Risk Assessment and
Risk Management Plan for
DIR 130
Limited and controlled release of wheat genetically
modified for improved grain quality
Applicant: Murdoch University
March 2015
PAGE INTENTIONALLY LEFT BLANK
DIR 130 – 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 130
Decision
The Gene Technology Regulator (the Regulator) has decided to issue a licence application for
the intentional 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 application
Application number
DIR 130
Applicant:
Murdoch University
Project Title:
Limited and controlled release of wheat genetically modified for improved
grain quality1
Parent organism:
Introduced genes and
modified traits:
Bread wheat (Triticum aestivum L.)

Dx5 and Dy10 genes from wheat (improved grain quality)

bar gene from Streptomyces hygroscopicus (selectable marker –
herbicide tolerance)
Proposed release dates:
May 2015 – December 2017
Proposed location:
One site in the local government area of Katanning, Western Australia
600 square metres each year
Proposed release size:
Primary purpose
To assess whether the introduction and expression of the genes will increase
the strength of dough
Risk assessment
The risk assessment concludes that there are negligible risks to the health and safety of people,
or the environment, from the proposed release. No additional controls are required to manage
these neglible risks beyond those proposed in the application.
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 and 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 title of the project as supplied by the applicant is ‘Field trial of glutenin transgenic wheat for grain quality
improvement’.
1
Summary
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DIR 130 – Risk Assessment and Risk Management Plan
Office of the Gene Technology Regulator
Credible pathways to potential harm that were considered included: unintended exposure to the
GM plant material; increased spread and persistence of the GM wheat relative to unmodified
plants; and transfer of the introduced genetic material to non GM wheat, 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 genes are
either identical, or similar, to those already existing in the environment (some of these genes
are present in unmodified wheat); and the proposed limits and controls effectively contain the
GMOs and their genetic material and minimise exposure.
Risk management plan
The risk management plan describes measures to protect the health and safety of people and to
protect the environment by controlling or mitigating risk. The risk management plan is given
effect through licence conditions.
As the level of risk is considered negligible, specific risk treatment is not required. However, as
this is a limited and controlled release, the licence includes limits on the size, location 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 site to ensure all
GMOs are destroyed.
Summary
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DIR 130 – Risk Assessment and Risk Management Plan
Office of the Gene Technology Regulator
Table of Contents
SUMMARY OF THE RISK ASSESSMENT AND RISK MANAGEMENT PLAN ................................................... I
Decision .......................................................................................................................................................................... I
The application................................................................................................................................................................ I
Risk assessment .............................................................................................................................................................. I
Risk management plan .................................................................................................................................................. II
TABLE OF CONTENTS ...............................................................................................................................................III
ABBREVIATIONS ......................................................................................................................................................... IV
CHAPTER 1
RISK ASSESSMENT CONTEXT.................................................................................................... 1
Section 1
Section 2
Section 3
3.1
3.2
5.4
Section 6
6.1
6.2
6.3
6.4
Section 7
7.1
7.2
Background .......................................................................................................................................... 1
Regulatory framework ......................................................................................................................... 1
The proposed dealings ......................................................................................................................... 2
The proposed limits of the dealings (size, location, duration and people) ........................................... 2
The proposed controls to restrict the spread and persistence of the GMOs and their genetic material
in the environment ............................................................................................................................... 3
The parent organism ............................................................................................................................ 4
The GMOs, nature and effect of the genetic modification ................................................................... 4
Introduction to the GMOs .................................................................................................................... 4
The introduced genes, encoded proteins and their associated effects .................................................. 5
Toxicity/allergenicity or other adverse effects upon health associated with the introduced genes,
their encoded proteins and associated products ................................................................................... 6
Characterisation of the GMOs ............................................................................................................. 7
The receiving environment .................................................................................................................. 8
Relevant abiotic factors ....................................................................................................................... 8
Relevant agricultural practices ............................................................................................................. 8
Presence of related plants in the receiving environment ...................................................................... 8
Presence of similar genes and encoded proteins in the environment ................................................... 8
Relevant Australian and international approvals ................................................................................. 9
Australian approvals ............................................................................................................................ 9
International approvals of GM wheat .................................................................................................. 9
CHAPTER 2
RISK ASSESSMENT ...................................................................................................................... 10
Section 1
Section 2
2.1
2.2
2.3
2.4
Section 3
Section 4
Introduction ....................................................................................................................................... 10
Risk Identification ............................................................................................................................. 11
Risk source ........................................................................................................................................ 11
Causal pathway .................................................................................................................................. 12
Potential harm .................................................................................................................................... 13
Postulated risk scenarios .................................................................................................................... 13
Uncertainty ........................................................................................................................................ 26
Risk evaluation .................................................................................................................................. 27
CHAPTER 3
RISK MANAGEMENT .................................................................................................................. 29
Section 1
Section 2
Section 3
3.1
3.2
Section 4
Section 5
Background ........................................................................................................................................ 29
Risk treatment measures for identified risks ...................................................................................... 29
General risk management .................................................................................................................. 29
Licence conditions to limit and control the release ............................................................................ 29
Other risk management considerations .............................................................................................. 32
Issues to be addressed for future releases .......................................................................................... 34
Conclusions of the RARMP .............................................................................................................. 34
REFERENCES
........................................................................................................................................................... 35
APPENDIX A
SUMMARY OF SUBMISSIONS FROM PRESCRIBED EXPERTS, AGENCIES AND
AUTHORITIES ............................................................................................................................... 44
APPENDIX B
SUMMARY OF SUBMISSIONS FROM THE PUBLIC ............................................................. 45
Section 4
Section 5
5.1
5.2
5.3
Table of Contents
III
DIR 130 – Risk Assessment and Risk Management Plan
Office of the Gene Technology Regulator
Abbreviations
APVMA
CCI
Australian Pesticides and Veterinary Medicines Authority
Confidential Commercial Information as declared under section 185 of the
Gene Technology Act 2000
DAFWA
DIR
FSANZ
GM
GMO
ha
HMW-GS
HMW-GSs
LGA
LMW-GS
LMW-GSs
m
NGNE
NLRD
OGTR
PC2
ppm
RARMP
Regulations
Regulator
TGA
the Act
Department of Agriculture and Food, Western Australia
Dealings involving Intentional Release
Food Standards Australia New Zealand
Genetically modified
Genetically modified organism
Hectare
High molecular weight glutenin subunit
High molecular weight glutenin subunits
Local government area
Low molecular weight glutenin subunit
Low molecular weight glutenin subunits
Metres
New Genes for New Environments
Notifiable low risk dealings
Office of the Gene Technology Regulator
Physical Containment level 2
Parts per million
Risk Assessment and Risk Management Plan
Gene Technology Regulations 2001
Gene Technology Regulator
Therapeutic Goods Administration
The Gene Technology Act 2000
Abbreviations
IV
DIR 130 – Risk Assessment and Risk Management Plan
Chapter 1
Office of the Gene Technology Regulator
Risk assessment context
Section 1 Background
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.
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).
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).
RISK ASSESSMENT CONTEXT
LEGISLATIVE REQUIREMENTS
(including Gene Technology Act and Regulations)
RISK ANALYSIS FRAMEWORK
OGTR OPERATIONAL POLICIES AND GUIDELINES
PROPOSED DEALINGS
Proposed activities involving the GMO
Proposed limits of the release
Proposed control measures
GMO
Introduced genes (genotype)
Novel traits (phenotype)
PREVIOUS RELEASES
PARENT ORGANISM
Origin and taxonomy
Cultivation and use
Biological characterisation
Ecology
RECEIVING ENVIRONMENT
Environmental conditions
Agronomic practices
Presence of related species
Presence of similar genes
Figure 1 Summary of parameters used to establish the risk assessment context
Section 2 Regulatory framework
Sections 50, 50A and 51 of the Act outline the matters that 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
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and authorities before preparation of the Risk Assessment and Risk Management Plan (RARMP;
see section 50 of the Act).
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. Five public submissions were received and
their considerations are summarised in Appendix B.
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.
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 the 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 dealings
Murdoch University proposes to release up to 35 lines2 of genetically modified (GM) wheat
into the environment under limited and controlled conditions.
The purpose of the trial is to assess whether the introduction and expression of the genes will
increase the strength of bread dough. Published research has shown that increasing the levels of
certain proteins by genetic transformation (ie introducing additional copies of the genes in order to
increase the quantity of their corresponding proteins) is a useful way of generating novel doughs for
characterisation. In addition, the release will allow the applicant to produce sufficient grain for
subsequent replicated trials.
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
possession, supply or use of the GMOs for any of the purposes above.
These dealings are detailed further below.
3.1 The proposed limits of the dealings (size, location, duration and people)
The applicant proposes to grow GM wheat plants between May 2015 and December 2017.
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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The GMOs are proposed to be planted in the New Genes for New Environment (NGNE)
facility located at Katanning in Western Australia. This facility is operated by the Department of
Agriculture and Food, Western Australia (DAFWA).
The maximum area of the trial in any year would be up to 0.06 hectares (ha).
Only trained and authorised staff are proposed to be permitted to deal with the GM wheat.
Any other visitors to the trial site would be accompanied by an authorised Murdoch University
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 environment
The applicant has proposed a number of controls to restrict the spread and persistence of the
GM wheat lines and the introduced genetic material in the environment, including:

locating the NGNE facility at least 50 m from the nearest waterway

surrounding the facility by a 1.8 m fence to exclude large animals and using a bird proof netting
(these are existing features of the NGNE facility)

implementing a rodent control program

surrounding the facility by a 10 m monitoring zone and 190 m isolation zone in which no
sexually compatible plants will be grown

cleaning any equipment used with the GM plants before removal from the site, and the disposal
of any material collected during cleaning in a manner approved by the Regulator

separating the GM wheat plants by a buffer of at least 4 m if other GM wheat DIRs are being
grown side by side in the same facility

