collaboration)
- UNIVERSITY OF CAMBRIDGE (Collaboration)
Publications
Atkinson N
(2016)
Introducing an algal carbon-concentrating mechanism into higher plants: location and incorporation of key components.
in Plant biotechnology journal
Atkinson N
(2019)
The pyrenoidal linker protein EPYC1 phase separates with hybrid Arabidopsis-Chlamydomonas Rubisco through interactions with the algal Rubisco small subunit.
in Journal of experimental botany
Atkinson N
(2017)
Rubisco small subunits from the unicellular green alga Chlamydomonas complement Rubisco-deficient mutants of Arabidopsis.
in The New phytologist
Dobrescu A
(2017)
A "Do-It-Yourself" phenotyping system: measuring growth and morphology throughout the diel cycle in rosette shaped plants
in Plant Methods
Donovan S
(2020)
CRISPR-Cas9-Mediated Mutagenesis of the Rubisco Small Subunit Family in Nicotiana tabacum
in Frontiers in Genome Editing
Khumsupan P
(2020)
Generating and characterizing single- and multigene mutants of the Rubisco small subunit family in Arabidopsis.
in Journal of experimental botany
Meyer MT
(2016)
Will an algal CO2-concentrating mechanism work in higher plants?
in Current opinion in plant biology
Description | Towards Aim 3A (Express further "known" components important for pyrenoid function): We have assembled a comprehensive vector database (see database section) for assembling all known and newly discovered CCM components (Jonikas/Griffiths labs) into single or multi-gene cassettes that can target algal and cyanobacterial CCM proteins to appropriate sub-cellular locations in plants. Successful introduction of several of these into higher plants was outlined in Atkinson et al. (2016, 2017). We have found that CAH6 does not localise to the chloroplast in Chlamydomonas or in tobacco. LCIB and LCIC both self-localised to the chloroplast and were able to interact in plants (shown by BiFC). Attempts to localise CAH3 to the chloroplast in higher plants have not yet been successful. This work is being continued by my PhD student Alex Vardakis. Instead, we focused on characterising the newly identified Rubisco linker protein EPYC1, which is required for pyrenoid formation (Mackinder et al. PNAS 2016). We found that EPYC1 interacts with the small subunit of Rubisco (SSU) and modified versions of higher plant Rubisco (e.g. the 1AMOD SSU (Atkinson, 2017)) are able to interact with EPYC1. We have now performed a detailed analysis of have the SSU-EPYC1 interaction, and have designed a quantitative yeast 2-hybrid assay to engineer EPYC1 and SSU variants with increased binding strengths. We have also expressed EPYC1 in higher plants and found thus far have found no negative affect on growth. Our work on EPYC1 characterisations and in planta expression has been submitted to The Plant Journal. Towards Aim 3B (Assess expression and location of untagged CCM proteins): Localisation of modified CCM transporters HLA3-GFP and LCIA-GFP to the chloroplast envelope has been confirmed by confocal imagery and immunoblotting against GFP. We have constructed new vectors lacking the GFP-marker and have now ordered antibodies for LCIA and HLA3 to track protein expression. We have also ordered antibodies for a new protein identified by the Jonikas lab during this project, EPYC1. New work by Mackinder et al. (Cell 2017) has revealed several new components required for pyrenoid assembly, including the new linker protein SAGA. We also have secured antibodies for this protein. Towards Aim 3C (Remove remaining native Rubisco small subunit (SSU)): We have employed a CRISPR/Cas9 approach to remove 3 of the 4 SSU genes located at a single chromosome locus. The approach has shown that we can generate a deletion event to in all 4 SSUs in Arabidopsis. My PhD student Panupon Khumsupan has now successfully generated a triple mutant that lacks all 3 major SSU isoforms (1A, 2B, 3B) and has complemented this line with a Chlamydomonas SSU. He is currently removing the final minor SSU isoform 1B using CRISPR/Cas9. Towards Aim 3D (Combine CCM characteristics in a single plant): We have developed multi-gene expression lines carrying between 2 and 4 gene expression cassettes and have transformed these into Arabidopsis. These