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TitleDIR 054/2004 - Office of the Gene Technology Regulator
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Table of Contents
                            EXECUTIVE SUMMARY
	Introduction
	The application
	The evaluation process
	Conclusions of the risk assessment
		Toxicity or allergenicity to humans and other organisms
		Weediness
		Transfer of introduced genes to other organisms
	The risk management plan (key licence conditions)
		Toxicity or allergenicity to humans and other organisms
		Weediness
		Transfer of introduced genes to other organisms
		General conditions
		Identification of issues to be addressed for future releases
		Monitoring and enforcement of compliance by the OGTR
	Further information
CHAPTER 1 BACKGROUND
	Section 1 The application
		Section 1.1 The proposed dealings
		Section 1.2 Parent organism
		Section 1.3 Genetic modifications and their effects
		Section 1.4 Method of genetic modification
	Section 2 Previous releases and international approvals
		Section 2.1 Previous Australian releases of similar GMOs
		Section 2.2 Approvals by other Australian government agencie
			2.2.1 Food Standards Australia New Zealand
		Section 2.3 International approvals
CHAPTER 2 SUMMARY OF RISK ASSESSMENT AND RISK MANAGEMENT PLA
	Section 1 Issues raised in Submissions on the Application an
	Section 2 Finalisation of the Risk Assessment and Risk Manag
	Section 3 Identification of Issues to be Addressed for Futur
	Section 4 Decision on the application
APPENDIX 1  INFORMATION ABOUT THE GMOS
	Section 1 Summary information about the GMOs
	Section 2 The parent organism
	Section 3 Method of genetic modification
		Section 3.1 Agrobacterium-mediated transformation
		Section 3.2  RNA interference (RNAi)
	Section 4 The introduced genes and their products
		Section 4.1  Effect of Silencing the SE genes
		Section 4.2  The nptII antibiotic resistance marker gene and
		Section 4.3  T-DNA border elements
	Section 5 Characterisation of the inserted genetic material
	Section 6 Expression of the introduced proteins
	Section 7 Pleiotropic effects of the genetic modification
	Section 8 Research requirements
APPENDIX 2 TOXICITY AND ALLERGENICITY TO HUMANS AND OTHER OR
	Section 1 Nature of the potential toxicity or allergenicity
	Section 2 Likelihood of the toxicity or allergenicity hazard
		2.7.1 Toxicity
			Silencing of an SE gene
				Alteration of starch composition
				NPTII protein
			2.7.2 Allergenicity
				Silencing of SE
				Alteration of starch composition
				NPTII protein
	Section 3 Conclusions regarding toxicity or allergenicity
	Section 4 Research Requirements
APPENDIX 3 WEEDINESS
	Section 1 Nature of the weediness hazard
	Section 2 Likelihood of the weediness hazard occurring
		Section 2.1 Inherent weediness of conventional non-GM wheat
		Section 2.2 Potential weediness of the GM wheat lines
		Section 2.3 Potential selective advantage conferred by the i
			2.3.2  Gene silencing construct
			2.3.2 Antibiotic resistance (NPTII) protein
			2.3.3 Silencing of the SE genes in combination with NPTII ex
		Section 2.4 Persistence of the GM wheat at the trial sites
		Section 2.5 Spread of GM wheat beyond the release site
	Section 3 Conclusions regarding weediness
	Section 4 Research Requirements
APPENDIX 4 TRANSFER OF INTRODUCED GENES TO OTHER ORGANISMS
	Section 1 Gene transfer from the GM wheat to other plants
		Section 1.1 Nature of the gene transfer hazard
		Section 1.2 Potential hazards from the introduced genes
			1.2.1 The gene silencing construct targeting the SE gene
			1.2.2 The nptII (antibiotic resistance) gene
			1.2.3 Promoters and other regulatory sequences
		Section 1.3 Likelihood of a hazard arising through gene tran
			1.3.1 Gene transfer to other Triticum aestivum plants
			1.3.2 Transfer to other Triticum, Aegilops and related speci
	Section 2 Gene transfer from the GM wheats to microorganisms
		Section 2.1 Nature of the gene transfer hazard
		Section 2.2 Potential hazards from the introduced genetic ma
			2.2.1 The gene silencing construct targeting an SE gene
			2.2.2 The nptII gene
			2.2.3 Promoters and other regulatory sequences
		Section 2.3 Other sources of the introduced genes in the env
			2.3.1 The gene silencing construct targeting the SE gene
			2.3.2 The nptII (antibiotic resistance) genes
		Section 2.4 Likelihood of a hazard arising through gene tran
			2.4.1 Bacteria
			2.4.2 Viruses
			2.4.3 Fungi
	Section 3 Gene transfer from the GM Wheat lines to animals,
		Section 3.1 Nature of the gene transfer hazard
		Section 3.2 Potential hazards from the introduced genetic ma
			3.2.1 The gene silencing construct targeting the SE gene
			3.2.2 The nptII (antibiotic resistance) gene
			3.2.3 Promoters and other regulatory sequences
		Section 3.3 Likelihood of a hazard arising through gene tran
	Section 4 Conclusions regarding gene transfer to other organ
		Section 4.1 Conclusions regarding gene transfer to non-GM wh
		Section 4.2 Conclusions regarding gene transfer to microorga
		Section 4.3 Conclusions regarding gene transfer to animals,
	Section 5 Research requirements
APPENDIX 5  LICENCE CONDITIONS
	SECTION 1  Interpretation and definitions
	SECTION 2  General conditions
		Duration of Licence
			Holder of Licence
			Project Supervisor
			No dealings with GMOs except as authorised by this licence
			GMOs covered by this licence
			Permitted dealings
			Persons covered by this GMO licence
			Informing people of their obligations
			Licence holder to notify of circumstances that might affect
			Additional information to be given to the Regulator
			People dealing with GMOs must allow auditing and monitoring
			Remaining an accredited organisation
	SECTION 3  Specific conditions
		Locations and size of trial
			Notice of planting
			Conditions relating to the Location
			Conditions about the Isolation Zone
			Inspections to be conducted in the Location while the GMOs a
			Inspections to be conducted in the Monitoring Zone while the
			Inspections to be conducted in the Isolation Zone while GMOs
			Notice of Harvest
			GMOs must be either harvested or Destroyed
			Seed and other Plant Material may be collected and stored
			Conditions in relation to the Cleaning of Location after GMO
			Notice of Cleaning
			General conditions in relation to the Cleaning of all other
			General conditions that apply wherever inspections must be u
			Restrictions during and after the GMOs are grown
			Transportation of the GMOs and Plant Material
			Contingency Plans
			Compliance Management Plan
			Reporting
			Testing methodology
			GMOs and Plant Material must not be consumed
APPENDIX 6 LEGISLATIVE REQUIREMENTS FOR ASSESSING DEALINGS I
	Section 1 The regulation of gene technology in Australia
	Section 2 The licence application
	Section 3 The initial consultation processes
	Section 4 The evaluation processes
	Section 5 Further consultation
	Section 6 Decision on licence
APPENDIX 7 SUMMARY OF PUBLIC SUBMISSIONS
APPENDIX 8 REFERENCES
                        
