Photoreceptor Engineering to Modulate Plant Growth

Lead Research Organisation: University of Glasgow
Department Name: College of Medical, Veterinary &Life Sci

Abstract

Light is critical for coordinating plant growth and development. Blue light (320-500 nm) in particular acts to regulate a wide range of responses that serve to promote growth. These processes include chloroplast relocation movements, leaf positioning and expansion, stomatal opening and phototropism, all of which influence a plant's photosynthetic competence by improving efficiency of light capture, reducing photodamage, and regulating gas exchange between leaves and the atmosphere. Collectively, these responses elicit dramatic effects on plant growth and are controlled by phototropin blue-light receptors. Therefore, manipulation of phototropin receptor activity offers additional opportunities to increase photosynthetic performance and promote growth under specific light conditions. Indeed, our recent results demonstrate the potential to alter plant growth through changes in phototropin reactivity. Hence, a major outcome of this work will be to establish a structural and functional blueprint for constructing engineered photoreceptors directed at optimising photosynthetic productivity under specific light conditions. This proposal therefore offers an additional approach to coordinate stepwise enhancements in photosynthetic performance with an aim to increasing yield that should ultimately offer new strategies to grow crops more efficiently. The work outlined in this proposal will also generate new photoreceptor components with tailored photochemical properties that will have utility in generating synthetic protein-based switches designed to regulate target cellular activities by light.

Technical Summary

Increasing plant biomass has the potential to offer solutions for the food, energy, and environmental challenges of the future. Enhancing photosynthetic capacity and efficiency are recognised in this regard as bottlenecks to raising plant productivity. Recent engineering strategies have targeted diverse aspects of photosynthesis. However, an additional approach would be to target physiological processes that serve to optimise photosynthetic productivity. These include phototropism, leaf flattening and positioning, solar tracking, chloroplast relocation and stomatal opening, all of which are regulated by phototropin blue light receptors. Thus, engineering phototropin function has the potential to modulate plant growth through changes in photosynthetic efficiency. By using a directed evolution approach, we have identified key amino acid changes that markedly increase or decrease phototropin reactivity. This proposal focuses on harnessing these findings to fine-tune photoreceptor reactivity with an aim to modulate plant growth through changes in photosynthetic performance in Arabidopsis thaliana. Given the high degree of sequence conservation between higher plant phototropins, the design principles established here should extend to plant species important for producing sustainable food supplies and alternative fuel sources. This project is therefore of central importance to understanding how light is integrated to control a variety of responses that collectively promote plant growth and will ultimately provide new strategies to manipulate yield for agronomic gain. Knowledge gained from this work will also have relevance in the design of photosensory modules for emerging optogenetic applications.

Planned Impact

Beneficiaries: Beneficiaries of the research will include: academic scientists interested in plant photobiology and plant biotechnology, synthetic biologists interested in the design of artificial photoreceptor systems, commercial organisations interested in developing new strategies to improve crop production, individuals (text books for teaching), and organisations involved in science communication to schools and to the wider public (e.g. Glasgow Science Centre). Phototropin research will also appeal to the general public who can relate to the early phototropism work of Charles Darwin and its impact on science and evolution.

Benefits: The impact of the research is derived from its relevance to understand how blue light coordinates a variety of processes that serve to optimise photosynthetic efficiency and promote plant growth and the potential relevance to crop improvement for agronomic gain. Phototropin blue light receptors elicit dramatic effects on plant growth by collectively regulating chloroplast relocation movements, leaf positioning and expansion, stomatal opening and phototropism, all of which influence a plant's photosynthetic competence by improving efficiency of light capture, reducing photodamage, and regulating gas exchange between leaves and the atmosphere. Thus, research on engineering phototropin function has the potential to generate new strategies to further modulate plant growth through changes in photosynthetic efficiency. Phototropin signalling pathways are also known to prime plant defence responses and potentiate pathogen resistance. Hence, the engineering strategies devised in this proposal could also prove useful towards improving pathogen tolerance. Phototropins are universal in higher plants. Therefore, this project has the potential, in the longer term, to create new avenues for crop improvement that could benefit farmers, consumers and the environment and contribute to the economic competitiveness of the UK. The staff assigned to the project will obtain knowledge and expertise that can be applied in related research fields or more widely in the commercial or public sectors. Career progression of the RA will also benefit directly from collaborative aspects of the research with Prof. Getzoff (The Scripps Research Institute, San Diego).

Activities: The project will be continually managed by the PI to engage potential beneficiaries. The PI will publish the research in high-impact scientific journals, write reviews and book chapters and inform the University Media Relations Office of research highlights. Discussions with relevant commercial organisations will be initiated when appropriate to promote exploitation. The PI will communicate the research to school and university students via visits and University open days, initiate discussions with the Glasgow Science Centre, and present lectures at national and international conferences, as well as within Universities throughout the UK. The PI's web site will be routinely updated to communicate the research to the general public.

