Metabolic engineering of triterpenoid pathways involved in plant defense in Arabidopsis and rice
Lead Research Organisation:
John Innes Centre
Department Name: UNLISTED
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
Plants produce a diversity array of natural products (secondary metabolites) that have important ecological functions. Triterpenes are one of the largest classes of plant-derived natural products. They protect plants against pests and diseases and are also important as drugs and anticancer agents. Arabidopsis and cereals such as rice do not make protective triterpene glycosides. These species are therefore excellent experimental systems for proof-of-concept experiments involving metabolic engineering of triterpene pathways into heterologous species. The aim of this proposal is to engineer triterpene metabolic pathways into Arabidopsis and rice by genetic modification, using recombineering and synthetic biology. At the end of this project we will have the necessary knowledge to devise strategies to introduce multiple genes into diverse plant species for control of plant pests and diseases based on synthesis of protective triterpenes.
Planned Impact
unavailable
Organisations
People |
ORCID iD |
| Anne Osbourn (Principal Investigator) |
Publications
Polturak G
(2022)
Defense-related phenylpropanoid biosynthetic gene clusters in rice.
in Science bulletin
| Description | This project was focused on the engineering of triterpene biosynthesis into model plant species (Nicotiana benthamiana, Arabidopsis and rice) using synthetic biology. Triterpenes are one of the largest classes of plant-derived natural products. These compounds protect plants against pests and diseases and they are also important as drugs and anticancer agents. Arabidopsis and cereals such as rice and maize do not make triterpene glycosides, which are associated with protection against pests and diseases. These plants were therefore excellent experimental systems for proof of concept experiments involving metabolic engineering of the triterpene pathway into heterologous species. Within this project we have established a customised toolkit of genes and enzymes for metabolic engineering of triterpene synthesis in plants and validated this toolkit by heterologous expression of single and multiple triterpene biosynthetic genes in Nicotiana benthamiana. We have also demonstrated the production of triterpene scaffolds in Arabidopsis and rice. We have developed sets of domesticated regulatory and coding sequences for plant engineering. Importantly, we have also shown that the promoters from a root-expressed pathway in oat can be used to drive the co-ordinate expression of multiple genes in roots of other plant species. A manuscript on co-ordinate regulation of these promoters is in preparation. |
| Exploitation Route | The triterpene toolkit that we have generated represents a valuable resource for engineering of high value compounds in plants. The transient plant expression technology developed at the John Innes Centre has proved very powerful in enabling us to produce hundreds of milligrams and even up to a gram of triterpene. This opens the way for engineering of high value compounds in plants using synthetic biology approaches. We have also published on a proposed common plant syntax for plant synthetic biology, i.e. a universal design format that will enable the exchange of parts, which will accelerate the ability of the community to engineer new plant metabolic pathways and other traits of commercial importance. |
| Sectors | Agriculture Food and Drink Chemicals Creative Economy Education Manufacturing including Industrial Biotechology Pharmaceuticals and Medical Biotechnology |
| URL | https://www.jic.ac.uk/directory/anne-osbourn/ |
| Description | This project has contributed to the generation of a customised toolkit of genes and enzymes for metabolic engineering of triterpene synthesis in plants. We have developed sets of domesticated regulatory and coding sequences for plant engineering. Importantly, we have also shown that the promoters from a root-expressed pathway in oat can be used to drive the co-ordinate expression of multiple genes in roots of other plant species. These promoters have been patented and licensed to academic researchers and to industry. They represent valuable tools for the engineering of multi-gene root-expressed traits such as nitrogen fixation into plants. A manuscript is in preparation from this project but more time is needed because the postdoc has returned to Spain following the end of her fellowship and has since been on maternity leave. We also need to complete further experiments that will furnish information on the regulators that govern promoter expression. |
| First Year Of Impact | 2016 |
| Sector | Agriculture, Food and Drink,Chemicals,Education,Manufacturing, including Industrial Biotechology |
| Impact Types | Societal Economic |
| Title | Establishment of a common syntax for plant synthetic biology |
