Betaine production in diatoms - engineering more nutritious and salt tolerant crops

Lead Research Organisation: University of East Anglia
Department Name: Graduate Office

Abstract

The quaternary nitrogen compound betaine is the most abundant osmolyte on Earth. Betaine production in organisms protects against osmotic, temperature and drought stresses and its release into the environment provides dissolved organic nitrogen. Also, it is desirable for crops to have high betaine levels due to its wide-ranging health benefits. Betaine synthesis is well understood in many organisms but there are no detailed studies on betaine synthesis in any marine eukaryotic phytoplankton - not a single synthesis gene is reported.

Using diatoms as model phytoplankton we have identified a betaine synthase gene that when cloned in Escherichia coli confers enhanced salt tolerance and the ability to produce betaine from glycine. With the need to maximize nutritional value and crop production in the modern world, science has successfully explored transgenic introduction of betaine synthesis genes into crop plants to enhance their stress tolerance and betaine production. Given that the diatom synthase has no significant identity to any characterised betaine synthesis enzyme, it is a good candidate for transgenic stress tolerance studies in higher plants.

This PhD aims to demonstrate the role of this betaine synthase in Thalassiosira pseudonana via overexpression and knocking down of the corresponding gene in this model diatom. We will study gene regulation and the physiology of transgenic diatoms under varied environmental conditions and monitor their osmolyte production by liquid chromatography/mass spectroscopy. To explore the potential of producing stress tolerant higher plants, the diatom betaine synthase will be introduced and expressed in Arabidopsis and wheat. Comprehensive training will be provided at UEA by Mock and Todd, and at JIC by Balk and Uauy.

Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M011216/1 01/10/2015 30/09/2023
1654293 Studentship BB/M011216/1 01/10/2015 30/09/2019 Ana Bermejo Martinez
 
Description Community Resource for Wheat Transformation
Amount £0 (GBP)
Organisation National Institute Of Agricultural Botany 
Sector Private
Country United Kingdom
Start 08/2016 
End 09/2017
 
Description Collaboration with JB Raina at the University of Technology Sydney, Australia 
Organisation University of Technology Sydney
Country Australia 
Sector Academic/University 
PI Contribution Myself and my research team (including Dr Carrion, Dr Curson and Dr Pinchbeck; and PhD students Ana Bermejo Martinez and Beth Williams) collaborate with Dr Raina to carry out the localisation of isotopes within microbes using NANOSIMS technology.
Collaborator Contribution We identified novel genes involved in the cycling of DMSP/DMS and of betaine in multiple different prokaryotic and eukaryotic microbes. We shared our data with Dr Raina who used his expertise to link our localisation work of key enzymes to the localisation of key labelled compounds within our model organisms.
Impact Our most recent Nature Microbiology paper Curson et al., (2018) DSYB catalyses the key step of dimethylsulfoniopropionate biosynthesis in many phytoplankton, was an out put from this collaboration. The work is multidisciplinary involving: Molecular biology, Molecular Ecology, bioinformatics, algal physiology, NANOSIMS and ImmunoGold localisation for example.
Start Year 2017
 
Description Microbial synthesis of dimethylsulphoniopropionate and dimethylsuphide (Prof. Xiao-Hua Zhang Ocean University, China) 
Organisation Ocean University of China
Country China 
Sector Academic/University 
PI Contribution Collaborating with Prof. Xiao-Hua Zhang (Ocean University, China) we are characterising the molecular genetic systems utilised by marine organisms in their catabolism and synthesis of dimethylsulphoniopropionate (DMSP).
Collaborator Contribution Prof. Xiao-Hua Zhang (Ocean University, China) has isolated thousands of marine microbes and is screening them for their ability to produce and catabolise dimethylsulphoniopropionate (DMSP). Any positive strains are shipped to UEA for molecular characterisation.
Impact There are no output as yet but a full NERC grant will be submitted on this subject in January 2015.
Start Year 2013
 
Description Relating denitrification to DMSP cycling 
Organisation University of East Anglia
Department Biomedical Research Centre
Country Unknown 
Sector Academic/University 
PI Contribution Through work on our current NERC grants, we collaborate with Prof. David Richardson, Dr Gary Rowley and Dr Andrew Gates on work relating the process of denitrification to DMSP cycling in important marine prokaryotes and eukaryotes. We provide expertise, bacterial and plankton model organisms and methods to characterise these processes in the marine environment. We cosupervise PhD students with these collaborators on this topic.
Collaborator Contribution Prof. David Richardson, Dr Gary Rowley and Dr Andrew Gates provide expertise in the denitrification process.
Impact 1 NERC EnvEast PhD studentiship and 1 UEA-funded PhD studentship.
Start Year 2016
 
