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Risk Assessment andRisk Management Plan for

DIR 128

Limited and controlled release of wheat and barley genetically modified for abiotic stress tolerance or

micronutrient uptake

Applicant: The University of Adelaide

August 2014

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DIR 128 – Risk Assessment and Risk Management Plan Office of the Gene Technology Regulator

Summary of the Risk Assessment and Risk Management Plan

for

Licence Application DIR 128DecisionThe Gene Technology Regulator (the Regulator) has decided to issue a licence for this application for a limited and controlled release of a genetically modified organism (GMO) into the environment. A Risk Assessment and Risk Management Plan (RARMP) for this application was prepared by the Regulator in accordance with requirements of the Gene Technology Act 2000 (the Act) and corresponding state and territory legislation, and finalised following consultation with a wide range of experts, agencies and authorities, and the public. The RARMP concludes that this field trial poses negligible risks to human health and safety and the environment and that any risks posed by the dealings can be managed by imposing conditions on the release.

The applicationApplication number DIR 128Applicant: The University of Adelaide

Project Title: Limited and controlled release of wheat and barley modified for abiotic stress tolerance or micronutrient uptake

Parent organisms: Bread wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.)Introduced genes and modified traits:

Thirty-three genes that are involved in salt tolerance, aluminium tolerance, drought tolerance (water use efficiency), nitrogen use efficiency, or micronutrient uptake.†

Two selectable marker genes from bacteriaProposed release dates: August 2014 – December 2019Proposed locations: Five sites, two in South Australia and three in Western AustraliaProposed release size: Total of 2.5 hectares per seasonPrimary purpose: To assess whether the introduction and expression of the specified group of

genes in plants affects yield potential under field conditions

Risk assessmentThe risk assessment concludes that risks to the health and safety of people, or the environment, from the proposed release are negligible.

The risk assessment process considers how the genetic modification and activities conducted with the GMOs might lead to harm to people or the environment. Risks are characterised in relation to both the seriousness and likelihood of harm, taking into account information in the application (including proposed limits and controls), relevant previous approvals, current scientific/technical knowledge, and advice received from a wide range of experts, agencies and authorities consulted on the RARMP. Both the short and long term potential harms are considered.

† The identities of some of the genes have been declared as Confidential Commercial Information (CCI) under section 185 of the Act.

Summary I


Credible pathways to potential harm that were considered included: unintended exposure to the GM plant material; increased spread and persistence of the GM wheat and barley relative to unmodified plants; and transfer of the introduced genetic material to non GM wheat or barley, or other sexually compatible plants. Potential harms associated with these pathways included toxicity to people and other animals, allergic reactions in people and environmental harms associated with weediness.

The principal reasons for the conclusion of negligible risks are that the introduced genetic modifications are unlikely to cause harm to human health or safety or to the environment, the introduced genes are similar to those already existing in the environment, and furthermore, the proposed limits and controls effectively contain the GMOs and their genetic material and minimise exposure.

Risk management planThe risk management plan concludes that risks posed by the proposed dealings can be managed so as to protect people and the environment by imposing conditions on the release.

Risk management is used to protect the health and safety of people and to protect the environment by controlling or mitigating risk. The risk management plan evaluates and treats identified risks, evaluates controls and limits proposed by the applicant, and considers general risk management measures. The risk management plan is given effect through licence conditions.

As the level of risk is assessed as negligible, specific risk treatment is not required. However, as this is a limited and controlled release, the licence includes limits on the size, locations and duration of the release, as well as controls including containment provisions at the trial site; prohibiting the use of GM plant materials in human food or animal feed; destroying GM plant materials not required for further studies; transporting GM plant materials in accordance with the Regulator’s guidelines; and conducting post-harvest monitoring at the trial sites to ensure all GMOs are destroyed.

Summary II


Table of ContentsSUMMARY OF THE RISK ASSESSMENT AND RISK MANAGEMENT PLAN...................................I

Decision............................................................................................................................. IThe application.................................................................................................................. IRisk assessment................................................................................................................ IRisk management plan..................................................................................................... II

TABLE OF CONTENTS..................................................................................................................................3

ABBREVIATIONS...........................................................................................................................................5

CHAPTER 1 RISK ASSESSMENT CONTEXT......................................................................................1

Section 1 Background...................................................................................................1Section 2 Regulatory framework...................................................................................1Section 3 The proposed dealings.................................................................................2

3.1 The proposed limits of the dealings (size, locations, duration and people)...............23.2 The proposed controls to restrict the spread and persistence of the GMOs and their

genetic material in the environment...........................................................................3Section 4 The parent organisms...................................................................................4Section 5 The GMOs, nature and effect of the genetic modification.............................4

5.1 Introduction to the GMOs..........................................................................................45.2 The introduced genes, encoded proteins and their associated effects........................95.3 Toxicity/allergenicity associated with the introduced genes, their encoded proteins

and associated products............................................................................................125.4 Characterisation of the GMOs..................................................................................13

Section 6 The receiving environment..........................................................................146.1 Relevant abiotic factors............................................................................................146.2 Relevant agricultural practices.................................................................................146.3 Presence of related plants in the receiving environment..........................................156.4 Presence of similar genes and encoded proteins in the environment.......................16

Section 7 Relevant Australian and international approvals.........................................167.1 Australian approvals.................................................................................................167.2 International approvals of GM wheat and barley.....................................................16

CHAPTER 2 RISK ASSESSMENT.........................................................................................................17

Section 1 Introduction.................................................................................................17Section 2 Risk Identification........................................................................................18

2.1 Risk source...............................................................................................................182.2 Causal pathway.........................................................................................................19

Table of Contents III


2.3 Potential harm...........................................................................................................202.4 Postulated risk scenarios..........................................................................................20

Section 3 Uncertainty..................................................................................................35Section 4 Risk evaluation............................................................................................36

CHAPTER 3 RISK MANAGEMENT.....................................................................................................38

Section 1 Background.................................................................................................38Section 2 Risk treatment measures for identified risks...............................................38Section 3 General risk management...........................................................................38

3.1 Licence conditions to limit and control the release..................................................383.2 Other risk management considerations....................................................................42

Section 4 Issues to be addressed for future releases.................................................44Section 5 Conclusions of the RARMP.........................................................................44

REFERENCES 45

APPENDIX A SUMMARY OF SUBMISSIONS FROM PRESCRIBED EXPERTS, AGENCIES AND AUTHORITIES.............................................................................56

APPENDIX B SUMMARY OF SUBMISSIONS FROM THE PUBLIC.....................................57

Table of Contents IV


AbbreviationsAPVMA Australian Pesticides and Veterinary Medicines AuthorityCaMV Cauliflower mosaic virusCCI Confidential Commercial Information as declared under section 185 of the

Gene Technology Act 2000DAFWA Department of Agriculture and Food, Western AustraliaDIR Dealings involving Intentional ReleaseFSANZ Food Standards Australia New ZealandGM Genetically modifiedGMO Genetically modified organismha Hectarehph Hygromycin phosphotransferaseLGA Local government aream MetresNGNE New Genes for New EnvironmentsNLRD Notifiable low risk dealingsnptII neomycin phosphotransferase IIOGTR Office of the Gene Technology RegulatorPC2 Physical Containment level 2PR Pathogenesis-relatedRARMP Risk Assessment and Risk Management PlanRegulations Gene Technology Regulations 2001Regulator Gene Technology RegulatorTGA Therapeutic Goods Administrationthe Act The Gene Technology Act 2000

Abbreviations V

PROPOSED DEALINGSProposed activities involving the GMOProposed limits of the releaseProposed control measures

PARENT ORGANISMOrigin and taxonomyCultivation and useBiological characterisationEcology

PREVIOUS RELEASES

GMOIntroduced genes (genotype)Novel traits (phenotype)

RISK ASSESSMENT CONTEXT

LEGISLATIVE REQUIREMENTS(including Gene Technology Act and Regulations)

RISK ANALYSIS FRAMEWORK

OGTR OPERATIONAL POLICIES AND GUIDELINES

RECEIVING ENVIRONMENTEnvironmental conditionsAgronomic practicesPresence of related speciesPresence of similar genes


Chapter 1 Risk assessment contextSection 1 Background

1. An application has been made under the Gene Technology Act 2000 (the Act) for Dealings involving the Intentional Release (DIR) of genetically modified organisms (GMOs) into the Australian environment.

2. The Act in conjunction with the Gene Technology Regulations 2001 (the Regulations), an inter-governmental agreement and corresponding legislation that is being enacted in each State and Territory, comprise Australia’s national regulatory system for gene technology. Its objective is to protect the health and safety of people, and to protect the environment, by identifying risks posed by or as a result of gene technology, and by managing those risks through regulating certain dealings with genetically modified organisms (GMOs).

3. This chapter describes the parameters within which potential risks to the health and safety of people or the environment posed by the proposed release are assessed. The risk assessment context is established within the regulatory framework and considers application-specific parameters (Figure 1).

Figure 1 Summary of parameters used to establish the risk assessment context

Section 2 Regulatory framework4. Sections 50, 50A and 51 of the Act outline the matters which the Gene Technology Regulator (the Regulator) must take into account, and who must be consulted with, in preparing the Risk Assessment and Risk Management Plans (RARMPs) that inform the decisions on licence applications. In addition, the Regulations outline further matters the Regulator must consider when preparing a RARMP. In accordance with section 50A of the Gene Technology Act 2000 (the Act), this application is considered to be a limited and controlled release application, as its principal purpose is to enable the applicant to conduct experiments and the applicant has proposed limits on the size, location and duration of the release, as well as controls to restrict the spread and persistence of the GMOs and their genetic material in the environment. Therefore, the Gene Technology Regulator (the Regulator) was not required to consult with prescribed experts, agencies

Chapter 1 – Risk assessment context 1


and authorities before preparation of the Risk Assessment and Risk Management Plan (RARMP; see section 50 of the Act).

5. Section 52 of the Act requires the Regulator to seek comment on the RARMP from the States and Territories, the Gene Technology Technical Advisory Committee, Commonwealth authorities or agencies prescribed in the Regulations, the Minister for the Environment, relevant local council(s), and the public. The advice from the prescribed experts, agencies and authorities and how it was taken into account is summarised in Appendix A. Thirteen public submissions were received and their considerations are summarised in Appendix B.

6. The Risk Analysis Framework (OGTR 2013) explains the Regulator’s approach to the preparation of RARMPs in accordance with the legislative requirements. Additionally, there are a number of operational policies and guidelines developed by the Office of the Gene Technology Regulator (OGTR) that are relevant to DIR licences. These documents are available from the OGTR website.

7. Any dealings conducted under a licence issued by the Regulator may also be subject to regulation by other Australian government agencies that regulate GMOs or GM products, including Food Standards Australia New Zealand (FSANZ), Australian Pesticides and Veterinary Medicines Authority (APVMA), Therapeutic Goods Administration (TGA), National Industrial Chemicals Notification and Assessment Scheme and Department of Agriculture. These dealings may also be subject to the operation of State legislation declaring areas to be GM, GM free, or both, for marketing purposes.

Section 3 The proposed dealings8. The University of Adelaide proposes to release up to 1262 lines‡ of genetically modified (GM) wheat and barley into the environment under limited and controlled conditions.

9. The purpose of the trial is to evaluate the candidate genes for yield potential under field conditions. In addition, the release will allow the applicant to produce sufficient grain for subsequent replicated trials.

10. The dealings involved in the proposed intentional release include:

conducting experiments with the GMOs

breeding the GMOs

propagating the GMOs

growing or culturing the GMOs

transporting the GMOs

disposing of the GMOs

and the possession, supply or use of the GMOs for the purposes of, or in the course of, any of the above. These dealings are detailed further below.

3.1 The proposed limits of the dealings (size, locations, duration and people)11. The applicant proposes to grow GM wheat and barley plants between June 2014 and December 2019.

12. The GMOs are proposed to be planted at five sites, two of which are in South Australia and three in Western Australia. The South Australian sites are located in the LGAs of Marion and Wakefield, and the Western Australian sites in the LGAs of Corrigin, Merredin, and Katanning. The

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


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latter two sites represent the New Genes for New Environment (NGNE) facilities operated by the Department of Agriculture and Food, Western Australia (DAFWA).

13. The collective area of the trial (over 5 sites) would be up to 2.5 hectares (ha) per growing season.

14. The applicant proposes that only trained and authorised staff would be permitted to deal with the GM wheat and barley. Any other visitors to the sites would be accompanied by an authorised University of Adelaide representative and would not deal with the GMOs.

3.2 The proposed controls to restrict the spread and persistence of the GMOs and their genetic material in the environment15. The applicant has proposed a number of controls to restrict the spread and persistence of the GM wheat and barley lines and the introduced genetic material in the environment, including:

locating the trial sites at least 200 m from other wheat and barley plants, unless such plants represent those (GM or non-GM) planted at another location as part of this trial or those approved for trial at the site under a separate DIR licence

surrounding each trial site with a fence, and controlling plant growth in a 10 m wide zone outside the fence by mowing, herbicide treatment and/or weeding

surrounding each location (in the fenced trial site) by a 2 m wide buffer where plant growth will be controlled by mowing, herbicide treatment and/or weeding

monitoring each location for volunteers, beginning two weeks prior to the expected start of flowering and continuing on a fortnightly basis until flowering has finished

during flowering, inspecting a 190 m wide zone surrounding the 10 m wide zone for wheat and barley (but see next point)

if there has been no cultivation or detection of wheat and barley in the 190 m wide zone in the previous two years, reducing inspection of that area to the 50 m wide area directly surrounding the 10 m wide zone

after harvest, monitoring locations for volunteers and related species at least once every 35 days for a period of two years; during this time period at least three irrigations will be performed to encourage germination of volunteers

destroying volunteers and related species

cleaning equipment used in harvesting, and threshing on site or transporting the heads to approved facilities for analysis or processing

storing seed remaining from analysis in an approved facility or destroying the seed by autoclaving or any other method approved by the Regulator

destroying waste material derived from harvest, and any stubble, by ploughing back into the soil, burning or burial

transporting and storing GM material in accordance with the Regulator’s Guidelines for the Transport, Storage and Disposal of GMOs (2011)

restricting access to trial sites to authorised staff

not allowing GM plant material or products to be used for human food or animal feed.

16. Figure 2 shows the proposed site layout including some of these controls. These controls, and the limits outlined above, have been taken into account in establishing the risk assessment context (this Chapter), and their suitability for containing the proposed release is evaluated in Chapter 3, Section 3.1.1.



Figure 2 Proposed trial layout, including some of the controls (not drawn to scale)

Section 4 The parent organisms17. The parent organisms are bread wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.), both exotic to Australia. Commercial wheat cultivation occurs in the wheat belt from south eastern Queensland through New South Wales, Victoria, Tasmania, southern South Australia and southern Western Australia. Barley is cultivated in these same areas, although a small amount is also grown in Tasmania.

18. The wheat cultivars used to generate the GM wheat lines are Bobwhite and Gladius, but some genes have been backcrossed into the commercial cultivars EGA Bonnie Rock, EGA-Burke, IGW-2971, Magenta, Frame and Drysdale. GM barley lines were generated in Golden Promise and WI4330, but some genes were backcrossed into the commercial cultivars Flagship, Gairdner, Commander.

19. Detailed information about the parent organisms are contained in the reference documents The Biology of Triticum aestivum L. em Thell (bread wheat) (OGTR 2008b) and The Biology of Hordeum vulgare L. (barley) (OGTR 2008a), which were produced to inform the risk assessment process for licence applications involving GM wheat and barley plants. These documents are available from the OGTR website.

Section 5 The GMOs, nature and effect of the genetic modification5.1 Introduction to the GMOs20. The applicant proposes to release up to 1262 lines of GM wheat and barley. The lines were produced using either Agrobacterium tumefaciens or biolistics mediated plant transformation. Information about these transformation methods can be found in the risk assessment reference document Methods of plant genetic modification available from the Risk Assessment References page on the OGTR website.

21. Each GM wheat and barley line has been transformed with a single gene from 33 genes of interest. On the basis of the trait that they are expected to induce, the genes can be divided into five groups: (i) drought tolerance; (ii) salt tolerance; (iii) aluminium tolerance; (iv) nitrogen use efficiency; and (v) micronutrient uptake. The GMOs also contain selectable marker genes, these


Location(planting area)

2 m wide bufferZone where the growth of plants is controlled

Fence

10 m wideMonitoring zone where the growth of plants is controlled

190 m wideIsolation zone inspected for wheat and barley during flowering of GMOs

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being the neomycin phosphotransferase II (nptII) gene and/or the hygromycin phosphotransferase (hptII) gene, both originating from E.coli.

22. A summary of these groups is presented in Table 1, with details provided in the following sections.

Table 1 Summary of GM wheat and barley lines proposed for release.

