Engineering ion flux of the stomatal complex for enhanced photosynthesis and water use efficiency
Lead Research Organisation:
John Innes Centre
Department Name: Crop Genetics
Abstract
Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
Technical Summary
We propose a concerted assessment of stomatal kinetics and its relevance to crop yields and water use. Our aims are (1) to demonstrate the feasibility of manipulating stomatal kinetics by translating knowledge of BLINK1 optogenetics in Arabidopsis into two crop models, and (2) to test the hypothesis of a two-cell, two-stroke 'pump' for solute transfer as a target for further enhancements in accelerating stomatal kinetics. In the latter case, we propose (i) a quantitative analysis of the ion transport characteristics of the surrounding cells and their coordination with stimuli known to trigger guard cell transport for stomatal opening and closing, and (ii) optogenetic and related manipulations of K+ flux of the surrounding cells in order to understand the transport coordination between surrounding and guard cells.
We will build on our recent success with the light-activated K+ channel BLINK1 in Arabidopsis in each case. Both the practical and fundamental challenges will take advantage of targeted optogenetic expression within the leaf epidermis and, additionally, on the development of new optogenetic tools with improved light sensitivities and altered light regulation. Experiments will follow methodologies similar to those used successfully to date, including voltage clamp, gas exchange and biomass studies. We will use extant knowledge of BLINK1 variants in extending the optogenetic tools, and we will draw on heterologous expression to identify and quantify the most promising of these prior to use. Finally, we will target expression between cell types to assess, in response to light, the component contributions from the surrounding and guard cells and to extract stomatal kinetics and their coupling to photosynthesis. We expect these studies to expand our fundamental understanding of stomatal mechanics and to establish their kinetics as a bona fide target for future efforts in crop improvement.
We will build on our recent success with the light-activated K+ channel BLINK1 in Arabidopsis in each case. Both the practical and fundamental challenges will take advantage of targeted optogenetic expression within the leaf epidermis and, additionally, on the development of new optogenetic tools with improved light sensitivities and altered light regulation. Experiments will follow methodologies similar to those used successfully to date, including voltage clamp, gas exchange and biomass studies. We will use extant knowledge of BLINK1 variants in extending the optogenetic tools, and we will draw on heterologous expression to identify and quantify the most promising of these prior to use. Finally, we will target expression between cell types to assess, in response to light, the component contributions from the surrounding and guard cells and to extract stomatal kinetics and their coupling to photosynthesis. We expect these studies to expand our fundamental understanding of stomatal mechanics and to establish their kinetics as a bona fide target for future efforts in crop improvement.
Planned Impact
This proposal is for a synergy in practical and fundamental research building on a core of ideas at the centre of the international plant photosynthesis and stomatal biology communities. The research will stimulate thinking around strategies for enhancing crop yields and reducing agricultural water consumption, and it should inform methodologies for approaching crop engineering. In the long term the research is expected to benefit fundamental researchers as well as agriculture and industry through conceptual developments as well as the introduction of new technologies relevant to plant productivity and water use efficiency. The research will feed into higher education training programmes through capacity building at the postgraduate and postdoctoral levels. Additional impact is proposed through public displays and the development of teaching resources building on the background work for this proposal. Finally the research will help guide future efforts in applications to agricultural/industrial systems. The applicants have established links with industrial/technology transfer partners and research institutes to take advantage of these developments. Further details of these, and additional impacts will be found in Part 1 of the Case for Support and in the attached Impact Pathways.