Document Text Contents
Page 1

Risk Assessment
and

Risk Management Plan





Application for licence for dealings involving an
intentional release into the environment






DIR 054/2004



Title: Field trial of genetically modified wheat
(Triticum aestivum L.) with altered grain starch






Applicant: CSIRO








April 2005

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DIR 054/2004 – Risk Assessment and Risk Management Plan Office of the Gene Technology Regulator

ABBREVIATIONS

antisense reverse (or backward) sequence
AOSCA Association of Official Seed Certifying Agencies
APVMA Australian Pesticides and Veterinary Medicines Authority
bp base pair
cm centimetre
CSIRO Commonwealth Scientifi c and Industrial Research Organisation
cv cultivar
DIR dealing involving intentional release
DNA deoxyribonucleic acid
dsRNA double stranded ribonucleic acid
EFSA European Food Safety Authority
EMBL European Molecular Biology Laboratory
endogenous derived from an organism
EPA Environmental Protection Agency of the United States of America
FAO Food and Agriculture Organisation of the United Nations
FDA Food and Drug Administration of the United States of America
FSANZ Food Standards Australia New Zealand
GM genetically modified
GMAC Genetic Manipulation Advisory Committee
GMO genetically modified organism
GTTAC Gene Technology Technical Advisory Committee
ha hectare
IgE immunoglobulin E
kD kilodalton
km kilometre
m metre
mRNA messenger ribonucleic acid
NLRD Notifiable Low Risk Dealing
nm nanometres
nos gene encoding nopaline synthase
nptII gene encoding NPTII
NPTII neomycin phosphotransferase II
OECD Organisation for Economic Cooperation and Development
OGTR Office of the Gene Technology Regulator
PCR polymerase chain reaction
PTGS Post-Transcriptional Gene Silencing
RARMP Risk Assessment and Risk Management Plan
RISC RNA-induced silencing complex
RNA ribonucleic acid
RNA ribonucleic acid
RNAi RNA interference
RNase III RNA degrading enzyme with specificity for dsRNA
sense coding sequence of DNA
siRNA small interfering RNA
spp species
T0….T5 generations after transformation
T-DNA transfer deoxyribonucleic acid
UK United Kingdom
US United States of America
US EPA United States Environmental Protection Agency

ABBREVIATIONS

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DIR 054/2004 – Risk Assessment and Risk Management Plan Office of the Gene Technology Regulator

interactions between this silencing and the antibiotic resistance marker gene or the metabolic
pathway in which this operates, and no reason to expect that this is likely to occur.

Section 2.4 Persistence of the GM wheat at the trial sites

172. Following harvest of the wheat from the trial sites, some remaining non viable plant
material would be incorporated into the soil. Some seed may fall to the ground at maturity
(seed shatter) and during harvest which may also be incorporated into the soil. However, this
would be minimised by the use of hand harvesting, which would be necessary to maintain the
purity of each GM wheat line. As explained in Appendix 1, wheat is not prone to shattering
and as indicated in Section 2.1 above, white wheats have very little seed dormancy.