GMO class

Gene of interest# Plant transformed Approx no. of lines

Drought tolerance

1 SIGNALLING PROTEIN 2 T. aestivum 18

2 TRANSCRIPTION FACTOR 1 T. aestivum 72





H. vulgare 36


8 PHOTOSYNTHESIS AND METABOLISM GENE 3 T. aestivum 18



11 CELL SPECIFICATION, PROLIFERATION AND DIVISION GENE 2

T. aestivum 36


T. aestivum 36

13 RNA METABOLISM PROCESSING GENE 1 H. vulgare 6

14 RNA METABOLISM PROCESSING GENE 2 H. vulgare 6

15 RNA METABOLISM PROCESSING GENE 3 T. aestivum 72

H. vulgare 72

16 RNA METABOLISM PROCESSING GENE 4 T. aestivum 72

H. vulgare 72

Salt tolerance

17 ScNHA1 (Saccharomyces cerevisiae) H. vulgare 40

T. aestivum 20

18 PpENA1 (Physcomitrella patens) H. vulgare 40

T. aestivum 20



GMO class

Gene of interest# Plant transformed Approx no. of lines

19 AtAVP (A. thaliana) H. vulgare 10

T. aestivum 5

20 AtCIPK16 (A. thaliana) T. aestivum 20

H. vulgare 20

Aluminium tolerance

21 TaALMT1 (T. aestivum) T. aestivum 14

H. vulgare 12

22 TaALMT1_minus_insert (T. aestivum) H. vulgare 9

23 ION TRANSPORTER 5 T. aestivum 9

H. vulgare 9

24 ION TRANSPORTER 6 T. aestivum 9

H. vulgare 9

25 ION TRANSPORTER 7C T. aestivum 9

H. vulgare 9

26 ScALMT1.M39.1_wt (Secale cereale) T. aestivum 8

H. vulgare 6

27 ScALMT1.M39.1_plus insert (Secale cereale) H. vulgare 6

28 HvAACT1 (H. vulgare) T. aestivum 2

H. vulgare 4

Nitrogen use efficiency



31 Aminotransferase T. aestivum 32

H. vulgare 32


T. aestivum 12

Micronutrient uptake

33 OsNAS2 (Oryza sativa) T. aestivum 6



# The identities of some of these genes have been declared as CCI. The applicant has assigned an identifier to these genes.

23. The vectors used for Agrobacterium mediated transformation are variants of a pMDC backbone, while the vectors used for biolistic transformation include variants of a pMDC backbone and those based on PHP. Promoters, genomic sequences, and terminators used in these vectors come from Agrobacterium tumefaciens, Cauliflower mosaic virus, barley, maize, potato, rice, sorghum, tobacco, durum wheat and bread wheat. They are listed in Tables 2, 3 and 4.

Table 2 Promoters used in constructs.

Name of promoter # Name of gene that promoter derived from# Source organism

CaMV35S Viral promoter Cauliflower mosaic virus

pAct OsAct O. sativa

pB4L TdHDZipl-4 T. durum

pB7L TdHDZipl-3 T. durum

pCor39 TdCor39 T. durum

pCor410b TdCor410b T. durum

pCor410H1(truncated) TdCor410H1 T. durum

pCor410H2 (truncated) TdCor410H2 T. durum

PROMOTER 1 GENE A T. durum

pDHN8 HvDHN8 H. vulgare

PROMOTER 2 GENE B T. durum

PROMOTER 3 GENE C T. durum

PROMOTER 4 GENE D T. durum

PROMOTER 5 GENE E O. sativa

PROMOTER 6 GENE F O. sativa

PROMOTER 7 GENE G O. sativa

pOsAnt1 Aldehyde dehydrogenase se1 O. sativa

PROMOTER 8 GENE H Z. mays

PROMOTER 9 GENE I T. durum

pUbi Polyubiquitin Z. mays

PROMOTER 10 GENE J T. durum

pWRKY71 (pJRCO189) OsWRKY71 O. sativa

PROMOTER 11 GENE K T. durum

PROMOTER 12 GENE L Z. mays



# The identities of some of the promoters and the genes from which they originate have been declared as CCI. The applicant has assigned an identifier to these promoters and genes.

Table 3 Genomic elements used in constructs.

Name of sequence# Description# Source organism#

Act intron (MOD1) intron O. sativa

APS amplification promoting sequences N. tabacum

Mini ALLSTOPS Stop codon -

STLS1 intron 2 intron S. tuberosum

Ubi intron intron Z. mays

Ubi1 5’UTR 5’ untranslated region Z. mays

UNTRANSLATED SEQUENCE 1

GENE M CCI

Ubi1 intron intron Z. mays

UNTRANSLATED SEQUENCE 2

GENE N CCI

# The identities of some of the genomic elements, the genes they come from and their source organisms have been declared as CCI. The applicant has assigned an identifier to these genomic elements and genes.

Table 4 Terminators used in constructs.

Name of terminator# Description# Source organism#

35S ter Viral terminator Cauliflower mosaic virus

TERMINATOR SEQUENCE 1

Terminator of the GENE A gene T. durum


Terminator of the GENE B gene T. durum


Terminator of the GENE O gene CCI


Terminator of the GENE P gene T. durum

Nos ter Nopaline synthase gene terminator A. tumefaciens

OsUBI ter Polyubiquitin gene terminator O. sativa


Terminator of the GENE Q gene CCI


Terminator of the GENE R gene CCI


Terminator of the GENE S gene CCI

TERMINATOR Terminator of the GENE T gene S. bicolor



Name of terminator# Description# Source organism#

SEQUENCE 8


Terminator of the GENE U gene A. tumefaciens

# The identities of some of the terminators, the genes they come from and their source organisms have been declared as CCI. The applicant has assigned an identifier to these terminators and genes.

5.2 The introduced genes, encoded proteins and their associated effects24. Four of the traits, drought tolerance, salt tolerance, aluminium tolerance and nitrogen use efficiency, can be grouped as abiotic stress tolerances. The natural mechanisms by which plants deal with abiotic stresses can be classified as either avoidance or tolerance, the former referring to those that obviously affect the morphology and/or physiology of a plant, and the latter to molecular changes that may have no visual effects (Howles & Smith 2013). The availability of water is the most important factor that limits crop yield globally (Castiglioni et al. 2008), but salinity and the presence of aluminium in acid soils likewise both have large negative impacts upon agriculture (Brunner & Sperisen 2013; Munns et al. 2012; Rengasamy 2010). Abiotic stress tolerances are multigenic traits, involving the interaction of genes where the protein products constitute different biochemical pathways. Further information on plant responses to abiotic stresses can be found in the RARMP for DIR 102 (available at <http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/ir-1>).

25. Nitrogen use efficiency is an important factor in crop plant productivity, this forming the impetus for research, both using conventional breeding and gene technology, to develop plants that absorb and use this element more efficiently. It can be described as the product of two components: uptake efficicency and utilisation efficiency (Witcombe et al. 2008). Increasing the efficiency of nitrogen use would reduce the need for fertilisers in agriculture and the pollution that excess nitrogen can cause in the environment. Further details of this trait are provided in the RARMP for DIR 094 (available at <http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/ir-1>).

26. The other trait that the applicant is attempting to engineer into plants is increased uptake of the micronutrient iron. Deficiencies in micronutrients affects more than 2 billion of the world’s population, with iron deficiency being the most common (Tulchinsky 2010; WHO 2014). Biofortification, the development of new crop varieties with increased levels of micronutrients, by either conventional breeding practices or genetic modification, provides an extremely promising approach to dealing with this problem (Nestel et al. 2006). In particular, it allows the staple food supplies of countries to be used and new germplasm to be spread internationally.5.2.1 Group 1 – Drought tolerance

27. The identities of all of the introduced genes linked to drought tolerance have been declared as CCI. The confidential information was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.

28. In reference to their general function in the cell, these genes are involved in either signalling, transcription, photosynthesis and metabolism, cell specification, proliferation and division, or RNA metabolism. Some involve the expression of microRNAs in plants, thus down-regulating the expression of genes by RNA interference (RNAi).5.2.2 Group 2 – Salt tolerance

ScNHA1

29. Some GM wheat and barley lines will contain the ScNHA1 gene encoding a Na+, K+/H+ antiporter (NHA) derived from Saccharomyces cerevisiae (yeast)(Prior et al. 1996). The gene belongs to the CPA2 (cation/proton antiporter) family of Na+/H+ antiporters in eukaryotes and


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resides in the subfamily NHA, which includes animal, plant, fungal and bacterial members (Brett et al. 2005).

30. Deletion of the ScNHA1 gene was found to increase the cytosolic pH of yeast cells suspended in water and acidic buffers, confirming a role of the protein in both regulating the cytoplasmic concentration of Na+/K+ and buffering the pH of cells (Sychrova et al. 1999). Other studies, investigating the growth of wild-type yeast and strains mutant with respect of this gene in media with varying concentrations of K+, have concluded that the protein can affect the main K+ influx system in this organism (Banuelos et al. 2002; Banuelos et al. 1998).

PpENA1

31. Some GM wheat lines and barley lines will contain PpENA1, a gene encoding an ENA-type (exitus natru) Na+ pumping ATPase derived from the moss Physcomitrella patens (Benito & Rodriguez-Navarro 2003). Eukaryotes have two types of Na+-pump ATPases, the Na+/K+-ATPases in animals and the ENA-type Na+-ATPases in fungi and mosses such as P. patens.

32. The PpENA1 protein has been shown to be able to complement a salt sensitive yeast strain deficient in Na+ and K+, as well as to act more generally as a Na+ pump in yeast (Benito & Rodriguez-Navarro 2003). Studies of the gene in the moss itself have demonstrated that its transcription is dramatically up-regulated in response to NaCl, and knockout mutants of the gene grow significantly slower than wild-type in media containing NaCl (Lunde et al. 2007).

AtAVP1

33. Some GM wheat lines and barley lines will contain the Arabidopsis gene AtAVP1, coding for a vacuolar H+-pyrophosphatase (H+-PPase).

34. H+-PPase proteins are proton pumps that use the energy gained from the breakdown of pyrophosphate to pump protons into the vacuoles of plant cells. This function of the proteins has been exploited in research aimed at enhancing the drought and salt tolerance of plants, the theory being that an increase in the activity of these proteins will generate a higher proton electrochemical gradient, thus energising secondary transporters, especially the Na+/H+ antiporters (Gaxiola et al. 2012; Pasapula et al. 2011).

35. Overexpression of the AtAVP1 gene in Arabidopsis increased the tolerance of plants to both drought and salt stress (Gaxiola et al. 2001), and similarly the overexpression of a homolog from Thellungiella halophile in maize and cotton improved tolerance to drought in the former and salinity and drought in the latter (Li et al. 2008; Lv et al. 2008; Lv et al. 2009). An unexpected consequence of overexpressing these genes in plants was increased proliferation of roots and shoots. Arabidopsis plants overexpressing AtAVP1 had enhanced leaf areas, root growth and dry weight compared to those observed in wild-type plants, while a loss of function mutant had leaves and roots of reduced size (Li et al. 2005). Overexpression of the Thellungiella halophile gene in cotton was accompanied by increased dry shoot and root masses regardless of whether the plants were grown in the presence or absence of salt (Lv et al. 2008). Another study has shown that maize plants overexpressing the Thellungiella halophile gene were more tolerant to phosphate deficit stress than wild-type plants, this being possibly due to their larger root systems (Pei et al. 2012).

AtCIPK16

36. Some GM wheat lines and barley lines will contain AtCIPK16, an Arabidopsis gene belonging to a family of calcineurin B-like (CBL) interacting protein kinases (CIPKs) that play a role in regulating plant cell responses to abiotic stress (Luan 2009). Calcium (Ca2+) serves as a second messenger during abiotic stress signalling, its release being detected by a number of sensors, including the CBL proteins. These latter proteins recruit the required CIPK to the cell membrane, where the kinase activates transporters and proteins involved in the stress response (Batistic & Kudla 2004; Luan 2009).

37. There are at least 25 AtCIPK genes in Arabidopsis thaliana, each CIPK interacting with one



or more members of the CBL family (Batistic & Kudla 2004). AtCIPK24 (also known as AtSOS2) was detected in a search for genes linked to salt tolerance in Arabidopsis, the mutant sos2 being hypersensitive to NaCl stress. Under salt stress, AtCBL4 (AtSOS3) recruits AtCIPK24 to the plasma membrane, where it activates (via phosphorylation) the Na+/H+ antiporter AtSOS1 to remove Na+ from the cell (Qiu et al. 2003; Qiu et al. 2002). Orthologs of CIPK24 have been identified in many plant species (Martinez-Atienza et al. 2007; Wang et al. 2004; Yu et al. 2007).

38. Map-based cloning has been used to identify AtCIPK16 as a gene linked to Na+ exclusion in Arabidopsis (Roy et al. 2013). Overexpression of the gene in Arabidopsis increased salt tolerance in both hydroponic and soil cultures, while conversely, use of a microRNA to suppress its expression in that plant resulted in elevated levels of Na+ in shoots grown in saline media. Overexpression of the gene in barley was also associated with increased salinity tolerance.5.2.3 Group 3 – Aluminium tolerance

TaALMT1, TaALMT1_minus_insert, ScALMT1.M39.1_wt, ScALMT1.M39.1_plus_insert

39. Some GM wheat lines and barley lines will contain one of seven genes coding for aluminium activated malate transporters (ALMTs). These genes come from wheat and rye (Secale cereale), some representing in vitro generated variants and mutations (single or double amino acid substitutions) of wild-type sequences. The excretion of malate has been linked to aluminium tolerance in plants, and the addition of malate to solutions containing aluminium can protect plants from toxic levels of this metal by forming a stable harmless complex (Delhaize & Ryan 1995; Delhaize et al. 1993). ALMTs are membrane localised proteins that facilitate the efflux of malate into the apoplast of cells (Sasaki et al. 2004; Yamaguchi et al. 2005), their heterologous expression in a range of organisms (including Xenopus laevis oocytes and the cells of various plants) being associated with an aluminium tolerance phenotype (Delhaize et al. 2004; Sasaki et al. 2004; Yamaguchi et al. 2005). In some cereals, such as rye, the number of ALTM1 genes at a certain chromosomal loci can vary (Collins et al. 2008).

40. The identities of three of the introduced genes linked to aluminium tolerance have been declared as CCI. The confidential information was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.

HvAACT1

41. Some GM wheat lines and barley lines will contain the barley gene HvAACT1 (HvMATE), coding for a Multidrug and Toxic Compound Extrusion (MATE) protein. Members of the MATE family of membrane transport proteins occur in both eukaryotes and prokaryotes. They are associated with the transport of a variety of organic molecules, including cationic drugs and plant secondary metabolites such as alkaloids (Omote et al. 2006). HvAACT1 is a citrate transporter which, in the presence of aluminium, facilitates the efflux of citrate from cells, this organic acid (in the manner of malate outlined above) chelating the metal and rendering it harmless.5.2.4 Group 4 – Nitrogen use efficiency

42. The identities of all of the introduced genes linked to nitrogen use efficiency have been declared as CCI. The confidential information was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.

43. In reference to their general function in the cell, these genes are involved in either transcription, primary metabolism, or cell proliferation and division.5.2.5 Group 5 – Micronutrient uptake

OsNAS2

44. Some GM wheat lines will contain a rice nicotianamine synthase gene (OsNAS2), the aim being to produce “biofortified” wheat (ie increasing the level of iron in wheat, thus being useful in regions of the world where wheat is a staple crop and iron deficiency amongst people is a problem).



45. Nicotianamine, a chelator and long distance transporter of transition metals such as iron, is biosynthesised by the trimerisation of S-adenosylmethionine, a reaction catalysed by the NAS enzyme; a by-product of the reaction is S-methyl-5’-thioadenosine. The mugineic acid family of phytosiderophores, molecules also involved in the acquistion of iron from the soil, are produced from nicotianamine by the subsequent activity of other enzymes (Higuchi et al. 1999; von Wiren et al. 2000). Amongst grasses, each species produces its own set of mugineic acids (Kim & Guerinot 2007).

46. The NAS gene family is largely plant specific, but examples of related genes are known from fungi and archaea (Herbik et al. 1999; Trampczynska et al. 2013). NAS genes have been isolated, or identified via bioinformatics in published genome sequences, from a number of plant species. Among the cereals, these include barley (Herbik et al. 1999; Higuchi et al. 1999), maize (Zhou et al. 2013), and rice (Higuchi et al. 2001).

47. Overexpression of NAS genes in plants has been shown to increase the levels of both nicotianamine and transition metals in cells. Overexpression of a barley NAS gene in both Arabidopsis and tobacco resulted in plants that showed improved tolerance to nickel, as well as accumulating large quantities of this metal in their shoots (Kim et al. 2005), while overexpression of the same gene in rice showed increased levels of both iron and zinc in seeds (Masuda et al. 2009). The combined expression in rice of an Arabidopsis NAS gene and genes coding for a ferritin and phytase led to a six-fold increase in the iron content of grain (Wirth et al. 2009). In another study in rice, three rice NAS genes were independently overexpresseed, in each case leading to increased levels of both iron and zinc (Johnson et al. 2011).5.2.6 Selectable marker genes

48. The vectors used to transform plant tissue contain one or both of two selectable marker genes. These are the neomycin phosphotransferase II (nptII) and hygromycin phosphotransferase (hpt) genes, both from E.coli. The proteins these genes encode inactivate kanamycin (as well as a range of structurally related antibiotics) and hygromycin B, respectively. More information on these genes, and selectable marker genes in general, can be obtained from the OGTR document Marker genes in GM plants, available on the OGTR website.