Description | One aim of this project was to test a selection of guard cell as well as epidermal/surrounding cell promoters in Brassica and barley. A range of constructs incorporating Guard cell and epidermal/surrounding cell promoters expressing GUS were generated, using golden gate methodology, to test their effectiveness in Brassica and barley. These promoters have now all been tested in the relevant crop and the most active identified. Results will be prepared for publication. In addition Brassica transgenic plants containing BLINK1 have been produced and some initial gas exchange and stomatal conductance measurements made. Initial analysis suggests differences in stomatal opening and closing in some transgenic plants. |
Exploitation Route | The funding could lead to crops with improved stress tolerance. |
Sectors | Agriculture Food and Drink |
Description | Member of the Food Standards Agency Advisory Committee on Novel Foods and Processes and their PGT sub-group. Ongoing. |
Geographic Reach | National |
Policy Influence Type | Participation in a guidance/advisory committee |
Impact | Contribution to the regulatory framework for products of precision breeding technologies |
Description | Precision Breeding Bill - Evidence provided during the committee stage |
Geographic Reach | National |
Policy Influence Type | Contribution to a national consultation/review |
Impact | Discussions with MPs, Members of the House of Lords, Defra and the Food Standards Agency have helped to inform the debates on the Precision Breeding Bill. |
Description | Took part in workshop with ARIA (Advanced Research and Invention Agency to further programme on food security and climate change |
Geographic Reach | National |
Policy Influence Type | Participation in a guidance/advisory committee |
Title | Characterised tissue specific promoters |
Description | Promoter::GUS lines for barley and Brassica (GC - guard cell, E - epidermal) In Brassica: -pKST1 (GC), pGC1 (GC), pMYB60 (GC), pAtML1 (E), pCER6 (E) and pGstA+WIR1 (E) expressed GUS in T0 leaves. Some of these promoters (pKST1, pGC1 and pAtML1) also expressed in brassica T1 homozygous lines, after ~7 hours of GUS staining. In barley: -Promoters that expressed GUS in guard cells and epidermal cells were pCST1 and pGstA+WIR1, respectively in T0 transgenics. pSNAC1 promoters showed mixed results. |
Type Of Material | Biological samples |
Year Produced | 2021 |
Provided To Others? | No |
Impact | Will shortly be prepared for publication |
Description | University of Glasgow |
Organisation | University of Glasgow |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | A new collaborative project was established with University of Glasgow where we will provide barley transformation and genome editing expertise. |
Collaborator Contribution | Our partners provide expertise in photoreceptor engineering. This expertise will be used to inform the design and assembly of constructs for barley transformation with the aim of increasing plant biomass. |
Impact | Project to start this year. |
Start Year | 2021 |
Description | ADHB strategic farmers group Scotland (Penny Hundleby) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | ADHB strategic farmers group Scotland - on-line talk on 'Genome Editing potential for pest control' |
Year(s) Of Engagement Activity | 2021 |
Description | Animal and Plant Health Welfare Directorate at Defra |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Policymakers/politicians |
Results and Impact | Fully day conference style event for Defra team to find out about JIC Science. Talk delivered on Gene Editing |
Year(s) Of Engagement Activity | 2023 |
Description | Church Farm Open day |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Public/other audiences |
Results and Impact | Open day at our field trial station |
Year(s) Of Engagement Activity | 2021 |
Description | FSA-N8 GM/GE horizon scanning workshop |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | The workshop was chaired by Prof Wendy Harwood and an introduction to genome editing provided. |
Year(s) Of Engagement Activity | 2020 |
Description | Farming Today Interview Penny Hundleby |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Interview on 'New plans for gene editing' |
Year(s) Of Engagement Activity | 2021 |
Description | Food Standards Agency workshop on Gene Editing |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Policymakers/politicians |
Results and Impact | Visit by FSA to update us on the recent consultation exercise and to talk through the secondary legislation stages (Precision breeding Act). Relationship building and informing |
Year(s) Of Engagement Activity | 2024 |
Description | Interview for Farming Today BBC radio 4 |
Form Of Engagement Activity | A broadcast e.g. TV/radio/film/podcast (other than news/press) |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Interview on Genome Editing for Farming Today |
Year(s) Of Engagement Activity | 2021 |
URL | https://www.bbc.co.uk/sounds/play/m000sq7b |
Description | Interview for Times Radio |
Form Of Engagement Activity | A broadcast e.g. TV/radio/film/podcast (other than news/press) |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Live interview on gene edited foods. |
Year(s) Of Engagement Activity | 2021 |
Description | New Scientist Live event |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | A presentation on genome editing was contributed to the event. |
Year(s) Of Engagement Activity | 2020 |
Description | Presentation at JIC Breeders Day on Genome editing |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Industry/Business |
Results and Impact | Presentation on genome editing to plant breeders, growers, agri-industries to raise awareness and understand challenges. |
Year(s) Of Engagement Activity | 2023 |
Description | Sense, Science and Sustainability |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | Sense, Science and Sustainability: Can genome editing and agroecology co-exist in the sustainable food and farming mix? Event hosted by A Bigger Conversation (a Beyond GM initiative) |
Year(s) Of Engagement Activity | 2020 |
Description | TV programme - Follow the Food |
Form Of Engagement Activity | A broadcast e.g. TV/radio/film/podcast (other than news/press) |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | Contribution to an episode of the series 'Follow the Food', filmed at the John Innes Centre. Focused on the use of genome editing for disease resistance. Episode 2 of series 2: Seeds of Life. Involved Dr Catherine Jacott, Prof Cristobal Uauy, Prof Wendy Harwood. |
Year(s) Of Engagement Activity | 2021 |
URL | https://www.bbc.com/reel/playlist/follow-the-food-series-2 |
Description | Visit from Minister of State for Food, Farming and Fisheries |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Policymakers/politicians |
Results and Impact | Visit my Rt Hon Mark Spencer MP to see Brassica Crop Transformation platform to discuss gene editing. Part of a wider site visit. Defra later issued a video from the visit |
Year(s) Of Engagement Activity | 2024 |
URL | https://www.jic.ac.uk/news/defra-minister-visits-norwich-research-park-to-observe-the-potential-of-p... |