| Description | Inventors in the field of mechanical and electronic engineering can access multitudes of components and, thanks to standardization, parts from different manufacturers can be used in combination with each other. The introduction of BioBrick standards for the assembly of characterized DNA sequences was a landmark in microbial engineering, shaping the field of synthetic biology. Here, we describe a standard for Type IIS restriction endonuclease-mediated assembly, defining a common syntax of 12 fusion sites to enable the facile assembly of eukaryotic transcriptional units. This standard has been developed and agreed by representatives and leaders of the international plant science and synthetic biology communities, including inventors, developers and adopters of Type IIS cloning methods. Our vision is of an extensive catalogue of standardized, characterized DNA parts that will accelerate plant bioengineering. To this end we have proposed a common syntax for exchange of DNA parts for plant synthetic biology. This is supported by the international community and has been published. |
| Type Of Material | Improvements to research infrastructure |
| Year Produced | 2015 |
| Provided To Others? | Yes |
| Impact | The establishment of a common plant syntax has paved the way for the introduction of an Open MTA for material transfer, now trialled and published (Kahl et al. 2018, Nature Biotechnology 36: 923), simple and open systems for recusrsive fabrication of DNA circuits (Pollak et al. 2018, New Phytologist: https://doi.org/10.1111/nph.15625) and for discussions with Addgene, the nonprofit global plasmid repository, archives and distributes plasmids for scientists. These advances are catalytic and we envisage that they will promote and accelerate plant biology research within the UK and internationally. |
| Description | Agritech Careers Day |
| 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 set up a day for sixth form science students to come to our research institute to learn about Agritech careers stemming from plant and microbial sciences. The students got to hear from a range of presenters from new PhD students to group leaders, spin out companies, industry and business. They also visited our field station to see large-scale crop trials out in the field. |
| Year(s) Of Engagement Activity | 2023 |
| Description | Built new image library for the Norwich Research Park |
| Form Of Engagement Activity | Engagement focused website, blog or social media channel |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Public/other audiences |
| Results and Impact | To help raise awareness of the high quality research carried out across the Norwich Research Park, we developed an open access image library to showcase images of NRP science accompanied by accessible legends suitable for a general audience. The image library has the potential to attract new traffic to the NRP institute websites and research group homepages and will allow groups such as the media, schools, the general public and other researchers to gain a glimpse of the great depth and diversity of research that is being carried out in Norwich. Although all the images are freely available to download and use, people are required to sign up to use the library and are asked to enter information at the time of download about their intended use of the image. This helps to capture who is using the library and where the images are being used. So far images have been downloaded 582 times and have been used in presentations, publications, on websites, for education and advertising. We ran an image competition to raise awareness across the NRP site and then assembled a judging panel to select the top 12 images which were used to create an NRP calendar for 2016. The calendar was sent to politicians, business leaders, industry and academics at institutions in the UK and internationally. The competition featured in the local paper, the Eastern Daily Press and an overall winner was picked and used to make a large canvas which is hanging in the new Centrum building. We have received many positive comments about the libraries ease of use for finding good quality, copyright free images to use in presentations from fellow researchers across the site. In October 2016 we put on a two-week, large scale image exhibition at the Forum in Norwich as part of the first Norwich Science Festival. The images attracted a lot of interest and several artists have been in touch with scientists whose research images were on display to set up new collaborations using science to inform artistic practice. We will post outcomes of these endeavours onto the image library website to widen our exposure to new audiences. |
| Year(s) Of Engagement Activity | 2015,2016 |
| URL | http://images.norwichresearchpark.ac.uk/ |
| Description | SAW Seminar - Sowing the seeds for science outreach |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Professional Practitioners |
| Results and Impact | Presented creative outreach projects and ways to capture impact at the department seminar to encourage scientists to be bold with their outreach plans and to demonstrate the value of documenting the process and the outcomes for reporting, reflection and to improve the method. |
| Year(s) Of Engagement Activity | 2020 |
| Description | SAW workshop at Barford Primary School 11 March 2014 |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Saw (Science, Art and Writing) project in Barford primary school together with Jenni Rant, Gemma Farré Martinez, James Reed, Chris Hann (artist) and Mike O'Driscoll (writer). We spent an entire day at school, explaining my project (Engineer wheat for "Take all" resistance) and opening a dialog about GMOs with 10 to 11 years old pupils. This project took place on the Tuesday 11th of March 2014. |
| Year(s) Of Engagement Activity | 2014 |