Exploiting seed coat properties to improve uniformity and resilience in brassica vigour

Lead Research Organisation: University of Exeter
Department Name: Physics

Abstract

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Technical Summary

Seed vigour is a multi-component trait that is critical for crop performance. Control of dormancy and germination rate are important components of vigour and are dependent on the interaction between plant genetics, the environment during seed production and seed applied agrochemicals. Seeds are highly sensitive to very small temperature changes during seed set, and these are sensed by the mother plant and used to control seed coat traits that affect seed dormancy and seed permeability. Climate change is increasing the unpredictability of vigour in commercial seed lots: therefore the next generation of crops will need high vigour that is uncoupled from effects of temperature variation during seed set. Our work shows that Arabidopsis mutants defective in seed coat development, tannin deposition and suberin deposition have exactly this phenotype. A new B. oleracea TILLING population and new genomic technologies will be exploited in a forward and reverse screening strategy to understand processes controlling seed coat properties in Brassicas, and to generate novel germplasm for breeding programs. Mutants identified from forward screens will be cloned by re-sequencing of bulk segregants and candidate genes confirmed using CRISPR or RNAi, and orthologues of genes known to affect seed permeability in Arabidopsis will be systematically knockout out in B. oleracea. These new alleles will be tested for vigour robustness in laboratory and field trials alongside commercial varieties at sites used for seed production for the UK market. A second strand to this work is the exploitation of seed permeability to enhance chemical uptake into seeds from commercial seed coatings. We will deploy SRS microscopy to analyse spatially, temporally and quantitatively how chemicals with different properties enter seeds with different permeability profiles, and to understand if seed coat permeability engineering has potential to improve chemical uptake into seeds.

Planned Impact

Impacts for the environment
Studies on seed uptake of agrochemicals are few, because of limitations of current technology described in this proposal. However, for many widely used compounds uptake is very poor and companies have responded to this simply by increasing dose. The result is that as little as 5% of the applied chemical actually enters the plant, with large quantities accumulating in agro-ecosystems, including soils and watercourses. It has now been shown in recent studies that invertebrate and bird diversity is being affected by seed-applied chemicals. It is becoming clear that the use of many seed-applied compounds may not be sustainable in the long term because these compounds have long half lives in the environment. This project aims to develop seeds that are highly permeable to compounds applied in industrial seed coatings and therefore to markedly improve uptake efficiency. This could lead to lower doses while maintaining effectiveness, and significantly reduce contamination of run-off into soils.

Impacts for seed companies
Seed companies frequently have problems with lot-to-lot variation of seed quality. This occurs because the seeds are produced at multiple sites around the world to ensure continuous supply of fresh seeds, or because stochastic temperature variation at production sites causes quality differences. In some cases this variation can be overcome by post-harvest seed enhancement, but this is expensive. Seed companies would benefit now from more predictability in quality, but are concerned that climate change is eroding their ability to reliably produce high quality seeds. The alleles we will isolate and characterise here have the potential to provide robust high quality, even in the face of environmental variation in the seed production conditions. After field trail (objective 3) these can be made available to our industrial collaborator for introgression into elite backgrounds (2-3 years). Because of this link we have a clear route to market and the potential to impact the seed market from 5 years after project end.

Impacts for growers
This project has the potential to impact growers by increasing the reliability of performance of seed they buy from seed companies. Germination of most varieties varies significantly from lot to lot. The new alleles produced in this project have the potential to eliminate much of this variation, improving the predictability and uniformity of seed raising. Increasing uniformity reduces the frequency with which harvest teams visit field sites and increases the proportion of the crop that attains the best grades, adding value for growers.

Training of seed technologists
Seed technologists are individuals trained in the analysing of seed vigour and seed enhancement. Currently there is a global shortage of seed technologists and little training available in the UK. Our program includes extensive experience in both genetics and seed vigour testing and a training secondment in the seed testing lab at Syngenta Vegetable Seeds. These secondments are essential to learn the analytical pipeline, use of software and the rules applied in accredited seed testing labs.

