Abstract
Exhaustive analysis of genetically modified crops over multiple decades has increased societal confidence in the technology. New Plant Breeding Techniques are now emerging with improved precision and the ability to generate products containing no foreign DNA and mimic/replicate conventionally bred varieties. In the present study, metabolomic analysis was used to compare (i) tobacco genotypes with and without the CRISPR associated protein 9 (Cas9), (ii) tobacco lines with the edited and non-edited DE-ETIOLATED-1 gene without phenotype and (iii) leaf and fruit tissue from stable non-edited tomato progeny with and without the Cas9. In all cases, multivariate analysis based on the difference test using LC-HRMS/MS and GC–MS data indicated no significant difference in their metabolomes. The variations in metabolome composition that were evident could be associated with the processes of tissue culture regeneration and/or transformation (e.g. interaction with Agrobacterium). Metabolites responsible for the variance included quantitative changes of abundant, well characterised metabolites such as phenolics (e.g. chlorogenic acid) and several common sugars such as fructose. This study provides fundamental data on the characterisation of gene edited crops, that are important for the evaluation of the technology and its assessment. The approach also suggests that metabolomics could contribute to routine product-based analysis of crops/foods generated from New Plant Breeding approaches.
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Data availability
Unprocessed data can be accessed at https://data.mendeley.com/drafts/djd85rr6z9.
Abbreviations
- ANOVA:
-
Analysis of variance
- Cas9:
-
CRISPR associated protein 9
- GC–MS:
-
Gas chromatography mass spectrometry
- LC-HRMS:
-
Liquid chromatography high resolution mass spectrometry
- PC:
-
Principal component
- PCA:
-
Principal component analysis
- PLS:
-
Partial least square
- RSD:
-
Relative standard deviation
- TCA:
-
Tricarboxylic acid
- UPLC-PDA:
-
Ultra performance liquid chromatography-photodiode array detector
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Acknowledgements
The authors thank Chris Gerrish and Kit Liew for excellent technical assistance.
Funding
This work was funded by European Union Funding for Research and Innovation “Horizon 2020—NEWCOTIANA—Project No. 760331” and in part by Biotechnology and Biological Sciences Research Council OPTICAR Project BB/P001742/1.
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PDF secured funding. MN, JA and ER designed constructs and created the edited plants. MN, GE, JA, MD and PDF designed the experiment. MD cultivated the plants, performed metabolite analysis and analysed the data. MD, JA, GE and PDF wrote the manuscript. All authors reviewed the manuscript.
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Drapal, M., Enfissi, E.M.A., Almeida, J. et al. The potential of metabolomics in assessing global compositional changes resulting from the application of CRISPR/Cas9 technologies. Transgenic Res 32, 265–278 (2023). https://doi.org/10.1007/s11248-023-00347-9
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DOI: https://doi.org/10.1007/s11248-023-00347-9