include HLA3, LCIA, and cyanobacterial bicarb transporters BicA and SbtA. Western analysis has shown that the proteins are being expressed. However, as outlined in Rae et al. (2017), all bicarb transporters from algal and cyanobacterial systems appear to be inactive when expressed in higher plants. New work by Mackinder (Cell 2017) has revealed several new components interacting with HLA3 and LCIA, which we believe are required for their activation. Recent funding secured from the Leverhulme Trust (3 years) will support further research to examine these new components, and will use the multi-gene expression lines generated in this study. Towards Aim 3E (Introduce novel pyrenoid components and chaperones): As new CCM components have been identified and verified by the Jonikas and Griffiths labs, we have commenced work with these in Arabidopsis. In particular, we are currently investigating the recently identified algal Rubisco linker protein EPYC1, and how it interacts with higher plant Rubisco (as outlined above). Working with Jonikas, we have recently uncovered the specific residues involved in the ineraction between EPYC1 and the SSU (a manuscript is currently being prepared). Our quantitative Y2H assay was also used to characterise the newly discovered pyrenoid component SAGA - a manuscript outlining SAGA is currently being prepared. Towards Aim 4 (Characterize Advanced Plant lines): Work to generate and characterise plants with clear evidence of Rubisco aggregation akin to a pyrenoid is ongoing. As in Atkinson (2017), we have generated pyrenoid-compatible Arabidopsis expressing the algal SSU required for pyrenoid assembly, and have shown that this form of Rubisco is near-equivalent to native Rubisco, in terms of functionality and growth complementation. Notably, we have provided new evidence that the Rubisco SSU plays an important role in the catalytic characteristics of Rubisco. We have expressed EPYC1 in lines expressing the algal SSU, and shown interaction between EPYC1 and Rubisco in planta by co-immunoprecipitation. In late 2018, in collaboration with Oliver Mueller Cajar (NTU), we have shown that hybrid Arabidopsis-Chlamydomonas Rubisco from plants can aggregate with EPYC1. We are no working to achieve the required ratio of EPYC1 and Chlamydomonas SSU expression in Arabidopsis to promote aggregation in planta. |
Exploitation Route | Academic routes: my research has significantly added to current state of the art of introducing CCM components from algae into higher plants to enhance photosynthesis. The project has led to 5 publications and at least 2 additional manuscripts that will be submitted in the next few months. The work has been presented at several invited seminar/conference events (e.g. Oxford University, York University, University of Neuchâtel (Switzerland), ETH Zurich (Switzerland), University of Cambirdge, University of Essex, University of Campinas (Brazil), University of Glasgow, the Royal Society and International Congress of Photosynthesis). We have publicized our work online with online interview with Garnet UK. Work on this project has led to three funded PhD students joining the lab, and has led to four further successful research grant applications (>80k each). Non-academic routes: We have developed a new, high-impact outreach educational tool that uses virtual reality to teach the public about Rubisco the importance and shortcomings of Rubisco, the primary carboxylating enzyme in photosynthetic organisms. We have participated in outreach for young school children (4-12) and their parents during the Scotland Midlothian science festival (http://midlothiansciencefestival.com/) and the Festival of Learning. Their objective was to teach the public about the importance and shortcomings of Rubisco, the primary carboxylating enzyme in photosynthetic organisms. |
Sectors | Agriculture Food and Drink Education Manufacturing including Industrial Biotechology |
URL | http://mccormick.bio.ed.ac.uk/ |
