Summary

The term “Cibus Canola“ comprises various herbicide tolerant plants which were created either by genome editing or by conventional breeding methods. The method as described in the article was developed for the detection of a specific point mutation in the genome of certain canola lines. New breeding methods such as genome editing as well as classical breeding and random biological processes during tissue culture cultivation of canola plants can all be the cause of such a mutation. Based on the information available, BVL concludes that the very point mutation described in this article was, in fact, not caused by a genome editing technique. The method described in the article can specifically detect this mutation. However, it cannot identify whether or not the mutation was induced in this canola line by genome editing. Therefore, based on food and feed regulations, unassailable evidence cannot be presented in a court of law for the identification of a gene edited canola plant, using this method in isolation.

In detail:

a) Information on the analysed material is contradictory to the alleged genome editing based provenance

The article states that canola lines C1511, C5507 and 40K as purchased and examined by the authors were produced using genome editing (ODM as technique). As a reference, the authors cite a published opinion of the Canadian health authorities (Health Canada) dated 2016 ². According to the authors, all three canola lines analysed are derived from canola line 5715.

In fact, ODM is mentioned in said opinion of Health Canada as part of the scientific description of canola line 5715. However, it makes no direct link to the point mutation studied by the authors. Yet another opinion of the Canadian competent authorities (Canadian Food Inspection Agency)³, published as early as 2013, describes the mutation studied in the article as being a spontaneous mutation and not being caused by ODM. Likewise, Cibus US LLC stated to the Canadian Authorities that the mutation in fact is the result of a spontaneous mutation (somaclonal variation) which appeared during the tissue culturing processing, and which is not caused by the ODM technique. Canola line 5715 is a cross between a conventionally produced canola line (Clearfield Canola Variety SP Cougar CL) and the line BnALS-57. Line BnALS-57, in fact, was produced within the framework of a genetic engineering project which used ODM as a genome editing tool. The mutation studied in the article, however, does not originate from ODM. In order to provide transparent information on this very complex matter to the public, BVL decided to include the information on canola line 5715 in the database EUgenius⁴.

Based on existing information of the manufacturer (see internet presentation of Cibus US LLC, dated 8 September2020) sulfonylurea-tolerant Falco^TM-canola lines, as presently traded in the US, are produced using conventional breeding methods⁵. From this information BVL concludes that, while the detection method presented in the article is able to detect herbicide tolerant canola lines displaying a certain point mutation, it cannot discriminate specifically whether or not the detected point mutation can be assigned to a canola line produced using ODM. It follows that the article describes a method to detect mutations, but not a method to discriminate between canola lines originating from the application of different breeding techniques.

b) No method for identification of genome editing in plants

Even after the ruling of the European Court of Justice (ECJ) dated 25 July 2018⁶, mutations caused by tissue culture are not legally considered genetic engineering. Based on the information given, BVL assumes that the point mutation mentioned in the article was not produced by genome editing technology (ODM).

In contrast to conventional genetic engineering, with the application of novel genomic techniques like CRISPR/Cas or ODM there usually is no insertion of foreign DNA into the genome of a plant. As a consequence, the identification of genome edited plants with just a point mutation is not possible using conventional event-specific methods that rely on the specific border sequence between the plant genome and the inserted foreign DNA. The detection of a specific point mutation has been technically possible for decades now and per se does not represent a novelty for science. Various publications on the possibilities and limitations of detection and identification methods clearly pointed out the fact that the real challenge is not the detection of a point mutation. Rather, the real challenge is the identification of organisms created with novel genomic techniques and to analytically prove that the mutations were indeed caused by genome editing ^7-10.

c) The analytics for the detection of a point mutation as presented in the article are valid, but not sufficient to identify genome edited canola

The technical evaluation of the article confirms that all parameters on sensitivity and quantification have been accurately described by the authors. The PCR-method presented is a valid method for the detection of a point mutation in a canola line. However, and contradictory to the authors‘ statement, this method cannot give unassailable court-proof evidence on how the mutation was created, i.e. by means of applying genome editing, by other technical means or by spontaneous natural mutation. In addition, the article does not provide any strategy on how to detect a genome edited-based mutation, if the manufacturer or breeder do not supply any information or a publication.

d) Efforts of BVL and Julius Kühn-Institut (JKI) to solve the problem of identification

In 2015, Cibus US LLC demanded a legally binding assessment of BVL to clarify whether point mutations, if created by genome editing (ODM in this case) would fall under the genetic engineering regulation. On 25 July 2018, ECJ decided in its ruling that, other than plants created by conventional methods of mutagenesis, plants modified by ODM are subjected to European genetic engineering law.