Impact for agrochemical development
Currently there are no good tools for monitoring agrochemical ingress into seeds and thus this is not often considered during development, screening and testing of potential new compounds for commercial seed coatings. SRS microscopy has great potential to be developed into a new tool for the agrochemical industry so that ingress into seeds with different properties can be monitored. In order to facilitate this we will hold a demonstration workshop at the end of the project. If chemicals can be monitored for uptake potential, leads can be screened for uptake potential, and modifications made that would improve uptake into seeds. In future SRS facilities could be built at agrochemical companies.
 
Description The main outcome of the Exeter lead part of the project (BB/L020637/1) was the development and application of a novel label-free, real-time imaging technique for visualising the distribution of compounds in the seed coat at the microscopic level. The ability to visualise the movement of unlabelled compounds into the seed coat on this scale represents a major advance in analytical capability for both plant biologists and agrochemical scientists.

The ability of a compound to permeate a seed coat depends greatly upon its chemical-physical properties. Therefore techniques that rely upon attachment of tracers (fluorophores and dyes) to visualise the distribution of compounds across the seed coat give a false result since they modify the transport kinetics of the compound under investigation. For this reason we have developed a label-free method for analysing uptake of AIs from seed imbibition media based on Stimulated Raman Scattering (SRS) microscopy. SRS is an optical microscopy that provides label-free quantitative imaging based on vibrational spectroscopy with sub-cellular spatial resolution. SRS has several key features that are ideal for imaging the entry of agrochemicals into seeds. (i) The near-infrared excitation permitts imaging through intact seed coat. (ii) It allows tracer-free mapping of small molecules such as agrochemicals, agrochemical formulations, water and endogenous seed coat polymers with high spatio-temporal resolution and no photobleaching. (iii) The SRS can be isolated from the background from heavily pigmented plant tissues. SRS has the potential to enable radically improved design and environmental profiles of existing and future seed-applied Active Ingredients by matching AIs to seed coats with tailored permeability profiles and to improve the environmental profiles of existing Active Ingredients.
Exploitation Route PI has received funding from project partner (Syngenta) to further investigate the potential of SRS for imaging the uptake of agrochemicals in-plants
Sectors Agriculture, Food and Drink,Environment,Healthcare,Pharmaceuticals and Medical Biotechnology

 
Description Impacts for the environment Studies on seed uptake of agrochemicals are few, because of limitations of current technology described in this proposal. However, for many widely used compounds uptake is very poor and companies have responded to this simply by increasing dose. The result is that as little as 5% of the applied chemical actually enters the plant, with large quantities accumulating in agro-ecosystems, including soils and watercourses. It has now been shown in recent studies that invertebrate and bird diversity is being affected by seed-applied chemicals. It is becoming clear that the use of many seed-applied compounds may not be sustainable in the long term because these compounds have long half lives in the environment. This project aims to develop seeds that are highly permeable to compounds applied in industrial seed coatings and therefore to markedly improve uptake efficiency. This could lead to lower doses while maintaining effectiveness, and significantly reduce contamination of run-off into soils. Impacts for seed companies Seed companies frequently have problems with lot-to-lot variation of seed quality. This occurs because the seeds are produced at multiple sites around the world to ensure continuous supply of fresh seeds, or because stochastic temperature variation at production sites causes quality differences. In some cases this variation can be overcome by post-harvest seed enhancement, but this is expensive. Seed companies would benefit now from more predictability in quality, but are concerned that climate change is eroding their ability to reliably produce high quality seeds. The alleles we will isolate and characterise here have the potential to provide robust high quality, even in the face of environmental variation in the seed production conditions. After field trail (objective 3) these can be made available to our industrial collaborator for introgression into elite backgrounds (2-3 years). Because of this link we have a clear route to market and the potential to impact the seed market from 5 years after project end. Impacts for growers This project has the potential to impact growers by increasing the reliability of performance of seed they buy from seed companies. Germination of most varieties varies significantly from lot to lot. The new alleles produced in this project have the potential to eliminate much of this variation, improving the predictability and uniformity of seed raising. Increasing uniformity reduces the frequency with which harvest teams visit field sites and increases the proportion of the crop that attains the best grades, adding value for growers. Training of seed technologists Seed technologists are individuals trained in the analysing of seed vigour and seed enhancement. Currently there is a global shortage of seed technologists and little training available in the UK. Our program includes extensive experience in both genetics and seed vigour testing and a training secondment in the seed testing lab at Syngenta Vegetable Seeds. These secondments are essential to learn the analytical pipeline, use of software and the rules applied in accredited seed testing labs. Impact for agrochemical development Currently there are no good tools for monitoring agrochemical ingress into seeds and thus this is not often considered during development, screening and testing of potential new compounds for commercial seed coatings. SRS microscopy has great potential to be developed into a new tool for the agrochemical industry so that ingress into seeds with different properties can be monitored. In order to facilitate this we will hold a demonstration workshop at the end of the project. If chemicals can be monitored for uptake potential, leads can be screened for uptake potential, and modifications made that would improve uptake into seeds. In future SRS facilities could be built at agrochemical companies.
First Year Of Impact 2017
Sector Agriculture, Food and Drink,Chemicals
Impact Types Societal,Economic