Description | 2015: publication of our work a high impact factor journal (Plant Biotechnology Journal) resulted in coverage on several science news websites (e.g. https://www.sciencedaily.com/releases/2015/11/151116112048.htm). 2015: Our publication (DOI: 10.1111/pbi.12497) and the broader scope of the project was discussed on an online interview with Garnet (http://blog.garnetcommunity.org.uk/arabidopsis-research-roundup-november-13th/) 2016: A new, high-impact educational tool and participated in outreach for young school children (4-12) and their parents during the Scotland Midlothian science festival (http://midlothiansciencefestival.com/). Their objective was to teach the public about the importance and shortcomings of Rubisco, the primary carboxylating enzyme in photosynthetic organisms. 2016: Spin off work, based on application of our vector database to other biological systems, has resulted in two funded projects (Phyconet and IBIOIC) with industrial partner Scottish Bioenergy to produce high value products in cyanobacteria. This work has led to three additional funded PhD students (1 CONACYT scholarship, 1 EastBio-CASE scholarship, 1 IBIOIC-CTP scholarship) joining the lab (2016-2017). 2017: online interview with Garnet discussing recent publication in New Phytologist journal Rubisco small subunits from the unicellular green alga Chlamydomonas complement Rubisco-deficient mutants of Arabidopsis (DOI: 10.1111/nph.14414). 2017: publication of review ( https://doi.org/10.1093/jxb/erx133) redefined the current challenges in introducing biophysical CCMs into higher plants. 2014-2018: Work on this project has led to four funded PhD students joining the lab (2 Darwin Trust scholarships, 2 EastBio scholarships). 2014-2018: Work on this project has led to four successful small research grant applications (>80k each) and one large Leverhulme Trust grant (£341k). 2019: Data generated in CAPP2 contributed to publication of recent work: "The pyrenoidal linker protein EPYC1 phase separates with hybrid Arabidopsis-Chlamydomonas Rubisco through interactions with the algal Rubisco small subunit" in JXB (https://doi.org/10.1093/jxb/erz275). 2019: Work on this project has led to a successful joint BBSRC-NSF research grant application - BB/S015531/1 (£699). |
First Year Of Impact | 2015 |
Sector | Agriculture, Food and Drink,Education,Manufacturing, including Industrial Biotechology |
Impact Types | Cultural Societal |
Description | BBSRC workshop to discuss new crop breeding technologies |
Geographic Reach | National |
Policy Influence Type | Contribution to a national consultation/review |
Impact | Led to publication of a position statement by the BBSRC on new and emerging techniques for crop improvement. |
URL | http://www.bbsrc.ac.uk/documents/genetic-crop-improvement-position-statement-pdf/ |
Description | Proposed Centre of Expertise in Plant Health - Stakeholder Workshop |
Geographic Reach | National |
Policy Influence Type | Contribution to a national consultation/review |
URL | http://scotland.landscapeinstitute.org/wp-content/uploads/2016/01/Scottish-Plant-Health-Strategy-dra... |
Description | Cryo-electron tomography of CO2-fixing pyrenoids to guide synthetic assembly |
Amount | £25,061 (GBP) |
Funding ID | BB/X004953/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 11/2022 |
End | 10/2023 |
Description | IBioIC Micro Accelerator grant |
Amount | £376,051 (GBP) |
Organisation | IBioIC |
Sector | Academic/University |
Country | United Kingdom |
Start | 02/2017 |
End | 12/2018 |
Description | Investigating inorganic carbon transport in globally important algal lineages |
Amount | £406,514 (GBP) |
Funding ID | BB/W009587/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 02/2022 |
End | 01/2025 |
Description | Leverhulme Trust award |
Amount | £349,148 (GBP) |
Organisation | The Leverhulme Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 02/2018 |
End | 01/2021 |
Description | PhycoNet outreach award |
Amount | £2,100 (GBP) |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 11/2017 |
End | 11/2018 |
Description | PhycoNet/BBSRC NIBB Proof of Concept funding award |
Amount | £82,898 (GBP) |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 03/2016 |
End | 12/2016 |
Description | Scottish Universities Life Sciences Alliance (SULSA) PEER Funding Award |
Amount | £30,760 (GBP) |
Organisation | Scottish Universities Life Sciences Alliance |
Sector | Academic/University |