5.3 Toxicity/allergenicity associated with the introduced genes, their encoded proteins and associated products49. The introduced genes originate from a range of “higher” plants (Arabidopsis, barley, maize, rice, rye, and wheat), the moss Physcomitrella patens and Saccharomyces cerevisiae (yeast). Other than Arabidopsis thaliana, all of the higher plants are widely consumed by people and animals, and as such people and animals have a long history of exposure to the proteins from these respective plants. P. patens is a common moss found around the peripheries of bodies of water, often exposed in late summer and early autumn. Isolates are available from North America, Europe, Africa, Japan and Australia (Prigge & Bezanilla 2010). There are no reports of this moss producing any toxic or allergenic compounds, but it should be appreciated that it is not part of the human diet. S. cerevisiae is a eukaryotic fungus that is commonly used in baking, brewing and wine making, as well as occurring on fruit and vegetables. As such, it has a long history of use by humans. There are no reports of any isolates of S. cerevisiae producing toxins that negatively affect the health of humans or animals (USEPA 1997).

50. A comprehensive search of the scientific literature yielded no information to suggest that the genes themselves, their protein products, or any associated products (except iron, see below) were toxic or allergenic to people, or toxic to other organisms. This includes homologues isolated from other species. However, no toxicity/allergenicity tests have been performed on any of the proteins. The protein encoded by one of the genes associated with drought tolerance (declared CCI) has an eight amino acid residue match with a known allergen. Matches of this length occur often when un-related amino acid sequences are compared, and is unlikely to imply that the protein will have


http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/riskassessments-1#risk%20assessment


allergenic properties (Goodman et al. 2008). Sequence homology searches may establish whether a protein has an allergenic potential, the length of homology being a balance between detecting meaningful information amongst false negative and false positive results (Codex Alimentarius Commission 2003). Further information that has been declared CCI and is pertinent to this gene was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.

51. Expression of the OsNAS2 gene in wheat is aimed at increasing the level of iron in that plant. Wheat has approximately 30g/g of iron, biofortification targeting a content in this plant of 52g/g (Bouis et al. 2011). Iron is absorbed through the digestive tract and transferred around the body by a bloodstream protein called transferrin (Balmadrid & Bono 2009). This ion plays a number of roles in the body, most prominently being part of the haem group in proteins such as haemoglobin, cytochrome C and catalase. It is hence essential to the transport of oxygen to cells, oxidative phosphorylation, and the decomposition of hydrogen peroxide, respectively. However, excessive iron in the diet quickly leads to saturation of transferrin, resulting in free iron in the blood, something which is directly toxic to organs (Balmadrid & Bono 2009).

52. Conditions such as thalassemia can be further complicated by iron overload, inducing endocrine diseases, hepatic failure and even death. This may at least in apart be due to inhibition of the expression of hepcidin, an iron regulating peptide (Nemeth 2010; Tanno et al. 2007). Other genetic disorders also lead to excessive iron uptake. For example, hereditary haemochromatosis is characterised by excessive absorption of iron, leading to symptoms such as lethargy, upper abdominal discomfort and loss of libido (Barlow-Stewart et al. 2007).

5.4 Characterisation of the GMOs5.4.1 Stability and molecular characterisation

53. All the genotypes of the GM plants are stable under glasshouse conditions. The copy number of introduced genes range from one to three in individual GM lines, but the genomic locations are not known for any of the introduced genes.5.4.2 Phenotypic characterisation

54. Preliminary phenotypic characterisation of the plants in glasshouse conditions has demonstrated that the introduced genes induce no major visible phenotypes or reduce viability. However, some of the lines have shown delayed flowering, this sometimes occurring over a month later than usually observed. Such delayed maturity may reduce yield in the field, with plants being forced to endure greater exposure to hot and dry conditions.

Section 6 The receiving environment55. The receiving environment includes: any relevant biotic/abiotic properties of the geographic regions where the release would occur; intended agricultural practices, including those that may be altered in relation to normal practices; other relevant GMOs already released; and any particularly vulnerable or susceptible entities that may be specifically affected by the proposed release (OGTR 2013).

56. The factors relevant to the growth, distribution and cultivation of commercial wheat and barley can be found in The Biology of Triticum aestivum L. em Thell (Bread Wheat) (OGTR 2008b) and The Biology of Hordeum vulgare L. (barley) (OGTR 2008a).

57. The proposed dealings involve two sites in South Australia and three sites in Western Australia.

58. Site 1, “Glenthorne farm”, is located close to the University of Adelaide’s Waite Campus at O’Halloran Hill. The site is fully fenced and access is managed by a designated laboratory manager.



59. Site 2, “Karawatha”, is part of a commercial farming operation located in a dryland agricultural area in the Pinery region, approximately 70 km north of Adelaide. Access to the site will be controlled by the owner.

60. Site 3 is a farm at Kunjin, near Corrigin, Western Australia. The site will be managed by a private company that provides research services.

61. Sites 4 and 5 are in NGNE facilities, respectively located at Katanning and Merredin, and operated by DAFWA.

6.1 Relevant abiotic factors62. As noted above, the release is proposed to take place at five sites. Two of these (O-Halloran Hill and Pinery) are located in South Australia, and three (Corrigin, Merredin and Katanning) are located in Western Australia.

O-Halloran Hill is typical of rain-fed wheat production environments in South Australia, while Pinery is located in a dryland agricultural area that is useful for the assessment of those GM lines that are engineered to show increased drought tolerance.

Corrigin is located in a region of Western Australia that has saline soils and is occasionally subjected to frosts. At present the level of salinity is such that the land is not subject to broad acre cropping. However, these environmental factors make the land useful for the trialling of plants with increased tolerances to salt and frost.

Katanning and Merredin are purpose built facilities, one (Katanning) representing the high rainfall environment used for growing wheat in Western Australia, and the other (Merredin), regions of that state where wheat is grown under low rainfall. The soil of Katanning is largely alkaline and sodic, while that of Merredin is a mixture of yellow sands, gravels, loamy earth and loamy duplex soils, but with also calcareous subsoils.

6.2 Relevant agricultural practices63. It is not anticipated that the agronomic practices for the cultivation of the GM wheat and barley by the applicant will be significantly different from conventional practices for these plants.

64. GM wheat and barley seeds would be planted in the trial sites in winter or early spring.

65. The proposed 2 m buffer zone and 10 m monitoring zone surrounding the trial site would be either mowed, herbicide treated or weeded to maintain vegetation at a height of less than 10 cm.

66. The applicant proposes to harvest all GM wheat and barley at maturity by hand or by machine (such as a custom Plot Harvester). Threshing of GM plants would occur on site, or heads transported to approved facilities for analysis and processing.

67. Seed that remains after harvest would be either stored in an approved facility for subsequent use or destroyed.

68. Volunteers would be removed by hand or killed by herbicide application.

6.3 Presence of related plants in the receiving environment69. The site of O’Halloran Hill is close to urban areas of Adelaide, the nearest commercial production areas for either wheat or barley being approximately 50 km distant. However, the other four sites are located in regions where cereals are grown as crops.

70. Barley and wheat are not known to hybridise with each other under natural conditions, but do hybridise with a range of other plants present in the Australian environment. This is discussed in the two OGTR documents, The Biology of Triticum aestivum L. em Thell (Bread Wheat) (OGTR 2008b) and The Biology of Hordeum vulgare L. (barley) (OGTR 2008a). A summary of the information contained in these documents is presented below.



71. Wheat (Triticum aestivum) is sexually compatible with a number of species within the tribe Triticeae that occur in Australia. Of particular importance are durum wheat (Triticum turgidum ssp. Durum), rye (Secale cereale), and Triticale. Hybridisation with durum wheat occurs readily (Wang et al. 2005), whereas that with rye (Dorofeev 1969; Leighty & Sando 1928; Meister 1921) and Triticale (Ammar et al. 2004; Kavanagh et al. 2010) is rarer. It also readily hybridises with Aegilops species, but although some specimens of this genus have been collected in Australia, presumably originating from seed accidently introduced or straying from that brought in for breeding programs (AVH 2012), no Aegilops species is considered to be naturalised.

72. Australasia possesses four native Triticeae genera – Australopyrum, Stenostachys, Anthosachne (Elymus), and Connorochloa (Barkworth & Jacobs 2011) – as well as a number of introduced species of Triticeae, such as Elytrigia repens (couch grass) and at least four Thinopyrum species (Bell et al. 2010). Thinopyrum ponticum (tall wheatgrass) has been used as a saltland pasture plant in Australia, and in some regions has come to be classified as a weed (Barrett-Lennard 2003; NYNRMP 2011). Although there has been no concerted investigation of natural hybridisation of these native and introduced Triticeae species with wheat, based on experience of hybridising wheat with most other members of the Triticeae, it is likely that it never occurs.

73. Barley is divided into three gene pools, the basis for this division being primarily the ability to form interspecific hybrids, and the use of data arising from molecular and cytogenetic studies (Zhang et al. 1999). The primary gene pool consists of Hordeum vulgare ssp vulgare (cultivated barley) and H. vulgare ssp spontaneum (the progenitor of cultivated barley), which are fully interfertile. H bulbosum constitutes the secondary pool, while the tertiary pool consists of approximately 30 Hordeum species. Hybridisation of H. vulgare and H. bulbosum usually results in the elimination of the genome of H. bulbosum and the formation of haploids of H. vulgare, but it has proven possible to produce partially fertile hybrids that can be backcrossed to H. vulgare (Pickering & Johnston 2005). Although hybrids can be formed between H. vulgare and members of the tertiary gene pool, due to infertility these have not proven useful in the introgression of germplasm into H. vulgare. However, at least in some cases, colchicine can be used to double chromosome number and lead to the production of hybrid seeds (Islam et al. 2007).

74. H. bulbosum has been grown as a pasture and forage grass in Australia (Knupffer 2009), and the tertiary pool species H. murinum ssp leporinum (barley grass), H. murinum ssp glaucum (blue barley grass) and H. marinum (sea barley grass) are also widespread in Australian pastures and the environment (Mallett & Orchard 2002; Smith 1968). However, the above described limitations on hybridisation between H. vulgare and plants from the secondary and tertiary gene pools implies there is negligible risk of gene transfer from the GM barley plants to these relatives. Hybridisation between barley and other species is virtually unknown in nature.

75. As the two NGNE facilities are multi-user facilities, it is possible that other GM and non-GM wheat and barley plants will be grown there in close proximity to those plants that are the subject of this application. Currently, GM wheat and barley plants from DIRs 099 and 112 may be present. Additionally, the other three proposed field trial sites currently have approval for GM wheat and barley from DIR 102 to be grown.

6.4 Presence of similar genes and encoded proteins in the environment76. All the introduced genes and other genetic elements are from plants and bacteria that are widespread and prevalent in the environment (see Section 5). Most of them are commonly consumed by people or people are naturally exposed to them.

77. The nptII and hph selectable marker genes are from E.coli, which is widespread in the environment.

78. Although some of the regulatory sequences are derived from plant pathogens, they comprise only small parts of the total genomes and cannot of themselves cause disease.



Section 7 Relevant Australian and international approvals7.1 Australian approvals7.1.1 Previous approval by the Regulator

79. Wheat and/or barley lines possessing some of the introduced genes have previously been approved for limited and controlled release by the Regulator under licence DIR 102:

AtAVP1

ScNHA1

PpENA1

AtCIPK16

TRANSCRIPTION FACTOR 6

Aminotransferase

80. Information on previous DIR licences can be found on the GMO record on the OGTR website. There have been no reports of adverse effects on human health or the environment resulting from any of these releases.7.1.2 Approval by other government agencies

81. No other approvals relating to human health and safety and environment are currently required from Australian government agencies for this GM wheat and barley trial. However, approvals may be required under some State governments’ legislation unrelated to human health and safety and the environment.

7.2 International approvals of GM wheat and barley82. Field trials of different GM wheat and barley plants have been approved internationally, including in the USA, Canada, Germany, Czech Republic, Denmark, Hungary, Iceland, Italy, Spain, Sweden and the United Kingdom. The traits that have been modified include: novel protein production, disease resistance, insect resistance, altered grain properties and herbicide tolerance§.

§< http://www.aphis.usda.gov/brs/status/relday.html>, <http://gmoinfo.jrc.ec.europa.eu/gmp_browse.aspx> accessed January 2014.


http://gmoinfo.jrc.ec.europa.eu/gmp_browse.aspx

http://www.aphis.usda.gov/brs/status/relday.html


RISK ASSESSMENT PROCESS *

Risk scenarios

Substantive Risks

Risk Evaluation

Consequence assessment

Likelihood assessment

Identification of substantive risks

Negligible risks

RISK IDENTIFICATION RISK CHARACTERISATION

Risk context

Postulation of risk

scenarios

* Risk assessment terms are defined in the Risk Analysis Framework 2013


Chapter 2 Risk assessmentSection 1Introduction

83. The risk assessment identifies and characterises risks to the health and safety of people or to the environment from dealings with GMOs, posed by or as the result of gene technology (figure 3). Risks are identified within the context established for the risk assessment (see Chapter 1), taking into account current scientific and technical knowledge. A consideration of uncertainty, in particular knowledge gaps, occurs throughout the risk assessment process.

Figure 3 The risk assessment process

84. Initially, risk identification considers a wide range of circumstances whereby the GMO, or the introduced genetic material, could come into contact with people or the environment. Consideration of these circumstances leads to postulating plausible causal or exposure pathways that may give rise to harm for people or the environment from dealings with a GMO (risk scenarios) in the short and long term.

85. Postulated risk scenarios are screened to identify substantive risks that warrant detailed characterisation. A substantive risk is only identified for further assessment when a risk scenario is considered to have some reasonable chance of causing harm. Pathways that do not lead to harm, or could not plausibly occur, do not advance in the risk assessment process.

86. A number of risk identification techniques are used by the Regulator and staff of the OGTR, including checklists, brainstorming, reported international experience and consultation (OGTR 2013). A weed risk assessment approach is used to identify traits that may contribute to risks from GM plants, as this approach addresses the full range of potential adverse outcomes associated with plants. In particular, novel traits that may increase the potential of the GMO to spread and persist in the environment or increase the level of potential harm compared with the parental plant(s) are used to postulate risk scenarios (Keese et al. 2013). In addition, risk scenarios postulated in previous RARMPs prepared for licence applications of the same and similar GMOs are also considered.

Chapter 2 – Risk assessment 17


87. Substantive risks (i.e. those identified for further assessment) are characterised in terms of the potential seriousness of harm (Consequence assessment) and the likelihood of harm (Likelihood assessment). The level of risk is then estimated from a combination of the Consequence and Likelihood assessments. Risk evaluation then combines the Consequence and Likelihood assessments to determine level of risk and whether risk treatment measures are required. The potential for interactions between risks is also considered.

Section 2Risk Identification88. Postulated risk scenarios are comprised of three components (Figure 4):

i. The source of potential harm (risk source).

ii. A plausible causal linkage to potential harm (causal pathway).

iii. Potential harm to an object of value, people or the environment.

Figure 4 Risk scenario

89. In addition, the following factors are taken into account when postulating relevant risk scenarios:

the proposed dealings, which may be to conduct experiments, develop, produce, breed, propagate, grow, import, transport or dispose of the GMOs, use the GMOs in the course of manufacture of a thing that is not the GMO, and the possession, supply and use of the GMOs in the course of any of these dealings

the proposed limits including the extent and scale of the proposed dealings

the proposed controls to limit the spread and persistence of the GMO

characteristics of the parent organism(s).

90. Additional information relevant to the risk assessment has been declared CCI. The CCI was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.

2.1 Risk source91. The source of potential harms can be intended novel GM traits associated with one or more introduced genetic elements, or unintended effects/traits arising from the use of gene technology.

92. As discussed in Chapter 1, each of the GM wheat and barley lines has been modified by the introduction of one of 33 genes for abiotic stress tolerance or micronutrient uptake. These introduced genes are considered further as potential sources of risk.

93. In addition, the GM lines contain either or both of the nptII and hph selection marker genes (see Chapter 1). However, these genes and their products have already been extensively characterised and assessed as posing negligible risk to human or animal health or to the environment by the Regulator as well as other regulatory agencies in Australia and overseas. As these genes have not been found to pose substantive risks to either people or the environment, their potential effects will not be further assessed for this application. More information on selectable


plausible causal linkage

potential harm to

an object of value

(people/environment)

source of

potential harm

(a novel GM trait)


marker genes can be obtained from the OGTR document Marker genes in GM plants, available on the OGTR website.

94. All of the introduced genes include regulatory sequences, which are derived from plants, a plant virus (Cauliflower mosaic virus), and a common soil bacterium (Agrobacterium tumefaciens). Regulatory elements performing the same functions are naturally present in plants, and the introduced elements are expected to operate in similar ways to endogenous ones. There is no evidence that regulatory sequences themselves have toxic or allergenic effects (EPA 1996). Although the viral sequence is derived from a plant pathogen, it only constitutes a small part of the genome and cannot itself cause disease. Hence, potential effects from the regulatory elements themselves will not be considered further. However, regulatory sequences, especially the promoters, control the levels of gene expression and hence the levels of the derived proteins in the GM plants. The effects of these protein levels on, in particular, the toxicity and allergenicity of these plants (or at least materials derived from them), will be discussed below.

95. The genetic modifications also have the potential to cause unintended effects in several ways including altered expression of endogenous genes by random insertion of introduced DNA in the genome, increased metabolic burden due to expression of the introduced proteins, novel traits arising out of interactions with non-target proteins and secondary effects arising from altered substrate or product levels in biochemical pathways. Unintended effects might result in adverse outcomes such as toxicity or allergenicity.