 
Description 21ENGBIO: Engineering targeted activation of fungicides at the plant-pathogen interface
Amount £83,471 (GBP)
Funding ID BB/W012936/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 01/2022 
End 01/2023
 
Description CONTRAST facility: clinical coherent Raman scattering facility
Amount £1,035,354 (GBP)
Funding ID EP/S009957/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 12/2018 
End 11/2020
 
Description Biological imaging of Zymoseptoria tritici wheat infection by stimulated Raman scattering microscopy 
Organisation Syngenta International AG
Country Switzerland 
Sector Private 
PI Contribution This is an industry funded project exploring the feasibility of using the CONTRAST facility to better understand how anti-fungal agrochemicals can be engineered with enhanced efficacy. My team are providing expertise in SRS imaging and undertaking all experiments carried out in the CONTRAST facility.
Collaborator Contribution Syngenta are providing funding (£25,000) for this project and expertise in the development of novel anti-fungal agrochemical agents.
Impact New understanding of the host-pathogen interaction during fungal infection of wheat
Start Year 2021
 
Description Chemical imaging by Coherent Raman Scattering (CRS) methods for com-pounds visualization in pesticidal target systems 
Organisation BASF
Country Germany 
Sector Private 
PI Contribution The evaluation of Coherent anti-Stokes Raman Scattering (CARS) and Stimulated Raman Scattering (SRS) to investigate uptake and distribution of pesticides in plant leaves after foliar application including co-localization of compounds with inner leaf structure (subcellular level), in roots, in seeds, in in-sects and microorganisms.
Collaborator Contribution Funding a post-Doctoral research for 12 months. Providing agrochemicals and agrochemical knowhow.
Impact None as yet. Project still running
Start Year 2021
 
Description Exploring Industry R&D Applications of Frequency Modulated SRS Imaging 
Organisation Unilever
Department Unilever UK R&D Centre Port Sunlight
Country United Kingdom 
Sector Private 
PI Contribution Feasibility study to explore the application of frequency modulated SRS for visualising the uptake of low molecular weight compounds in to human hair
Collaborator Contribution Financial support of postDoctoral salary for 6 months
Impact academic publication in preparation
Start Year 2015
 
Description Next Generation Optical Analysis for Agrochemical Research & Development 
Organisation Syngenta International AG
Department Syngenta Ltd (Bracknell)
Country United Kingdom 
Sector Private 
PI Contribution Translation of SRS techniques developed at Exeter into an analytical tool for agrochemical R&D
Collaborator Contribution Access to state of the art standard analytical tools that are not available at Exeter
Impact none yet.
Start Year 2015