Description Subcellular localisation of DMSP and DMS-producing enzymes in eukaryotes and prokaryotes 
Organisation University of Barcelona
Country Spain 
Sector Academic/University 
PI Contribution Collaborating with Dr. Elena Mercade (University of Barcelona, Spain) we are growing eukaryotes and prokaryotes that produce DMSP and/or DMS and providing specific antibodies against the enzymes involved to localise them at a subcellular level.
Collaborator Contribution Dr. Elena Mercade (University of Bacelona, Spain) has prepared samples for immunogold labelling with the specific antibodies provided by us and used Transmission Electronic Microcoscopy to localise the enzymes involved in DMSP and/or DMS synthesis in the cells.
Impact The collaboration has successfully allowed the localisation of key enzymes involved in DMSP and/or DMS production in diatoms and bacteria. These results will be included in the forthcoming publications.
Start Year 2014
 
Description The importance of bacterial DMSP production in coral reef environments. 
Organisation Australian Government
Department Australian Institute of Marine Science
Country Australia 
Sector Public 
PI Contribution We provide knowledge and expertise on bacterial DMSP production to enable an environmental microbiological study of the importance of bacteria in DMSP production in coral reefs. This involved us designing enrichment experiments for the isolation of DMSP-producing bacteria. We provided ratified gene probes and techniques to study the expression and abundance of key genes involved in DMSP production in environmental samples. Furthermore, we provided expertise in the analysis of metagenomic data generated from coral reef environmental samples.
Collaborator Contribution Dr David Bourne and Dr Jean-Baptiste Raina obtained samples from coral reefs and conducted experiments to study the importance of bacteria in DMSP production under the guidance of our group. This involved the isolation of high quality DNA and RNA which is being analysed for the abundance and diversity of bacterial DMSP synthesis genes. Dr Raina is localising DMSP in model phytoplankton for our project using nanoSIMS.
Impact None yet.
Start Year 2015
 
Description The importance of bacterial DMSP production in coral reef environments. 
Organisation University of Technology Sydney
Country Australia 
Sector Academic/University 
PI Contribution We provide knowledge and expertise on bacterial DMSP production to enable an environmental microbiological study of the importance of bacteria in DMSP production in coral reefs. This involved us designing enrichment experiments for the isolation of DMSP-producing bacteria. We provided ratified gene probes and techniques to study the expression and abundance of key genes involved in DMSP production in environmental samples. Furthermore, we provided expertise in the analysis of metagenomic data generated from coral reef environmental samples.
Collaborator Contribution Dr David Bourne and Dr Jean-Baptiste Raina obtained samples from coral reefs and conducted experiments to study the importance of bacteria in DMSP production under the guidance of our group. This involved the isolation of high quality DNA and RNA which is being analysed for the abundance and diversity of bacterial DMSP synthesis genes. Dr Raina is localising DMSP in model phytoplankton for our project using nanoSIMS.
Impact None yet.
Start Year 2015
 
Description The production of betaine by marine phytoplankton (Dr Ruth Airs PML) 
Organisation Plymouth Marine Laboratory
Country United Kingdom 
Sector Academic/University 
PI Contribution During our NERC grant (NE/J01138X/1) we identified candidate genes involved in the synthesis of betaine. We have shown that model diatoms make betaine and that our candidate genes when cloned and expressed confer the ability to produce betaine and confer salt tolerance to Escherichia coli. In collaboration with Dr Ruth Airs we have established liquid chromatography mass spectrometry methods to detect betaine and dimethylsulphoniopropionate (DMSP) from biological samples.
Collaborator Contribution Dr Ruth Airs at PML helped us to develop and establish liquid chromatography mass spectrometry methods to detect betaine and dimethylsulphoniopropionate (DMSP) from biological samples.
Impact My postdoctoral researcher Research technicians on the NERC Grant (NE/J01138X/1) have presented our findings at the research conferences detailed below: -The Molecular Life of Diatoms, Paris, Dr Andrew Curson (Tues 25th June) -MMEG Molecular Microbial Ecology Group 2013, University of Essex, Ana Bermejo Martinez (Tues 17th Dec 2013) -6th International symposium on biological and environmental chemistry of DMS(P) and related compounds, ICM-CSIC, Barcelona, Ana Bermejo Martinez (26th - Fri 30th May 2014) As a result of this work we put together a NERC ENVEAST PhD proposal with Dr Todd as PI, and Dr Ruth Airs and Prof, Mock as CoIs. The work is multi-disciplinary as it involves phytoplankton physiology, biogeochemistry, microbiology, biochemistry, molecular genetics, molecular ecology and analytical biology.
Start Year 2013
 