Country | United Kingdom |
Start | 07/2015 |
End | 08/2016 |
Description | The University of Edinburgh Innovation Initiative Grant |
Amount | £3,235 (GBP) |
Organisation | University of Edinburgh |
Sector | Academic/University |
Country | United Kingdom |
Start | 12/2015 |
End | 12/2016 |
Description | The University of Edinburgh Innovation Initiative Grant |
Amount | £2,512 (GBP) |
Organisation | University of Edinburgh |
Sector | Academic/University |
Country | United Kingdom |
Start | 05/2017 |
End | 06/2018 |
Description | The University of Edinburgh Innovation Initiative Grant |
Amount | £1,746 (GBP) |
Organisation | University of Edinburgh |
Sector | Academic/University |
Country | United Kingdom |
Start | 05/2014 |
End | 06/2015 |
Description | The York Physics of Pyrenoids Project (YP3): Nanostructured Biological LLPS:Next-Level-Complexity Physics of CO2-fixing Organelles |
Amount | £2,488,444 (GBP) |
Funding ID | EP/W024063/1 |
Organisation | Engineering and Physical Sciences Research Council (EPSRC) |
Sector | Public |
Country | United Kingdom |
Start | 03/2022 |
End | 03/2025 |
Description | Wellcome Trust Institutional Strategic Support Fund (ISSF) |
Amount | £112,105 (GBP) |
Organisation | Wellcome Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 05/2014 |
End | 06/2015 |
Title | CyanoGate Toolkit |
Description | The CyanoGate kit contains 117 parts and acceptor vectors for generating multi-part assemblies in either integrative (suicide) or self-replicating plasmid vectors for working in cyanobacteria. The core parts of the kit are supplied on a 96 well plate (from Addgene), while some additional components (i.e. 18 terminators and 3 variants of the Pcpc560 promoter) are available for separate order (from Addgene). The kit is compatible with the plant Modular Cloning (MoClo) syntax (also known as PhytoBricks). You require the backbone acceptor vectors in the MoClo Plant Parts Kit (#1000000044) (Engler et al., 2014, ACS SynBio) with this kit to progress from level 0 to level T assemblies. The CyanoGate kit includes 45 promoters, 5 minimal (up to the TSS) promoters, 21 terminators, 20 flanking sequences for HR-mediated genomic integration, 5 selectable marker cassettes, 4 level T (for Transformation) acceptor vectors, and parts for building CRISPRi- and sgRNA-based systems. |
Type Of Material | Biological samples |
Year Produced | 2019 |
Provided To Others? | Yes |
Impact | The kit is high-throughput Golden Gate cloning system for cyanobacterial research so will have a significant impact on this community. |
URL | http://www.addgene.org/kits/mccormick-cyanogate/ |
Title | Biophysical carbon concentrating mechanism vector database |
Description | We have developed a detailed and comprehensive database of Gateway- and Golden Gate-based vectors containing plant-domesticated parts of the biophysical Chlamydomonas and cyanobacterial carbon concentrating mechanisms (CCMs). Currently our database contains 742 vectors / glycerol stocks and 364 Agrobacterium glycerol stocks. |
Type Of Material | Database/Collection of data |
Year Produced | 2014 |
Provided To Others? | Yes |
Impact | The progression from a gateway-based system to Golden Gate has accelerated our capacity to build multigene-vectors for expressing and testing many gene combinations and building CCMs in planta and in other heterologous systems (e.g. yeast, Xenopus). |
Description | 14-PSIL Combining Algal and Plant Photosynthesis (CAPP2) - ANU |
Organisation | Australian National University (ANU) |
Country | Australia |
Sector | Academic/University |
PI Contribution | The partnership was set up to enhance the outputs from CAPP2. Prof Price (Australia National University) works on the RIPE project with Prof Steve Long (UIUC Champaign Urbana) and has significant expertise in the cyanobacterial CCM and strategies to introduce this into higher plants. We have worked with ANU to produce a review. |
Collaborator Contribution | Griffiths (Cambridge) set up the CCM9 meeting in 2016, which resulted in this collaboration to write a review in 2017. We have continued to collaborate in terms of knowledge exchange. |
Impact | Produced manuscript: Rae et al. 2015 Journal of Experimental Botany (https://doi.org/10.1093/jxb/erx133) |