monitoring the planted locations at least once every fortnight during the flowering of the GM
plants

post-harvest monitoring of the trial site (once every 35 days) and destruction of any volunteer
wheat for at least 24 months

destroying any plant material collected during cleaning by autoclaving, hammer-milling,
incineration, burial, or any other method approved by the Regulator

ploughing back waste material and stubble from harvesting into the soil

grain/dough testing in the PC2 laboratory at Katanning, all laboratory equipment being
subjected to cleaning both before and after use

transporting and storing GM material in accordance with the Regulator's Guidelines for the
Transport, Storage and Disposal of GMOs (2011)
not allowing GM plant material or products to be used for human food or animal feed.
Figure 2 shows the proposed site layout at the Katanning NGNE facility, 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.
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Office of the Gene Technology Regulator
190 m wide
Isolation Zone,
inspected for
wheat during
the flowering
of GMOs
2 m wide
Buffer
Zone where
the growth
of plants is
controlled
10 m wide
Monitoring
Zone, where
the growth of
plants is
controlled
NGNE site
Fence
Planting
area where
GM wheat
is planted
Figure 2 Proposed trial layout, including some of the controls (not drawn to scale)
Section 4 The parent organism
The parent organism is bread wheat (Triticum aestivum L.), which is 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.
All lines were initially in the cultivar Bobwhite, but were backcrossed into the cultivars
Calingiri, Wyalkatchem, Westonia, and IGW2836 (NLRD 2341/2007).
Detailed information about the parent organism is contained in the reference document The
Biology of Triticum aestivum L. em Thell (bread wheat) (OGTR 2008), which was produced to
inform the risk assessment process for licence applications involving GM wheat plants. This
document is available from the OGTR website.
Section 5 The GMOs, nature and effect of the genetic modification
5.1
Introduction to the GMOs
The applicant proposes to release up to 35 lines of GM wheat. The lines were originally
produced in the wheat cultivar Bobwhite by biolistics mediated plant transformation. Information
about this transformation method can be found in the document Methods of plant genetic
modification available from the Risk Assessment References page on the OGTR website.
Each GM wheat line has been transformed with a construct containing one of two genes of
interest, or a hybrid of these genes. The GM plants also contain the selectable marker gene bar,
originating from Streptomyces hygroscopicus.
The expression of the individual Dx5 and Dy10 genes in the GM wheat lines are controlled by
their native 5’ and 3’ regulatory sequences. For the Dy10-Dx5 hybrid gene, the expression is
controlled by the Dy10 promoter and the Dx5 terminator. The bar gene is driven by the promoter
consisting of the 5’ untranslated exon and first intron of the maize ubiquitin (ubi-1) gene
(Christensen & Quail 1996; Christensen et al. 1992), and followed by the nos terminator from
Agrobacterium tumefaciens. A summary of these genes is presented in Table 1.
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Table 1 Summary of genes and regulatory elements introduced into the GM wheat plants.
Promoter (origin)
Dx5 (wheat)
Dy10 (wheat)
Dy10 (wheat)
Ubi-1 (maize)
Gene of interest (origin)
Dx5 (wheat)
Dy10 (wheat)
Dy10-Dx5 hybrid (Sequences of wheat genes Dx5 and Dy10)
bar (Streptomyces hygroscopicus)
Terminator (origin )
Dx5 (wheat)
Dy10 (wheat)
Dx5 (wheat)
nos (Agrobacterium tumefaciens)
5.2
The introduced genes, encoded proteins and their associated effects
5.2.1
Glutenin genes
The application involves a field trial of GM wheat plants containing either of two wheat
glutenin genes, Dx5 and Dy10 (Anderson & Greene 1989; Anderson et al. 1989), or a hybrid gene
consisting of a single open reading frame of sequences derived from both Dx5 and Dy10 (Blechl &
Anderson 1996). The hybrid gene codes for a mature protein that consists of amino acids 1-124
from Dy10 fused N-terminal to amino acids 130-848 from Dx5 (Blechl & Anderson 1996).
Glutenins are seed storage proteins that act in germinating seeds as a supply of elements such
as carbon, nitrogen and sulphur. They are divided into two classes: low molecular weight glutenin
subunits (LMW-GSs) and high molecular weight glutenin subunits (HMW-GSs) (Akagawa et al.
2007; Battais et al. 2008; Wieser 2007).
The production of wheat dough is dependent upon the structure and composition of the
glutenin and gliadin seed storage proteins, which in the presence of water crosslink to form a
matrix, gluten. In particular, the HMW-GSs, such as Dx5 and Dy10, play a critical role in
determining the viscoelastic properties that are crucial for the formation of bread dough (Shewry et
al. 2003). Bread wheat has three loci, designated Glu-A1, Glu-B1 and Glu-D1, that contain HMWGS genes, these being located on the long arms of chromosomes 1A, 1B and 1D, respectively
(Shewry et al. 2003; Shewry & Tatham 1997). Each locus contains two genes, one gene encoding
an x-type and the other a y-type subunit. The amino acid sequences of these subunits are
characterised by highly repeated blocks of sequence. Both x -type and y -type subunits contain
hexapeptide and nonapeptide motifs, the major difference between these two types of subunits
being that the former also contains tripeptide motifs (Shewry et al. 1992; Shewry et al. 2003).
Each gene has alleles. As specfic x-type and y-type alleles usually occur together at a locus,
their separation by recombination is difficult; this gene pair hence constitute what can be regarded
as a single allele. The Dx5 and Dy10 genes occur together as an ‘allele’ (Glu-D1d) of the locus
located on chromosome 1D. Other ‘alleles' at this locus include Dx2+Dy12 (Glu-D1a) and
Dx4+Dy12 (Glu-D1b) (Zheng et al. 2011). In many plants, certain HMW-GS genes are expressed at
low levels or silent (Forde et al. 1985; Wieser & Zimmermann 2000). As such, a plant commonly
produces a smaller number of HMW-GS proteins than may be theoretically predicted (Anjum et al.
2007; Shewry et al. 2003).
The properties of dough that are linked to glutenin have been investigated by a number of
studies that have involved the transformation into wheat of glutenin genes (Blechl et al. 2007).
Many of these experiments have involved the co-bombardment of immature embryos with high
copy number plasmids containing the genes of interest and a plasmid with a herbicide tolerant gene
to enable selection of transformants. In one study, aimed at evaluating agronomic performance of
bread wheat lines transformed with glutenin genes, the introduction of Dx5 and Dy10 did not show
significant changes in traits associated with performance (Bregitzer et al. 2006). Other research with
varieties of bread wheat transformed with the Dx5 and Dy10 genes has shown that both these genes
affect the mixing properties of dough, but in different ways (Blechl et al. 2007). Increasing the level
of the Dy10 protein over five times above that normally found in wheat plants did not prevent
dough mixing. However, smaller increases in the Dx5 protein led to greater dough strength,
problems with mixing, higher amounts of polymeric protein and lower sedimentation values. It has
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been suggested that these differences between these two HMW-GSs is possibly due to variations
between the sizes and composition of their repetitive domains, and their abilities to form both
intermolecular and intramolecular disulphide bonds (Blechl et al. 2007).
The studies described above were conducted in a wheat background containing endogenous
Dx5 and Dy10 genes. Other work has involved separately transforming the Dx5 and Dy10 genes
into a genetic background that does not contain these genes, thus enabling the individual effects of
these two genes on bread quality to be more accurately evaluated (Leon et al. 2009). Expression of
Dy10 alone resulted in lines with greater dough strength than those expressing Dx5. Conventional
crossing of these lines demonstrated that a combination of Dx5 and Dy10 produced a dough with
the greater strength and better dough mixing properties than some other subunit combinations that
were evaluated (Leon et al. 2010).
Varieties of durum wheat (Triticum turgidum L. ssp. durum) have also been co-transformed
with the wheat genes Dx5 and Dy10 (Gadaleta et al. 2008). The mixing times and peak resistances
of dough from the transformed lines were likewise greater than those recorded in the wild-type
parents, indicating that expression of the inserted genes was having a positive effect upon strength.
With respect to the wild-type parent varieties, yields from the transformed lines were similar.
In another line of research, durum wheat varieties separately expressing the HMW-GS 1Ax1
gene and the Pina gene linked to grain hardness were generated, and then conventional breeding
used to combine the two genes (Li et al. 2010). Analysis of these lines suggested that overexpression of both these genes can enhance dough strength. The effects of changing the levels of
gliadins on dough properties has also been investigated, the down-regulation of these genes being
mediated by RNAi (Gil-Humanes et al. 2012; Gil-Humanes et al. 2008).
Examination of the protein complement of wheat plants that have been transformed via
biolistics with HMW-GS genes has revealed that there are frequently HMW-GS proteins that are
either larger or smaller than the predicted sizes (Blechl & Vensel 2013). These are likely the result
of transformation events that alter the sequence of the genes or result in tandem repeats.
5.2.2
Selectable marker genes
All the GM wheat lines contain the selectable marker gene bar, derived from Streptomyces
hygroscopicus. This gene codes for the enzyme phosphinothricin N-acetyltransferase (PAT), which
provides tolerance to the broad spectrum herbicide phosphinothricin, also known as glufosinate
ammonium. This gene has been used extensively as a selectable marker gene in the production of
GM plants. More information on selectable marker genes in general, can be obtained from the
OGTR document Marker genes in GM plants, available on the OGTR website.
The plants may also contain the bacterial ampicillin resistance (bla) gene (Sutcliffe 1979),
which codes for the enzyme beta-lactamase. The promoter of this gene is specific for prokaryotic
organisms; thus the ampicillin protein will not be expressed in plants.
5.3
Toxicity/allergenicity or other adverse effects upon health associated with the
introduced genes, their encoded proteins and associated products
The introduced genes of interest, Dx5 and Dy10, originate from wheat. This plant is widely
consumed by people and animals, and as such people and animals have a long history of exposure
to its proteins.
There is no evidence that wheat has any toxic properties. However, a minority of people suffer
from allergies to wheat (and other cereals). The two most well characterized allergies to wheat are
baker’s asthma (induced by the inhalation of wheat flour during grain processing) and exercise
induced anaphylaxis (a reaction that occurs if someone undergoes physical activity soon after
consumption of wheat) (Hischenhuber et al. 2006; Tatham & Shewry 2008). These allergies are
likely due to a number of compounds in cereals, although the most important triggers are believed
to be the -amylase inhibitors and gluten (glutenin and gliadin) seed storage proteins (Battais et al.
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2008; Salcedo et al. 2011; Tatham & Shewry 2008). Rashes, wheezing, swelling and abdominal