96. However, the range of possible unintended effects produced by genetic modification is not likely to be greater than that from accepted conventional breeding techniques (Bradford et al. 2005; Committee on Identifying and Assessing Unintended Effects of Genetically Engineered Foods on Human Health 2004; The GM Science Review Panel 2003). Conventional methods of plant breeding may also induce unanticipated changes in plants (Haslberger 2003a), but new varieties produced by such techniques have rarely had traits that are undesirable for human health, safety or the environment (Hajjar & Hodgkin 2007)**. Therefore, unintended effects resulting from the process of genetic modification will not be considered further.

2.2 Causal pathway97. The following factors are taken into account when postulating plausible causal pathways to potential harm:

routes of exposure to the GMOs, the introduced gene(s) and gene product(s) potential effects of the introduced gene(s) and gene product(s) on the properties of the

organism potential exposure to the introduced gene(s) and gene product(s) from other sources in the

environment the environment at the site(s) of release agronomic management practices for the GMOs spread and persistence (invasiveness) of the GM plant, including

o establishmento reproductiono dispersal by natural means and by people

tolerance to abiotic conditions (eg climate, soil and rainfall patterns) tolerance to biotic stressors (eg pest, pathogens and weeds)

** More detail on potential for unintended effects as a result of the process of genetic modification can be found in the document Methods of plant genetic modification available from the Risk Assessment References page on the OGTR website.


http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/reference-3/$FILE/marker-genes-ref-1.pdf


tolerance to cultivation management practices gene transfer to sexually compatible organism gene transfer by horizontal gene transfer (HGT) unauthorised activities.

98. Although all of these factors are taken into account, some have been considered in previous RARMPs or are not expected to give rise to substantive risks.

99. The potential for horizontal gene transfer (HGT) and any possible adverse outcomes has been reviewed in the literature (Keese 2008) as well as assessed in many previous RARMPs. HGT was most recently considered in the RARMP for DIR 108. This and other RARMPs are available from the GMO Record on the OGTR website or by contacting the OGTR. No risk greater than negligible was identified due to the rarity of these events and because the wild-type gene sequences are already present in the environment and available for transfer via demonstrated natural mechanisms. Therefore, HGT will not be assessed further.

100. The potential for unauthorised activities to lead to an adverse outcome has been considered in previous RARMPs. The Act provides for substantial penalties for non-compliance and unauthorised dealings with GMOs. The Act also requires the Regulator to have regard to the suitability of the applicant to hold a licence prior to the issuing of a licence. These legislative provisions are considered sufficient to minimise risks from unauthorised activities, and no risk greater than negligible was identified in previous RARMPs. Therefore, unauthorised activities will not be considered further.

2.3 Potential harm101. Potential harms from GM plants include:

harm to the health of people or desirable organisms, including toxicity/allergenicity reduced establishment of desirable plants, including having an advantage in comparison to

related plants reduced yield of desirable vegetation reduced products or services from the land use restricted movement of people, animals, vehicles, machinery and/or water reduced quality of the biotic environment (eg providing food or shelter for pests or

pathogens) or abiotic environment (eg negative effects on fire regimes, nutrient levels, soil salinity, soil stability or soil water table).

reduced biodiversity through harm to other organisms or ecosystems.

102. These harms are based on those used to assess risk from weeds (Standards Australia 2006). Judgements of what is considered harm depend on the management objectives of the land where the GM plant is expected to spread to and persist. A plant species may have different weed risk potential in different land uses such as dryland cropping or nature conservation.

2.4 Postulated risk scenarios103. Six risk scenarios were postulated and screened to identify substantive risk. These scenarios are summarised in Table 5 and more detail of these scenarios is provided later in this Section. Postulation of risk scenarios considers impacts of the GM wheat and barley or their products on people undertaking the dealings, as well as impacts on people and the environment if the GM plants or genetical material were to spread and/or persist.

104. In the context of the activities proposed by the applicant and considering both the short and long term, none of the six risk scenarios gave rise to any substantive risks that could be greater than negligible.



Table 5 Summary of risk scenarios from dealings with GM wheat and barley genetically modified for abiotic stress tolerance or micronutrient uptake

Risk scenario

Risk source Causal pathway Potential harm Substantive risk?

Reason

1 Introduced abiotic stress tolerance and micronutrient uptake genes

Growing GM plants at the sites

Expression of genes in GM plants

Exposure of people who specifically deal with the GM plant material or other organisms that come into contact with the GM plant material in the trial sites

Allergic reactions in people or toxicity in people and other organisms

No The limited scale, short duration and other proposed limits and controls minimise exposure of people and other organisms to the GM plant material.

Plant material from the GMOs would not be used for human food or animal feed.

Most encoded proteins occur naturally in the environment and are not known to be toxic or allergenic (the proteins have a history of safe use).


Dispersal of GM seed outside trial limits

Growth of GM plants


Spread and persistence of populations of GM plants outside trial limits

Exposure of people or other organisms to GM plant material

Allergic reactions in people or toxicity in people and other organisms, andreduced establishment and yield of desirable plants, reduced biodiversity

No The limited scale, short duration and other proposed limits and controls minimise the likelihood that GM plant material would leave a trial site.

Abiotic stress tolerance may enable the GM plants to grow in a wider range of environments or perform better under stress compared to non-GM wheat and barley. However, it is unlikely that this would lead to weediness or greater harm to the environment than that found with non-GM plants of these species.


Dispersal of GM pollen outside trial limits

Vertical transfer of introduced genes to other sexually compatible plants, such as commercial varieties of wheat and barley

Expression of genes in plants




Most encoded proteins occur naturally in the environment and are not known to be toxic or allergenic to people or toxic to other organisms (the proteins have a history of safe use).





Spread and persistence of populations of GM plants outside a trial site

Reduced establishment and yield of desirable plants, reduced biodiversity


Abiotic stress tolerance may enable the GM plants to grow in a wider range of environments or perform better under stress compared to non-GM wheat and barley. However, it is unlikely that such spread would cause greater harm to the environment than that found with non-GM plants



Risk scenario

Risk source Causal pathway Potential harm Substantive risk?

Reason

of these species.


Dispersal of GM pollen within a site

Hybidisation of GM plants of this trial with GM plants (including volunteers) of another trial

Expression of genes in stacked GM plants



No The limited scale, short duration and other proposed limits and controls minimise exposure of people and other organisms to the GM plant material.

Most encoded proteins occur naturally in the environment and are not known to be toxic or allergenic to people or toxic to other organisms (the proteins have a history of safe use).

The stacking of genes from different GM plants is unlikely to increase the toxicity or allergenicity of plants.



Hybidisation of GM plants of this trial with GM plants (including volunteers) of another trial

Dispersal of plants or viable plant material containing stacked genes outside a site





GM plants that are produced by such gene transfer may grow in a wider range of environments or perform better under stress compared to non-GM wheat and barley. However, it is unlikely that such spread would cause greater harm to the environmental than that found with the GM plants or non-GM plants of these species.

The stacking of genes from different GM plants is unlikely to increase the weediness of plants.

2.4.1 Risk scenario 1Risk source Causal pathway Potential harmIntroduced abiotic stress tolerance and micronutrient uptake genes

Growing GM plants at the sites


Exposure of people who specifically deal with the GM plant material or other organisms that come into contact with the GM plant material in the trial sites


Risk source

105. The source of potential harm for this postulated risk scenario is the introduced abiotic stress tolerance and micronutrient uptake genes.

Causal pathway

106. The abiotic stress tolerance and micronutrient uptake genes are expressed in the plant tissues. People who are involved in the breeding, cultivating, harvesting, transporting and processing of the GM wheat and barley may be exposed to its products through contact (including inhalation of pollen). This would be expected to mainly occur in the trial sites, but could also occur anywhere the



GM plant material was transported or used for experimental analysis. Organisms that may be present in the trial sites, including birds, rodents and invertebrates, may be exposed to the GM plant material.

107. The proposed limits and controls of the trial would minimise the likelihood that people or other organisms would be exposed to GM plant material. Although people may directly handle the GM plant material, it is not to be used as human food. Further, as the trial is limited to five sites totalling a maximum of 2.5 ha, only a small number of people would deal with the GM plant material and a small number of organisms are likely to be exposed to it. GM plant material is not to be used as animal feed.

108. Fences surrounding each of the trial sites will exclude livestock and other large animals, while rodent control measures will be used to reduce the number of these animals. The two NGNE facilities are covered by bird netting.

109. The applicant has also proposed a series of measures, such as the monitoring and inspecting of sites, together with the cleaning of equipment, the storing of seed and the disposal of waste material that will together help reduce exposure of people to GM plant material in sites and the adjacent areas.

Potential harm

110. People exposed to the proteins expressed from the introduced genes or their associated products may show toxic or allergenic reactions, while organisms may show toxic reactions.

111. Proteins are not generally associated with toxic effects. As opposed to small molecular weight chemicals (eg pesticides), proteins have a number of properties that limit their ability to produce toxic effects upon ingestion, these including their likely digestion in the gastrointestinal tract and difficulties they encounter in traversing plasma membranes (Hammond et al. 2013). However, a small number of proteins have been shown to be toxic to humans and mammals, these originating mainly from animals (eg snakes, scorpions) and bacteria (Henkel et al. 2010; Karalliedde 1995). Lectins and protease inhibitors are plant proteins that have toxic properties, but for most people the level of exposure and response to the majority of these compounds is such that they are often classified as anti-nutrients (Delaney et al. 2008). The most well known plant proteins that are definitely toxic to humans are those lectins that consist of a ribosome-inactivating peptide (RIP) linked to a carbohydrate binding peptide, examples being ricin, abrin and modeccin (de Virgilio et al. 2010; Stirpe 2005).

112. All known food allergens are proteins, those derived from plants coming chiefly from peanut, tree nuts, wheat and soybean (Delaney et al. 2008; Herman & Ladics 2011). The structural and functional properties of plant food allergens can be used to classify them into approximately 30 families, these then being grouped into a small number of superfamilies (Hauser et al. 2008; Radauer & Breiteneder 2007; Salcedo et al. 2008). The major superfamiles are the prolamins, cupins, pathogenesis-related (PR) proteins, profilins and protease inhibitors.

113. Chapter 1, Section 5.3 presents a review of the potential toxic and allergenic properties of the proteins encoded by the introduced genes. It was concluded that none of the introduced proteins were likely to be toxic to people or other organisms, or allergenic to people. In respect of the general information on toxic and allergenic plant proteins outlined above, none of the introduced plant proteins (including that from the moss Physcomitrella patens) can be classified as a lectin or protease inhibitor (ie a likely toxin), or a prolamin, cupin, PR protein etc (ie a likely allergen). Although some strains of the yeast S. cerevisiae can produce toxins that kill sensitive strains of this organism (Schmitt & Breinig 2006), no strain is known to produce a toxin that is active against humans or animals, and the introduced yeast protein is not related to any of the known yeast allergens.



114. Non-GM wheat flour can produce allergic and other immune responses in susceptible individuals on inhalation or ingestion. Several types of allergic and immune reactions to wheat products have been recorded, with baker’s asthma and celiac disease being the best characterised. Bakers asthma is a respiratory allergy to inhaled flour and dust from grain processing, which is one of the most important occupational allergies in many countries (reviewed by Arts et al (2006) and Tatham & Shewry (2008)). Celiac disease is an autoimmune (genetic) disorder that is triggered by the consumption of gluten (Denham & Hill 2013; Hischenhuber et al. 2006). It is characterised by the immune system attacking the small intestine and inhibiting the absorption of nutrients into the body, likely leading to permanent tissue damage. These undesirable properties of cereals are not expected to be altered by expression of the introduced genes in the GM wheat and barley lines proposed for release.

115. For the evaluation of protein safety, the ILSI International Food Biotechnology Committee has collaborated with a group of experts to produce a two tiered method to assess the safety to health of proteins (Delaney et al. 2008; Hammond et al. 2013). The first tier examines five issues: (i) history of safe use of the protein; (ii) bioinformatics analysis; (iii) mode of action; (iv) in vitro digestibility and stability; and (v) expression level and dietary intake. Only if potential safety issues were identified in this evaluation would a second tier assessment be recommended, a prime example of such an assessment being a dose toxicology study.

116. The introduced proteins were examined against the first tier criteria:

(i) History of safe use. All of the plant proteins come from plants that do not raise any toxicological concerns and, other than Arabidopsis and P. patens, have a history of largely safe use in human diets (the major exceptions being allergenic reactions in a minority of people to some cereal proteins and nitrate in wheat being converted to toxic nitrites when consumed in large amounts by ruminants). Yeast has been used in the human diet for millennia, its allergenic properties well known and characterised. Six proteins (TRANSCRIPTION FACTOR 7, TaALMT1_minus_insert, ION TRANSPORTER 5, ION TRANSPORTER 6, ION TRANSPORTER 7C, and ScALMT1.M39.1_plus insert) contain amino acid substitutions or insertions/deletions of sequence; although uncertainty exists in regards to the effects such changes will have on the the toxicity and allergenicity of these proteins, there is no reason to expect these changes will affect such properties.

(ii) Bioinformatic analysis. None of the introduced proteins are members of protein classes that are known to have members with toxic or allergenic properties.

(iii) Mode of action. The functional categories of all but one of the genes (PHOTOSYNTHESIS AND METABOLISM GENE 4 from Arabidopsis) are known . Other than OsNAS2 (discussed below), the predicted roles of these proteins do not present any noteworthy concern. The mutations in the above mentioned six proteins may affect their activity, but not their biochemical functions.

(iv) In vitro digestibility and stability. No data.

(v) Expression level and dietary intake. No data.

117. Except for the six proteins with mutations and the one protein from Arabidopsis for which the function is unknown, all the proteins successfully pass criteria (i), (ii) and (iii). However, as noted above, for the six proteins with mutations, it is unlikely that their toxic or allergenic properties will be any different than those of the corresponding wild-type proteins.

118. Issues relating to points (iv) and (v) may need to be addressed prior to a commercial release of any of the GM wheat and barley lines.

119. Introduction of the OsNAS2 gene is aimed at increasing the level of iron in wheat. As outlined in Chapter 1, Section 5.6, while iron is an essential micronutrient, elevated levels of iron in a diet can be toxic. The majority of iron poisonings that occur are due to excessive consumption of dietary



suppliments (eg multivitamin tablets) that contain iron (Balmadrid & Bono 2009). The recommended adult daily allowance for iron in the United States is between 8-20 mg (CDC 2011), but it is estimated that a typical resident of that country may consume 15 to 40 mg of iron per day (Fine 2000). If 40 mg (the upper figure) is consumed by a 80 kg individual, this represents 0.5 mg/kg. Although the minimum toxic dose of iron is not known, it is suggested that a single ingestion of 20-40 mg/kg will produce signs of mild toxicity, but only a level above 60 mg/kg will lead to severe toxicity (Balmadrid & Bono 2009). The applicant is aiming to double the content of iron in wheat at most, a level that in an average diet is unlikely to produce any symptoms of toxicity.

120. Organisms exposed to the proteins expressed from the introduced genes or their associated products may show toxic reactions. The information presented above regarding the potential toxicity of the proteins to humans may well be applicable to other mammals; in the case of other organisms, there is no direct information. However, it is likely that only a small number of organisms will ever feed on the GM plant material. Moreover, as the genes are derived from yeast, moss and a range of plants that are common in the environment, it is likely that most organisms that may enter the trial sites and feed on the GM plants, have had prior exposure to these proteins or their homologues in other plants.

121. Gene technology has the potential to cause unintended effects in several ways, including altered expression of an endogenous gene by random insertion of an introduced DNA in the genome, increased metabolic burden due to higher expression of the introduced protein, novel traits arising out of interactions with non-target proteins and secondary effects arising from altered substrate or product levels in biochemical pathways. Such an effect could lead to elevation of the concentration of a normally benign wheat or barley compound to a level where it induces a toxic or allergenic reaction if consumed in an average diet. It is also possible that an entirely novel compound could be produced with such a reaction. In this context, it is important to note that changes of this nature, such as the unexpected increase in the level of an endogenous toxin, can also be induced in plants by conventional methods of plant breeding (Haslberger 2003b). As noted above, the experience of conventional breeding is that the molecular constitution of a new variety produced by such techniques has rarely been such that it has been considered undesirable for human health. The implication is that the movement into wheat and barley of any of the genes that are the subject of this application, none of which belong to any known class of toxins or allergens, is unlikely to result in the production (directly or indirectly) of a novel toxin or allergen (Steiner et al. 2013; Weber et al. 2012). This includes the production of such a compound via the production of a fusion protein.

122. Eight of the introduced genes code for transcription factors. Although the applicant hopes that these genes will play a role in inducing either drought tolerance or nitrogen use efficiency, it is possible that their expression could influence other biochemical processes, potentially leading to a plant with toxic or allergenic properties.

123. In wheat and barley, the synthesis of endogenous allergens, such as gluten, is likely regulated primarily at the transcriptional level. Activation and down-regulation of the expression of their respective genes probably involves a number of different transcription factors binding to each other, and either directly or indirectly to DNA promoter elements. Transcription factors that have been associated with gluten genes include SPA (storage protein activator), a member of the Opaque2 (O2) class of basic leucine zipper (bZIP) factors (Albani et al. 1997; Ravel et al. 2009) and DOF (DNA binding with one finger) proteins (Dong et al. 2007; Romeuf et al. 2010). Only one of the transcription factors in this application belongs to any such class, but within that class it is not a member of the same group as that protein associated with gluten biosynthesis. The identity of this introduced gene has been declared as CCI. The confidential information was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.