Start Year | 2016 |
Description | 14-PSIL Combining Algal and Plant Photosynthesis (CAPP2) - Cambridge |
Organisation | University of Cambridge |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Currently collaborating on introduction of novel Chlamydomonas components into higher plants to modify photosynthetic efficiency. We have introduced the algal CCM components studied by Cambridge into higher plants for further characterisation. |
Collaborator Contribution | Cambridge brings specific expertise in CCM and photosynthetic physiology. We have shared access of data, protocols and biological materials between labs. Information provided by Cambridge has strongly impacted on the specific components used to build a pyrenoid in higher plants. |
Impact | Produced manuscript: Atkinson et al. 2015 Plant Biotechnology Journal (doi: 10.1111/pbi.12497), Atkinson et al. 2017 New Phytologist (doi:10.1111/nph.14414). |
Start Year | 2014 |
Description | 14-PSIL Combining Algal and Plant Photosynthesis (CAPP2) - JIC |
Organisation | John Innes Centre |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Currently collaborating on introduction of novel Chlamydomonas components into higher plants to modify photosynthetic efficiency. We have progressed on research initially started in JIC (Smith Lab) in 2008 by introducing novel CCM components into higher plants for further characterisation. |
Collaborator Contribution | JIC brings specific expertise in plant metabolism and photosynthesis. We have shared access of data, protocols and biological materials between labs. |
Impact | Produced manuscript: Atkinson et al. 2015 Plant Biotechnology Journal (doi: 10.1111/pbi.12497) |
Start Year | 2014 |
Description | 14-PSIL Combining Algal and Plant Photosynthesis (CAPP2) - Princeton |
Organisation | Carnegie Institution for Science (CIS) |
Country | United States |
Sector | Charity/Non Profit |
PI Contribution | Currently collaborating on introduction of novel Chlamydomonas components into higher plants to modify photosynthetic efficiency. We have introduced the algal CCM components discovered by Prof. Martin Jonikas in Princeton (was in Carnegie) into higher plants for further characterisation. |
Collaborator Contribution | Princeton brings specific expertise/data of novel components involved in the CCM. We have shared access of data, protocols and biological materials between labs. Information provided by Princeton has strongly impacted on the specific components used to build a pyrenoid in higher plants. |
Impact | Produced manuscript: Atkinson et al. 2015 Plant Biotechnology Journal (doi: 10.1111/pbi.12497), Atkinson et al. 2017 New Phytologist (doi:10.1111/nph.14414). |
Start Year | 2014 |
Description | 14-PSIL Combining Algal and Plant Photosynthesis (CAPP2) - UIUC Champaign Urbana |
Organisation | University of Illinois at Urbana-Champaign |
Country | United States |
Sector | Academic/University |
PI Contribution | The partnership was set up to enhance the outputs from CAPP2, since Dr Justin McGrath and Prof Steve Long already developed a model for cyanobacteria; the goal was to develop the model for the algal (chlamydomonas) system, and potential operation in a higher plant cell. We will provide Dr McGrath with physiology data from plants expressing Chlamydomonas CCM components as needed. |
Collaborator Contribution | Dr McGrath visited UK for one week and worked at Cambridge on developing the model; a co-ordination meeting was also held during his visit to allow all UK partners to contribute to model development. Originally we had planned for one of our US collaborators to be present, but this was not possible at the last minute. |
Impact | Currently working on a joint output as a draft of a peer-reviewed paper |
Start Year | 2014 |
Description | 14-PSIL Combining Algal and Plant Photosynthesis (CAPP2) - York |
Organisation | University of York |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | This collaboration with Dr Luke Mackinder (York) was set up following Mackinder's appointment as a PI (Mackinder was a postdoc in the Jonikas lab (Princeton). We are currently collaborating on introduction of novel Chlamydomonas components into higher plants to modify photosynthetic efficiency. We have introduced the algal CCM components studied by York into higher plants for further characterisation. |
