Periodic Reporting for period 1 - PhytoENM (Applying novel analytical methods to elucidate bio-nano interactions at individual cell level in plants)
Período documentado: 2021-10-12 hasta 2023-10-11
In recent years, the agronomic application of Engineered NanoMaterials (ENMs) in plants (phytonanotechnology) has emerged to revolutionize conventional plant production systems for increased disease resistance, nutrient utilization, and crop yield. Along with the groundbreaking potential of such techniques, one should be careful about their trophic transfer to plants. This topic has been rarely studied, compared to the toxic effect of ENMs in human systems. However, assessment of ENM toxicity to plant cells is critical to the
implementation of nanotechnology in agriculture and support of global sustainability and the EC's European Green Deal. The proposed project addresses potential key cellular processes involved in the delivery of ENMs to plants and the safe use and social acceptance of phytonanotechnology, assessing potential adverse effects, including the risks associated with the transfer of ENMs through the food chain. To do so, novel analytical techniques are essential that could study metal-based ENMs and their distribution within individual cells. The development of Single Particle ICP-MS (SP-ICP-MS) is a new area of research which allows rapid detection and analysis of ENMs in a variety of matrices and applications, as it allows discrete pulses of positively charged ions to be detected and measured in a time resolved manner using microsecond data acquisition rates. PhytoENM will go beyond, applying the concept of Single Cell ICP-MS (SC-ICP-MS), where individual cells are rapidly analyzed for their particulate & ionic content, something that has not been done before in plants. The project will also use novel stable-isotope labelling techniques pioneered by the applicant team. This would allow the understanding of the interaction of metal-based ENMs in single cells spatially, and the related mechanistic pathways, leading to information on their distribution and potential toxicity.
The overall aim of the project is to elucidate fundamental plant-ENM interactions, by systematically evaluating the translocation and fate within plant cells of a reference group of ENMs, chosen for their relevance to phytonanotechnology. The work is focused on staple food plants (rice and wheat), to ensure agricultural relevance. In the proposed work, plant cellular uptake and translocation will be linked to the intrinsic physicochemical properties of ENMs, something that has been scarcely studied to date given a lack of appropriate analytical tools. With the very recent arrival of SC-ICP-MS, however, a novel analytical method pioneered by the supervisory partnership (PerkinElmer, supported by UoB), a unique opportunity has emerged. PhytoENM will, for the first time, test cell line and multi-cellular plant material to assess quantitatively uptake (dissolved vs. particulate) in individual single cell populations. The SC-ICP-MS technique has so far been tested on unicellular organisms (algae) and mammalian cell lines, but has yet to be tested comprehensively in plants and thus realize its potential commercialization and impact on agricultural practices. Plant cells are, however, discrete from algae and mammalian cells and dedicated methods are needed. The proposed work is highly novel and will open up the field of phytonanotechnology and facilitate its acceptance.
implementation of nanotechnology in agriculture and support of global sustainability and the EC's European Green Deal. The proposed project addresses potential key cellular processes involved in the delivery of ENMs to plants and the safe use and social acceptance of phytonanotechnology, assessing potential adverse effects, including the risks associated with the transfer of ENMs through the food chain. To do so, novel analytical techniques are essential that could study metal-based ENMs and their distribution within individual cells. The development of Single Particle ICP-MS (SP-ICP-MS) is a new area of research which allows rapid detection and analysis of ENMs in a variety of matrices and applications, as it allows discrete pulses of positively charged ions to be detected and measured in a time resolved manner using microsecond data acquisition rates. PhytoENM will go beyond, applying the concept of Single Cell ICP-MS (SC-ICP-MS), where individual cells are rapidly analyzed for their particulate & ionic content, something that has not been done before in plants. The project will also use novel stable-isotope labelling techniques pioneered by the applicant team. This would allow the understanding of the interaction of metal-based ENMs in single cells spatially, and the related mechanistic pathways, leading to information on their distribution and potential toxicity.
The overall aim of the project is to elucidate fundamental plant-ENM interactions, by systematically evaluating the translocation and fate within plant cells of a reference group of ENMs, chosen for their relevance to phytonanotechnology. The work is focused on staple food plants (rice and wheat), to ensure agricultural relevance. In the proposed work, plant cellular uptake and translocation will be linked to the intrinsic physicochemical properties of ENMs, something that has been scarcely studied to date given a lack of appropriate analytical tools. With the very recent arrival of SC-ICP-MS, however, a novel analytical method pioneered by the supervisory partnership (PerkinElmer, supported by UoB), a unique opportunity has emerged. PhytoENM will, for the first time, test cell line and multi-cellular plant material to assess quantitatively uptake (dissolved vs. particulate) in individual single cell populations. The SC-ICP-MS technique has so far been tested on unicellular organisms (algae) and mammalian cell lines, but has yet to be tested comprehensively in plants and thus realize its potential commercialization and impact on agricultural practices. Plant cells are, however, discrete from algae and mammalian cells and dedicated methods are needed. The proposed work is highly novel and will open up the field of phytonanotechnology and facilitate its acceptance.
