17-ERACoBioTech - SUSPHIRE - Sustainable Bioproduction of Pheromones for Insect Pest Control in Agriculture

Lead Research Organisation: Earlham Institute
Department Name: Research Faculty

Abstract

The aim of this project is to enable bio-based manufacturing of insect pheromones in plants and fungi for the sustainable control of insect pests of agriculture and horticulture. This project builds on a proof-of-concept in which moth sex-pheromones were produced in plants.

Some of the most aggressive pests of agriculture are insect larvae. Semiochemicals are chemicals emitted by insects for communication. The most widely known of these are sex pheromones, produced by virgin females to attract mates of the same species. Dispensing insect sex pheromones in plant production environments is used to trigger sexual confusion in the target species and prevent breeding, thereby providing a highly species-specific control method. This presents a sustainable alternative to conventional pesticides, the use of which are progressively being restricted due to concerns about their non-specificity and negative impacts on biodiversity. Insect sex pheromones are already used as a pest-control strategy; however, chemical synthesis is currently the only approach for manufacturing and the use of toxic ingredients and the creation of toxic by-products is inevitable for some pathways. Further, the unusual chemical characteristics of many insect pheromones mean that chemical synthesis strategies are not cost effective. For example, Coccoidea species (scale insects and mealybugs) are aggressive pests of agriculture and horticulture and better control methods are highly desirable. However, their sex pheromones have unusual structures for which chemical synthesis is both difficult and expensive.

Previous studies by SUSPHIRE partners demonstrated that it is possible to engineer N. benthamiana plants to produce high quantities of moth sex pheromones. The SUSPHIRE project will improve on this initial proof-of-concept and will also identify and validate the genes in the Coccoidea pheromone biosynthetic pathway. We aim to optimize bioproduction of insect pheromones in plants and fungi and to evaluate the pheromones produced in these species in laboratory and field conditions.

The SUSPHIRE project aims to provide a sustainable, low-cost manufacturing platform for the commercial production of insect pheromones and reduce the cost of production of pheromones that are currently commercially non-viable. The introduction of these biotechnology approaches to pheromone production will expand the use of sex pheromones for sustainable pest control in agriculture, reducing its current environmental impact and providing sustainable manufacturing platforms.

Technical Summary

Previously we have demonstrated that it is possible to engineer N. benthamiana plants to produce high quantities of moth sex pheromones. The SUSPHIRE project will improve on this initial proof-of-concept and will also identify and validate the genes in the Coccoidea pheromone biosynthetic pathway. We aim to optimize bioproduction of insect pheromones in plants and fungi and to evaluate the pheromones produced in these species in laboratory and field conditions

SUSPHIRE will develop optimal genetic constructs for inducible heterologous biosynthesis of moth and Coccoidea pheromones in plants and fungi. We will develop and test tuneable, orthogonal regulatory elements. We will produce and analyse the yield, purity and efficacy of moth pheromones heterologously produced in plants and fungi, obtaining systems data and using modelling to inform a metabolic engineering strategy with the aim of increasing metabolic flux to the pathway and/or reducing expression of genes from competing pathways. We will generate expression data from Coccoidea species and use a comparative transcriptomics approach to identify irregular terpenoid core-structure forming enzymes as well as decorating enzymes. Genes will be identified, expression constructs assembled and delivered to plants and fungi, and the yield, purity and efficacy of heterologously produced mealy bug pheromones will be assessed.

We will maximize the impact of project results, by conducting cost-analyses, life-cycle analyses, scoping exercises for the commercialisation of the project outcomes together with a responsible research program as well as a comprehensive set of activities for communication and dissemination. Our aim is to understand the potential impacts and respond to the needs of stakeholders, including our intended market (growers), as well as other potential beneficiaries (e.g. vendors and consumers) that are not represented within our project consortium of ecologists, molecular biologists and chemists.

Planned Impact

The total use of insecticides and acaricides in the EU has risen to 3.078 tons/year (Eurostat 2014). Several pesticides have been at the heart of campaigns against large-scale agriculture facing criticism for environmental damage and negative impacts on biodiversity. In contrast, pheromones are precisely targeted to the pest species as, in general, each insect species has its own chemically distinct pheromone composition. Pheromones represent a more environmentally friendly approach to Integrated Pest Management; the pheromone market had an estimated value of $1385 million in 2013 and is projected to reach $2450 million in 2020. However, the production of some pheromones by chemical synthesis, as well as being costly, utilises toxic chemicals. SUSPHIRE aims to contribute to the reduction of pesticide applications in agriculture, by enabling the biosynthesis of pheromones that are difficult or impossible via chemical synthesis and increasing the sustainability of current manufacturing platforms, which often use hazardous chemicals or result in hazardous waste. SUSPHIRE will contribute to the goal of a circular economy reducing unrecyclable chemical waste and employing self-sustained CO2-fixing live bioreactors.

The insect pest control market was valued at $11.89 billion in 2015 and it is projected to reach $17.35 billion by 2022. The biotechnological approach proposed here could overcome current limitations in pheromone production, establishing Europe as a leader in this area. Sex-pheromones, considered the most effective compounds for mating disruption, account for 65% of the total pheromone market. The advances in innovation produced in SUSPHIRE will constitute an added value for the European research space, with the possibility of claiming IP in the technological applications. With increasing restrictions on pesticide use and the growth of the pheromone market, SUSPHIRE is poised to support a highly positive medium-term economic impact in the European agrochemical business market.

Many of the materials developed in this project have clear potential for wider impact in other biotechnology fields e.g. molecular pharming. We seek to encourage greater enterprise in industrial biotechnology by facilitating access to basic tools and enabling technologies on which we will not make IP claims. Our IP strategy is, therefore, to protect information related to the bioproduction of pheromones but to encourage greater enterprise in industrial biotechnology through facilitating access to tools and enabling technologies for plants and fungi.

Finally, SUSPHIRE will reinforce the current EU leadership in plant synthetic biology. EU groups are pioneering the design of genetic circuits and the use of programmable DNA binding proteins in plants (e.g. CRISPR/Cas9) for the development of non-food plant crops for bio-production of high-value products such as proteins, peptides and bioactive secondary metabolites, particularly for pharmaceutical and medical/veterinary, diagnostic, agricultural and industrial applications. In this regard, SUSPHIRE research efforts will be synergistic (and contemporary) with H2020 research projects aimed to develop plant biofactories for industrial applications using classic genetic engineering (BB-07-2017) and new plant breeding techniques (BIOTEC-07-2017).

Publications

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Vazquez-Vilar M (2018) DNA assembly standards: Setting the low-level programming code for plant biotechnology. in Plant science : an international journal of experimental plant biology