Bioceres announces commercial release of GMO wheat in Brazil

Bioceres Crop Solutions announced that the National Technical Biosafety Commission (CTNBio) completed the safety assessment of HB4 wheat and approved its commercialization and cultivation in the country. The request was presented by Tropical Melhoramento e Genética (TMG), which has a partnership with Bioceres. The content of the partnership was not explained by the companies.

Although the conclusion was included in the minutes of the CTNBio meeting (image above), to date there has been no publication in the Official Gazette of the Union, a legally necessary milestone to publicize the decision.

Initially, the request had been for the commercial release of genetically modified wheat flour (event IND-00412-7) "for exclusive use in food, feed or derived or processed products". This request was granted at the 246th CTNBio meeting.

On November 9, 2022, there was an addition. The claim now includes “cultivation, production, handling, transportation, transfer, commercialization, import, export, storage, release and disposal of this GMO”. The decision reported here included the amended request.

According to the company's statement, HB4 technology is a key tool in adapting agricultural systems to a more extreme climate, such as in cases involving severe water stress.

Brazil is the second country where regulatory agencies have approved HB4 wheat for cultivation, after Argentina, and completes the approval processes for the company's target wheat markets in Latin America.

The extract from the opinion reads more information: "[…] wheat genetically modified for tolerance to drought and herbicide, event IND- 00412-7 and its derivatives, containing the HaHB4 gene, coming from Sunflower, and pat, coming from Streptomyces viridochromogenes. […] CTNBio exempts the applicant from post-commercial release monitoring based on the first paragraph of article 18 of CTNBio Normative Resolution 32, which provides that GMOs and their Risk Class 1 derivatives released for commercial use will be exempt from monitoring post-commercial release".

Executive summary

To understand the issue in greater depth, see the text below. It comes from the executive summary that TMG presented to CTNBio when requesting commercial release for the cultivation of HB4 Wheat:

"The genetically modified event of Wheat IND-ØØ412-7 or Wheat HB4, was developed by the Argentine company Bioceres. In Brazil, Bioceres and Tropical Melhoramento e Genética (TMG) signed a partnership to enable the use of this genetically modified event in the country. Through of this partnership, TMG had access to studies carried out in other countries, and was responsible for the biosafety report on Wheat IND-ØØ412-7, which was presented to CTNBIO and approved by the Commission, for the purpose of human and animal nutrition (Process CTNBIO 01250.014650/2019-71, Technical Opinion CTNBIO 7795/2021). Now, through LPMAs and containment studies carried out in Brazil, TMG obtained the data and wrote the risk assessment to the environment and is requesting commercial release for cultivation of Wheat HB4.

The data presented here comes from studies carried out in Argentina, combined with the LPMAs and containment studies carried out in Brazil. Wheat IND-ØØ412-7 (also designated as “Wheat HB4”), conventional parental control (non-genetically modified) and commercial varieties (Reference Standards) were field tested.

The HaHB4 gene, from Sunflower (Helianthus annuus), gives wheat significant potential for increasing productivity in environments with low water availability, without compromising the production ceiling in optimal environmental conditions for the crop. This contribution has been demonstrated over several years and in test sites in Argentina and now in Brazil.

Wheat HB4 was generated using microparticle bombardment methodology, and contains the following introduced genes: 1) the sunflower HaHB4 transcription factor gene, 2) the bar gene from Streptomyces hygroscopicus that confers resistance to glufosinate. The HaHB4 gene (Helianthus annuus Homeobox 4), naturally present in sunflower, is expressed at very low levels and activated when plants are subjected to conditions of water stress, saline stress, darkness or insect attacks. Thus, the HAHB4 protein is already a natural component of human and animal nutrition, as well as in environments where conventional sunflower cultivation occurs.

Wheat HB4 has one complete copy of HaHB4 with functional regulatory elements, and two complete copies of bar with its regulatory elements in the correct position. The HAHB4 protein in Wheat HB4 belongs to the HD-Zip family of transcription factors, characterized by the presence of two functional domains: the Homeodomain (HD), responsible for binding to DNA, and a “Leucine Zipper” (LZ) motif, involved in protein-protein interaction and dimerization. The protein does not have any insecticidal or herbicide tolerance function and acts mainly in response to abiotic stress, mainly in ethylene-signaled pathways. The PAT protein in Wheat HB4 confers on plants the phenotype of tolerance to herbicides based on glufosinate ammonium, through the expression of the protein phosphinothricin N-acetyl-transferase.