pains are all symptoms of wheat allergy, while pathogenic mechanisms of induction include IgEmediated and cell-mediated (Hischenhuber et al. 2006). Wheat allergy is often a temporary
condition of young children that is outgrown by the age of five (Pietzak 2012).
Both HMW-GSs and LMW-GSs have been implicated in wheat allergy. Experiments have
suggested that HMW-GS proteins are allergens in exercise induced anaphylaxis (Matsuo et al.
2005; Morita et al. 2009; Yokooji et al. 2013). Recombinant expressed proteins have been shown to
react with sera from patients with wheat allergies (Eriksson et al. 2012; Maruyama et al. 1998).
Screening of a wheat cDNA expression library with antisera from wheat allergenic patients
identified clones coding for HMW-GS, three likely representing a Dx gene and three Bx7, other
experiments identifying the latter as an important allergen (Baar et al. 2014). Three specific
sequences in HMW-GSs (which occur in both the Dx5 and Dy10 proteins (Anderson et al. 1989;
Sugiyama et al. 1985; Thompson et al. 1985)) have been identified as IgE-binding epitopes (Matsuo
et al. 2005).
In the case of LMW-GS proteins, a proteomic study of wheat allergens identified nine
members of this class as important allergens, but could not confirm previously reported examples
classified as HMW-GS (Akagawa et al. 2007). Techniques such as screening of a wheat cDNA
library with sera from patients with wheat allergy have also suggested a connection of LMW-GS
proteins to this condition (Baar et al. 2012).
Coeliac (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. People can also suffer from gluten intolerance (gluten
sensitivity), an adverse reaction to wheat that is neither an allergenic reaction or mediated by the
immune system (Pietzak 2012; Sapone et al. 2012).
Distinct varieties of wheat often have different quantities (per grain) of glutenin and gliadin
proteins, the quantities in any variety also being influenced by the environment in which the plant is
grown (Plessis et al. 2013). From a molecular viewpoint, the synthesis of these proteins 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 protein levels include members of the Opaque2 subfamily of basic
leucine zipper (bZIP) factors (storage protein activators) (Albani et al. 1997; Ravel et al. 2009) and
DOF (DNA binding with one finger) proteins (Dong et al. 2007; Romeuf et al. 2010). The levels of
such transcription factors, and hence seed storage proteins, may be linked to biochemical issues,
including the assimilation of nitrogen and the availability of amino acids. Quantitative trait loci
linked to the levels of gluten proteins have also been identified and mapped (Zhang et al. 2011).
5.4
Characterisation of the GMOs
5.4.1
Stability and molecular characterisation
Based on experiments in the United States, the genes are stably inserted into the genome of
the cultivar Bobwhite. There is no reason to expect that conventionally breeding of these genes into
other cultivars will affect their stability. However, the sites of insertion and copy numbers of the
inserted genes are not known. The proposed field trial will evaluate the expression of the genes in a
number of Australian cultivars.
5.4.2
Phenotypic characterisation
Preliminary phenotypic characterisation of the plants in glasshouses has demonstrated that the
introduced genes induce no major visible phenotypes or reduce viability.
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Section 6 The receiving environment
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).
The factors relevant to the growth, distribution and cultivation of commercial wheat can be
found in The Biology of Triticum aestivum L. em Thell (Bread Wheat) (OGTR 2008).
It is proposed that the dealings be conducted in the NGNE facility, operated by DAFWA,
located at Katanning in Western Australia.
6.1
Relevant abiotic factors
The Katanning NGNE facility is purpose built for the trialling of GM plants. Katanning
represents the high rainfall environment used for growing wheat in Western Australia. The soil of
Katanning is largely alkaline and sodic.
6.2
Relevant agricultural practices
It is not anticipated that the agronomic practices for the cultivation of the GM wheat by the
applicant will be significantly different from conventional practices for these plants.
GM wheat seeds would be planted in the trial site in winter or early spring. Seed that remain
after harvest would be either stored in an approved facility for subsequent use or destroyed.
Volunteers would be removed by hand or killed by herbicide application.
6.3
Presence of related plants in the receiving environment
Wheat (Triticum aestivum L.) 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. Wheat also readily hybridises with
Aegilops species, but no Aegilops species are considered to be naturalised in Australia. Any
specimens of Aegilops that have been collected in Australia presumably originate from seed
accidently introduced amongst wheat seed, or straying from that brought in for breeding programs
(AVH 2012).
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 unlikely that it occurs.
As the Katanning NGNE facility is designed to be accommodate more than one trial (which
may reflect more than one licence holder), it is possible that other GM and non-GM wheat plants
will be grown there in close proximity to those plants that are the subject of this application.
Currently, GM wheat plants from DIR 128 which are genetically modified for abiotic stress
tolerance or micronutrient uptake may be present.
6.4
Presence of similar genes and encoded proteins in the environment
The introduced genes and other genetic elements are from a plant (wheat) that is widespread
and prevalent in the environment (see Section 5). It is commonly consumed by people who are
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therefore exposed to the proteins. Other cereals (that are consumed by people) have glutenin
proteins (Shewry & Halford 2002).
The bar selectable marker gene comes from Streptomyces hygroscopicus, which is widespread
in the environment.
Section 7 Relevant Australian and international approvals
7.1
Australian approvals
7.1.1
Approval by the Regulator
Wheat lines possessing the introduced genes have not previously been approved by the
Regulator.
Information on previous DIR licences for GM wheat can be found on the GMO Record on the
OGTR website.
7.1.2
Approval by other government agencies
No other approvals are currently required for this GM wheat trial.
7.2 International approvals of GM wheat
Field trials of the GM wheat lines in the Bobwhite genetic background have been carried out
in the USA in 2002 and 2003 (information provided by the applicant).
Field trials of different GM wheat 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 tolerance3.
3
USDA release permit applications, EU GMO Register, accessed November 2014.
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Chapter 2
Office of the Gene Technology Regulator
Risk assessment
Section 1 Introduction
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.
RISK ASSESSMENT PROCESS *
Postulation
of risk
scenarios
Risk
context
Identification of
substantive risks
Consequence
assessment
Substantive
Risks
Risk
scenarios
Risk
Evaluation
Likelihood
assessment
Negligible risks
RISK IDENTIFICATION
RISK CHARACTERISATION
* Risk assessment terms are defined in the Risk Analysis Framework 2013
Figure 3 The risk assessment process
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.
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.
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. 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.
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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). 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 2 Risk Identification
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.
source of
potential harm
potential harm to
plausible causal linkage
(a novel GM trait)
an object of value
(people/environment)
Figure 4 Risk scenario
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).
2.1 Risk source
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.
As discussed in Chapter 1, each of the GM wheat lines has been modified by the introduction
of one of two HMW-GS genes, or a hybrid of both, that are expected to confer improvement in
grain quality. These introduced genes are considered further as potential sources of risk.
The GM lines also contain the bar selection marker gene (see Chapter 1, Section5.2.2). This
gene and its product has 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. Toxicity feeding study expermiments have failed to establish
any deleterious effects of the PAT protein upon animals (Herouet et al. 2005; MacKenzie et al.
2007; Merriman 1996). More information on selectable marker genes can be obtained from the
OGTR document Marker genes in GM plants, available on the OGTR website. As this gene has not
been found to pose substantive risks to either people or the environment, its potential effects will
not be further assessed for this application.
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In addition, the GM plants may also contain the bla gene (see Chapter 1, Section5.2.2). There
is no evidence that the protein encoded by this gene has toxic or allergenic properties. The
RARMPs for DIRs 070/2006 and 071/2006 concluded that the risks posed from the expression of a
prokaryotic promoter driven bla gene in GM plants were negligible. Even if the bla gene was
expressed in the wheat, human exposure to the beta-lactamase protein is routine as ampicillin
resistant bacteria occur widely in the environment. For example, E.coli with such a trait is common
in the normal human intestine (EFSA 2004; EFSA 2009). Therefore, this gene will not be assessed
further for this application.
The HMW-GS genes include endogenous wheat regulatory sequences while the promoter of
the bar gene is derived from a maize ubiquitin gene. There is no evidence that regulatory sequences
themselves have toxic or allergenic effects (EPA 1996); such effects for these sequences will not be
further assessed for this application. 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.
2.2 Causal pathway
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 establishment
o reproduction
o 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)
tolerance to cultivation management practices
gene transfer to sexually compatible organisms
gene transfer by horizontal gene transfer (HGT)
unauthorised activities.
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.
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.
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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 harm
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.
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 scenarios
Five risk scenarios were postulated and screened to identifiy substantive risk. These scenarios
are summarised in Table 2 and more detail of these scenarios is provided later in this Section.
Postulation of risk scenarios considers impacts of the GM wheat or their products on people
undertaking the dealings, as well as impacts on people and the environment if the GM plants or
genetic material were to spread and/or persist.
In the context of the activities proposed by the applicant and considering both the short and
long term, none of the five risk scenarios gave rise to any substantive risks.
Table 2 Summary of risk scenarios from dealings with GM wheat genetically modified for improved
grain quality
Risk
Risk source
scenario
1
Introduced
genes for
improved
grain quality
Chapter 2 – Risk assessment
Causal pathway
Potential harm
Growing GM plants at the site