Collaborator Contribution | Mackinder was a lead postdoc on the CAPP2 project in Jonikas's lab. Mackinder has contributed to generating a step-change in understanding of the components of the algal pyrenoid and CCM processes. McCormick and Mackinder continue to collaborate closely through a funded Leverhulme Trust project (Feb 2018). |
Impact | Produced manuscript: Atkinson et al. 2015 Plant Biotechnology Journal (doi: 10.1111/pbi.12497), Atkinson et al. 2017 New Phytologist (doi:10.1111/nph.14414). |
Start Year | 2017 |
Description | Alistair McCormick |
Organisation | University of Edinburgh |
Department | Centre for Synthetic and Systems Biology (SynthSys) |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Gene discovery and functional characterisation to guide plant engineering to improve photosynthesis |
Collaborator Contribution | Plant engineering of functionally characterised genes by my team. |
Impact | Joint funding from Leverhulme Trust (RPG-2017-402). Joint funding BBSRC/NSF-Bio (BB/S015337/1) |
Start Year | 2013 |
Description | York Physics of Pyrenoids Project |
Organisation | University of York |
Department | Department of Physics |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | The research is based on preliminary data generated by my lab across multiple UKRI supported projects. We provide the biological expertise within the consortium. |
Collaborator Contribution | They provide theoretical modelling, biophysics and biochemistry expertise to enable a multidisciplinary approach to understand pyrenoid evolution, structure and function. |
Impact | This is multi-disciplinary including biology, biophysics and soft matter physics. |
Start Year | 2020 |
Title | RUBISCO-BINDING PROTEIN MOTIFS AND USES THEREOF |
Description | Described herein are chimeric polypeptides that include one or more Rubisco-binding motifs (RBMs) and a heterologous polypeptide. Additional aspects of the present disclosure provide genetically altered plants having a chimeric polypeptide including one or more Rubisco-binding motifs (RBMs) and a heterologous polypeptide. Further aspects of the present disclosure relate to genetically altered plants having a stabilized polypeptide including two or more RBMs and one or both of an algal Rubisco-binding membrane protein (RBMP) and a Rubisco small subunit (SSU) protein. Other aspects of the present disclosure relate to methods of making such chimeric polypeptides and plants, as well as cultivating these genetically altered plants. |
IP Reference | WO2021025962 |
Protection | Patent granted |
Year Protection Granted | 2021 |
Licensed | No |
Impact | We are in current talks with the Bill and Melinda Gates foundation about further funding related to this patent. |
Title | Plant Magic software for leaf rosette area analysis |
Description | Plant Magic is a software tool for analysing images of plants and calculating the leaf/rosette area under both night and day conditions. An associated hardware tool is used for night imaging. |
Type Of Technology | New/Improved Technique/Technology |
Year Produced | 2016 |
Impact | The software was developed by a Masters student (Andrei Dobrescu). Andrei has now been accepted as a PhD student co-supervised by McCormick and Sotirios Tsaftaris (UoE, Eng) to continue work on plant image analysis. The software is currently used by several labs in UoE and will shortly be released on the open Plant Image Analysis website (http://www.plant-image-analysis.org/). An additional hardware tool (a low cost far-red LED array with a raspberry Pi camera) was developed to capture images throughout the diel cycle. This method for building this setup is being prepared for publication. |
Description | Article in Infinity Magazine, pg 12 Issue 15 (2016-17) |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | Magazine article in Infinity magazine (pg 12 Issue 15,2016-17) outlining a spin-off project - collaboration between academics Boajun Wang (UoE), Chris Howe (Cambridge) and an industrial partner (Scottish Bioenergy) to develop synthetic biology tools for producing high value products in cyanobacteria. |
