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An Integrated Multi-Omics Analysis of the NK603 Roundup-Tolerant GM Maize Reveals Metabolism Disturbances caused by the Transformation Process

corn cob, snow, winter

Abstract

Glyphosate tolerant genetically modified (GM) maize NK603 was assessed as ‘substantially equivalent’ to its isogenic counterpart by a nutrient composition analysis in order to be granted market approval. We have applied contemporary in depth molecular profiling methods of NK603 maize kernels (sprayed or unsprayed with Roundup) and the isogenic corn to reassess its substantial equivalence status. Proteome profiles of the maize kernels revealed alterations in the levels of enzymes of glycolysis and TCA cycle pathways, which were reflective of an imbalance in energy metabolism. Changes in proteins and metabolites of glutathione metabolism were indicative of increased oxidative stress. The most pronounced metabolome differences between NK603 and its isogenic counterpart consisted of an increase in polyamines including N-acetyl-cadaverine (2.9-fold), N-acetylputrescine (1.8-fold), putrescine (2.7-fold) and cadaverine (28-fold), which depending on context can be either protective or a cause of toxicity. Our molecular profiling results show that NK603 and its isogenic control are not substantially equivalent.

Introduction

The application of genetic engineering (GE) to modify edible crops is often advocated as one of the most important scientific advances to improve farming systems and feed the world in a more sustainable manner1. GE has been used to create crops adapted to abiotic stress, resistant to pathogens, with a longer shelf life, or with enhanced nutritional properties. However, commercialization of these traits is currently minor. Agricultural genetically modified (GM) crops are dominated by plants engineered to tolerate application of a herbicide or/and to produce their own insecticides2. A total of 180 million hectares of GM crops are currently cultivated worldwide on around 1.5 billion hectares constituting approximately 10% of global arable land3. Approximately 80% of GM crops have been modified to tolerate application of and thus accumulate glyphosate-based herbicide residues without dying in order to facilitate weed management.

Regulations for the release of genetically modified organisms (GMOs) of any kind in a country are covered by the national biosafety regulations of that nation. Guidance on risk assessment (RA) aim at identifying and avoiding adverse effects by early detection and proper evaluation of intended and potential unintended changes in a GMO. These should be detected and identified at early stages of RA, often referred to as “hazard identification”. Hazard identification is essential to the RA process as it sets the foundation of what is considered or observed in later steps in the risk assessment process4. In the US, the Food and Drug Administration considers GM technology as an extension of conventional breeding and GMO crops are deregulated once nutritional and compositional “substantial equivalence” is demonstrated5. The set of parameters and analyses necessary to declare a GMO as substantially equivalent to its conventional counterpart is still vague and focuses on a restricted set of compositional variables, such as the amounts of protein, carbohydrate, vitamins and minerals. GMOs are then declared substantially equivalent when sufficient similarities appear for those selected variables6. Remarkably, while a majority of GMO crops have been modified to withstand and thus accumulate a herbicide without dying, analysis for residues for such pesticides are neglected in compositional assessment7.

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