Investigation and characterisation of diamide insecticide resistance conferred by target-site mutations in the ryanodine receptor of lepidopteran pest
Lead Research Organisation:
CARDIFF UNIVERSITY
Department Name: School of Medicine
Abstract
Diamide insecticides, such as flubendiamide and chlorantraniliprole, act as conformation sensitive activators of insect ryanodine receptors (RyR), releasing calcium from intracellular stores in insect muscle, and causing an irreversible paralysis. These compounds are the main active ingredients in diamide formulations used to control a diverse range of herbivorous insects, particularly lepidopterans such as the diamondback moth, Plutella xylostella, a notorious global pest on cruciferous crops. This common pest has recently developed widespread resistance to diamides through specific mutations in the trans-membrane domain of the RyR. Additional diamide insecticides such as cyclaniliprole, broflanilide and tetraniliprole are currently under development and will further increase the selection pressure on a number of target pests. It is therefore a key priority to investigate these resistance mechanisms by means of detailed biochemical and molecular studies. This project will characterise their impact on diamide insecticide action and RyR function, using state of the art approaches including cell-line expression of recombinant proteins, radioligand binding studies, electrophysiological recordings, calcium imaging and CRISPR/Cas9 guided drosophila transgenesis, and will be key to providing a fundamental understanding of diamide insecticide binding to allelic RyR variants. This knowledge will further impact future resistance management strategies for diamides and the discovery pipeline for novel insecticidal RyR modulators. You will be trained in standard biochemical, molecular and genetic techniques that are widely applicable and will also benefit from learning more specialised techniques such as insect transgenesis. You will also be required to spend 3 months on placement with the industry partner Bayer CropScience in Monheim, Germany.
Publications
Troczka BJ
(2017)
Rapid selection for resistance to diamide insecticides in Plutella xylostella via specific amino acid polymorphisms in the ryanodine receptor.
in Neurotoxicology
Richardson E
(2020)
Diamide resistance: 10 years of lessons from lepidopteran pests
in Journal of Pest Science
Homem RA
(2020)
Evolutionary trade-offs of insecticide resistance - The fitness costs associated with target-site mutations in the nAChR of Drosophila melanogaster.
in Molecular ecology
Richardson E
(2021)
Chimeric Investigations into the Diamide Binding Site on the Lepidopteran Ryanodine Receptor.
in International journal of molecular sciences
Richardson E
(2022)
Diamide insecticide resistance in transgenic Drosophila and Sf9-cells expressing a full-length diamondback moth ryanodine receptor carrying an I4790M mutation.
in Pest management science
Studentship Projects
Project Reference | Relationship | Related To | Start | End | Student Name |
---|---|---|---|---|---|
BB/N504075/1 | 26/10/2015 | 24/10/2019 | |||
1653661 | Studentship | BB/N504075/1 | 01/01/2016 | 31/12/2019 | Ewan Richardson |
Description | The stated purpose of the studentship was to determine the impact of ryanodine receptor mutations upon diamide efficacy. To that end, candidate mutations identified in the field were inserted into the ryanodine receptor of an otherwise diamide-susceptible moth, species Plutella xylostella. The biology of mutated ryanodine receptors was investigated both in-vitro and in-vivo. Key outcomes of the work: - Creation of a resistance-model fly strain which expresses the ryanodine receptor (and mutants there-of) of a moth in place of its own. - Quantitative and qualitative characterisation of two field-derived mutations, in terms of their impact upon diamide efficacy, in moth cell lines. Additionally, characterisation of one of these mutations in vivo, by use of the afore-mentioned fly model. |
Exploitation Route | Findings may be used by manufacturers and farmers to predict diamide control failures in the field Additionally, the developed fly model may vastly accelerate repeated investigations of this kind. |
Sectors | Agriculture Food and Drink Chemicals Environment Pharmaceuticals and Medical Biotechnology |
Description | Genetics Society Training Grant |
Amount | £1,000 (GBP) |
Organisation | The Genetics Society |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 07/2017 |
End | 09/2017 |
Description | PhD - Investigation and characterisation of diamide insecticide resistance due to target site mutations in the ryanodine receptor of lepidopteran pests |
Organisation | Bayer |
Department | Bayer CropScience Ltd |
Country | United Kingdom |
Sector | Private |
PI Contribution | Main PhD supervisor. Co-supervisor. |
Collaborator Contribution | Co-supervisors. Access to equipment & facilities |
Impact | Work in progress |
Start Year | 2015 |
Description | PhD - Investigation and characterisation of diamide insecticide resistance due to target site mutations in the ryanodine receptor of lepidopteran pests |
Organisation | Cardiff University |
Department | School of Medicine |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Main PhD supervisor. Co-supervisor. |
Collaborator Contribution | Co-supervisors. Access to equipment & facilities |
Impact | Work in progress |
Start Year | 2015 |
Description | Genetics society article: Injection steps for CRISPR/Cas9 |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | A written article for the Genetics Society Magazine, detailing best practice for CRISPR/Cas9 oocyte injection in Drosophila melanogaster. The article was prompted by a visit made, under funding from the society, to a specialist genomic engineering faculty in Maryland, USA. |
Year(s) Of Engagement Activity | 2018 |
Description | Hack.Agtech |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Professional Practitioners |
Results and Impact | 50 engineers, scientists and students from diverse backgrounds attended a three-day meeting in order to make partnerships towards developing solutions for boosting food security in the developing world |
Year(s) Of Engagement Activity | 2017 |