124. The possibility of a harm to human health eventuating from expression of a transcription factor can be reviewed against the above discussed experience of conventional breeding. Many of the unknown genes that have been moved into plants by conventional breeding must code for transcription factors, and the process of crop domestication provides a number of examples of new phenotypes that have been specifically linked to genetic changes involving these proteins (Doebley et al. 2006; Sang 2009), but none of these has been associated with the generation of a plant that has induced one or more harms to human or animal health.

Conclusion: Risk scenario 1 is not identified as a substantive risk, due to the likely limited exposure of humans to the expressed proteins, and the predicted lack of significant toxicity or allergenicity of the introduced proteins to humans and other organisms. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.2.4.2 Risk scenario 2Risk source Causal pathway Potential harmIntroduced abiotic stress tolerance and micronutrient uptake genes

Dispersal of GM seed outside trial limits

Growth of GM plants


Spread and persistence of populations of GM plants outside trial limits


Allergic reactions in people or toxicity in people and other organisms, andreduced establishment and yield of desirable plants, reduced biodiversity

Risk source

125. The source of potential harm for this postulated risk scenario is the introduced abiotic stress tolerance and micronutrient genes.

Causal pathway

126. The abiotic stress tolerance and micronutrient genes are present in plant tissues. If seed was dispersed outside any of the trial sites, this seed could germinate and give rise to plants expressing the introduced genes. These plants could spread and persist in the environment outside the trial limits and people and other organisms may be exposed to GM plant materials.

127. Dispersal of GM plant material outside the limits of any trial site could occur through the activity of people (including the use of agricultural equipment), the activity of animals such as rodents, herbivores and birds, or through extremes of weather such as flooding or high winds. Wheat and barley lack seed dispersal characteristics such as stickiness, burrs and hooks, which can contribute to seed dispersal via animal fur (Howe & Smallwood 1982). The intended introduced traits of the GM plants, abiotic stress tolerance and micronutrient uptake, are not expected to alter these characteristics of seeds.

128. Seed dispersal for wheat and barley through endozoochory (the ingestion and excretion of viable seeds) has not been reported. Nevertheless, it cannot entirely be discounted that wheat and barley seeds could be dispersed and germinate after passage through the digestive system of some mammals or birds. For example, viable wheat seeds have been detected in cattle dung (Kaiser 1999). Seeds which survive chewing and digestion by animals are typically small and dormant (Malo & Suárez 1995). Corellas were shown to excrete some viable wheat seeds, although the proportion is extremely low (Woodgate et al. 2011).

129. Kangaroos, rabbits and rodents are known pests of wheat and barley crops, and cattle or sheep may graze cereals. Each site will be fenced, limiting the possibility of seed dispersal by any large animals such as cattle and sheep. Rabbits favour soft, green, lush grass (Myers & Poole 1963) and select the most succulent and nutritious plants first (Croft et al. 2002). Although viable seeds from a variety of plant species have been found in rabbit dung, viable wheat seeds were not among them (Malo & Suárez 1995). Barley seeds were not recorded in this study. Other studies have shown that



generally very few viable seed are obtained from rabbit dung (Welch 1985; Wicklow & Zak 1983). Rodents are opportunistic feeders and their diets include seeds and other plant material (Caughley et al. 1998). They may not only eat and destroy seed at the seed source but also hoard seeds (AGRI-FACTS 2002), which increases the possibility of seed dispersal. Similar to other licences issued for field trial of GM plants including wheat and barley, the applicant is proposing a 10 m monitoring zone around the GM planting area to be maintained in a manner that does not attract or harbour rodents (such as keeping it bare or mown) and implementation of rodent control measures if rodents are detected. Only a very limited amount of rodent activity has been observed at trial sites under these other GM field trial licences. These standard measures minimise the potential for seed dispersal by rodents.

130. Characteristics that influence the spread (dispersal of the plant or its genetic material) and persistence (establishment, survival and reproduction) of a plant species determine the degree of its invasiveness. These characteristics include the ability to establish in competition with other plants, to tolerate standard weed management practices, to reproduce quickly, prolifically and asexually as well as sexually, and to be dispersed over long distances by natural and/or human means.

131. Baseline information on the weediness of wheat and barley, including factors limiting the spread and persistence of non-GM plants of these species, is given in The Biology of Triticum aestivum L. em Thell (Bread Wheat) (OGTR 2008b) and The Biology of Hordeum vulgare L. (barley) (OGTR 2008a). In summary, both wheat and barley have some characteristics of invasive plants, such as being capable of wind-pollination (although wheat is predominantly self-pollinating) and the ability to germinate or to produce seed in a range of environmental conditions. However, these cereals lack most of the characteristics that are common to invasive plants, such as the ability to produce a persisting seed bank, rapid growth to flowering, continuous seed production as long as growing conditions permit, high seed output, high seed dispersal and long-distance seed dispersal (Keeler 1989). In addition, both cereals have been bred to avoid seed shattering, and white wheat cultivars have little seed dormancy (OGTR 2008a; OGTR 2008b).

132. The expected phenotypic differences between the GM wheat and barley and their non-GM progenitors are either tolerance to an abiotic stress or increased uptake of iron. These introduced traits are not expected to alter the reproductive or dispersal characteristics of the GM plants. In reference to competitive ability, there is the potential for the GM plants to have an increased distribution in the natural environment and agricultural settings, especially as it is hoped that the introduction of the genes will make the GM plants more tolerant to conditions of abiotic stress. Although the performance of the GM plants in the field is yet to be determined (which will act as an indication of their performance in natural environments), abiotic stress tolerance per se cannot be interpreted as a cue for the plants to increase their invasiveness. Due to the complexity of environmental conditions, the traits are not expected to have a significant effect on the invasiveness of the GM plants. However, there is uncertainty around how tolerance to abiotic stresses will impact on the potential survivial of the GM plants outside of the agricultural setting, in particular what level of improved tolerance will lead to a significant change to persistence.

133. The techniques of conventional breeding (eg selection of plants amongst available germplasm, wide crosses, mutagenesis) have been used to produce varieties of wheat and barley that possess various abiotic stress tolerances. The experience of conventional breeding is a useful backdrop against which to view the potential invasiveness of the abiotic stress tolerant GM plants in this application.

134. Crosses with wild relatives have been used to transfer useful genes to crop plants (Goodman et al. 1987; Hajjar & Hodgkin 2007; Maxted & Kell 2009; Prescott-Allen & Prescott-Allen 1998). Wheat is a crop where such transfer has been most fruitful. Wild relatives of wheat that have been exploited as sources of genes include a whole range of Aegilops species (eg Ae. tauschii, Ae. speltoides, Ae. squarrosa), Triticum species (eg T. turgidum subsp. dicoccoides and T. monococcum) and Thinopyrum bessarabicum. Although the transfer of genes conferring disease and



pest resistance has been most common, a survey of published reports of the use of wild relatives in breeding found that 13% concerned abiotic stress tolerance (Maxted & Kell 2009).

135. Mutagenesis has also been used to generate a number of varieties of cereals (and many other crops) that have abiotic stress tolerances (Ahloowalia et al. 2004; Cheema et al. 1999; Tomlekova 2010)( http://mvgs.iaea.org/). With respect to its progenitor, the gamma radiation induced barley variety Golden Promise has been shown to possess both enhanced drought and salt tolerances (Forster 2001). Aluminium tolerant lines of barley have been identified amongst four varieties of barley exposed to chemical mutagenesis (Nawrot et al. 2001).

136. Importantly, no abiotic stress tolerant wheat or barley plant that has been generated by any form of conventional breeding has been demonstrated to have increased invasiveness, and the introduced genes are not expected to increase the potential invasiveness of GM plants relative to non-GM plants subjected to conventional breeding.

137. De-domestication, the evolutionary loss of traits gained under domestication, is frequently found amongst the small number of known examples of the evolution of invasiveness amongst existing crop plants, this being most obviously reflected in the acquisition of the ability to more readily disperse seed or fruits (Ellstrand et al. 2010). The only known case of invasiveness in wheat is in Tibet, where ‘semi-wild’ wheat (presumably the result of de-domestication) has shattering heads (brittle rachis) and hulled seeds (toughened glumes preventing free threshing) (Ayal & Levy 2005). Both these traits are absent from domesticated wheat, underlining their importance in marking the boundary between cultivated and weedy forms of this plant; shattering allows easy seed dispersal in the wild, while the hull around grains protects the grain from abiotic and biotic stresses. Although the de-domestication of barley has not been recorded, compared to its wild progenitor, domesticated barley possesses a non-brittle rachis, with some modern varieties being hulled and some hull-less, the latter being preferred for human food due to the ease to isolate grain (Azhaguvel & Komatsuda 2007; Taketa et al. 2008).

138. However, there is no reason to believe that the introduction of the genes into wheat and barley is likely to lead to a reversion of either the non-shattering or hull-less traits. The obvious rarity of the natural loss of the non-shattering and hull-less traits in conventionally bred commercial wheat and barley varieties indicate their genetic stability. Further, none of the introduced genes are amongst those known to be associated with shattering or the toughness of the glumes (Sang 2009).

139. Genes that have been associated with abiotic stresses have frequently been shown to have pleiotropic effects, a phenomenon that is likely due to most such genes playing important roles in genetic regulation. Not only can a gene provide tolerance to multiple individual abiotic stress (eg both cold and salinity), but can even impart both abiotic and biotic stress tolerances (Howles & Smith 2013). However, as outlined above, although there is uncertainty about the abilities of the GM plants to spread and persist, in the natural setting it is unlikely that these abilities are significantly changed compared to the non-GM counterparts. If a plant acquires a particular biotic stress tolerance (eg increased fungal resistance), then that would likely be regarded as a bonus in an agricultural situation but may not provide an advantage in a natural setting such as a native reserve.

140. The proposed limits and controls of the trial would minimise the likelihood of spread and persistence of the GM plants. The small size (up to 2.5 ha per year) will limit the potential for dispersal of GM plant material and exposure to this material. Each of the proposed trial sites will be surrounded by a fence and only approved staff with appropriate training will have access to these sites, which will minimise potential for dispersal of seed by grazing livestock and people. This will reduce inadvertent access by humans and prevent grazing livestock from entering any site, thus minimising dispersal of GM plant material and exposure to this material. Dispersal of GM plant material by authorised people entering the proposed trial site would be minimised by a standard condition of DIR licences which requires the cleaning of all equipment used at the trial site, including clothing. All GM plant material will be transported in accordance with the Regulator’s transport guidelines, which will minimise the opportunity for its dispersal. Limits and controls are



further discussed in Chapter 3. Furthermore, extra conditions associated with growing the GM plants in the NGNE facilities would also reduce the likelihood of plant material being mixed between different GM trials conducted concurrently in these facilities and subsequently spread.

Potential harm

141. As discussed in Section 2.3 of this Chapter, all plants have the potential to lead to harm in certain environments. Harms that may arise from a certain plant species in a particular environment include adverse effects on the health of people, animals or the environment, reducing establishment, yield and/or quality of desired plants, and restricting physical movement.

142. As discussed in risk scenario 1, the introduced gene products are not expected to be toxic or allergenic to people, or toxic to other organisms. This would apply even if the GM wheat and barley plants established beyond the trial limits.

143. However, GM wheat and barley plants established beyond the trial limits could potentially impact the environment by reducing the establishment of desired plants. In turn, this could lead to the fragmentation of the habitats of other plants, decreasing the probability of these plants (and the animals that live amongst these plants) maintaining effective breeding populations. As such, there may be a reduction in the biodiversity in regions where the GM plants grew. In reference to native habitats, it must nevertheless be appreciated that there would have to be large numbers of GM plants before the establishment of native plants was affected.

144. In particular, if GM wheat and barley lose the trait of non-shattering heads, they would be expected to spread and persist to a greater degree than that of non-GM commercial varieties of these species, their grain being naturally lost and less dependant on human intervention for dispersal. Such GM plants would likely reduce the number of domesticated plants growing successfully to maturity and setting seed in fields, hence reducing the yield of these cereals. This is an essential facet of the problem with Ae. cylindrica in the United States, where the heads of this plant typically shatter prior to wheat harvest, generating a seed bank in the soil that persistently gives rise to these plants amongst the desired cereals (Colquhoun & Fandrich 2003; Yenish et al. 2009). In the case of wheat, prior to late maturity, it could possibly be difficult, if not impossible, to distinguish between shattering and non-shattering varieties.

145. The potential of these harms can be evaluated against the experience of conventional breeding. No commercially released variety of wheat or barley that is the product of any form of conventional breeding has been recorded to have negatively impacted the environment (or the health of humans and/or animals), beyond that normally associated with these cereals, and subsequently flagged as an environmental weed. In this context, it is especially relevant to remember, as noted above, that a number of these conventional varieties have been bred for abiotic stress tolerance, and none have been recorded as causing harms to the environment.

146. Therefore, the introduction into wheat and barley of any of the genes that are the subject of this application is unlikely to result in the GM plants possessing a trait that leads them to being classified as a weed (National Research Council 1989).

Conclusion: Risk scenario 2 is not identified as a substantive risk as none of the engineered traits are associated with weediness, it is unlikely that any of the characteristics associated with weeds will occur in the GM plants, and because of the limits and controls proposed for the field trial. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.2.4.3 Risk scenario 3

Risk source Causal pathway Potential harmIntroduced abiotic stress tolerance and micronutrient



Allergic reactions in people or toxicity in people and other



Risk source Causal pathway Potential harmuptake genes



organisms

Risk source


Causal pathway

148. The abiotic stress tolerance and micronutrient genes are present in plant tissues. Pollen from the GM plants could be transferred outside of a trial site (eg via wind) and fertilise sexually compatible plants, whether they be non-GM wheat or barley, or plants from another species. Alternatively, if seed was dispersed outside any of the trial sites, plants expressing the introduced genes may grow and subsequently disperse pollen. Hybrid plants possessing the introduced genes may form the basis for the spread of these genes in other varieties of wheat or barley, or other plant sexually compatible species. People and other organisms could be exposed to the proteins expressed from the introduced genes through contact with (including inhalation of pollen) or consumption of GM plant material deriving from the plants to which the genes have been transferred.

149. Baseline information on vertical gene transfer associated with non-GM wheat and barley plants can be found in The Biology of Triticum aestivum L. em Thell (Bread Wheat) (OGTR 2008b) and The Biology of Hordeum vulgare L. (barley) (OGTR 2008a).

150. It should be noted that vertical gene flow per se is not considered an adverse outcome, but may be a link in a chain of events that may lead to an adverse outcome.

151. The proposed limits and controls of the trial (discussed in risk scenarios 1 and 2) would minimise the likelihood of the dispersal of pollen, seed and exposure to GM plant material.

152. Wheat plants and barley plants are predominantly self-pollinating and the chances of natural hybridisation occurring with commercial crops or other sexually compatible plants are low and decreases with distance to the GM plants. Outcrossing rates decline significantly over distance, with most pollen falling within the first few metres. Rates are also influenced by the genotype of the variety, and environmental conditions, such as wind direction and humidity.

153. Wheat is sexually compatible with many species within the genus Triticum, and in closely related genera such as Aegilops, Secale (rye) and Elytrigia (Chapter 1, Section 6.3). Durum wheat (other than bread wheat, the only Triticum species present in Australia) can cross with wheat, although there are no reports of gene flow beyond 40 m (Matus-Cadiz et al. 2004). Hybrids between wheat and Secale cereale are sterile, but treatment with colchicine doubles the chromosome number and results in a fertile plant, the commercialised Triticale (Knupffer 2009). Natural hybridisation between wheat and Triticale rarely occurs (Ammar et al. 2004; Kavanagh et al. 2010), at least partly due to both species being largely self-fertilising (Acquaah 2007). Elytrigia repens does occur as an introduced plant in Australia, but a review of possible means of pollen-mediated gene flow from GM wheat to wild relatives in Europe concluded that there was a minimal possibility of gene flow from wheat to Elytrigia spp. (Eastham & Sweet 2002). Species of Aegilops are not known in Australia. Although specific data is lacking, it is likely that hybridisation of wheat and barley with the four native Australasian Triticeae genera never occurs under natural conditions. Limitations on hybridisation between H. vulgare and plants from the secondary and tertiary gene pools implies there is negligible risk of gene transfer from the GM barley plants to these relatives (OGTR 2008a).

154. The applicant proposes to control related species within a 10 m monitoring zone around the trial site and to prevent cultivation of other wheat, barley and sexual compatible plant species within 200 m of the trial site. Isolation from related species and other wheat and barley cultivation will greatly restrict the potential for pollen flow and gene transfer. In addition, the applicant proposes to



perform post-harvest monitoring and to destroy any volunteer plants found at the site to ensure that no GM wheat and barley remain that could then hybridise with sexually compatible plants.

Potential harm

155. People who are exposed to the proteins expressed from the introduced genes or their associated products through contact or consumption of GM plant material may show toxic or allergenic reactions, while organisms may show toxic reactions from consumption of GM plant material.