Description 'DMSP production in marine bacteria and algae: idenitification of novel synthesis genes' research talk at Microbiology Society Annual Conference in Belfast April 2019 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Research talk at Microbiology Society annual conference, presenting a summary of research work in Todd's lab.
Year(s) Of Engagement Activity 2019
 
Description 'Marine Microbial DMSP Synthesis - from genes to microbes to pathways' at GEOMAR in Kiel, Germany in October 2018. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited research presentation at GEOMAR in Kiel Germany, presenting a summary of research work in Todd's lab.
Year(s) Of Engagement Activity 2018
 
Description 'Marine Microbial DMSP Synthesis - from genes to microbes to pathways' at the Marine Microbes GRC 2018, Lucca, Italy, 1-6th July 2018. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited research talk at the Gordon Research Conference Marine Microbes conference, covering all research work from the Todd group.
Year(s) Of Engagement Activity 2018
 
Description 'Surface marine sediments are factories for DMSP and the climate active gas DMS' at the ASM Microbe meeting, June 2019, San Francisco. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited reesearch presnetation at ASM conference in San Francisco, summarising the reseacrh from Todd's lab.
Year(s) Of Engagement Activity 2019
 
Description BIO Research Colloquium 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Other audiences
Results and Impact I presented my research as a poster in the BIO Research Colloquium, a one day conference where PIs from the School of Biological Sciences present their research and posgraduates present posters. I obtained a second prize for poster presentation.
Year(s) Of Engagement Activity 2017
 
Description European Nitrogen Cycle meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact I presented my research as a poster at the European Nitrogen Cycle meeting. Experienced as well as early carreer researchers from around Europe attend to this conference.
Year(s) Of Engagement Activity 2016
 
Description Jonathan Milner Talk 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Other audiences
Results and Impact I took part in the organization and chaired a free lecture given by Jonathan Milner, CEO of ABCAM. Undergraduates, posgraduates and PIs from University of East Anglia from all backgrounds (predominantely business and sciences) attended.
Year(s) Of Engagement Activity 2016
 
Description Microbiology Seminar 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact I presented my research as a guest speaker in the microbiology seminar held at the Kenyan Medical Research Centre-Wellcome Trust, Kilifi, Kenya.
Year(s) Of Engagement Activity 2017
 
Description Molecular Microbial Ecology Group (MMEG) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact I gave an oral presentation about my research to the Molecular microbial ecology group. Around 100 early carreer researchers and PIs attend to this meeting from around the UK.
Year(s) Of Engagement Activity 2017
 
Description Molecular Microbiology seminar series 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact Formal talk about my research to the Molecular Microbiology department at the School of Biological Sciences, University of East Anglia. Posgraduate students and PI from the department, as well as other interested audience from the university attend to this seminar series.
Year(s) Of Engagement Activity 2016
 
Description Nature Microbiology Community blog post 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact A blog covering the research published in Nature Microbiology 2018 published in Nature Microbiology community blog.
Year(s) Of Engagement Activity 2018
URL https://naturemicrobiologycommunity.nature.com/users/84434-andrew-curson/posts/30656-dmsp-synthesis-...
 
Description Norwich Science Festival 2018 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Norwich Science Festival is a big event organised by the Forum Trust focused on Science outreach to general public. In 2018, the Todd lab had a stand showcasing the research don in the lab with explanations and activities catered to all ages. During the day >100 people visited the stands and activities showing great engagement and interest.
Year(s) Of Engagement Activity 2018
 
Description Pint of Science 
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 I was "Our Body" Event Manager for Pint of Science, a three day event with talks from researchers from Norwich Research Park to a general audience (delivered in a local pub).
Year(s) Of Engagement Activity 2016
 
Description Posgraduate Colloquium 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact I presented a talk as part of the posgraduate colloqiuum held by the School of Biological Sciences. All posgraduate students from the school as wells as some PIs were present.
Year(s) Of Engagement Activity 2016
 
Description Present in the Pub 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact The talks was an engaging and informal presentation of the research carried our as part of my PhD. It took place in an informal setting at the Graduate Bar at the University of East Anglia. Attendees included posgraduate students from very different areas from humanities to sciences.
Year(s) Of Engagement Activity 2015
 
Description Press cover 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Publication in Nature Microbiology, 2017, was covered by Spanish regional newspaper "La Voz de Galicia" with a reach of thousands of people.
Year(s) Of Engagement Activity 2017
URL https://www.lavozdegalicia.es/noticia/sociedad/2017/02/22/bacterias-marinas-producen-gas-influye-cli...
 
Description Seminar & Journal Club 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact I gave a talk as part of a seminar series at the university of Kilifi, Kenya. This activity was aimed to undergraduate students from the Biotecnology and the Biology departments in order to expose them to international research.
Year(s) Of Engagement Activity 2017