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 site
 Allergic
reactions in
people or
toxicity in
people and
other
organisms
Substantive
risk?
No
Reason
 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.
13
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Risk
Risk source
scenario
Causal pathway
Office of the Gene Technology Regulator
Potential harm
Substantive
risk?
2
Introduced
genes for
improved
grain quality
Dispersal of GM seed outside
trial limits

Growth of GM plants

Expression of genes in 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
 Reduced
establishment
and yield of
desirable
plants
 Reduced
biodiversity
No
3
Introduced
genes for
improved
grain quality
Dispersal of GM pollen outside
trial limits

Vertical transfer of introduced
genes to other sexually
compatible plants, such as
commercial varieties of wheat

Expression of genes in plants

Exposure of people or other
organisms to GM plant material
 Allergic
reactions in
people or
toxicity in
people and
other
organisms
 Reduced
establishment
and yield of
desirable
plants
 Reduced
biodiversity
No
4
Introduced
genes for
improved
grain quality
Dispersal of GM pollen within
the NGNE facility

Hybridisation of GM plants of
this trial with GM plants
(including volunteers) of
another trial

Expression of genes in stacked
GM plants

Exposure of people or other
organisms to GM plant material
 Allergic
reactions in
people or
toxicity in
people and
other
organisms
No
Chapter 2 – Risk assessment
Reason
 The introduced proteins are
not known to be toxic.
Although they have been
associated with
allergenicity, there is
uncertainty whether they
will affect the allergenic
threshold in individuals.
 The limited scale, short
duration and other
proposed limits and
controls minimise the
likelihood that GM plant
material would leave a trial
site.
 The introduced proteins are
not known to be toxic.
Although they have been
associated with
allergenicity, there is
uncertainty whether they
will affect the allergenic
threshold in individuals.
 The introduced genes are
not expected to increase
the ability of the GM plants
to spread and persist.
 The limited scale, short
duration and other
proposed limits and
controls minimise the
likelihood that GM plant
material would leave a trial
site.
 The introduced proteins are
not known to be toxic.
Although they have been
associated with
allergenicity, there is
uncertainty whether they
will affect the allergenic
threshold in individuals.
 The introduced genes are
not expected to increase
the ability of the GM plants
to spread and persist.
 The limited scale, short
duration and other
proposed limits and
controls minimise exposure
of people and other
organisms to the GM plant
material.
 The introduced proteins are
not known to be toxic.
Although they have been
associated with
allergenicity, there is
uncertainty whether they
will affect the allergenic
14
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Risk
Risk source
scenario
5
2.4.1
Introduced
genes for
improved
grain quality
Office of the Gene Technology Regulator
Causal pathway
Potential harm
Dispersal of GM pollen within
NGNE site

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 the trial
limits

Expression of genes in stacked
GM plants

Spread and persistence of
populations of GM plants
outside a trial site
 Reduced
establishment
and yield of
desirable
plants
 Reduced
biodiversity
Substantive
risk?
No
Reason
threshold in individuals.
 The stacking of genes from
different GM plants is
unlikely to increase the
toxicity or allergenicity of
the hybrid GM plants.
 The limited scale, short
duration and other
proposed limits and
controls minimise the
likelihood that GM plant
material would leave a trial
site.
 The introduced genes are
not expected to increase
the ability of the GM plants
to spread and persist.
 The stacking of genes from
different GM plants is
unlikely to increase the
weediness of the hybrid
GM plants.
Risk scenario 1
Risk source
Introduced genes
for improved
grain quality
Causal pathway
Growing GM plants at the site
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 site
Potential harm
 Allergic
reactions in
people or
toxicity in
people and
other
organisms
Risk source
The source of potential harm for this postulated risk scenario is the introduced genes for
improved grain quality.
Causal pathway
The grain quality improvement genes are expressed in the plant tissues. People who are
involved in the breeding, cultivating, harvesting, transporting and experimenting of the GM wheat
may be exposed to its products through contact (including inhalation of pollen). This would be
expected to mainly occur in the trial site, but could also occur anywhere the GM plant material was
transported or used for experimental analysis. Organisms that may be present in the trial site,
including birds, rodents and invertebrates, may be exposed to the GM plant material.
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 a single site
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of 0.06 ha, only a small number of people would deal with the GM plant material and a small
number of other organisms are likely to be exposed to it. GM plant material is not to be used as
animal feed.
A fence surrounding the NGNE facility will exclude livestock and other large animals, while
rodent control measures will be used to reduce the number of these animals. Further, the facility has
bird netting.
The applicant has also proposed a series of measures, such as the monitoring and inspecting
of the site, 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 the trial site and
the adjacent areas.
Potential harm
People exposed to the proteins expressed from the introduced genes or their associated
products may show toxic or allergic reactions, while other organisms may show toxic reactions
(Chapter 2, Section 2.3). 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 the 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; Wu & Sun 2011). Other plant proteins,
found to be toxic at least to mice, include the urease-like-protein canatoxin from jack beans
(Follmer et al. 2001) and an acidic protein and a glycoprotein from soybean (Morais et al. 2010;
Vasconcelos et al. 2008; Vasconcelos et al. 1994).
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 superfamilies are the prolamins,
cupins, pathogenesis-related (PR) proteins, profilins and protease inhibitors. Beyond toxicity and
allergenicity, specific proteins and chemicals have been associated with autoimmune diseases and
food intolerances (ASCIA 2014; Barragan-Martinez et al. 2012; Selmi et al. 2012).
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. In respect of the general information on
toxicity and plant proteins outlined above, none of the introduced plant proteins can be classified as
a lectin or protease inhibitor (ie a likely toxin).
The glutens are prominent members of the prolamin family that have been associated with
allergenicity, and, as noted in Chapter 1, Section 5.3, the HMW-GS proteins have been associated
with negative effects on the health of people. These effects have been grouped as allergies (eg the
consumption of glutenin can induce baker’s asthma or exercise induced anaphylaxis), autoimmune
diseases (eg coeliac disease) and ‘gluten sensitivity’ (Sapone et al. 2012). Specific studies that have
used the Dx5 and Dy10 proteins, or epitope peptides based on their sequences, have implicated
these HMW-GSs in exercise induced anaphylaxis (Matsuo et al. 2005; Morita et al. 2009; Yokooji
et al. 2013).
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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 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.
The proteins Dx5 and Dy10 were examined against the first tier criteria:
(i) History of safe use. The introduced plant proteins come from a plant (wheat) that does not
raise any toxicological concerns and has a history of largely safe use in human diets. A
minority of people have allergenic reactions to wheat, possess coeliac disease or have ‘gluten
sensitivity’.
(ii) Bioinformatic analysis. The introduced proteins do not have any sequence similarity with
known toxins. However, they are members of a protein class, the prolamins, that is known to
have members with allergenic properties.
(iii) Mode of action. Plants are responsible for the production of a wide range of small
molecular weight compounds, usually designated as secondary metabolites, many of which
are toxic to humans and herbivore animals (Wink 2009; Wink & Van Wyk 2008). From the
perspective of the plant, they help defend the plant against the predatory activities of these
groups. The metabolites of greatest concern are the neurotoxins, followed by cytotoxins and
compounds that act as poisons in particular organs. The introduced proteins are considered as
storage proteins for amino acids that can be used in germination and seedling growth (Shewry
& Halford 2002). They do not have enzymatic functions. As such, they are not expected to
give rise to any (enzymatic) small molecular weight products that could have toxic properties.
(iv) In vitro digestibility and stability. No data.
(v) Expression level and dietary intake. No data.
The hybrid gene codes for a protein that consists of the 124 N-terminal amino acids from the
mature Dy10 protein fused N terminal to the C terminal 719 amino acids from Dx5 (Blechl &
Anderson 1996). Such a protein does not have a history of safe use, but the experience of
conventional breeding is that the spurious fusing of gene sequences does not lead to proteins that
are of health or environmental concern (Steiner et al. 2013; Weber et al. 2012). Further, as
discussed above, the sequences of Dx5 and Dy10 themselves have not been associated with toxicity,
and it is not expected that a fusion protein of these sequences will have a mode of action that is
different from its progenitors.
Although the Dx5 and Dy10 proteins are endogenous to wheat, it is possible that increased
quantities of one or both of these proteins may affect the allergenic threshold level of wheat with
respect to gluten. The threshold level of a food can be defined as the maximum quantity of a food,
containing one or more allergenic proteins, that can be tolerated without producing any adverse
(allergenic) reaction. It can be measured by a number of methods, including epidemiological
studies, analytical procedures and anecdotal evidence (FDA 2006). In these circumstances, data
from points (iv) and (v) may take on greater importance.
The threshold for eliciting an allergic reaction by a single allergen has rarely been determined,
varies between individual people, and can vary in any one individual over time due to factors such
as stress, exercise and the use of medications (Fernandez et al. 2013). Further, there have been few
studies of the concentrations of endogenous food allergens across different cultivars of one species.
When such studies have been conducted, allergen concentrations have been discovered to vary
widely between cultivars, and a range of environmental factors can affect the level in any individual
cultivar (Fernandez et al. 2013; Herman & Ladics 2011). No mechanisms are presently in place to
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evaluate and segregate ‘low’ and ‘high’ allergenic cultivars of plants that constitute our food supply
(Panda et al. 2013). With such a background, it is not possible to accurately answer the question of
how much of an increase in an endogenous allergen would lead to a measurable increase in the level
of harm.
Nevertheless, it is possible that an elevation in the level of HMW-GSs in GM wheat could
increase the spectrum of people who cannot tolerate food derived from this plant. Although people
with food allergies largely avoid the offending foods altogether (gluten-free food being defined by
standard 1.2.8, clause 16, of the Australia New Zealand Food Standards Code as ‘no detectable
gluten’ (http://www.comlaw.gov.au/Details/F2012C00218)), there are individuals with moderate
allergies who know, through experience, they can consume limited amounts of the these foods. The
consequence to such individuals of an increase in allergen levels in GM food could be an elicitation
of a severe allergenic response (Fernandez et al. 2013). However, as the GM wheat will not be used
for human food, no harm as a result of humans consuming the GM wheat will occur.
Experiments with the promoter of the Dx5 gene fused to the GUS reporter gene have
demonstrated that this promoter drives expression of a gene in the endosperm of seeds, with no
expression being detected in leaf, root inflorescence or floret tissues (Lamacchia et al. 2001).
Similar results have been obtained when other HMW-GS promoters have been used to drive
reporter genes (Furtado et al. 2009; Weibo et al. 2009). As glutenins are classified as seed storage
proteins, these patterns of expression are not unexpected. Thus, in relation to the GM wheat of this
application, there is unlikely to be a risk of a pollen-induced allergic reaction, but a possible route
that people handling plant material may be exposed to the proteins is through contact with seed.
Nevertheless, there is uncertainty as to whether the expression of Dx5 and Dy10 would affect the
allergenicity of the GM wheat proposed for release.
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 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 2003). However, even though conventional
breeding can involve the movement of hundreds and even thousands of genes into a plant, there has
never been a report of a completely novel toxin or allergen appearing in a new line of a plant
produced by such techniques (Steiner et al. 2013; Weber et al. 2012). The implication is that the
movement into wheat of any of the genes that are the subject of this application, none of which
belong to any known class of toxin, is unlikely to result in the production (directly or indirectly) of
a novel toxin. This includes the production of such a compound via the site of insertion or the
production of a fusion protein. In reference to allergenicity, the introduced glutenin proteins are in a
protein class containing allergens which is already present in non-GM wheat. However, the GM
wheat will not be used for human food.
Conclusion: Risk scenario 1 is not identified as a substantive risk due to the proposed limits
and controls designed to minimise exposure of people and other organisms to the GM plant material
and the lack of known toxicity of the introduced proteins. Therefore, this risk could not be greater
than negligible and does not warrant further detailed assessment.
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2.4.2
Office of the Gene Technology Regulator
Risk scenario 2
Risk source
Introduced genes
for improved
grain quality
Causal pathway
Dispersal of GM seed outside trial limits