156. In the rare event of vertical transfer of the introduced genetic material from the GM plants to non-GM wheat and barley plants or sexually compatible species, the genetic material is expected to behave in similar ways as in the GM wheat and barley. As discussed in risk scenario 1, the introduced gene products are not expected to be toxic or allergenic to people, or toxic to other organisms and there is no reason to expect the production of an associated compound with a toxic or allergenic property.

157. The traits that have been engineered into the GM plants of this application could become, via vertical gene transfer, combined with traits possessed by other non-GM commercially cultivated wheat and barley plants. However, as discussed above (risk scenario 1), plants that are the product of hybridisation between two varieties are unlikely to possess a level of toxicity or allergenicity greater than that of either parent.

Conclusion: Risk scenario 3 is not identified as a substantive risk, due to the predicted lack of toxicity or allergenicity of the introduced proteins to humans or other organisms, there being no reasonable expectation that toxicity or allergenicity will be a problem in any plants that are the products of the GM plants and other plants, and the nature of the limits and controls proposed for the field trial to restrict gene flow. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.2.4.4 Risk scenario 4

Risk source Causal pathway Potential harmIntroduced abiotic stress tolerance and micronutrient uptake genes






Risk source


Causal pathway

159. The abiotic stress tolerance and micronutrient genes are present in plant tissues including pollen. Pollen from the GM plants could be transferred outside of a trial site (eg via wind) and fertilise sexually compatible plants, whether they be non-GM wheat or barley, or plants from another species. Alternatively, if seed was dispersed outside any of the trial sites, plants expressing the introduced genes may grow and disperse pollen. Hybrid plants possessing the introduced genes may form the basis for the spread of these genes in other (initially non-GM) varieties of wheat or barley, or other plant species. These plants could persist in the environment, displacing other, desirable plants.

160. The proposed limits and controls of the trial (discussed in risk scenarios 1 and 2) would minimise the likelihood of the dispersal of pollen and seed. Vertical transfer was reviewed in risk scenario 3. The proposed limits and controls of the trial would minimise the possibility that seed



would leave any trial site, or that pollination would occur to plants outside a trial site. Wheat and barley are predominantly self-pollinating, but are capable of crossing with a number of plant species that exist in Australia.

Potential harm

161. If the vertical transfer of the introduced genes from the GM plants causes the recipient species to spread and persist in the environment to a degree greater than normally found amongst these species, they may produce one or more harms. These potential harms were summarised in risk scenario 2. In particular, the GM plants may act to reduce the establishment and yield of desired plants and subsequently reduce biodiversity.

162. Risk scenario 2 summarises the reasons that the introduced genes are unlikely to make the GM wheat and barley lines more weedy, particularly as they are not associated with the key weediness traits of shattering heads and hulled seeds, although there is some uncertainty about whether the traits could potentially improve the persistence of the GM plants outside of the agricultural setting.These reasons are likely to be applicable to any plants to which the genes are transferred.

163. The traits that have been engineered into the GM plants of this application could become, via vertical gene transfer, combined with traits possessed by other non-GM commercially cultivated wheat and barley plants. However, as discussed above (risk scenario 2), plants that are the product of hybridisation between two varieties are unlikely to possess a level of weediness greater than that of either parent.

Conclusion: Risk scenario 4 is not identified as a substantive risk, as the introduced genes are unlikely to lead to increased weediness in the GM plants themselves or any other plants to which they are transfered, and because of the nature of the limits and controls proposed for the field trial. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.2.4.5 Risk scenario 5



Hybridisation of GM plants of this trial with GM plants (including volunteers) of another trial




Risk source


Causal pathway

165. The abiotic stress tolerance and micronutrient genes are present in plant tissues. The NGNE facility is a multiuser facility, where at any one time, more than one licence holder may be conducting trials of different GM plants. Pollen from the GM plants of one trial could inadvertently fertilise other sexually compatible GM plants, including volunteers, inside a site. This pollen could be the result of transfer by an agent (eg wind, insects) from one planting area to another, or derived from volunteer plants from either trial. In early 2014, the only licence that is authorised to trial sexually compatible GM plants in either of these facilities is DIR 112, a licence held by CSIRO for the growth of GM wheat and barley with altered grain composition or nutrient utilisation efficiency (abiotic stress tolerance) at Merredin. DIR 099, also held by the CSIRO, authorises the growth of GM plants with the same traits as DIR 112 at Merredin, but the period for plantings has finished and there has been no application for an extension of time. In the case of the non-NGNE sites, all



three sites have been previously used for field trials of GM plants under DIR 102, volunteers of which could still arise and hybridise with the plants of this application. Currently, DIR 102 authorises the growth of GM wheat and barley at the three non-NGNE sites of this application until December 2015. The traits of the DIR 102 plants are classified as abiotic stress tolerances and some of the genes are identical to the genes in this application. As volunteer numbers from DIR 102 are expected to be low, stacking between volunteers from DIR 102 and DIR 128 is less likely to occur than stacking between GM plants from DIR 128. Even if unintentional stacking did occur, it would still result in plants with abiotic stress tolerance.

166. People working with any trial could be exposed to the hybrid plants.

167. Vertical gene transfer associated with non-GM wheat and barley plants was discussed in risk scenario 3. In summary, these two cereals are predominantly self-pollinating, but are capable of crossing with a number of plant species that exist in Australia.

168. The applicant proposes to surround each planting area in any trial site by a 2 m wide buffer zone where plant growth will be controlled by mowing, herbicide treatment and/or weeding. Further, the applicant has requested permission to grow GM wheat and barley from different licences next to each other providing they are separated by buffer zones of at least 4 m. The RARMP for DIR 094 considered the possibility of hybidisation occurring over these distances, and concluded that it would be minimal.

169. The licence for DIR 112 specifies that if the GM plants of that trial are grown in the Merredin NGNE facility at the same time as sexually compatible GM and non-GM plants from another licence, seed from any plants of DIR 112 must not be used for the development of commercial cultivars. Such a measure would help minimise the likelihood that the genes of this application would be spread to other plants and being inadvertently grown outside a trial site, leading to people being inadvertently exposed to GM material.

170. The proposed limits and controls of the trial would minimise the possibility that seed would leave any trial site. These are discussed under risk scenario 2. Therefore exposure would be restricted to people working at the trial sites. All GM seed leaving trial sites will be transported in accordance with the Regulator’s transport guidelines, which will minimise the opportunity for its dispersal and exposure of other people.

Potential harm

171. People who are exposed to the proteins expressed from the introduced genes or their associated products through contact or consumption of GM plant material may show toxic or allergenic reactions, while organisms may show toxic reactions from consumption of GM plant material.

172. As discussed in risk scenario 1, the introduced gene products of this application are not expected to be toxic or allergenic to people, or toxic to other organisms and there is no reason to expect the production of an associated compound with a toxic or allergenic property. The toxicity and allergenicity associated with the introduced genes of other sexually compatible GM plants growing in a NGNE facility would be the subject of the RARMP(s) associated with their licence(s). According to the RARMPs for DIR 099, 102 and 112, there is no information to suggest that the proteins encoded by the introduced genes of those applications are likely to be toxic to people or other organisms, or allergenic to people. As discussed above (risk scenario 1), plants that are the product of hybridisation between two GM varieties are unlikely to possess a level of toxicity or allergenicity greater than that of either parent (ie the stacking of genes from different GM plants will be unlikely to generate a plant with a higher level of toxicity or allergenicity than the individual GM parents).

Conclusion: Risk scenario 5 is not identified as a substantive risk, due to the predicted lack of significant toxicity or allergenicity of the introduced proteins to humans or other organisms, and the



nature of the limits and controls proposed for the field trial. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.2.4.6 Risk scenario 6



Hybridisation of GM plants of this trial with GM plants (including volunteers) of another trial

Dispersal of plants or viable plant material containing stacked genes outside a site




Risk source


Causal pathway

174. The abiotic stress tolerance and micronutrient genes are present in plant tissues. As discussed in Risk Scenario 5, the NGNE facility is a multiuser facility, where at any one time, more than one licence holder may be conducting trials of different GM plants. Pollen from the GM plants of one trial could inadvertently fertilise other sexually compatible GM plants, including volunteers, inside a site. GM wheat and barley plants with altered grain composition or nutrient utilisation efficiency (abiotic stress tolerance) from DIR 112 and 099 may be present at Merredin. Additionally, the non-NGNE sites may have GM wheat and barley plants or volunteers with abiotic stress tolerances from the DIR 102 field trial.

175. If hybridisation occurred between the GM plants of this application and those of DIR 112, 102 or 099, the progeny would have the traits of abiotic stress tolerance or micronutrient uptake stacked with additional abiotic stress tolerances, altered grain composition or altered nutrient utilisation efficiency.

176. If plant material, which unknowingly was the product of hybridisation between the GM plants of this trial and other GM plants in any site, was purposely taken outside of a trial site, or was inadvertently dispersed outside of a trial site, GM hybrid plants could arise from the germination of seed. These plants then could spread and persist in the environment. However, it is unlikely that the stacking of the traits of the GM plants of DIRs 099, 102 and 112 (altered grain composition and abiotic stress tolerance) with those of this application (abiotic stress tolerance and micronutrient uptake) will significantly influence the invasiveness of the plants, but this is an area of uncertainty. As discussed above (scenario 2), the combination of such traits is not likely to produce a plant that is more invasive than the individual parents but it is uncertain whether the persistence of these GM plants outside of the agricultural setting may be improved.

177. Risk scenario 5 reviews the growing of GM plants from different trials adjacent to each other, such as possibly may occur in the NGNE facilities, noting the applicant proposes using buffer zones and structuring the licence similar to other licences that have concerned the growing of different GM plants close to each other.

178. The proposed limits and controls of the trial would minimise the possibility that seed would leave any trial site. These are discussed under risk scenario 2. All GM seed will be transported in accordance with the Regulator’s transport guidelines, which will minimise the opportunity for its dispersal.



Potential harm

179. If the vertical transfer of genes from the GM plants causes the recipient species to spread and persist in the environment to a degree greater than normally found amongst these species, they may produce one or more harms. These harms were summarised in risk scenario 2. In particular, the GM plants may act to reduce the establishment and yield of desired plants and biodiversity.

180. As discussed in risk scenario 2, the introduced gene products of this application are unlikely to cause the GM wheat and barley lines to have a greater impact on the environment than non-GM wheat and barley, these reasons being applicable to any plants to which the genes are transferred. The weediness associated with the introduced genes of other sexually compatible GM plants growing in a NGNE facility or at the other field trial sites would be the subject of the RARMP(s) associated with their licence(s). According to the RARMPs for DIR 099, 102 and 112, there is no information to suggest that the proteins encoded by the introduced genes of those applications are likely to induce weediness in GM wheat and barley. Based upon the above discussed experience of conventional breeding (risk scenario 2), plants that are the product of hybridisation between two varieties are unlikely to have a greater impact on the environment than that of either parent (ie the stacking of genes from different GM plants will be unlikely to generate a plant more harmful than the individual GM parents).

Conclusion: Risk scenario 6 is not identified as a substantive risk, as the introduced genes of this application are unlikely to lead to increased weediness of GM wheat or barley, it is unlikely that the stacking of genes from different GM trials will lead to a weedy plant, and because of the nature of the limits and controls proposed for the field trial. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.

Section 3Uncertainty181. Uncertainty is an intrinsic part of risk analysis††. There can be uncertainty about identifying the risk source, the causal linkage to harm, the type and degree of harm, the change of harm occurring or the level of risk. In relation to risk management, there can be uncertainty about the effectiveness, efficiency and practicality of controls.

182. Risk analysis can be considered as part of a first tier uncertainty analysis, namely a structured, transparent process to analyse and address uncertainty when identifying, characterising and evaluating risk. However, there is always some residual uncertainty that remains. If the residual uncertainty is important and critical to decision making, then this residual uncertainty may be subjected to further analysis (=second tier uncertainty analysis), such as building ‘worst case’ scenarios, or by using meta-analysis where results from several studies are combined.

183. There are several types of uncertainty in risk analysis (Bammer & Smithson 2008; Clark & Brinkley 2001; Hayes 2004). These include:

uncertainty about facts:

– knowledge – data gaps, errors, small sample size, use of surrogate data

– variability – inherent fluctuations or differences over time, space or group, associated with diversity and heterogeneity

uncertainty about ideas:

– description – expression of ideas with symbols, language or models can be subject to vagueness, ambiguity, context dependence, indeterminacy or under-specificity

– perception – processing and interpreting risk is shaped by our mental processes and social/cultural circumstances, which vary between individuals and over time.

†† A more detailed discussion of uncertainty is contained in the Regulator’s Risk Analysis Framework available from the Risk Assessment References page on the OGTR website or via Free call 1800 181 030.




184. For DIR 128, uncertainty is noted particularly in relation to the characterisation of:

Potential increases in toxicity or allergenicity as a result of the genetic modifications

Potential for increased survival of the GMOs, including in land uses outside of agriculture

The effect of stacking of genes of this application with genes belonging to other sexually compatible GM plants grown in the two NGNE facilities or at the other field trial sites.

185. Additional data, including information to address these uncertainties, may be required to assess possible future applications for a larger scale trial, reduced containment conditions, or the commercial release of these GM wheat or barley lines if they are selected for further development.

186. Chapter 3, Section 4, discusses information that may be required for future releases.

Section 4Risk evaluation187. Risk is evaluated against the objective of protecting the health and safety of people and the environment to determine the level of concern and, subsequently, the need for controls to mitigate or reduce risk. Risk evaluation may also aid consideration of whether the proposed dealings should be authorised, need further assessment, or require collection of additional information.

188. Factors used to determine which risks need treatment may include:

risk criteria level of risk uncertainty associated with risk characterisation interactions between substantive risks.

189. Six risk scenarios were postulated whereby the proposed dealings might give rise to harm to people or the environment. The level of risk for each scenario was considered negligible in relation to both seriousness and likelihood of harm, in the context of the control measures proposed by the applicant, and considering both the short and long term. The principal reasons for these conclusions are summarised in Table 5 and include:

limits on the size, location and duration of the release proposed by The University of Adelaide

controls proposed by The University of Adelaide to restrict the spread and persistence of the GM wheat and barley plants and their genetic material

the genetic modifications are unlikely to give rise to adverse effects on human health and safety or the environment

widespread presence of the same and similar genes, proteins and associated products in the environment and lack of evidence of harm from them

limited ability and opportunity for the GM wheat and barley plants to transfer the introduced genes to commercial wheat or barley crops or other sexually related species

none of the GM plant materials or products will enter human food or animal feed supply chains.

190. The Risk Analysis Framework (OGTR 2013a), which guides the risk assessment and risk management process, defines negligible risks as insubstantial with no present need to invoke actions for their mitigation. Therefore, no controls are required to treat these negligible risks. Therefore, the Regulator considers that the dealings involved in this proposed release do not pose a significant risk to either people or the environment.



Chapter 3 Risk managementSection 1 Background

191. Risk management is used to protect the health and safety of people and to protect the environment by controlling or mitigating risk. The risk management plan addresses risks evaluated as requiring treatment, evaluates controls and limits proposed by the applicant, and considers general risk management measures. The risk management plan informs the Regulator’s decision-making process and is given effect through licence conditions.

192. Under section 56 of the Act, the Regulator must not issue a licence unless satisfied that any risks posed by the dealings proposed to be authorised by the licence are able to be managed in a way that protects the health and safety of people and the environment.

193. All licences are subject to three conditions prescribed in the Act. Section 63 of the Act requires that each licence holder inform relevant people of their obligations under the licence. The other statutory conditions allow the Regulator to maintain oversight of licensed dealings: section 64 requires the licence holder to provide access to premises to OGTR inspectors and section 65 requires the licence holder to report any information about risks or unintended effects of the dealing to the Regulator on becoming aware of them. Matters related to the ongoing suitability of the licence holder are also required to be reported to the Regulator.

194. The licence is also subject to any conditions imposed by the Regulator. Examples of the matters to which conditions may relate are listed in section 62 of the Act. Licence conditions can be imposed to limit and control the scope of the dealings. In addition, the Regulator has extensive powers to monitor compliance with licence conditions under section 152 of the Act.

Section 2 Risk treatment measures for identified risks195. The risk assessment of risk scenarios listed in Chapter 2 concluded that there are negligible risks to people and the environment from the proposed field trial of GM wheat and barley. These risk scenarios were considered in the context of the scale of the proposed release (Chapter 1, Section 3.1), the proposed containment measures (Chapter 1, Section 3.2), and the receiving environment (Chapter 1Section 6), and considering both the short and the long term. The Risk Analysis Framework (OGTR 2013), which guides the risk assessment and risk management process, defines negligible risks as insubstantial with no present need to invoke actions for their mitigation. Therefore, there are no licence conditions to treat these negligible risks.

Section 3 General risk management196. The limits and controls proposed in the application were important in establishing the context for the risk assessment and in reaching the conclusion that the risks posed to people and the environment are negligible. Therefore, to maintain the risk context, licence conditions have been imposed to limit the release to the proposed size, locations and duration, and to restrict the spread and persistence of the GMOs and their genetic material in the environment. The conditions are detailed in the licence and summarised in this Chapter.