Several analyses, evaluations and studies were carried out in the field and in containment with Wheat HB4, in Argentina and Brazil, to prove its safety for human and animal consumption and during its cultivation and presence in the environment.

Extensive analysis of the HAHB4 protein confirms its food, nutritional and environmental safety. This conclusion is based on evidence from multiple sources: origin of the gene from a vegetable already used in human food and history of use and exposure; evaluation of the digestibility and thermal stability of the HAHB4 protein in an in vitro test; comparisons of the amino acid sequence with known toxins, allergens and allergenic sequences, through bioinformatics analyses; glycosylation status; oral toxicity study of the HAHB4 protein; and HAHB4 protein level in forage and HB4 wheat grains.

In vitro simulated gastric fluid (SGF) assay revealed rapid degradation of HAHB4 protein. No protein fragments were observed after the first 30 seconds of digestion. The HAHB4 protein did not fragment during exposure to prolonged heat cycles, according to SDS-PAGE analyses, without any significant changes. Bioinformatics research revealed no homology of HAHB4 with known allergenic or toxic proteins. The results indicate that the HAHB4 protein is unlikely to cause an allergic reaction in humans or that it is toxic to humans or animals and is therefore safe for human and animal consumption.

An oral toxicity study of HAHB4 protein was conducted in rats at a dose of 3822 mg of HAHB4 protein per kg of body weight. All mice survived and did not show any adverse effects caused by the protein tested during the study. All these results suggest that the maximum tolerated dose of HAHB4 protein without observing adverse effects is greater than 3822 mg/kg BW and, therefore, the No Observed Adverse Effect Level (NOAEL) is greater than 3822 mg/kg BW. For this reason, it is concluded that acute oral toxicity must be very low, since no mortality was observed in rats, as well as no adverse effects were observed from the consumption of the HAHB4 protein.

The PAT protein is expressed in numerous genetically modified crops that have been approved for commercial release, in Brazil by CTNBio and by other agencies in other countries around the world. These approvals included abundant information and data on the physicochemical characteristics of the PAT protein that confirm the lack of allergenic or toxic properties, including its digestibility, thermal sensitivity and absence of toxicity. The absence of toxic actions attributable to the PAT protein has been widely verified over almost 20 years of consumption of several genetically modified crops that have the gene and express the PAT protein.

HAHB4 protein expression levels were determined in wheat seeds and leaf tissue collected from field trials in Argentina (2012-2013). A specific and sensitive LC-MS/MS method was developed and validated to detect the expected low levels of this transcription factor. As expected for a transcription factor, HAHB4 protein expression levels in Wheat HB4 were below the lower limit of quantification (LLOQ). This suggests that the IND-ØØ412-7 Wheat event does not contain levels of genetically modified protein that could negatively affect non-target organisms, as HaHB4 gene expression in IND-ØØ412-7 Wheat is extremely low. Thus, the HAHB4 protein does not have the potential to produce adverse effects on field exposure levels on representative species of beneficial invertebrates.

The compositional analysis of Wheat HB4 was carried out following the OECD consensus document. 45 components (nutrients, micronutrients, vitamins, minerals and antinutrients) were determined in grain and forage samples from Wheat HB4, conventional parental control, and a set of commercial reference varieties, in field trials carried out in six different locations . Considering the natural variability that the composition of the wheat crop presents, the composition of Wheat HB4 was substantially equivalent to the composition of its conventional parental control and/or was within the natural variability of commercial reference varieties and/or within the range reported in the literature .

The agronomic performance of the genetically modified IND-ØØ412- 7 wheat event was evaluated in comparison with conventional parental control and commercial reference varieties. The trials were conducted in several locations in Argentina and Brazil. Characteristics measured included: seed germination; agronomic and phenotypic evaluations; and ecological assessments, including damage caused by diseases and insects, with no significant difference being found between HB4 Wheat and its non-genetically modified conventional, or even between commercial cultivars. A study of pollen morphology and pollen fertility was carried out in Argentina. The resulting data supports the conclusion that HB4 Wheat does not differ from its conventional parental control and commercial reference varieties beyond its intended effect. The results demonstrated that HB4 Wheat has plant characteristics similar to those of commercial reference varieties and does not represent an environmental risk compared to conventional parental control.