Growth of GM plants

Expression of genes in GM plants

Spread and persistence of populations of GM plants outside trial limits

Exposure of people or other organisms to GM plant material
Potential harm
 Allergic reactions
in people or
toxicity in people
and other
organisms
 Reduced
establishment
and yield of
desirable plants
 Reduced
biodiversity
Risk source
The source of potential harm for this postulated risk scenario is the introduced genes for
improved grain quality.
Causal pathway
The grain quality improvement genes are expressed in plant tissues. If seed was dispersed
outside the trial site or persisted at the site, 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.
Dispersal of GM plant material outside the limits of the 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, through extremes of weather such as flooding or high winds, or
persistence at the site once the trial has finished.
Wheat lacks seed dispersal characteristics such as stickiness, burrs and hooks, which can
contribute to seed dispersal via animal fur (Howe & Smallwood 1982). The intended introduced
trait of improved grain quality is not expected to alter these characteristics of seeds.
Seed dispersal for wheat through endozoochory (the ingestion and excretion of viable seeds)
has not been reported. Nevertheless, it cannot entirely be discounted that wheat 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).
Kangaroos, rabbits and rodents are known pests of wheat crops, and cattle or sheep may graze
cereals. The site is fenced, limiting the possibility of seed dispersal by any large animals such as
cattle, sheep and kangaroos. 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). Other studies have shown that generally very few viable seeds are obtained
from rabbit dung (Welch 1985; Wicklow & Zak 1983). Rodents are opportunistic feeders and their
diet 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. Only a very limited amount of rodent activity has been observed at
trial sites under other GM field trial licences.
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
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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.
Baseline information on the weediness of wheat, 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 2008). In summary, wheat has some characteristics of invasive plants,
such as being capable of out-crossing (although it is predominantly self-pollinating) and the ability
to germinate or to produce seed in a range of environmental conditions. However, it lacks 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, wheat has been bred to avoid seed shattering, and white wheat cultivars have little seed
dormancy (OGTR 2008).
The expected phenotypic difference between the GM wheat lines and their non-GM
progenitors is improved grain quality. This introduced trait is 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. 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), the trait of improved
grain quality cannot readily be interpreted as a cue for the GM plants to increase their invasiveness.
The techniques of conventional breeding (eg selection of plants amongst available germplasm,
wide crosses, mutagenesis) have been used to produce varieties of wheat that possess a range of
traits. This experience is a useful backdrop against which to view the potential invasiveness of the
GM plants with improved grain quality in this application. 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. Mutagenesis has
also been used to generate a number of varieties of cereals (and many other crops) that have a
various traits (Ahloowalia et al. 2004; Cheema et al. 1999; Tomlekova 2010)(FAO/IAEA database
of mutation enhanced technologies for agriculture).
An increase in the quantity of seed storage proteins in seeds could increase the fitness of the
seeds, this in turn increasing the ability of the GM wheat plants to spread and persist. However, no
wheat plant that has been generated by any form of conventional breeding has been demonstrated to
have increased invasiveness. On the basis of this experience, the genetic modifications are not
expected to increase the potential invasiveness of GM plants relative to non-GM plants.
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.
If the GM wheat were to lose the trait of non-shattering heads, it would be expected to spread
and persist to a greater degree than non-GM commercial varieties, its seed being lost before harvest
and contributing greater amounts of seed to a seed bank. As a result, it would be less dependant on
human intervention for dispersal. However, there is no reason to believe that the introduction of the
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genes into wheat 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 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, the latter also
being a marker of domestication (Sang 2009).
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 0.06 ha per year) will limit the potential for
dispersal of GM plant material and exposure to this material. The proposed trial site is surrounded
by a fence and only approved staff with appropriate training will have access, this minimising
potential for dispersal of seed and exposure to plant material by grazing livestock and people.
Dispersal of GM plant material by authorised people entering the proposed trial site would be
further 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. Furthermore, extra conditions associated with growing the GM plants in the multiuser
NGNE facility would also reduce the likelihood of GM plant material being spread. Monitoring of
the trial site for volunteers once the GM wheat has been harvested is expected to minimise the
likehihood of persistence. Limits and controls are further discussed in Chapter 3.
Potential harm
If GM wheat plants were to establish beyond the trial limits, they could potentially cause one
or more of the harms outlined in Section 2.3 of this chapter. As discussed in risk scenario 1, the
introduced gene products are not expected to be toxic to humans or other organisms. Although
glutenins have been associated with allergenicity, there is uncertainty whether or not their
expression in wheat (the organism from which they derive) would affect the endogenous
allergenicity of that plant.
With respect to the environment, spread and persistence of the GM plants could reduce 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. In an agricultural setting, a persistent seed bank of GM wheat could
reduce the establishment and yield of subsequent crops.
The only expected phenotypic difference between GM wheat and non-GM wheat is altered
levels of glutenin. There is no reason to believe that such a trait will have any effect upon the ability
of the GM plants to spread and persist, thereby producing any environmental harms. As discussed
in Chapter 1, no phenotypic differences were observed between GM wheat and non-GM wheat
when grown in glasshouses, and a standard condition of a licence for a field trial would be that the
applicant immediately notify the OGTR of any unintended effects.
It is important to note that no commercially released variety of wheat that is the product of any
form of conventional breeding has been recorded to have negatively impacted the environment (or
the health of animals and/or animals), beyond that normally associated with these cereals, and
subsequently flagged as an environmental weed. These conventionally generated varieties represent
a wide range of traits (Goodman et al. 1987; Hajjar & Hodgkin 2007; Maxted & Kell 2009;
Prescott-Allen & Prescott-Allen 1998). Therefore, the introduction into wheat of any of the genes
that are the subject of this application is unlikely to produce a plant that poses a risk to the
environment that is any different from the many wheat plants produced by conventional breeding
(National Research Council 1989).
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Conclusion: Risk scenario 2 is not identified as a substantive risk due to the proposed limits
and controls designed to restrict dispersal of the GM wheat and to minimise exposure of people and
other organisms to the GM plant material, and the lack of known toxicity of the introduced proteins.
Further, the engineered trait is not associated with weediness. Therefore, this risk could not be
greater than negligible and does not warrant further detailed assessment.
2.4.3
Risk scenario 3
Risk source
Introduced
genes for
improved grain
quality
Causal pathway
Dispersal of GM pollen outside trial limits

Vertical transfer of introduced genes to other sexually compatible plants,
such as commercial varieties of wheat