3.1 Licence conditions to limit and control the release3.1.1 Consideration of limits and controls proposed by The University of Adelaide

197. Sections 3.1 and 3.2 of Chapter 1 provide details of the limits and controls proposed by The University of Adelaide in their application. These are discussed in the risk scenarios postulated for the proposed release in Chapter 2. Many of these proposed control measures are considered

Chapter 3 – Risk management 37


standard for GM crop trials and have been imposed by the Regulator in previous DIR licences. The appropriateness of these controls is considered further below.

198. The duration of the field trial would be confined to six growing seasons. The trial would be limited to five sites, with a collective area of 2.5 ha per season. The small size and duration of the trial would limit the potential exposure of humans and other organisms to the GMOs (risk scenario 1).

199. Only authorised personnel with appropriate training would be permitted to deal with the GMOs. This measure would limit the potential exposure of humans to the GMOs (risk scenario 1).

200. The applicant proposes to monitor for the presence of rodents by placing rodent baits inside the fenced areas. Combined with the use of a monitoring zone (below), these measures should both limit exposure of rodents to the GMOs (risk scenario 1) and minimise potential dispersal of GMOs outside the trial sites by rodents (risk scenario 2). A licence condition requires that for the period while GMOs are being grown, and until a trial site has been cleaned, measures must be implemented to control rodents within a site.

201. Birds are known to cause damage to cereal crops mostly during germination in autumn, but may feed on the crop at different times including during grain ripening (Temby & Marshall 2003). An extensive search of the literature did not identify any reports of birds other than emus transporting and dispersing wheat seed (eg through the digestive tract or taking panicles containing viable seed) or seedlings from wheat crops. The white wheat varieties have a thin seed coat (Hansen 1994) and are readily digested by birds (Yasar 2003). Therefore it is considered appropriate that no measurements are proposed to be taken to prohibit the access of birds to the trial site. However, in this respect it should be noted that both the NGNE facilities at Merredin and Katanning are covered by bird netting.

202. Each trial site is to be surrouned by a fence. This will minimise the potential exposure of livestock and other large animals to the GMOs (risk scenario 1) and the potential dispersal of the GMOs by livestock and other large animals (risk scenario 2).

203. The applicant proposes to surround each trial site with a 2 m buffer zone and a 10 m monitoring zone. It is a standard requirement of GM wheat and barley licences that the monitoring zone is maintained in a manner that does not attract or harbour rodents, such as keeping the area either free of vegetation or planted with vegetation mown to a height of less than 10 cm. This would serve the following purposes:

reduce rodent activity

facilitate detection of plants or related species that might hybridise with GM wheat or barley

facilitate detection of GM plant material that has been dispersed during sowing or harvesting.

204. The applicant proposes that the monitoring zone would be surrounded with a 190 m isolation zone where no sexually compatible species will be grown. In addition, inspections are to be conducted for related species in the planting area, monitoring zone and isolation zone, and any found destroyed prior to flowering.

205. The potential for pollen movement and gene flow between GM wheat and other sexually compatible species has been addressed at some length in DIR 100, DIR 102, DIR 111 and DIR 112. On the basis of the evidence detailed there, including scientific literature on gene flow, international containment measures for GM wheat trials, and the rules for producing basic and certified seed, 200 m isolation is considered adequate to minimise gene flow from the GM wheat



plants to other wheat plants or other sexually related species outside the release site. Therefore the combination of a 10 m monitoring zone plus a 190 m isolation zone will mimimise gene flow to other wheat or barley crops and related species (risk scenario 4).

206. However, at sites where there has been no cultivation or detection of wheat or barley volunteers in the isolation zone for the previous 2 years, the applicant proposes that the inspection area be reduced to 50 m. This is similar to conditions in licence DIR 102, which provide for the Regulator to give approval for the reduced inspection area, and the appropriateness of a distance of 50 m for inspection was discussed in the RARMP for DIR 102. However, the presence of related species in the isolation zone is not solely dependent on past crops or volunteers, as seed can be introduced inadvertently as contaminants in stock feed or seed for sowing. Therefore the option for a reduced inspection area is not included in the licence.

207. The applicant proposes to plant at the same sites that DIR 102 GM wheat and barley (modified for abiotic stress tolerance) have been grown. As discussed in Chapter 2, risks associated with stacking of abiotic stress tolerance genes from DIR 128 and 102 were considered negligible. It is considered that the containment measures, in particular the monitoring requirements, would still minimise the chance of spread and persistence of any unintended stacked GMOs (risk scenarios 5 and 6).

208. The two NGNE facilities are multi-user facilities. As there is hence the possibility that separate trials of other sexually compatible GM plants (most obviously wheat and barley) will concurrently take place in these facilities, it is important to consider the issues of vertical gene transfer and mixing of seeds between plants of different trials. The applicant mentions that DIRs 092, 093, 094, which allow the growth of plants from these three different DIRs in close proximity, requires a minimum of 4 m between plants grown under each licence (corresponding to a 2 m buffer zone for each DIR). The licence for DIR 112 specifies that other GM plants under a separate licence may be grown within the Merredin NGNE facility provided they are not grown in the Location or Buffer Zone of DIR 112, the latter being 2 m wide. A minimum distance of 4 m (which may include buffer zones) should separate the GM plants of this application and any other sexually compatible GM plants that are grown in the NGNE facilities.

209. If any GM plants in this application are grown in one of the NGNE facilities at a time when sexually compatible GM plants authorised by another licence are concurrently being grown, seed from the GM plants of this application cannot be used in the future development of cultivars for commercial release. These measures would minimise the likelihood of mixing between seed from different trials (risk scenario 2) or vertical gene transfer (risk scenarios 5 and 6), and subsequent spread of, or exposure to, this GM material.

210. The applicant has proposed a number of measures to minimise the persistence of any GM wheat and barley plants and seeds in the seed bank at the release site after harvest of the trial. These measures include post-harvest tillage to the depth of the original planting and irrigation to promote germination of remaining seed. After harvest, locations would be monitored at least once every 35 days for a period of 2 years and three irrigations applied. Volunteer plants that emerge would be destroyed before flowering.

211. There is a difference in germination rates between buried grain and grain lying on the surface; grains remaining on the surface, for example following shallow tillage after harvest, can generally easily germinate and become established (Ogg & Parker 2000). Shallow tillage after harvest, combined with irrigation, will germinate much of the seed lying on the surface (Ogg & Parker 2000). However, deep cultivation in certain soil types can reduce seed viability but can also encourage prolonged dormancy in seeds as a result of a cool, moist low oxygen environment (Ogg & Parker 2000; Pickett 1989).



212. It is therefore considered that under Australian conditions, a 2 year time period during which tillage and a number of irrigations are performed, and monitoring conducted on a regular basis with the failure to detect volunteers for a minimum of 6 months prior to the end of the time period, would effectively manage survival and persistence of viable wheat and barley seeds in the soil. It is appropriate the area receive at least 3 irrigations, at intervals of at least 28 days, with the last required irrigation occurring at a time that would promote germination of volunteers within the final volunteer-free period. A period of natural rainfall may be taken as irrigation only with the agreement of the Regulator. Evidence (such as rainfall measurements, photos etc.) that the rainfall has been sufficient to promote germination needs to be provided. Additionally, prior to the last irrigation the area must be tilled. These treatments will ensure seeds are exposed to sufficient moisture and placed at an appropriate depth for germination, as well as encouraging the microbial decomposition of any residual seed. These measures will minimise the persistence of the GMOs in the environment and are included as licence conditions.

213. In considering potential for spread and persistence of the GMOs, it is important to consider the potential dispersal of grain during sowing and harvesting (mechanical dispersal). This is most likely to result in dispersal of grain into the area immediately around the trial. The applicant has proposed a buffer zone of 2 m around the area where the GMOs are planted, which would be subject to the same post-harvest management as the GMO planting area. The licence requires that a 2 m buffer zone and any other areas where GM material has been dispersed, including during harvest or threshing, must be monitored to manage the possibility of mechanical dispersal of seed from the trial location and its persistence after the trial. All equipment used in connection with cultivating and harvesting the GMOs are required to be cleaned on site prior to removal.

214. The applicant has stated that any plant material taken off-site for experimental analysis will be transported according to the Regulator’s Guidelines for the transport, storage and disposal of GMOs. These are standard protocols for the handling of GMOs to minimise exposure of people and other organisms to the GMOs (risk scenario 1), dispersal into the environment and gene flow/transfer (risk scenarios 2, 3 and 4). This is included as a licence condition.

215. The applicant does not propose using any of the plant material for human or animal consumption. FSANZ conducts mandatory premarket assessments of GM products in human foods. As the GM wheat and barley has not been assessed by FSANZ, a condition in the licence prohibits material from the trial from being used for human food or animal feed.3.1.2 Summary of licence conditions to be implemented to limit and control the release

216. A number of licence conditions have been imposed to limit and control the release, based on the above considerations. These include requirements to:

limit the release to a total area of up to 5 sites, two in South Australia (O-Halloran Hill and Pinery) and three in Western Australia (Corrigin, Merredin and Katanning) between July 2014 and December 2019

limit the maximum area of any site to 0.5 ha, and thus collectively the area of the trial in any season to 2.5 ha in total

surround the area where GMOs are grown with a 2 m buffer zone

implement measures to control rodents within the planting area

surround the trial site with a 10 m monitoring zone, maintained in a manner that does not attract or harbour rodents, and in which related species must be prevented from flowering

surround the monitoring zone with a 190 m isolation zone, in which no other crops of wheat or barley may be grown, and where growth of related species are controlled



harvest the GM wheat and barley plant material separately from other crops

clean the areas and equipment after use

apply measures to promote germination of any wheat or barley seeds that may be present in the soil after harvest, including irrigation and tillage

monitor for at least 24 months after harvest, and destroy any wheat or barley plants that may grow, until no volunteers are detected for a continuous 6 month period

destroy all GM plant material not required for further analysis or future trials

transport and store GM material in accordance with the Regulator’s guidelines

not allow GM plant material to be used for human food or animal feed.

3.2 Other risk management considerations217. All DIR licences issued by the Regulator contain a number of conditions that relate to general risk management. These include conditions relating to:

applicant suitability

contingency plans

identification of the persons or classes of persons covered by the licence

reporting structures

a requirement that the applicant allows access to the trial sites and other places for the purpose of monitoring or auditing.

3.2.1 Applicant suitability

218. In making a decision whether or not to issue a licence, the Regulator must have regard to the suitability of the applicant to hold a licence. Under section 58 of the Act, matters that the Regulator must take into account include:

any relevant convictions of the applicant (both individuals and the body corporate)

any revocation or suspension of a relevant licence or permit held by the applicant under a law of the Commonwealth, a State or a foreign country

the capacity of the applicant to meet the conditions of the licence.

219. On the basis of information submitted by the applicant and records held by the OGTR, the Regulator considers The University of Adelaide suitable to hold a licence.

220. The licence includes a requirement for the licence holder to inform the Regulator of any circumstances that would affect their suitability.

221. In addition, any applicant organisation must have access to a properly constituted Institutional Biosafety Committee and be an accredited organisation under the Act.3.2.2 Contingency plan

222. The University of Adelaide is required to submit a contingency plan to the Regulator before planting the GMOs. This plan would detail measures to be undertaken in the event of any unintended presence of the GM wheat outside of the permitted areas.

223. The University of Adelaide is also required to provide a method to the Regulator for the reliable detection of the presence of the GMOs or the introduced genetic materials in a recipient



organism. This instrument is required before conducting any of the licenced dealings with the GMOs.3.2.3 Identification of the persons or classes of persons covered by the licence

224. The persons covered by the licence are the licence holder and employees, agents or contractors of the licence holder and other persons who are, or have been, engaged or otherwise authorised by the licence holder to undertake any activity in connection with the dealings authorised by the licence. Prior to growing the GMOs, The University of Adelaide is also required to provide a list of people and organisations who will be covered by the licence, or the function or position where names are not known at the time.3.2.4 Reporting requirements

225. The licence obliges the licence holder to immediately report any of the following to the Regulator:

any additional information regarding risks to the health and safety of people or the environment associated with the trial

any contraventions of the licence by persons covered by the licence

any unintended effects of the trial.

226. A number of written notices would also be required under the licence that would assist the Regulator in designing and implementing a monitoring program for all licensed dealings. The notices would include:

expected and actual dates of planting

details of areas planted to the GMOs

expected dates of flowering

expected and actual dates of harvest and cleaning after harvest

details of inspection activities.3.2.5 Monitoring for Compliance

227. The Act stipulates, as a condition of every licence, that a person who is authorised by the licence to deal with a GMO, and who is required to comply with a condition of the licence, must allow inspectors and other persons authorised by the Regulator to enter premises where a dealing is being undertaken for the purpose of monitoring or auditing the dealing. Post-release monitoring continues until the Regulator is satisfied that all the GMOs resulting from the authorised dealings have been removed from the release site.

228. If monitoring activities identify changes in the risks associated with the authorised dealings, the Regulator may also vary licence conditions, or if necessary, suspend or cancel the licence.

229. In cases of non-compliance with licence conditions, the Regulator may instigate an investigation to determine the nature and extent of non-compliance. The Act provides for criminal sanctions of large fines and/or imprisonment for failing to abide by the legislation, conditions of the licence or directions from the Regulator, especially where significant damage to health and safety of people or the environment could result.



Section 4 Issues to be addressed for future releases230. Additional information has been identified that may be required to assess an application for a large scale or commercial release of these GM wheat or barley lines, or to justify a reduction in containment conditions. This includes:

additional molecular and biochemical characterisation of the GM wheat and barley lines, particularly with respect to production of potential toxins or allergens

additional phenotypic characterisation of the GM wheat and barley lines, particularly with respect to traits that may contribute to weediness.

Section 5 Conclusions of the RARMP231. The risk assessment concludes that this proposed limited and controlled release of GM wheat and barley poses negligible risks to the health and safety of people or the environment as a result of gene technology, and that these negligible risks do not require specific risk treatment measures.

232. However, conditions have been imposed to limit the release to the proposed size, locations and duration, and to restrict the spread and persistence of the GMOs and their genetic material in the environment, as these were important considerations in establishing the context for assessing the risks.



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Zhou, M.-L., Qi, L.-P., Pang, J.-F., Zhang, Q., Lei, Z., Tang, Y.-X., Zhu.X.M., Shao, J.-R., Wu, Y.-M. (2013). Nicotianamine synthase gene family as central components in heavy metal and phytohormone response in maize. Functional and Integrative Genomics 13: 229-239

References 54


Appendix A Summary of submissions from prescribed experts, agencies and authorities‡‡

Advice received by the Regulator from prescribed experts, agencies and authorities on the consultation RARMP is summarised below. 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.Abbreviations: GM: Genetically Modified; RARMP: Risk Assessment and Risk Management Plan.

Sub.No:

Summary of issues raised Comment

1 Sees no problem with the application and has no comments on the RARMP.

Noted

2 Notes that the licence prohibits the use of material from the trials for human or animal consumption. Has no further comments on the licence application.

Noted.

3 Agrees with the overall conclusions of the RARMP and the proposed limits and controls.

Noted.

The Regulator should consider further acknowledging uncertainty regarding potential for the introduced genes for abiotic stress tolerance and micronutrient uptake to result in increased survival of the GMOs or increases in toxicity or allergenicity, including in the summary table [Table 5].

Additional text has been added to the risk scenarios acknowledging uncertainty about the potential for improved persistence of the GM plants outside of the agricultural setting. Table 5 was not modified as this is a brief summary of the scenarios.

This application involves a large number of lines, and suggests the Regulator consider the upper limits of what qualifies as a limited and controlled release.

This issue was considered as part of requirements under Section 50A of the Act, prior to preparation of a RARMP. The Regulator will continue to consider the appropriateness of limits and controls in determining if an application meets the requirements of section 50A.

The Regulator should consider seeking information on the results of the trial to inform future applications for release of any of these GM lines, with respect to the potential risks to human health and the environment.

Chapter 3 lists additional information that may be required to assess future applications for larger scale trials, reduced containment measures or commercial release.

The Regulator should consider clarifying the text in the RARMP regarding other government approvals, as approval for trials in South Australia would be required as part of the South Australian moratorium.

The South Australian moratorium on GM crops does not relate to protection of human health or the environment, and is a matter for the SA government and not the Regulator. Wording associated with the statement has been modified.

Commends Table 5 in the RARMP as a useful device to summarise causal pathways

Noted.

4 Has no concerns regarding the potential for harm to humans or the environment, and accordingly has no objection to the granting of the licence for DIR 128.

Noted.

‡‡ Prescribed agencies include GTTAC, State and Territory Governments, relevant local governments, Australian Government agencies and the Minister for the Environment.

Appendix A 55


Sub.No:

Summary of issues raised Comment

5 Given the biology and ecology of wheat and barley, and the limited scale and duration of the release, the environmental risks posed by the trial are likely to be low and manageable.

Noted.

Both wheat and barley have been listed as minor weeds in Australia and California. The traits (including any stacking) may have the potential to enhance growth and survival of the GM wheat and barley and sexually compatible species in non-agricultural areas. Therefore, the RARMP (in particular Risk Scenarios 2, 4 and 6) could be improved by including more detailed discussion about this.

Additional text has been added to these risk scenarios to describe the uncertainty about whether the GM traits may increase the potential for persistence outside an agricultural setting.

New sexually compatible species may be introduced into Australia in the future (eg perennial wheat). Even though there are only low rates of out-crossing, hybridisation between these new species and the GM plants may lead to weediness.

