Dir 111 Limited and controlled release of wheat and barley genetically modified for altered grain composition, nutrient utilisation efficiency, disease resistance or stress tolerance Applicant

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Summary of issues raised in submissions received from prescribed experts, agencies and authorities¹⁷ on the consultation RARMP for DIR 111

The Regulator received several submissions from prescribed experts, agencies and authorities on the consultation RARMP. All issues raised in submissions that related to risks to the health and safety of people and the environment were considered in the context of the currently available scientific evidence and were used in finalising the RARMP that formed the basis of the Regulator’s decision to issue the licence. The submissions received that raised issues relating to risks to the health and safety of people and the environment are summarised below.

Summary of issues raised	Comments
Any risk resulting from gene stacking should be addressed in more detail in the risk assessment.	More detail added in Risk Scenarios 1 and 3.
Consideration should be given in the risk assessment (Section 2.2) to the potential for weediness as a result of the presence of the bar gene in the GMOs.	A paragraph has been added in Risk Scenario 3 to address this issue and corresponding text in Chapter 1, Section 6.3 has also been amended.
The distance of the isolation zone needs to be clarified.	The apparent inconsistency in isolation distance has been clarified, including that any plantings of GMOs will be at least 200 m from the nearby salt tolerance wheat trial.
Advises that more discussion about siRNA target sequences in humans as well as the potential for accumulation of plant siRNAs sequences in blood and possible effects should be included in the RARMP.	More detail added in Risk Scenarios 1 and 2.
Suggests that the RARMP acknowledges in paragraph (258) that there is uncertainty in relation to the characterisation of risk scenario 2 as well as risk scenario 1, regarding potential increase in toxicity and allergenicity as a result of the introduced genes	Text has been amended to this effect.
Given the proposed licence authorises dealings with the GM wheat and barley that may occur in different states, the licence should be explicit that:
all such dealings should comply with relevant state laws	The licence contains a preamble, which makes it clear that the licence is issued by the Regulator in accordance with the Gene Technology Act 2000 and, as applicable, Corresponding State Law. It also notes that dealings permitted by the licence may be subject to the operation of State legislation declaring areas to be GM, GM free, or both, for marketing purposes.
any animal experiments occur only as authorised through a duly constituted animal research ethics committee any GM material be transported according to the OGTR Guidelines for Transport, Storage and Disposal.	Licence conditions require approval of animal experiments by an animal ethics committee, and transport of GM plant material according to the Regulator’s guidelines.
the GMOs covered include wheat or barley containing any combination of introduced genes and regulatory sequences in the relevant table.	The GMOs covered by the licence are listed in the appendix to the licence, which specifies the genes and regulatory sequences authorised.
One reason Risk scenario 2 is not identified as a risk that warrants further assessment is because the CSIRO Human Research Ethics committee is required to undertake a full assessment of potential risk to human volunteers. It is unclear how this assessment would differ from the assessment of risk to the health and safety of people the OGTR is required to do under the Act and how this reason contributes to Risk scenario 2 being identified as a risk that does not warrant further assessment.	Section 4(b) of the Act acknowledges that the Act provides a regulatory framework in conjunction with other Commonwealth and State regulatory schemes. The Human Research Ethics Committee (HREC) will consider potential adverse health effects and any other ethical issues for volunteers, and may have access to additional information generated from animal feeding studies conducted as part of this trial.