Expression of genes in plants

Exposure of people or other organisms to GM plant material
Potential harm
 Allergic
reactions in
people or
toxicity in
people and
other
organisms
 Reduced
establishment
and yield of
desirable plants
 Reduced
biodiversity
Risk source
The source of potential harm for this postulated risk scenario is the introduced genes for
improved grain quality.
Causal pathway
The grain quality improvement genes are expressed in the plant tissues. Pollen from the GM
plants could be transferred outside of the trial site (eg via wind) and fertilise sexually compatible
plants, whether they be non-GM wheat, or plants from another sexually compatible species.
Alternatively, if seed was dispersed outside the trial site, 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 other sexually compatible
plant 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.
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. Baseline information on
vertical gene transfer associated with non-GM wheat plants can be found in The Biology of Triticum
aestivum L. em Thell (Bread Wheat) (OGTR 2008).
Wheat 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.
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
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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 with the four
native Australasian Triticeae genera never occurs under natural conditions.
The proposed limits and controls of the trial would minimise the likelihood of the dispersal of
pollen and exposure to GM plant material. For example, the applicant proposes to control related
species within 200 m of the trial site. Isolation from related species and other wheat 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 remain that could then hybridise with sexually compatible plants.
Potential harm
If the vertical transfer of the introduced genes from the GM plants caused the resulting plants
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 are summarised in Section 2.3 of this chapter.
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. The
GM plants may act to reduce the establishment and yield of desired plants and subsequently reduce
biodiversity.
In the rare event of the vertical transfer of the introduced genetic material from the GM plants
to non-GM wheat plants or sexually compatible species, it is expected that this material in the
recipient will have the properties that it possesses in the GM wheat parent. As discussed in risk
scenario 1, the introduced gene products are not expected to be toxic to humans or other organisms.
Although they have been associated with allergenicity, there is uncertainty whether their expression
in wheat (the organism from which they derive) would affect the endogenous allergenicity of that
plant. There is no reason to expect these gene products to have toxic or allergenic properties in a
recipient arising from hybridisation with sexually compatible species that are any different from
those found in the GM wheat. The uncertainty with respect to allergenicity would apply to any
plants that were the product of hybridisation. Risk scenario 2 summarises the reasons that the
introduced genes are unlikely to make the GM wheat lines more weedy, these reasons likewise
being applicable to any plants to which the genes are transferred via hybridisation.
The traits that have been introduced into the GM plants of this application could become, via
vertical gene transfer, combined with traits of other non-GM commercially cultivated wheat plants.
However, there is no reason to believe that the resulting plants would possess a level of toxicity or
allergenicity greater than that of either parent, or a level of weediness greater than that of either
parent.
Conclusion: Risk scenario 3 is not identified as a substantive risk due to the proposed limits
and controls designed to restrict dispersal of pollen flow from the GM wheat and to minimise
exposure of people and other organisms to the GM plant material, and the lack of known toxicity of
the introduced proteins. Further, the introduced trait is not associated with weediness, and it is
unlikely that any of the characteristics associated with weeds will occur in hybrids with the GM
plants. Therefore, this risk could not be greater than negligible and does not warrant further detailed
assessment.
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2.4.4
Office of the Gene Technology Regulator
Risk scenario 4
Risk source
Introduced
genes for
improved grain
quality
Causal pathway
Dispersal of GM pollen within the NGNE facility

Hybridisation of GM plants of this trial with GM plants (including volunteers)
of another trial

Expression of genes in stacked GM plants

Exposure of people or other organisms to GM plant material
Potential harm
 Allergic
reactions in
people or
toxicity in
people and
other
organisms
Risk source
The source of potential harm for this postulated risk scenario is the introduced genes for
improved grain quality.
Causal pathway
The grain quality improvement genes are expressed in the 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 late 2014, the only licence that is authorised to trial GM plants
in the Katanning NGNE facility that are sexually compatible with those of this application is DIR
128, held by The University of Adelaide. This latter licence authorises the growth of GM wheat and
barley with either abiotic stress tolerance or micronutrient uptake.
People working in or visiting the NGNE facility could be exposed to the GM hybrid plants or
material from the GM hybrid plants, such as pollen. Any animals accessing the site may also be
exposed to hybid GM plants.
The applicant has requested permission to grow GM wheat from this licence, and any other
licence, next to each other provided they are separated by buffer zones of at least 4 m (2m for this
trial and 2m for other trial). The RARMP for DIR 094 considered the possibility of hybridisation
between wheat plants occurring over these distances, and concluded that it would be minimal.
The licence for DIR 128 specifies that if the GM plants of that trial are grown at the
Katanning NGNE facility at the same time as sexually compatible GM and non-GM plants from
another licence, seed from any plants of DIR 128 must not be used for the development of
commercial cultivars. Imposing such a measure on the proposed trial of this application would help
minimise the likelihood that the improved grain quality genes would be transferred to other plants
and that people and animals would inadvertently be exposed to GM material.
The proposed limits and controls would minimise the possibility that any hybrid seed would
leave the trial site. These are discussed in risk scenario 2. Therefore, exposure would be restricted to
people working at or visiting 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 and animals.
Potential harm
Exposure to the proteins expressed from the introduced genes or their associated products
through contact or consumption of GM plant material may cause toxicity or allergenicity in
humans, or toxicity in other organisms.
As discussed in risk scenario 1, the introduced gene products are not expected to be toxic to
humans or other organisms. Although they have been associated with allergenicity, there is
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uncertainty whether their expression in wheat (the organism from which they derive) would affect
the endogenous allergenicity of that plant.
The toxicity and allergenicity associated with the introduced genes of other sexually
compatible GM plants growing in the NGNE facility would be the subject of the RARMP(s)
associated with their licence(s). According to the RARMP for DIR 128, there is no information to
suggest that the proteins encoded by the introduced genes of that application are likely to be toxic to
people or other organisms, or allergenic to people. 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 4 is not identified as a substantive risk due to the proposed limits
and controls designed to minimise exposure of people and other organisms to the GM plant material
and the lack of known toxicity of the introduced proteins. There is no reasonable expectation that
the stacking of genes from different GM trials will lead to plant with an increased level of toxicity.
Therefore, this risk could not be greater than negligible and does not warrant further detailed
assessment.
2.4.5
Risk scenario 5
Risk source
Introduced
genes for
improved grain
quality
Causal pathway
Dispersal of GM pollen within the NGNE facility

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 the trial limits

Expression of genes in stacked GM plants

Spread and persistence of populations of GM plants outside a trial site
Potential harm
 Reduced
establishment
and yield of
desirable plants
 Reduced
biodiversity
Risk source
The source of potential harm for this postulated risk scenario is the introduced genes for
improved grain quality.
Causal pathway
The grain quality improvement genes are expressed in the plant tissues. As discussed in risk
scenario 4, 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 plants with abiotic stress tolerance and/or micronutrient uptake from DIR 128 may
be present at the Katanning NGNE facility.
If hybridisation occurred between the GM plants of this application and those of DIR 128, the
progeny would have the traits of abiotic stress tolerance, micronutrient uptake stacked with
improved grain quality.
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, considering the
nature of the traits in DIR 128 and this application, there is no reason to expect that their stacking
will significantly increase the invasiveness of the hybrid GM plants beyond those intrinsic to the
parent GM plants.
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Risk scenario 4 reviews the growing of GM plants from different trials adjacent to each other,
such as possibly may occur in the NGNE facility, noting the applicant proposes using buffer zones
and structuring the licence similar to other licences that have sought to minimise outcrossing
between different GM plants growing in close proximity.
The proposed limits and controls of the trial would minimise the possibility that seed would
leave any trial site or persist at the site once the trial is completed. These are discussed under risk
scenario 2. All GM seed would be transported in accordance with the Regulator’s transport
guidelines, which would minimise the opportunity for its dispersal.
Potential harm
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 Section 2.3 of this chapter and risk
scenario 2. In particular, the GM plants may act to reduce the establishment and yield of desired
plants and biodiversity.
As discussed in risk scenario 2, the introduced gene products are unlikely to cause the GM
wheat lines to have adverse impacts on the environment greater than those normally associated with
non-GM wheat, 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
the NGNE facility would be the subject of the RARMP(s) associated with their licence(s).
According to the RARMP for DIR 128, there is no information to suggest that the proteins encoded
by the introduced genes of that application are likely to increase the weediness in GM wheat. 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 5 is not identified as a substantive risk due to the proposed limits
and controls designed to restrict persistence and dispersal of the GM wheat. There is no reasonable
expectation that the stacking of genes from different GM trials will lead to plant with a level of
weediness greater than that of the progenitor GM plants. Therefore, this risk could not be greater
than negligible and does not warrant further detailed assessment.
Section 3 Uncertainty
Uncertainty is an intrinsic part of risk analysis4. There can be uncertainty about identifying the
risk source, the causal linkage to harm, the type and degree of harm, the chance 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.
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.
There are several types of uncertainty in risk analysis (Bammer & Smithson 2008; Clark &
Brinkley 2001; Hayes 2004). These include:

uncertainty about facts:
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.
4
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
Office of the Gene Technology Regulator
–
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.
For DIR 130, uncertainty is noted particularly in relation to the characterisation of:
Potential increases in toxicity or allergenicity as a result of the genetic modification
Potential for increased survivial of the GMOs, including in land uses outside of agriculture,
and
Potential for an increase in the level of risk as a result of stacking of the introduced genes
with those in other sexually compatible GM plants grown in the NGNE facility.
Additional data, including information to address the uncertainties identified above, may be
required to assess possible future applications for a larger scale trial, reduced containment
conditions, or the commercial release of these GM wheat lines if they are selected for further
development.
Chapter 3, Section 4, discusses information that may be required for future release.
Section 4 Risk evaluation
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.
Factors used to determine which risks need treatment may include:




risk criteria
level of risk
uncertainty associated with risk characterisation
interactions between substantive risks.
Five risk scenarios were postulated whereby the proposed dealings might give rise to harm to
people or the environment. In the context of the control measures proposed by the applicant, and
considering both the short and long term, none of these scenarios were identified as substantive
risks that could be greater than negligible. The principal reasons for these conclusions are
summarised in Table 2 and include:
limits on the size, location and duration of the release proposed by Murdoch University
controls proposed by Murdoch University to restrict the spread and persistence of the GM
wheat 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 and proteins in the environment and lack
of evidence of harm from them
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limited ability and opportunity for the GM wheat plants to transfer the introduced genes to
commercial wheat crops or other sexually related species
none of the GM plant materials or products will enter human food or animal feed supply
chains.
Therefore, risks to the health and safety of people, or the environment, from the prosed release
of the GM wheat plants into the enviromnet are considered to be negligible. 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. Hence, the Regulator considers
that the dealings involved in this proposed release do not pose a significant risk to either people or
the environment.
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Risk management
Section 1 Background
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 and considers controls and limits proposed by the applicant, as well as
general risk management measures. The risk management plan informs the Regulator’s decisionmaking process and is given effect through licence conditions.
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.
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.
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 risks
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. 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), the receiving environment (Chapter 1,
Section 6), and considering both the short and the long term. The risk evaluation concluded that no
additional controls are required to treat these negligible risks.
Section 3 General risk management
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, location 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 release
3.1.1
Consideration of limits and controls proposed by Murdoch University
Sections 3.1 and 3.2 of Chapter 1 provide details of the limits and controls proposed by
Murdoch University 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 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.
The duration of the field trial would be confined to three growing seasons. The trial would be
limited to a single site with a maximum area of 0.06 ha per year. The small size and duration of the
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trial would limit the potential exposure of humans and other organisms to the GMOs (risk scenario
1).
Only authorised personnel would be permitted to deal with the GMOs. A standard licence
condition requires all people dealing with the GMOs to be informed of relevant licence conditions.
These measures would limit the potential exposure of humans to the GMOs (risk scenario 1).
The Katanning NGNE trial site is surrounded 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).
A rodent control program is a standard feature of the NGNE facility. Combined with the use
of a monitoring zone (below), this measure should both limit exposure of rodents to the GMOs (risk
scenario 1) and minimise potential dispersal of GMOs outside the trial site 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 the site.
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
measures are needed to prohibit the access of birds to the trial site. However, in this respect it
should be noted that the NGNE facility at Katanning is covered by bird netting.
The applicant proposes to surround the trial site with a monitoring zone of 10 m width within
which the growth of plants should be controlled. This zone is a standard requirement of GM wheat
licences. Such an area is also an unfavourable habitat for rodents. In summary, the monitoring zone
would fulfill the following purposes:

to facilitate detection of plants or related species that might hybridise with GM wheat

to reduce rodent activity in the NGNE facility.
The applicant proposes that the fence around the Katanning NGNE facility would be
surrounded by 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 GMO planting area, monitoring
zone and isolation zone, and any found must be destroyed prior to flowering.
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, a
width of 200 m 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, surrounded by a 190 m isolation zone, will manage gene flow to other
wheat crops and related species (risk scenario 3).
The NGNE facility is designed to accommodate more than one user. Hence, as there is the
possibility that separate trials of other sexually compatible GM plants (most obviously wheat) may
concurrently take place in the Katanning facility, 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 each (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. The more recent licence DIR 128 does not exclude the
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growing of other GM wheat plants in the two NGNE facilities. A minimum distance of 4 m (which
corresponds to the 2m buffer zones from each trial) should separate the GM plants of this
application and any other sexually compatible GM plants that are grown in the Katanning NGNE
facility. If any GM plants in this application are grown in the Katanning NGNE facility 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 or vertical gene transfer (risk scenarios 3, 4 and 5), and subsequent spread of, or
exposure to, this GM material.
The applicant has proposed a number of measures to minimise the persistence of any GM
wheat 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, area planted to the GM wheat 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.
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).
It is therefore considered that under Australian conditions, a 2 year time period during which
shallow 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 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.
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 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.
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 4 and 5). This is included as a licence condition.
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.
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As the GM wheat 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
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 the NGNE facility of Katanning, WA, between May 2015 and December
2017

limit the maximum area of planting to 0.06 ha in any year

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
may be grown, and where growth of related species are controlled

harvest the GM wheat plant material separately from other crops

clean the areas and equipment after use

apply measures to promote germination of any wheat 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 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

not use any seed for further planting if other trials of sexually compatible plants are
occurring in the NGNE facility concurrently