In 2019, a working group within BVL started a pioneering project to clarify the problem of detection and identification using canola lines (experimental lines). For these lines, appropriate and well characterized reference material is available. These lines contain an identical point mutation but, whilst in some of the lines this mutation was created by conventional mutagenesis (somaclonal variation), in others the same mutation was caused by genome editing. Preliminary results indicate that with the customary PCR method the detection, but not the differentiation of these lines is possible. Therefore, the way these lines were produced could not be identified. In order to foster scientific research on this problem, the Federal Ministry of Food and Agriculture (BMEL) in conjunction with BVL and JKI has launched a call on a scientific research project featuring a ”feasibility study on detection and identification methods for genome edited plants and plant products“. The study aims to investigate the problem described above in more detail and shall further evaluate if and how evidence presented in a court of law for methods for detection and identification can be established for official controls of food and feed.

The national reference laboratory on GMOs which is located within the BVL, will test the method described in the article regarding its performance characteristics using available reference material.

References:

1. Chhalliyil, P.; Ilves, H.; Kazakov, S.A.; Howard, S.J.; Johnston, B.H.; Fagan, J. (2020). A Real-Time Quantitative PCR Method Specific for Detection and Quantification of the First Commercialized Genome-Edited Plant. Foods. 9:1245.
2. Health Canada Novel Food Information, https://www.canada.ca/en/health-canada/services/food-nutrition/genetically-modified-foods-other-novel-foods/approved-products/novel-food-information-cibus-canola-event-5715-imidazolinone-sulfonylurea-herbicide-tolerant.html
3. DD 2013-100: Determination of the Safety of Cibus Canada Inc. (Incorporated)´s Canola (Brassica napus L. (Linnaeus)) Event 5715, https://www.inspection.gc.ca/plant-varieties/plants-with-novel-traits/approved-under-review/decision-documents/dd-2013-100/eng/1427383332253/1427383674669
4. https://euginius.eu/euginius/pages/gmo_detail.jsf?gmoname=5715
5. https://www.falcoseed.com/technology
6. Urteil des Europäischen Gerichtshofs (Große Kammer) vom 25. Juli 2018 in der Rechtssache C‑528/16, http://curia.europa.eu/juris/document/document.jsf?text=&docid=204387&pageIndex=0&doclang=DE&mode=lst&dir=&occ=first&part=1&cid=929085#download=1
7. Wissenschaftlicher Bericht der Ressortforschungseinrichtungen BVL, JKI, FLI, TI, MRI und BfR im Auftrag des BMEL, https://www.bmel.de/DE/themen/landwirtschaft/gruene-gentechnik/neue-molekularbiologische-techniken.html
8. European Network of GMO Laboratories (ENGL) Detection of Food and feed Plant Products obtained by new Mutagenesis Techniques (2019), https://gmo-crl.jrc.ec.europa.eu/ENGL/ENGL.html
9. Grohmann L.; Keilwagen J.; Duensing N.; Dagand E.; Hartung F.; Wilhelm R.; Bendiek J.; Sprink T. (2019) Detection and Identification of Genome Editing in Plants: Challenges and Opportunities. Front Plant Sci. 10:236. https://www.frontiersin.org/articles/10.3389/fpls.2019.00236/full
10. Broll H.; Braeuning A.; Lampen A. (2019). European Court of Justice decision for genome editing: Consequences on food/feed risk assessment and detection. Food Control. 104:288.

Year of issue 2020
Date 09.09.2020