Year(s) Of Engagement Activity | 2016 |
URL | https://issuu.com/edinburghresearchandinnovation/docs/infinite-magazine-2016 |
Description | Festival of Learning - virtual reality Rubisco |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Undergraduate students |
Results and Impact | We applied our outreach tool developed in 2017 to reach a wider audience through the UoE Festival of Learning week. The virtual reality game teaches about the importance and shortcomings of Rubisco, the primary carboxylating enzyme in photosynthetic organisms. Gamification of Rubisco catalysis allowed engagement with the public on the importance of Rubisco and to discuss relatively complex biochemistry. |
Year(s) Of Engagement Activity | 2017,2018 |
URL | http://www.festivalofcreativelearning.ed.ac.uk/event/photosynthesis-virtual-reality |
Description | High yield crops a step closer in light of photosynthesis discovery |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Media (as a channel to the public) |
Results and Impact | Press release was taken up by a variety of online science sites. |
Year(s) Of Engagement Activity | 2015 |
URL | https://www.sciencedaily.com/releases/2015/11/151116112048.htm |
Description | Hosted the Plastid Preview 2016 |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | The McCormick Lab hosted the annual Plastid Preview (2016) in Edinburgh. The Plastid Preview is an EU/UK conference for postgraduate students and young researchers to meet, discuss their work and foster new collaborations. |
Year(s) Of Engagement Activity | 2016 |
URL | https://www.eventbrite.co.uk/e/the-plastid-preview-2016-tickets-24974428187 |
Description | Invited Talk (Oxford University) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Professional Practitioners |
Results and Impact | Gave a research presentation to the Oxford Plant Biology department. ~70 people attended. Exposed me to research at Oxford and potential future collaboration opportunities. |
Year(s) Of Engagement Activity | 2021 |
Description | Invited seminar, University of York, UK (presentation). |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Postgraduate students |
Results and Impact | Invited seminar by Mike Hayden (met at the FAPESP-British Council Workshop) to discusscurrent progress in the CAPP project. |
Year(s) Of Engagement Activity | 2015 |
Description | Invited seminar. University of Oxford, UK. |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Postgraduate students |
Results and Impact | Invited Departmental Seminar series for the Department of Plant Sciences in Oxford. |
Year(s) Of Engagement Activity | 2017 |
URL | https://www.plants.ox.ac.uk/event/seminar-dr-alistair-mccormick-university-edinburgh |
Description | Midlothian Science Festival - virtual reality Rubisco |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | We developed a new, high-impact educational tool and participated in outreach for young school children (4-12) and their parents during the Scotland Midlothian science festival (http://midlothiansciencefestival.com/). Their objective was to teach the public about the importance and shortcomings of Rubisco, the primary carboxylating enzyme in photosynthetic organisms. Gamification of Rubisco catalysis allowed engagement with the public on the importance of Rubisco and to discuss relatively complex biochemistry, even with four year olds! |
Year(s) Of Engagement Activity | 2016 |
URL | http://mccormick.bio.ed.ac.uk/amccormi/sites/sbsweb2.bio.ed.ac.uk.amccormi/files/MSF%202016%20Collag... |
Description | Online interview discussing recent paper (DOI: 10.1111/nph.14414) |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Media (as a channel to the public) |
Results and Impact | Online interview with Garnet discussing recent publication in New Phytologist journal Rubisco small subunits from the unicellular green alga Chlamydomonas complement Rubisco-deficient mutants of Arabidopsis (DOI: 10.1111/nph.14414). |
Year(s) Of Engagement Activity | 2017 |
URL | http://blog.garnetcommunity.org.uk/arabidopsis-research-roundup-january-26th/ |