Toxicity should be considered arising from unexpected changes to the metabolome upon combination of traits, eg altered nutrient utilisation with altered grain composition.	More detail added to the RARMP. Combination of traits and their potential to increase toxicity were considered and amended in Risk Scenario 1. Unexpected changes in the GMOs as a result of the introduction of individual genetic elements or their combination are considered in Risk Scenario 6.
Advises that some genetic modifications are aimed at changing the characteristics of the GM wheat or barley that restrict their potential for persistence. Therefore, these characteristics should not be named as justification for the limited potential for spread and persistence of the GMOs. Additionally, altered phenotypes have not been assessed in current commercial varieties.	The purpose of the genetic modification is to produce plants that are more productive in agricultural environments than non-GM wheat or barley plants. These changes could also result in greater persistence in agricultural and other land uses. Risk Scenario 3 and the section on uncertainty have been amended to reflect this. The limits and controls of the trial will restrict spread and persistence of the GM plants.
Suggests that subtle unexpected effects may affect risk profiles, particularly considering the large number of genetic modifications and the potential for gene stacking. Proposes that unexpected effects may arise from the introduction of transcription factors.	One of the purposes of the trial is to observe agronomic performance in a field situation. More detail has been added to Risk Scenario 6. The limits and controls will restrict spread and persistence of the GM plants regardless of the source of potential unexpected effects.
The following additional data requirements for any larger scale or commercial release of these GM wheat and barley lines are suggested: Data on relative growth rates of the GM wheat and barley lines including tolerance to environmental stresses, such as drought, heat, salt, cold etc. this should include a consideration of early vigour of the GM plants. Data on the biomass accumulation for each GM wheat and barley line together with the harvest index. Data on the effect of the introduced traits on GM wheat and barley tillering, grain size, altered root structure and root depth. Data on the seed dormancy of the GM wheat and barley. Data on the seedling germination rate for each wheat and barley line. Data on the disease resistance of the wheat and barley Data on the characteristics which are assumed to limit the potential weediness of wheat and barley, such as head shattering, seed dispersal or seed longevity. Data allowing comparison of the potential weediness of GM strains relative to the existing elite strains grown commercially in Australia. More comprehensive information on the potential effects of modifications to genes related to improved water use efficiency, carbon assimilation and photosynthesis. It is recommended that the effect of trait-plus-trait analysis on the data collection requirements be examined in greater detail prior to finalising the RARMP	The section on uncertainty was changed to be somewhat more specific. The RARMPs provide a broad guidance on data requirements for possible future larger scale or commercial releases. The OGTR also strongly encourages potential applicants to discuss details of data required prior to submission of such applications. If sufficient data is not presented in a future application, the application may not be accepted, more data may be requested from the applicant, risk treatment measures imposed or a licence refused. The applicant has been advised of these suggestions.
Recommends that the buffer zone of 2 m and the monitoring zone of 10 m be extended to 20 and 50 m respectively for rodent control. Also recommends that registered bait be applied whenever rodent activity is detected.	A licence condition has been added requiring additional rodent control, such as baiting or trapping, to be implemented if rodent activity is detected. The literature regarding rodent control was reviewed, and this together with the 2 m buffer zone and 10 m monitoring zone, maintained so as to not attract or harbour rodents, is considered adequate.