One of the key steps for the project was to synthesise and characterise ENMs. Hence, Gold (Au) nanoparticles were synthesised and characterised. Then experiments were designed for method development and validation using SC-ICP-MS to analyse the ENM in collaboration with PerkinElmer. Even though we struggled a lot initially due to instrument malfunctioning, however, we made some progress to test synthesised Au-ENM with this. However, we were not able to test real samples as the plant cell protoplasting was progressing slowly. The next stage was In vitro plant cell experiments using plant cell lines and single cells, to establish the transport, distribution and toxicity of ENMs. Not much significant progress was made with this as we faced analytical challenges while working on the plant cell protoplasting. However, all these challenges helped us learn a lot about this completely unique and novel technique. These information si going to help us explore the SC-ICP-MS technique further to unravel it's future capabilities. We also tried simpler plants (Arabidopsis thaliana) first to standardise the method, however, results were not consistently encouraging, possibly due to lack of cell stability caused by higher osmotic pressure post enzymatic digestion. As the in vitro plant cell experiments were not completed, hence, the In vivo validation could not be performed. Premature termination of the project was another reason that we could not continue to try and achieve this goal. Additionally, we could not proceed with the policy implication studies due to lack of overall scientific experimental data.
Dissemination of the results to academic audiences was done, based on whatever progress we could have made after facing the technical challenges through participating in seminars and conferences. Even though we did not manage to generate enough dataset to present in a conference, however we did participate in the BNASS 2022 (Manchester, UK) and also in one workshop seminar at the PerkinElmer R&D headquarters in Seer Green, Beaconsfield, UK to share our results. I also presented the research and results internally at UoB in the PERCAT as well as Environmental Nanoscience Research Group meetings.
Due to lack of significant scientific dataset, we haven’t publish any scientific article based on the project yet. However, while working on the MSCA project, I published one open access article as detailed below (the project Reference H2020-MSCA-IF-2020-101031051 was duly acknowledged):
Mandal D, Aghababaei M, Das SK, Majumder S, Chatterjee D, Basu A. Isolation and Identification of Arsenic Hyper-Tolerant Bacterium with Potential Plant Growth Promoting Properties from Soil. Minerals. 2022; 12(11):1452.
We are also preparing a draft manuscript based on the PhytoENM project and when published, it will include reference to the EU funding and will be in line with the funder’s open access publishing policy.
Majumder et al. (in prep.) Understanding the future of single cell analysis for nansafety – a perspective from agro-economics and global food security. Target journal: Nature Perspectives.
Dissemination of the results to academic audiences was done, based on whatever progress we could have made after facing the technical challenges through participating in seminars and conferences. Even though we did not manage to generate enough dataset to present in a conference, however we did participate in the BNASS 2022 (Manchester, UK) and also in one workshop seminar at the PerkinElmer R&D headquarters in Seer Green, Beaconsfield, UK to share our results. I also presented the research and results internally at UoB in the PERCAT as well as Environmental Nanoscience Research Group meetings.
Due to lack of significant scientific dataset, we haven’t publish any scientific article based on the project yet. However, while working on the MSCA project, I published one open access article as detailed below (the project Reference H2020-MSCA-IF-2020-101031051 was duly acknowledged):
Mandal D, Aghababaei M, Das SK, Majumder S, Chatterjee D, Basu A. Isolation and Identification of Arsenic Hyper-Tolerant Bacterium with Potential Plant Growth Promoting Properties from Soil. Minerals. 2022; 12(11):1452.
We are also preparing a draft manuscript based on the PhytoENM project and when published, it will include reference to the EU funding and will be in line with the funder’s open access publishing policy.
Majumder et al. (in prep.) Understanding the future of single cell analysis for nansafety – a perspective from agro-economics and global food security. Target journal: Nature Perspectives.
As the project was terminated early due to technical as well as professional reasons, the larger societal impacts could not be achieved. Due to insufficient dataset, the policy framework analysis regarding the long term impact on future food security as well as the environmental and public health based on the regulatory reviews and experimental outcomes could not be executed.