Description | Online interview discussing recent paper (DOI: 10.1111/pbi.12497) |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Media (as a channel to the public) |
Results and Impact | Online interview with Garnet discussing recent publication in Plant Biotechnology Journal "Introducing an algal carbon-concentrating mechanism into higher plants: location and incorporation of key components" (DOI: 10.1111/pbi.12497). |
Year(s) Of Engagement Activity | 2015 |
URL | http://blog.garnetcommunity.org.uk/arabidopsis-research-roundup-november-13th/ |
Description | Participation and presentation at the FAPESP-British Council Workshop |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Attended the FAPESP-British Council Workshop, Campinas, Brazil to talk about current progress in the CAPP project and to discuss collaborations with UK and Brazilian researchers. |
Year(s) Of Engagement Activity | 2015 |
Description | Participation in an activity, workshop or similar - RSB Science Week - virtual reality Rubisco |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Schools |
Results and Impact | We applied for funding from the RSB to host a co-ordinated exhibition at the Midlothian Science Festival during RSB Science Week in 2018, using our VR outreach tool developed in 2017 . The virtual reality game teaches about the importance and shortcomings of Rubisco, the primary carboxylating enzyme in photosynthetic organisms. Gamification of Rubisco catalysis allowed engagement with the public on the importance of Rubisco and to discuss relatively complex biochemistry. |
Year(s) Of Engagement Activity | 2018 |
URL | https://www.rsb.org.uk/get-involved/biologyweek |
Description | Presentation and poster at the 9th International Symposium on Inorganic Carbon Utilization by Aquatic Photosynthetic Organisms, Cambridge, UK |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Invited presentation at the 9th International Symposium on Inorganic Carbon Utilization by Aquatic Photosynthetic Organisms. University of Cambridge, UK. |
Year(s) Of Engagement Activity | 2016 |
URL | http://www.plantsci.cam.ac.uk/research/howardgriffiths/ccm9 |
Description | Presentation at the Chloroplast Metabolism and Photosynthesis Conference, University of Neuchâtel, Switzerland. |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Invited presentation at the Chloroplast Metabolism and Photosynthesis Conference (2017). University of Neuchâtel, Switzerland. |
Year(s) Of Engagement Activity | 2017 |
URL | https://www.unine.ch/files/live/sites/dp-biol/files/shared/DPbiol/courses/2017/abstract_booklet_chlo... |
Description | Presentation at the Plant Transport meeting, University of Glasgow, UK. |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Invited presentation of current progress in the CAPP project at the Plant Transport meeting, University of Glasgow, UK. |
Year(s) Of Engagement Activity | 2017 |
URL | http://ptm.psrg.org.uk/ |
Description | Presentations at the Plastid Preview meeting. University of Essex, UK |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Invited presentation of current progress in the CAPP project at the Plastid Preview meeting. University of Essex, UK. Two presentations were presented, one by myself and one by my research assistant Nicky Atkinson |
Year(s) Of Engagement Activity | 2015 |
URL | http://photocomm.ku.dk/trainingevents/2015_plastidpreview/Plastid_Preview_-_Programme.pdf |
Description | School visit - Chapter One Nurseries: Science Expo |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | Outreach activity for school children and teachers. Children were provided with digital USB light microscopes and lab samples to view plants and discuss. They were then encouraged to collect their own samples from the class room or garden for viewing and discussion. |
Year(s) Of Engagement Activity | 2015,2016 |
Description | School visit - The Apple Tree Nursery: Science Expo |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | Outreach activity for school children and teachers. Children were provided with digital USB light microscopes and lab samples to view plants and discuss. They then collected and drew pictures to document their own samples . |
Year(s) Of Engagement Activity | 2017 |