Summary of issues raised in submissions received from the public on the consultation RARMP for DIR 111

The Regulator received three submissions from the public on the consultation RARMP. These submissions, summarised in the table below, raised issues relating to human health and safety and the environment. These were considered in the context of currently available scientific evidence in finalising the RARMP that formed the basis of the Regulator’s decision to issue the licence.

View (general tone): n = neutral; x = do not support; y = support

Issues raised: AR: Antibiotic resistance; E: Environmental risk; GS: Gene stacking; H: Human health; RA: Risk analysis; Res: research; T: Toxicity; U: Uncertainty; W: Weediness

Other abbreviations: Act: Gene Technology Act 2000; Ch: Chapter; FSANZ: Food Standards Australia New Zealand; GM: Genetically Modified; GMO: Genetically Modified Organism; LC: Licence Conditions; RAF: Risk analysis framework; RARMP: Risk Assessment and Risk Management Plan.

Type: I: individual; NGO: Non-government organisation.

Sub. No:	Type	View	Issue	Summary of issues raised	Comment
1	I	x	U, H, E, RA	Considers that trials like this are not adequate for ensuring the effects of the GM living organisms in the environment are known and believes that there are sure to be ramifications that are not knowable from such limited trials. Objects to humans and ecosystems being eventually the testing ground for the release or artificial life forms.	The current application is not for a large scale or commercial release of GM wheat or barley. The RARMP for this release considered all relevant information in the context of the limited scale of the proposed release, and concluded that risks to the environment are negligible. One of the purposes of the trial is to evaluate the agronomic performance and analyse any changes in grain composition of the GMOs so that more knowledge about the GMOs can be obtained from the trial.
2	I	x	U, H, Res	As part of the CSIRO development of their GMOs. Early, prudent, controlled trials should be undertaken to determine suitability of GM products for the food chain. In addition, the effects of gene stacking should be assessed before exposing humans to the GM grain.	This limited and controlled trial, including the animal and human nutritional studies, will contribute towards this. It should be noted that the nutritional trials under this licence do not included stacked GMOs. The GMOs from the trial will not be used in commercial human food or animal feed. Further information would be required for an application for commercial release of any GMOs included in this trial. FSANZ approval would also be required before any product from these GMOs could be used commercially in human food.
3	NGO	x	RA	Advises that the application fails to satisfy the requirement of the Risk Analysis Framework by inadequately responding to the questions posed by the application form. Advises the RARMP is also deficient in its treatment of key matters [detailed below] and does not satisfy the requirements in the Risk Analysis Framework. Recommends that the application be rejected. Recommends that the RARMP be substantially revised.	Consistent with the RAF, the RARMP for this release considered information provided in the application and the currently available scientific information, in the context of the limited scale of the proposed release, and concluded that risks to human health and the environment are negligible.
			AR, H	Section 4 (aa) of the Act requires the application of the precautionary principle. In the interests of human and animal health, the OGTR and other regulators should apply the principle to antibiotic resistance marker genes, by requiring their removal from all commercial GM food crops and the food products derived from them. Advises that the RARMP does not assess any risks scenarios to do with the presence of the ampicillin resistance gene present in the vector backbone. Recommends that the applicants be required to develop and submit a feasibility plan to the OGTR and FSANZ, prior to any licence being granted, for the complete removal of antibiotic resistance marker genes from all GM wheat and barley lines for which the applicants may subsequently apply for commercial release.	Discussion of the ampicillin resistance gene, which is not expressed in the GM plants, has been included in the RARMP. The antibiotic resistance marker genes in these GMOs have been considered in detail in other RARMPs, by regulatory bodies in other countries and by international bodies, and have not been found to pose a risk to people or the environment. No adverse effects from their use in field trial or commercial release of GMOs have been recorded in Australia or elsewhere.
			U, Res	States that complete characterization, at both molecular and phenotypic levels, of GMOs proposed for release should be required prior to licensing. But section 5.7 of the RARMP discloses that groups 2 and 4 do not satisfy this condition. Recommends that the applicants be required to reapply for these components of the present application, after the mature plantlets have been fully characterized. It appears odd that: “no phenotypic screens are currently available” (application page 8) for wheat seed in Group #2 which is derived from trials licensed under DIRs 094 and 099 and from glasshouse grown material. Recommends that all this information supplied in a timely manner prior to any field testing be a prerequisite for the licensing of these lines.	The current application is not for a large scale or commercial release of GM wheat or barley. Both the application and RARMP for this release considered the relevant information in the context of the limited scale of the release and the proposed control measures, and concluded that risks to the environment are negligible.
			U, Res	To produce scientific data useful for the evaluation of the environmental and public health safety, it is unsatisfactory that the applicants propose to trial only one control wheat genotype (the parent line) and up to 14 negative segregants. Recommends that the OGTR require some other well-performing wheat and barley varieties to also be trialled so that robust data is available for comparison.	The RARMPs provide broad guidance on data requirements for possible future larger scale or commercial releases. The OGTR also strongly encourage potential applicants to discuss details of data required prior to submission of such applications. If sufficient data is not presented in a future application, the application may not be accepted, more data may be requested from the applicant, risk treatment measures imposed or a licence refused.