Publications

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Agliassa C (2018) Geomagnetic field impacts on cryptochrome and phytochrome signaling. in Journal of photochemistry and photobiology. B, Biology

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Buckley AM (2015) LOV-based reporters for fluorescence imaging. in Current opinion in chemical biology

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Cosentino C (2015) Optogenetics. Engineering of a light-gated potassium channel. in Science (New York, N.Y.)

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Fankhauser C (2015) Plant phototropic growth. in Current biology : CB

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Homans RJ (2018) Two photon spectroscopy and microscopy of the fluorescent flavoprotein, iLOV. in Physical chemistry chemical physics : PCCP

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Petersen J (2017) Functional characterization of a constitutively active kinase variant of phototropin 1. in The Journal of biological chemistry

 
Description We have successfully engineered photoreceptors to improve photosynthetic productivity and plant growth. Publications have been published in Science and PNAS and a patent filed in connection with PBL for one of the approaches used.
Exploitation Route Further patent application/funding being discussed/pursued along with transfer of the technology to crop relevant species.
Sectors Agriculture, Food and Drink,Education

 
Description Research was publicised via poster displays at the Glasgow Science Centre for the general public. Interest in photobiological research was stimulated. In 2017, my group participated in a widely and well received public outreach event as a part of the Glasgow Botanic Gardens Bicentenary celebrations by presenting their research to the general public through fun activities and by unveiling a booklet that showcasing the rich history and diverse cutting-edge research carried out by the different plant science groups at Glasgow. Furthered in 2018 by showcasing plant science research at the Glasgow Science Festival. This was also continued in 2019.
First Year Of Impact 2014
Sector Agriculture, Food and Drink,Education
Impact Types Societal

 
Title BLINK1 
Description Light regulated potassium channel that has potential in neural silencing. Now demonstrated utility in plants (Science paper pending). Patent has been filed in collaboration with PBL. 
Type Of Material Technology assay or reagent 
Year Produced 2015 
Provided To Others? Yes  
Impact Published in Science and awaiting further examination of its utility for optogenetics. 
 
Description BLINK Patent 
Organisation Plant Bioscience Limited Technology
Country United Kingdom 
Sector Private 
PI Contribution Data that has now secured a patent filing.
Collaborator Contribution Patent filing.
Impact Patent filed.
Start Year 2019
 
Description Justyna Labuz 
Organisation Jagiellonian University
Country Poland 
Sector Academic/University 
PI Contribution We have successfully engineered the activity of phototropin receptors to modulate plant growth.
Collaborator Contribution Justyna Labuz has characterised chloroplast movement responses.
Impact A manuscript based on our work has been submitted to PNAS.
Start Year 2017
 
Description Matt Jones 
Organisation University of Essex
Department School of Biological Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution Collaborative research on engineered photoreceptors.
Collaborator Contribution Collaborative research on engineered photoreceptors.
Impact Initial data obtained and studies being extended to contribute to publication.
Start Year 2017
 
Description Mike Blatt 
Organisation University of Glasgow
Country United Kingdom 
Sector Academic/University 
PI Contribution Collaboration has capitalised on using synthetic photosensory tools to manipulate plant physiology. The work produced has just been accepted for publication in Science. A patent application has been filed.
Collaborator Contribution Electrophysiological and physiological characterisation of transgenic Arabidopsis.
Impact The work produced has just been accepted for publication in Science.
Start Year 2017
 
Description NMR 
Organisation Southern Methodist University
Country United States 
Sector Academic/University 
PI Contribution Collaborative work using NMR spectroscopy for protein structure.
Collaborator Contribution Collaborative work using NMR spectroscopy for protein structure. Hosted visiting PhD student.
Impact New data that should contribute to publication.
Start Year 2016
 
Description Noriyuki Suetsugu 
Organisation University of Kyoto
Country Japan 
Sector Academic/University 
PI Contribution This works is focussed on characterising NPH3/RPT2 proteins in plants and their contribution to optimising photosynthetic light capture.
Collaborator Contribution Dr Suetsugu visited and worked with us as a JSPS research fellow.
Impact One review has been published and another manuscript submitted to Plant Physiology.
Start Year 2016
 
Description Protein Crystallography 
Organisation Southern Methodist University
Country United States 
Sector Academic/University 
PI Contribution Provided constructs for protein expression/purification
Collaborator Contribution Protein Crystallography
Impact In progress
Start Year 2016
 
Description iLOV 
Organisation Tianjin Institute of Industrial Biotechnology
Country China 
Sector Charity/Non Profit 
PI Contribution Newton fellowship for IBioIC student to visit Tianjin to explore the utility of the iLOV FP in industrial bioprocessing.
Collaborator Contribution Hosting PhD student and training.
Impact N/A
Start Year 2017
 
Title LOV-based optogenetics for plants 
Description Artificial light-activated K+ channel that has been used to improve carbon assimilation, water use and growth in plants. 
Type Of Technology New/Improved Technique/Technology 
Year Produced 2019 
Impact Science paper pending and patent filed. 
 