Based on the information already presented in process 01250.014650/2019-71, the data and information presented in this document and the approval for human and animal consumption by CTNBIO (Technical Opinion 7795/2021), it is concluded that Wheat HB4 does not represent a risk to human consumption, animal consumption and does not cause an environmental impact when compared to conventional wheat, that is, a negligible risk. With this document, CIBio da TMG SA. requests the commercial release of HB4 Wheat for the purpose of cultivation, production, handling, transportation, transfer, commercialization, import, export, storage, release and disposal of this GMO."

UPDATE on 07-04-2023

Due to questions about the safety of this cultivar, CTNBio issued a technical note on 30/03/2023. Among other considerations, the body informed that:

"CTNBio rejects the technical questions raised about the biosafety assessment carried out. A point-by-point response was prepared by an expert in the commercial release process for planting IND-ØØ412-7 (HB4) wheat, of which we highlight:

A. the DNA sequences inserted into the wheat plant genome were evaluated for the potential to generate undesirable effects from a biosafety point of view. No significant risks to the environment were identified. On the contrary, IND-ØØ412-7 wheat plants were phenotypically similar to conventional wheat, differing only in the characteristics that were introduced via transgenics in all experimental tests;

B. phenomena of deletion, insertion and inversions of DNA segments also occur in the conventional breeding process. The literature is rich in reports of these phenomena within the same species, particularly in polyploid species, such as wheat;

C. the data provided by the applicant proved, based on scientifically appropriate experiments, the stability of the inserts in successive generations of wheat. Furthermore, the genes HaHb4 (which confers greater tolerance to water stress) and bar (which confers tolerance to glufosinate ammonium) are inherited en bloc (cosegregate) in a Mendelian manner;

D. the evaluation of inserted genes carried out by Southern Blot and DNA sequencing – using the PACBIO platform, today the gold standard for characterizing and “deciphering” the genomes of different organisms – brought information that covers the requirements of risk analysis regarding the characterization of the regions of the genomes where the two insertions occurred;

E. Data on the behavior of wheat in Brazil were generated in planned releases into the environment installed in representative regions for wheat cultivation, Cambé/PR and Uberlândia/MG, in two different growing seasons. The field experiments were carried out in accordance with the provisions of the Normative Resolutions for Planned Releases into the Environment (RN06, RN35). Complete data from studies carried out were presented.

F. It is essential to state that in addition to the data provided by TMG, CTNBio's decision was also based on scientific literature and information provided by the public hearing.

CTNBio reiterates what it stated in its opinion: that IND-ØØ412-7 (HB4) wheat is as safe for the environment, human health and animal health as its conventional equivalent."

In the same document, the agency added:

"CTNBio did not request the CNBS analysis, mainly for the reasons we highlighted:

1. The inserted characteristic of tolerance to water stress may be important to minimize losses due to droughts, which become more frequent due to climate changes resulting from global warming. It can also help in the expansion of wheat cultivation in the country, notably in those regions where winter has low rainfall, and can help the country, a large importer, to become self-sufficient;

2. The country has experience in the cultivation of genetically modified vegetables, cultivating them on a large scale for more than 15 years with no proven problems to human health, animal health or the environment;

3. The same genes inserted in wheat IND-ØØ412-7 (HB4) are present in genetically modified soybeans IND-00410-5 released for commercial use since 2019;

4. Cultivation in a neighboring country, in regions with edaphoclimatic similarities, can lead to seed smuggling and illegal cultivation in the country if there is an impediment to cultivation in Brazil. A similar case was observed with RR soybeans, the so-called Maradona soybeans, in which a large contingent of farmers in the southern region of Brazil planted and harvested GM soybeans illegally. This fact generated the need for the government to issue Provisional Measure 133/2003, converted into Law no. 10.688/2013.

5. All biosafety assessment standards were complied with by the National Biosafety Technical Commission;

6. The administrative process was strictly observed with wide publicity through official channels;

7. The risk analysis carried out by the Commission did not identify any element that indicated an increased risk of HB-4 Wheat in relation to conventional wheat.

The complainants do not present elements or information that would require a new biosafety assessment by CTNBio. The decision on the convenience and opportunity of adopting the technology now approved by the Commission is within the scope of the powers of the National Biosafety Council - CNBS, and the provision of this appeal is not recommended by this advisory body."

In the link below you can read the full opinion that analyzed the request for release, minutes of the body's meeting and the statement, excerpts of which are transcribed above.