			GS, U	The applicant’s intention to stack genes and its potential impacts are inadequately addressed in sections 177 and 182 of the RARMP. Recommends that the GMOs with stacked traits proposed by the applicant should be the subject of separate applications to the OGTR as the absence of specificity about what is proposed and the complete lack of characterisation of the GMOs make this part of the application impossible to assess and therefore unacceptable.	Risk Scenarios 1, 3 and 6 were amended to give greater consideration of gene stacking. The proposed limits and controls were considered adequate to manage any risks from the proposed release.
			GS,W, Res	Gene stacking may significantly increase the capacity of wheat or barley to survive in the environment and to become weedy. Even in the absence of robust data the RARMP asserts a lack of ‘significant’ harm. Weed management already directly costs the Australian economy around $4-5 billion pa. Commonwealth regulators and publicly funded researchers in this field have a responsibility to actively prevent the deliberate or inadvertent introduction of new, less-manageable weeds. Recommends that the applicant be required to manage these trials so that data is gathered to show the extent to which wheat and barley, especially those varieties with GM altered survival traits, may persist in the Australian environment and become weedy.	Risk Scenario 3 and the section on uncertainty have been amended to better reflect the potential for increased persistence. Additional phenotypic characterisation of the GM wheat and barley lines, particularly with respect to traits that may contribute to weediness, is noted as information that may be required to justify a larger scale or commercial release (Ch3, section 4). The limits and controls of the trial will restrict spread and persistence of the GM plants.
			RA	The applicant seeks approval to decommission parts of the whole trial site as the trials in that location end and monitoring is discontinued (Section 7.4, Page 7). But this implies a questionable assumption that such trial plots will be unaffected by continuing adjacent trials. The whole site should be managed as a single entity for the duration of the licence. Recommends that this request not be approved.	Persistence of the GMOs at each trial location within the larger site is managed by post-harvest inspection requirements. An area is not released from post-harvest requirements until the Regulator is satisfied that this is justified.
			RA	The application and RARMP cite studies which include authors with links to industry. Recommends that the OGTR not rely exclusively on sources of advice, data or research results from sources that are tainted by conflicts of interest.	The Regulator does not rely solely on information supplied by the applicant. A range of available information is considered in the risk assessment. The Regulator takes into account the quality and relevance of each piece of information and considers the weigh of evidence as described in the RAF.
			RA	Section 80 of the RARMP is cavalier in its dismissal of a potential allergy problem with glutamates. The web link at footnote 10 in the RARMP does not exist. Claims that the naturally occurring bound form of glutamate may have different effects than its free form and the RARMP ignores this nuance. Recommends that the OGTR and FSANZ give more serious and sustained attention to the specific problem of increased free glutamates in GM foods and related issues of altered levels of metabolites in GM plants and their food products.	The RARMP notes that studies on the toxicity and allergenicity of the GMOs may be required if approval was sought for use of the GMOs in commercial foods in Australia. The web link at footnote 10 in the RARMP was corrected.
			RA	Criticises that the RARMP for, on a number of occasions, not providing evidence to support conclusions that negative outcomes are unlikely, considering only likely scenarios when it is often unexpected events that create catastrophic impacts, and not considering worst case scenarios. The RARMP discounts the cited evidence (Zhang et al., 2011) that natural plant miRNAs can be absorbed by mammals through food intake, and have the potential to modulate gene expression in animals, which directly challenges one of our regulators’ core assumptions. Recommends that the OGTR and other regulators critically re-evaluate their numerous assumptions to bring more scientific rigour into their assessments. Scientific critics and the interested public, as well as pro-GM advocates, should be included in their deliberations.	The application was assessed in the context of the limited scale of the release and the proposed control measures. The introduced genes were originally derived from wheat and barley (with the exception of the introduced marker genes, which are discussed above) and are therefore unlikely to lead to production of novel toxins or allergens. More detail has been added to Risk Scenario 1, including in relation to miRNA. The Act prescribes broad consultation on RARMPs, including with experts, relevant government agencies, local government areas and the public.
			U, Res, RA	Asserts that application deficiencies were ignored and that the application should be rejected. Recommends that trait stability and the number and position of all transgenic inserts should be required to be determined before an application is accepted. Advises that genetic manipulation techniques are lacking in precision that GMOs are unpredictable and pose safety risks while acknowledging that the GMOs are not to be commercialised in the current DIR.	Uncertainty is an intrinsic property of risk assessments. In the context of the limited scale of the release and the proposed control measures, and acknowledging knowledge gaps, the dealings proposed were assessed to pose negligible risk to people or the environment.