Description 6th and 7th Edition Plant Physiology and Development 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact Worked with other editors and authors to formulate chapters for the teaching text book Plant Physiology.

Edition still in press so difficult to gage at present but should provide an instrumental guide for school pupils and students.
Year(s) Of Engagement Activity 2015,2016,2017
 
Description Botanics bicentenary 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact My group participated in a widely and well received public outreach event as a part of the Glasgow Botanic Gardens Bicentenary celebrations by presenting their research to the general public through fun activities and by unveiling a booklet that showcasing the rich history and diverse cutting-edge research carried out by the different plant science groups at Glasgow.
Year(s) Of Engagement Activity 2017
URL https://www.ugplantsci.org
 
Description Chair of the GRC on Photosensory Receptors and Signal Transduction 
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 I organised this biennial meeting which was attended by academics, students and from Industry as well as NASA. The meeting was highly successful based on questionnaire feedback and will run again in two years time.
Year(s) Of Engagement Activity 2016
 
Description DFG Roundtable Discussion on Photoreceptors 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Invited to attend DFG discussion group evaluating status and future scope of photoreceptor research.
Year(s) Of Engagement Activity 2017
 
Description Gatsby Summer School 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact I gave a plenary lecture and participated in the summer school to encourage teachers and students to consider plant science and photo biological research.
Year(s) Of Engagement Activity 2015
URL http://www.slcu.cam.ac.uk/outreach/gatsbyplants
 
Description Glasgow Science Festival: Science Sunday, June 2018 
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 Contributed to and participated in activities for the Glasgow Science Festival: Science Sunday, June 2018
Year(s) Of Engagement Activity 2018
 
Description Interaction with James Hutton Institute, Dundee. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Study participants or study members
Results and Impact Visited to initiate collaborative research. Collaborations now secured through a joint PhD student.
Year(s) Of Engagement Activity 2018
 
Description Invited participant at the New Phytologist Next Generation Scientist meeting. John Innes Conference Centre, Norwich Research Park, Norwich, UK, July 2017. 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Invited participant at the New Phytologist Next Generation Scientist meeting. John Innes Conference
Centre, Norwich Research Park, Norwich, UK, July 2017.Myself (Editor, New Phytologist), Chris Surridge
(Chief Editor, Nature Plants), Anne Knowlton (Senior Editor, Current Biology), Adam Wheeler (Senior
Publisher, Wiley) and Ashlynne Merrifield (Publisher, Taylor & Francis) sat on an expert panel and offered
advice on the processes of communicating science and publishing, as well as an Q&A sessions.
Year(s) Of Engagement Activity 2017
 
Description Invited seminar speaker, Freie Universitat Berlin 
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 Invited as one of two plenary speakers to a workshop on photobiology organised by PhD students at the Freie Universitat Berlin. Participated in discussions of future research directions
Year(s) Of Engagement Activity 2015
 
Description Invited speaker and session chair, International Congress of Light and Light Barcelona, August 2019. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Study participants or study members
Results and Impact Invited speaker and session chair, International Congress of Light and Light Barcelona, August 2019. Presented two talks in different session on opotgenetic strategies to improve plant growth.
Year(s) Of Engagement Activity 2019
 
Description Invited speaker and session chair, International Symposium on Plant Photobiology, Barcelona, June 2019. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Invited speaker and session chair, International Symposium on Plant Photobiology, Barcelona, June 2019. Presented research on how to engineer photoreceptor to improve plant growth.
Year(s) Of Engagement Activity 2019
 
Description Invited speaker, SEB Meeting - From Proteome to Phenotype (Dec, 2017). Edinburgh, UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Study participants or study members
Results and Impact Invited speaker, SEB Meeting - From Proteome to Phenotype (Dec, 2017). Edinburgh, UK. Presented research on how to engineer photoreceptor to improve plant growth.
Year(s) Of Engagement Activity 2017
 
Description Invited speaker, Symposium in Honour of Winslow Briggs (April, 2018). Stanford, USA. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Study participants or study members
Results and Impact Invited speaker, Symposium in Honour of Winslow Briggs (April, 2018). Stanford, USA. Prof. Briggs sadly passed away at the age of 90 in Feb. 2019. Fund raising has been established to provide support/honour for plant scientists in his recognition.
Year(s) Of Engagement Activity 2018
 
Description Nagoya Institute of Technology International Symposium on Frontier Materials 
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 Invited speaker and workshop participant
Year(s) Of Engagement Activity 2016
 
Description Royal Society 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Other audiences
Results and Impact Invited to serve on the Royal Society University Research Fellowship (URF) Biological Panel as of 2015.

Yet to implement.
Year(s) Of Engagement Activity 2015
 
Description Summer students 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact Host summer students to gain laboratory experience and educate them about plant science.

Can be effective in changing students views towards plant science and help make them decide to pursue this as a degree or career.
Year(s) Of Engagement Activity Pre-2006,2006,2007,2008,2011,2012,2013,2014