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 considerations
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 site and other places for the
purpose of monitoring or auditing.
3.2.1
Applicant suitability
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)
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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.
On the basis of information submitted by the applicant and records held by the OGTR, the
Regulator considers Murdoch University suitable to hold a licence.
The licence includes a requirement for the licence holder to inform the Regulator of any
circumstances that would affect their suitability. 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
Murdoch University 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.
Murdoch University would also be 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 would be 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
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, Murdoch University is also be 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
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.
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
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
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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.
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.
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 releases
Additional information has been identified that may be required to assess an application for a
large scale or commercial release of these GM wheat lines, or to justify a reduction in containment
conditions. This includes:
additional molecular and biochemical characterisation of the GM wheat lines, particularly
with respect to production of potential toxins or allergens
additional phenotypic characterisation of the GM wheat lines, particularly with respect to
traits that may contribute to weediness.
Section 5 Conclusions of the RARMP
The risk assessment concludes that this proposed limited and controlled release of GM wheat
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.
However, conditions have been imposed to limit the release to the proposed size, location 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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DIR 130 – Risk Assessment and Risk Management Plan
Office of the Gene Technology Regulator
Appendix A Summary of submissions from prescribed
experts, agencies and authorities5
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.
Summary of issues raised
Comment
Agrees with the overall conclusions of the RARMP.
Noted.
Should consider further data on allergenicity that may
be required for any future commercial release
application.
Future data requirements identified in Section 4, Chapter 3 lists
potential allergens produced by the GM wheat lines.
Agrees with the Regulator’s assessment that this trial
poses negligible risks to human health and safety or
the environment under the proposed conditions of the
RARMP.
Noted.
Satisfied with the conclusions of the RARMP and has
no technical comments.
Noted.
No objection to the issue of a licence for DIR 130.
Noted.
Notes that the licence will prohibit the use of material
from the trials for human or animal consumption. No
further comments on the licence application at this
stage.
Noted
5
Prescribed agencies include GTTAC, State and Territory Governments, relevant local governments, Australian
Government agencies and the Minister for the Environment.
Appendix A
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DIR 130 – Risk Assessment and Risk Management Plan
Office of the Gene Technology Regulator
Appendix B Summary of submissions from the public
The Regulator received five 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.
Abbreviations:
Issues raised: AT: Alternative technology; C: Containment; CC: Consumer choice; CP:
Consultation process; D: Detection of GMOs; E: Environment; Ec: Economic issues; F: Food
safety; FL: Food labelling; H: Human health; L: liability; LC: Licence condition; M: Monitoring;
MT: Marketing and trade; R: Research; RA: Risk analysis; S: Segregation
Other abbreviations: Act: the Gene Technology Act 2000; APVMA: Australian Pesticides and
Veterinary Medicines Authority; Ch: Chapter; FSANZ: Food Standards Australia New Zealand;
GM: Genetically modified; GMO: Genetically modified organism; Regulator: the Gene
Technology Regulator.
Submission Issue
number
1
Appendix B
Summary of issues raised
Comment
C
Opposed to all trials of DIR 130 GM wheat
unless they are in a totally confined area
from which no pollen can escape.
Wheat plants are predominantly self-pollinating
and the chances of natural hybridisation occurring
with commercial crops or other sexually
compatible plants are low. Licence conditions
require isolation from other wheat plants to
minimise the opportunity of pollen flow.
H
Claims that numerous studies (eg Seralini,
Carman) have demonstrated with animals
the serious possible health effects from
eating GM food. More and more doctors in
the US are advising patients to avoid all GM
foods. Consumers, once they have
recognised the effect GM food can have on
their gut and consequently their health, will
never accept GM food. No GM foods have
been proven safe despite the claims of the
industry.
No products from this GM wheat trial will enter the
human food supply. The RARMP concluded that
the limited and controlled release of the GM wheat
lines included in this application poses negligible
risks to the health and safety of people.
FSANZ has critically evaluated the studies cited as
evidence of adverse effects from GM foods and
concluded that these studies provided no grounds
to revise its conclusions on the safety of food
derived from the previously approved GM crops
(details available on the FSANZ website).
C, Ec,
MT, FL
Claims that GM food will always only be
used for industrial purposes or animal feed
with lower prices due to strong consumer
resistance.
Concerned that the whole wheat harvest in
Australia would have to be labelled as GM
due to contamination if GM wheat is
released. WA wheat is already at risk due to
testing of GM wheat in the open
environment.
Also claims that the only reason for the
continued imposition of GM food on the
general public is either corporations or
governments which believe they will make a
lot of money.
Strict licence conditions have been imposed to
minimise the likelihood of the GM wheat mixing
with non-GM wheat.
Issues of marketing and trade implications are
outside the scope of the Regulator’s assessment
required by the Act. These issues are the
responsibility of the State Governments and the
industry. FSANZ is responsible for labelling of GM
foods.
45
DIR 130 – Risk Assessment and Risk Management Plan
Submission Issue
number
Office of the Gene Technology Regulator
Summary of issues raised
Comment
E
GM canola is spreading down the roads in
areas of the wheat belt in WA and this is
also a serious problem in the US. GM wheat
will make the weed problem worse.
The DIR 130 licence imposes strict controls that
restrict the spread and persistence of the GMOs in
the environment, including monitoring of the trial
site to ensure no GMOs remain following
completion of the trial.
AT
Claims that selective breeding and marker
assisted breeding have been proven to be
far more successful than any GM breeding
and commodities from such technologies
would be welcomed by the public.
The comparison and choice between technologies
are outside the scope of assessment by the
Regulator.
2
CP
Requests that the trial not proceed and that
all GM trials be specifically approved by the
citizens and residents of the council
governing the area in which the trial GM
wheat will be planted.
As required by the Act, the Regulator consulted with
the Local Government Area where the GM wheat is
proposed to be planted Consultation with the public
was also undertaken, which involved advertising in
the Australian and the WA Farm Weekly
newspapers, the Australian Government gazette
and on the OGTR website, as well as emailing
people on OGTR’s client register. The Regulator
considered the issues raised in the submissions
related to human health and safety and the
environment before making the decision to issue the
licence.
3
H, E, R,
M, PU
Urges the OGTR to reject the DIR 130
application.
Advocates a GM freeze - no new GM foods
approved; no new GM crop approvals and
no new GM crop growing areas. During the
GM freeze, the safety of GM crops on sale
must be re-assessed and data on yield,
pesticide use and effects on biodiversity
must be investigated and the results of
these studies made public. Before any more
open field trials are approved, thorough
testing, monitoring, surveillance, problem
reporting and recall mechanisms are
needed.
The Regulator has prepared a comprehensive
RARMP, in accordance with the requirements of
the Act, and includes a comprehensive and critical
assessment of data supplied by the applicant,
together with a thorough review of other relevant
national and international scientific literature. The
RARMP concluded that risks to human health and
safety and the environment, including effects on
biodiversity are negligible as a result of this limited
and controlled release of GM wheat. No products
from this GM wheat trial will enter the human food
supply.
The DIR 130 licence imposes conditions for
monitoring the trial site for 24 months after field
trials have completed to ensure no GMOs remain
before the site can be signed off. Prior to
conducting any dealings, the licence holder is also
required to prepare a contingency plan for
measures to be taken in the event of the
unintended presence of the GMOs outside an area
that must be inspected, including reporting,
recovering, destroying and inspecting any of the
GMOs. The licence requires that any risks to
health and safety of people or the environment or
unintended effects be reported. The Regulator is
able to suspend, cancel or vary a licence if risks
associated with the field trial are identified.
The regulation of agricultural chemicals in
Australia is the responsibility of APVMA.
Appendix B
46
DIR 130 – Risk Assessment and Risk Management Plan
Submission Issue
number
Appendix B
Office of the Gene Technology Regulator
Summary of issues raised
Comment
F
Claims that there is a growing body of
evidence which questions the safety of GM
food and the American Academy of
Environmental Medicine recommends
avoiding GM foods when possible.
No products from this GM wheat trial will enter the
human food supply. FSANZ is responsible for
human food safety assessment, including GM
food.
C, L
It remains unresolved who is responsible for
the control and clean-up of the spread and
spill of GM seed and crops. There is little or
no education and training for non-GM
farmers, government instrumentalities and
the general public.
Claims that escapes from GM trials have
occurred and attempts to eradicate the
resulting GM weeds post-trial is likely to
continue for many years, potentially forever.
For limited and controlled release, the licence
holder is responsible for the control and clean-up of
the spread or spill of GM material outside the trial
sites. Prior to conducting any dealings, the licence
holder is required to prepare a contingency plan for
measures to be taken in the event of the unintended
presence of the GMOs outside an area that must be
inspected, including reporting, recovering,
destroying and inspecting any of the GMOs.
Monitoring and management of GM wheat
volunteers beyond the trial period are discussed in
Chapter 3 of the RARMP. The licence requires at
least two years of monitoring the trial site and
destroying volunteers before the site can be
considered for sign-off. This would effectively
manage survival and persistence of viable wheat
seeds in the soil.
In the case of commercial release of GM crops,
matters relating to segregation and coexistence of
different farming systems are the responsibility of
the States, Territories and industry.
C
OGTR "Buffer zones" of 200 metres around
the crops are inadequate to stop the spread
of transgenic material. Contamination has
also occurred during transport of GM
material.
Based on scientific literature, a 200 m isolation
zone is considered in the RARMP as adequate to
limit gene flow to non-GM wheat. This has been
shown to be an effective containment measure for
GM wheat trials under previously issued licences.
The licence holder is required to comply with DIR
130 licence conditions for safe transport of GM
material.
C
Claims that GM crops are proven to
contaminate with examples from: 1) In 2013,
unapproved GM wheat from trials years
earlier was found growing in an Oregon field
in the USA, which resulted in rejections of
US wheat shipments to a number of Asian
destinations; 2) China intercepted &
destroyed shipments of GM corn seed that
attempted to bypass Chinese environmental
& food safety tests; 3) most of the GM
canola trial sites in Tasmania have still not
been cleaned after 10 years.
As this application is for a limited and controlled
release (field trial), strict licence conditions are
imposed to restrict the spread and persistence of
the GMOs and their genetic material in the
environment. These include conditions to isolate
the trial site from other wheat and crops, cleaning
of equipment used with GM plant materials,
secure transport and storage of GM plant
materials, and post-harvest monitoring at the trial
site to ensure all GM plants are destroyed. Similar
conditions have been effective for other GM wheat
trials conducted in Australia.
C
Concern has been raised about the
maintenance of GM wheat research at
Merredin, WA, following an ABC report in
June 2014.
The OGTR is aware of the claims made in the
media and has investigated them. The Regulator
was satisfied that the issue did not represent a risk
to people and the environment and work with
GMOs undertaken in the Merredin facility is
managed according to licence conditions. The
licence holder intends to plant at Katanning facility
and not at Merredin.
47
DIR 130 – Risk Assessment and Risk Management Plan
Submission Issue
number
4
Appendix B
Office of the Gene Technology Regulator
Summary of issues raised
Comment
MT
GM wheat is not approved or grown
commercially anywhere in the world. Claims
that there is no apparent market for GM
wheat products and the WA Farmers
Association Grains Group has commented
that GM wheat would do more damage
to WA wheat markets than good. A petition
to stop Australian wheat trials also
happened in New Caledonia.
See comments for Submission 1 regarding issues
relating to marketing and trade.
S, MT
The Marsh v Baxter appeal is set to be
heard in the Court of Appeal of the WA
Supreme Court March 23-25,
2015. Assessment of this application must
be delayed until after the appeal judgement
as it could contribute significantly to the risk
assessment.
The Marsh vs Baxter legal case relates to
commercially approved GM crops and involves
segregation and marketing issues, not health and
safety issues, and as such is outside the scope of
the Regulator’s assessment required by the Act.
C
Based on a recent ABC report, regulatory
See comments for Submission 3 regarding this
oversight, surveillance and monitoring at the issue.
NGNE is poor, creating a significant risk of
escape of GM organisms.
D
The community must be educated on the
trials before release so that the community
can be vigilant about escapes. Gene test
kits must be available so that GM
contamination outside the trial area can be
tested.
This is a limited and controlled field trial with strict
licence conditions to minimise the likelihood of
escape from the trial limits. Field trial sites where
GMOs are planted are published on the OGTR
website.
Prior to conducting any dealings with the GMOs,
the licence holder must provide to the Regulator
written methodology to reliably detect the GMOs.
M
History proves there is a high risk of
escapes such as the unapproved GM wheat
found in a wheat field in Oregon causing
importing countries to place immediate
embargoes on GM wheat. This occurred
many years after the trial was
completed. The DIR130 RARMP must
reflect the ongoing surveillance, testing and
monitoring beyond the trial period.
See comments for Submission 3 regarding
containment of GM wheat trials in Australia.
Monitoring and management of GM wheat
volunteers beyond the trial period are discussed in
Chapter 3 of the RARMP and the licence imposes
strict measures to minimise the spread and
persistence of the GM wheat.
48
DIR 130 – Risk Assessment and Risk Management Plan
Submission Issue
number
5
Summary of issues raised
Comment
H
Long-term animal feeding studies with
human health end-points must be
conducted in a confined controlled setting
before intentional release of the GM
organisms to the environment. This has not
been done and a theoretical argument of
“substantial equivalence” cannot safely be
assumed until such testing has been done
to determine unintended consequences and
to quantify risks of side-effects. Harm is
indicated from GM organisms and
associated pesticides and herbicides, for
example:
 the American Academy of
Environmental Medicine
recommends avoiding GM foods
when possible
 A peer-reviewed paper reported
harm to pigs fed GM feed
 Glyphosate has been found in
mother’s breast milk, urine and
babies’ feeding tubes.
FSANZ is responsible for human food safety
assessment, including GM food. An analysis of the
pig feeding study has been published by FSANZ
(http://www.foodstandards.gov.au/consumer/gmfoo
d/Pages/Response-to-Dr-Carman%27s-study.aspx).
Issues relating to herbicide use are outside the
scope of the Regulator’s assessments. The
APVMA has regulatory responsibility for
agricultural chemicals, including herbicides, in
Australia.
CC
The world has already rejected GM wheat
from earlier attempts in the US and Canada
in 2004 to commercialise it. Consumers do
not want to place their families at
risk. Application DIR 130 must be rejected.
Consumer choice is outside the scope of
responsibility of the Regulator. This is a GM wheat
field trial and not a commercial release. No
products from this field trial will enter the human
food supply.
MT
Strongly opposed to this high risk trial and
calls for a GM freeze. It jeopardizes our
major export market, and undermines
Australia's standing and credibility overseas
and at home.
Issues of marketing and trade implications are
outside the scope of the Regulator’s assessment
required by the Act. These issues are the
responsibility of the State Governments and the
industry.
R, H, E
Claims that the OGTR and FSANZ are
inextricably linked and share a joint
accountability. Neither can be relied on
because the government has forced them to
be reliant on GM company researchers for
safety testing. Thorough, peer reviewed,
long term epidemiological studies by
independent medical researchers remain to
be done on human health and safety, and
environmental safety.
When preparing a RARMP, the Regulator
considers not only information provided by the
applicant, but also published scientific literature,
and advice received from a range of Australian
government authorities, agencies, experts and the
public. Only when satisfied that any risks to human
health and the environment can be managed, the
Regulator will consider issuing a licence for
intentional release of the assessed GMOs into
Australian environment.
F
A dedicated national surveillance system for
potential health effects of GM foods needs
to be put in place immediately.
FSANZ is responsible for human food safety
assessment, including GM food.
All risks associated with GM crops,
eg increased use of herbicides and
pesticides, must be incorporated into the
risk analysis framework
Risks to the health and safety of people and the
environment as a result of the release of the GM
wheat are considered by the Regulator. Issues
related to the use of agricultural chemicals are the
responsibility of APVMA.
RA
Appendix B
Office of the Gene Technology Regulator
49