2 More information on the process for assessment of licence applications to release a genetically modified organism (GMO) into the environment is available from the Office of the Gene Technology Regulator (OGTR) (Free call 1800 181 030 or at <http://www.ogtr.gov.au/>), and in the Regulator’s Risk Analysis Framework (OGTR 2009) at .

3The term ‘line’ is used to denote plants derived from a single plant containing a specific genetic modification made by one transformation event.

4 More information on the process for assessment of licence applications to release a genetically modified organism (GMO) into the environment is available from the Office of the Gene Technology Regulator (OGTR) (Free call 1800 181 030 or at <http://www.ogtr.gov.au/>), and in the Regulator’s Risk Analysis Framework (OGTR 2009) at .

5The term ‘line’ is used to denote plants derived from a single plant containing a specific genetic modification made by one transformation event.

6 More information on Australia’s integrated regulatory framework for gene technology is contained in the Risk Analysis Framework available from the Office of the Gene Technology Regulator (OGTR). Free call 1800 181 030 or at <http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/riskassessments-1>.

7The term ‘line’ is used to denote plants derived from a single plant containing a specific genetic modification resulting from a single transformation event.

8 Isoform: any of several different forms of the same protein as a result of changes in the DNA sequence, or different forms of a protein may also be produced from related genes, or may arise from the same gene by alternative splicing.

9 Triple null mutants: A null mutant lacks a functional gene product for a particular gene. Wheat, a hexaploid, has three genomes each with two copies of each resident gene. Since many genes will be repeated in each of the three genomes, to have no functional gene product, null mutations must be present in all three genomes – or triple null.

10 Source: http://www.fda.gov/Food/FoodIngredientsPackaging/GenerallyRecognizedasSafeGRAS/GRASSubstancesSCOGSDatabase/default.htm, accessed 25 January 2012

11 The iodometric determination of amylose content is based on measuring the color change induced when iodine binds to linear regions of α-1,4 glucan against a standard curve generated using known concentrations of purified potato amylose and amylopectin (Regina et al. 2006).

12 Gelatinisation. The temperature at which the grain swells up irreversibly. It is determined by the amylopectin chain length distribution, amylose content and the amount of starch structural order. A reduction in swelling power with increasing amylose content is consistently noted for all starch types (Tan 2006).

13 More information on Australia’s integrated regulatory framework for gene technology is contained in the Risk Analysis Framework available from the Office of the Gene Technology Regulator (OGTR). Free call 1800 181 030 or at <http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/riskassessments-1>.

14< http://www.aphis.usda.gov/brs/status/relday.html>, <http://gmoinfo.jrc.ec.europa.eu/gmp_browse.aspx> accessed 20 October 2011.

15 Pleiotropy is the effect of one particular gene on other genes to produce apparently unrelated, multiple phenotypic traits (Kahl 2001).

16 A more detailed discussion is contained in the Regulator’s Risk Analysis Framework available at <http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/riskassessments-1> or via Free call 1800 181 030.

17 GTTAC, State and Territory Governments, Australian Government agencies, LGAs and the Minister for the Environment.