Risk scenarios 3 and 4 consider outcrossing to sexually compatible plants currently in the areas around the trial sites. There were no substantive risks identified, mainly due to the controls being proposed and the short duration of the trial. If new sexually compatible plants were introduced into Australia and were identified near the sites while the GMOs are growing, the licence conditions would minimise gene flow to these new species.

Recommends more guidance on future data requirements, especially in regards to traits that may lead to weediness. Suggests the collection of a variety of data, including growths rates, plant morphology, seed dormancy and germination, disease resistance, head shattering, seed dispersal, seed longevity, and comparison with existing elite cultivars.

Chapter 3 lists additional information that may be required to assess future applications for larger scale trials, reduced containment measures or commercial release. Applicants/licence holders liaise with the OGTR about collection of appropriate data, and the DIR application forms provide guidance on the information required. Nevertheless, the applicant will be provided with these suggestions for data collection.

6 Supports the conclusion that DIR 128 poses negligible risks of harm to human health and the environment assuming that the below ambiguities are considered and rectified.

Noted.

Paragraph 13 on page 3 of the RARMP states that visitors to the sites would be accompanied by authorised University of Adelaide staff. However, the licence allows any person to access a site as long as these persons are aware of the licence conditions.

Text in paragraph 13 has been modified to indicate that this is what the applicant proposes. However, the licence does not restrict access to sites but requires the licence holder to inform people of licence conditions prior to allowing them to conduct dealings with the GMOs.

Paragraphs 59 and 60 of the RARMP mentions the locality Kunjin for one site but the locality is not mentioned for the other two sites.

The text of the RARMP has been amended to more clearly define the locations of the trial sites. Once GMOs are planted at a site, GPS coordinates will be posted on OGTR’s website.

Paragraph 204 of the RARMP states that GM plant material must be destroyed if grown near other sexually compatible GM plants but the licence (condition 25) only states that the GM seed must not be used for future development. Suggest both statements refer to either plant material or seed.

Paragraph 204 (now paragraph 209 in the final RARMP) has been modified to indicate that seed must not be used for future development.

Appendix A 56


Appendix B Summary of submissions from the public

The Regulator received 13 submissions from the public on the consultation RARMP. The issues raised in these submissions are summarised in the table below. All issues raised in the submissions that related to risks to the health and safety of people and the environment 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.Issues raised: AT: Alternative technology; C: containment, E: Environment; EC: Economic issues; GT: gene transfer; F: food; FL: food labelling; H: Health; L: liability; LC: Licence condition; M: Marketing; P: Pesticides; R: Research; RA: Risk analysis; S: Segregation.

Other abbreviations: Act: the Gene Technology Act 2000; APVMA: Australian Pesticides and Veterinary Medicines Authority; FSANZ: Food Standards Australia New Zealand; GM: Genetically modified; GMO: Genetically modified organism.

Sub. No:

Issue Summary of issues raised Comment

1 GT When a similar "Trial" of GE Canola was undertaken, the researchers seemed to have no knowledge of "The Birds And The Bees". Cross-pollination occurred with 10,000Acres of normal canola.

GM crop field trials (‘limited and controlled releases’) licenced by the Regulator are subject to strict conditions to restrict the spread and persistence of the GMOs and their genetic material in the environment. Conditions imposed on this field trial include measures to minimise pollen transfer to non-GM wheat and barley crops by isolating the field trial by 200m from sexually compatible species. These measures have been effective in previous GM wheat and barley trials.

H We have no idea about how ingestion of such abnormal proteins may induce aberrant immune responses, possibly inducing CJD-like human disease. What precautions are in place?

All the introduced genes are derived from plants and bacteria that are widespread and prevalent in the environment. Most of them are commonly consumed by people or people are naturally exposed to them. None of the introduced proteins are members of protein classes that are known to have members with toxic or allergenic properties. No GM material from the trial will enter the human food supply or be used for animal feed.

2 E, M, RA

The public does not want GMO crops or food grown in SA and hence that is why there is a moratorium in SA. However, the trials will go ahead due to corporate agribusiness pressure being applied to our governments and regulators, who are supposed to be there to protect people’s health and the environment. OGTR appears to be ‘a rubber stamp’, doing no independent testing of its own. Other countries have banned GMOs. We do not want our natural biodiversity contaminated with GM organisms.

States have the ability to introduce moratoria on growing GM crops for marketing and trade purposes, not for health and safety reasons. The Regulator is required to assess GMO applications in accordance with the Act, the object of which is to the protect the health and safety of people and the environment. Comprehensive RARMPs include a thorough and critical assessment of data supplied by the applicant, together with a review of other relevant national and international scientific literature. The RARMP concluded that risks to human health and safety of people, and to the environment, from this limited and controlled release of GM wheat and barley are negligible. Nevertheless, strict conditions have been imposed to restrict the spread and persistence of the GMOs and their genetic material in the environment.

AT GM wheat and barley are not needed as there are excellent traditional varieties from which seed can be saved. If there was better food distribution, instead of mega profits for giant agribusiness then gene technology would not be needed.

These matters do not relate to risks to human health and safety and the environment and are outside the scope of assessments conducted by the Regulator.

Appendix B 57


Sub. No:


3 GT Concerned about wind mediated dispersal of GM pollen to any nearby wheat and barley crops and whether this could end up being a legal minefield for both the University and the federal government.

Licence conditions impose strict controls that minimise the potential for pollen transfer to non-GM wheat and barley crops due to wind by requiring an isolation distance of at least 200 metres. These measures have been effective in previous GM wheat and barley trials.

4 E, R Objects to trials for yield before trials for ecological impact are exhaustively pursued.

The RARMP concluded that risks to human health and safety of people, and to the environment, from this limited and controlled release of GM wheat and barley are negligible. Data relating to ecological impacts would be required for a future application for a commercial release of any of these GMOs. Relevant data may be collected during this field trial.

5 GT Controls should include not planting in area where organic wheat or barley is the surrounding crop even if outside 200m buffer zone.

Wheat and barley plants are predominately self-pollinating and most pollen falls within the first few metres. Review of the literature indicates that isolating the GM wheat and barley by at least 200m will effectively minimise the likelihood of pollen transfer to any nearby wheat and barley crops, including organic crops. These measures have been effective in previous GM wheat and barley trials.

6 F, AT, EC

Wants food to stay free from GMO. Other means of farming produce food just as nutritious. Wants open air trials and commercial cultivation banned and research on sustainable farming systems supported. GM crops are not accepted worldwide and countries buy Australian wheat because it is GM free.

These matters do relate to risk to human health and safety and the environment and are outside the scope of assessments conducted by the Regulator. Marketing and trade issues are the responsibility of States and industry.

GT Because so many crops are wind-pollinated, it is almost impossible to keep areas GM free.

Licence conditions impose strict controls that minimise the potential for pollen transfer via wind to sexual compatible plants by requiring an isolation distance of at least 200 metres.

7 EC, AT Australian governments must create a secure future for Australian wheat by adopting and implementing policies to: ban open air GM wheat field trials; reject commercial cultivation of GM wheat; and support research on sustainable farming systems.

Alternative agricultural practices and marketing and trade issues are outside the scope of assessments conducted by the Regulator. Marketing and trade issues are the responsibility of States and industry.

C, S The GM wheat trials and commercialisation may contaminate conventional and organic wheat. This could occur at any point from seed to spoon via cross-pollination, misreading of labels, mistakes in handling, illegal planting, poor machinery cleaning, spillages and ineffective segregation.

The application is for a limited and controlled release (field trial). Strict licence conditions have been imposed to minimise spread and persistence of the GM wheat and barley. These include conditions to isolate trial sites from other wheat and barley crops, cleaning of equipment used with GM plant materials, secure transport and storage of GM plant materials, and post-harvest monitoring at trial sites to ensure all GM plants are destroyed.

EC A GrainGrowers’ report [What the World Wants From Australian Wheat, 2011] states that wheat importers in SE Asia, north Asia, Middle East and Europe (which accounts to about 80% of exported Australian wheat) will not buy GM wheat now or in the foreseeable future. The report also states the domestic and feed millers do not believe Australian consumers would accept GM wheat. International food companies also reject foods with GM. GM commercialisation poses an unacceptable risk to the industry.

Marketing and trade issues are outside the scope of assessments conducted by the Regulator. These issues are the responsibility of States and industry.

8 - Wants DIR 128 open air trial of GM wheat and barley in WA (and SA) rejected.

Appendix B 58


Sub. No:


R, H, F GM wheat is not grown anywhere in the world, and we in Australia do not want to be used for risky experimentation. GM products should be tested for safety and health impacts by an independent panel of experts before being put on the market. Findings should be made public.

The Regulator uses information provided by applicants as well as published scientific literature to assess GMO applications. The detailed RARMPs are publicly available on the OGTR website. This trial has limits on scale and duration, and has strict controls to minimise spread and persistence of the GM wheat and barley. The RARMP concluded that this trial poses negligible risks to the health and safety of people and the environment. Further information related to the potential for toxicity, allergenicity or weediness has been identified that may be required to assess an application for larger scale trials or commercial release of any of these GMOs. If any of the GMOs were proposed to be used in human food, FSANZ would conduct a pre-market safety assessment, which would also be subject to public consultation and made available on their website.

EC There is a ban on the commercial growing of GM crops in WA and SA and this needs to be respected.

States have the ability to introduce moratoria on growing GM crops for marketing and trade purposes, not for health and safety reasons. The Regulator is required to assess GMO applications in accordance with the Act, the object of which is to protect the health and safety of people and the environment. People dealing with GMOs must comply with State requirements as well as conditions imposed by the Regulator.

FL Consumers need the right to be able to choose GM free food. The current labelling laws are full of loopholes making it impossible to eat GM free, unless eating organic.

Labelling of GM foods is the responsibility of FSANZ.

P GM crops are increasingly found to need more (not less) chemicals to grow. This has severe negative impacts on soil and water, our environment, human health and animal health.Glyphosate resistant crops lead to glyphosate resistant weeds.

The regulation of agricultural chemicals, including herbicide resistance management, in Australia is the responsibility of APVMA. The APVMA will not register a chemical product unless satisfied that its approved use would not be likely to have an effect that is harmful to people or the environment.

S Evidence from around the world shows there is a high risk that GM crops will contaminate conventional and organic varieties.Cites recent case from WA Supreme Court (Marsh vs Baxter) case, and suspended imports due to unapproved GM wheat in Oregon, USA.

Marketing and trade issues, including matters relating to segregation and coexistence of different farming systems, are the responsibility of the States and industry, not the Regulator.Note that the Marsh vs Baxter case, in which an organic farmer claimed damages from a neighbouring GM grower for loss of organic certification, related to commercially approved GM canola, not to GM material from any limited and controlled release authorised by the Regulator.The current application is a limited and controlled release (field trial) of GM wheat and barley. Strict licence conditions have been imposed to restrict and spread and persistence of the GMOs. There has been no documented loss of containment from any field trial authorised by the Regulator.

EC Raises similar issues as submission number 7 in relation to the GrainGrowers’ report.

See response to submission 7.

9 S, EC Raises the same issues regarding the GrainGrowers report as in submission 7, and regarding GM crops contaminating conventional and organic varieties as in submission 8.

See relevant elements of responses to submissions 7 and 8.

Appendix B 59


Sub. No:


10 C, S, EC Wants DIR 128 open air trial of GM wheat and barley rejected.The ban on the commercial growing of GM crops in WA and SA needs to be respected. No one gains anything from the GM crops, except for the GMO producing companies.Concerned about the damage to organic wheat suppliers due to the possibility of contamination from GM crops. As there are only a handful of organic grain suppliers in WA, it is imperative that these trials are aborted as our industry cannot afford to lose any more suppliers as per the Steve Marsh case.

Marketing and trade issues, including matters relating to segregation and coexistence of different farming systems, are the responsibility of the States and industry, not the Regulator. States have the ability to introduce moratoria on growing GM crops for marketing and trade purposes. People dealing with GMOs must comply with State requirements as well as conditions imposed by the Regulator.As noted in response to submission 8, the Marsh vs Baxter case related to commercially approved GM canola, not to GM material from any limited and controlled release authorised by the Regulator.The current application is a limited and controlled release (field trial) of GM wheat and barley. Strict licence conditions have been imposed to restrict and spread and persistence of the GMOs.

M, L Raises the same issues regarding the, GrianGrowers’ report as submission 7.

See responses to submission 7.

RA, E, H GM products should be tested for safety and health impacts by an independent panel of experts before being put on the market. Findings should be made public.

The Regulator uses information provided by applicants as well as published scientific literature to assess GMO applications. The detailed RARMPs are publicly available on the OGTR website. This trial has limits on scale and duration, and has strict controls to minimise spread and persistence of the GM wheat and barley. The RARMP concluded that this trial poses negligible risks to the health and safety of people and the environment. Further information related to the potential for toxicity, allergenicity or weediness has been identified that may be required to assess an application for larger scale trials or commercial release of any of these GMOs. If any of the GMOs were proposed to be used in human food, FSANZ would conduct a pre-market safety assessment, which would also be subject to public consultation and made available on their website.

11 - Objects to the approval of DIR 128 GM wheat and barley trial.

C, S There have been documented examples of so-called ‘trials’ of GM crops contaminating the environment. Cites 3 examples: unapproved GM wheat in Oregon, USA (2013), GM rice in US (2006) and GM canola Topas 19/2 in Australia (2005).

Strict licence conditions have been imposed to restrict the spread and persistence of the GMOs in the environment, including isolation from sexually compatible species, and cleaning and post-harvest monitoring of trial sites. Similar conditions have been effective for other GM wheat and barley trials conducted in Australia.

EC GM wheat and barley is not grown commercially anywhere in the world. Market rejection of GM wheat and barley means that no country wants to import it. Wheat is Australia’s most important agricultural commodity. Two thirds of our wheat is exported. Australia’s clean paddocks are being used for a GM experiment for which there is no market.


Appendix B 60


Sub. No:


RA Given the past record of the OGTR in approving GM "trials", we have no expectation that the precautionary principle will prevail or that you will take our arguments into consideration.

As required by the Gene Technology Act, the Regulator has prepared a comprehensive RARMP, including a thorough and critical assessment of data supplied by the applicant, together with a review of other relevant national and international scientific literature. The risk assessment process, outlined in the OGTRs Risk Analysis Framework (April 2013. All comments received on the consultation RARMP that were relevant to health and safety of people and the environment were taken into consideration in finalising the RARMP and making a decision on issuing a licence. Information on how submissions have been considered is presented in the appendix. Strict licence conditions have been imposed on this field trial to restrict the spread and persistence of the GMOs and their genetic material in the environment.

H, E The health and safety of people or the environment are not served by approval of experimental GM wheat and barley constructs into the open paddocks of Australia.

The RARMP concluded that this limited and controlled field trial poses negligible risks to the health and safety of people and the environment. Strict licence conditions have been imposed on this field trial to restrict the spread and persistence of the GMOs and their genetic material in the environment.

12 S, EC There are many unresolved issues of liability and unacceptable risks associated with GM contamination worldwide. Current laws and regulations, surveillance and monitoring are inadequate to deal with segregation demands of different markets and public concerns over health and environmental safety. Cites Marsh vs Baxter case in WA. Calls for a freeze of new approvals of GM crops, including open-air trials until thorough testing.

Marketing and trade issues, including matters relating to segregation and coexistence of different farming systems, are the responsibility of the States and industry, not the Regulator. The RARMP concluded that this limited and controlled field trial poses negligible risks to the health and safety of people and the environment.

C Who is responsible to the control and cleanup of the spread and spill of GM seed and crops as the GM industry is turning their backs on these problems? Escapes for GM trials have occurred and attempts to eradicate the resulting GM weeds post-trial is likely to continue for many years. Cites 2013 examples: unapproved GM wheat in Oregon, China intercepting and destroying shipments of GM corn and seed that attempted to bypass Chinese environmental and food safety tests, GM canola failing to germinate in WA and harm to pigs fed GM soy and GM corn. Also mentions trial sites in Tasmania that still have GM canola weeds after 10 years.

As this application is for a limited and controlled release (field trial), strict licence conditions have been imposed to restrict the spread and persistence of the GMOs and their genetic material in the environment. These include conditions to isolate trial sites from other wheat and barley crops, cleaning of equipment used with GM plant materials, secure transport and storage of GM plant materials, and post-harvest monitoring at trial sites to ensure all GM plants are destroyed. Similar conditions have been effective for other GM wheat and barley trials conducted in Australia.

M The world has already rejected GM wheat from earlier attempts in the US and Canada in 2004 to commercialise it. People do not want to eat GM wheat & barley and Australia’s established markets must not be placed at avoidable risk.


13 S, AT Objects to the GM wheat and barley trials taking place in WA. Despite buffer zones around GM canola, even their property has been affected. Believes that GM is not the solution and that farming for children’s future and health is of great priority.

The RARMP concluded that this limited and controlled field trial poses negligible risks to the health and safety of people and the environment. Strict licence conditions have been imposed to restrict the spread and persistence of the GMOs and their genetic material in the environment.Marketing and trade issues, including matters relating to segregation and coexistence of different farming systems, are the responsibility of the States and industry. They are not the responsibility of the Regulator.

Appendix B 61


Appendix B 62

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