173. Tillage of the release site post-harvest is not required. The parent wheat NB1 is a white
wheat and is not expected to possess dormancy factors. Hence irrigation without tillage of the
release site is likely to encourage germination of volunteers while treatment with herbicides
will eliminate any volunteers.

174. The applicant originally proposed a 12 month post-harvest monitoring period following
the two year trial. However due to reports that viable wheat seeds can persist in soil for
considerably longer than 12 months (16 -24 months, Anderson & Soper 2003) and based on
advice received, the Regulator decided that the post harvest monitoring period be extended to
24 months. The applicant proposes to promote germination of volunteer wheat by irrigation
of the trial area at least twice following harvest in December or January and to destroy any
volunteers by application of a non-selective herbicide. No crops would be planted at the site
between the winter wheat growing seasons.

175. Licence conditions have been imposed that require post harvest monitoring for 24
months at the conclusion of the final season and destruction of wheat volunteers to ensure
that the GM wheat does not persist in the environment at the release site.

Section 2.5 Spread of GM wheat beyond the release site

176. The size of the trial site, combined with the limited duration of two growing seasons,
would minimise the likelihood of the GM wheat lines establishing as weeds outside of the
proposed trial sites. In addition the parental cultivar, NB1 from the UK is not adapted to
Australian conditions, which is likely to limit its ability to persist in the Australian
environment. Licence conditions impose a 500 m isolation zone (from any other wheat)
which includes a 10 m monitoring zone in which the presence of volunteer wheat and
sexually compatible species will be monitored.

177. The fencing around the trial site would limit the movement of plant materials away
from the trial site through the exclusion of large animals. These GM wheat lines are in a
white wheat parental background, which have a thin seed coat and are therefore easily broken
down in the digestive system of mammals and birds. Wheat is widely used as a feed for birds
because it is nutritious and easily digested (Yasar 2003). While the high amylose trait may
make these GM wheat lines less nutritious, as the starch is less available (see Appendix 1), it
is unlikely to affect utilisation of the other components of the wheat grain and hence unlikely
to result in excretion of viable seed by birds. Even if birds or animals were to enter the site,
they would be more likely to eat the GM wheat or grain on site rather than carry it elsewhere
for storage or consumption. In the event that grain was dispersed by birds, it is unlikely that

APPENDIX 3 WEEDINESS 42

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DIR 054/2004 – Risk Assessment and Risk Management Plan Office of the Gene Technology Regulator

the grains would germinate and develop as weeds. Therefore there is no requirement to deter
birds by employing whistling tape or bird-proof netting.

178. Workers moving within the GM wheat would need to ensure that GM wheat seeds are
not transported to other wheat growing sites on their clothing. There is also the possibility of
dispersal occurring due to the activities of rodents and insects, which may be able to access
the sites.

179. The risk of small animals, such as mice, gaining access to the trial sites and removing
plant material and seeds would depend greatly on the density of these animals in the trial area
at the time of the trial. Mouse numbers fluctuate every three to four years on average in
Australia and after a mouse plague can remain very low for up to two years (Brown &
Singleton 2002). Periods of between four and seven years are typical between mouse plagues
in a particular region (Brown & Singleton 2002). Rodents are opportunistic feeders and their
diet can include seeds, the pith of stems and other plant materials (Caughley et al. 1998). If
any grain is removed from the trial sites, it is likely to be consumed by the rodents (see
Appendix 2 for an assessment of the potential toxicity of the GM wheat lines). Rodent
plagues do not occur frequently in the Australian Capital Territory and no rodent species are
considered to be pest animals (Caughley et al. 1998). The likelihood of rodents inhabiting the
trial site can be minimised by removal of material capable of attracting or harbouring them.

180. If not consumed, the risk of seed germination and growth to reproductive maturity
would be low as wheat has poor competitive ability against other plants (Keeler 1989; Keeler
et al. 1996; Lemerle et al. 2001). The average territory size of mice varies between breeding
and non-breeding seasons, from 0.015 to 0.2 hectares respectively (Caughley et al. 1998). A
circle with a radius of 25 m has an area of just over 0.2 hectares, suggesting that the 500 m
isolation zone would prevent rodents moving GM wheat seeds to unmonitored growing
environments. In the event that grain was dispersed by rodents, it is unlikely that the grains
would germinate and develop as weeds.

181. A variety of insects are likely to feed on the crop, however it is unlikely that most of
these would contribute to the dispersal of material from the GM wheat plants beyond the trial
sites. It is possible that ants may remove seeds for underground storage but to depths where
germination is highly unlikely. No data is available on the species of ants present at the trial
sites, so typical territory size and seed storage behaviour is unknown.

182. Gene flow via cross-pollination with non-GM wheat plants would contribute to the
potential spread and persistence of the GMOs in the environment outside the release sites.
This issue is discussed in detail in Appendix 4, Section 1.3.

183. The proposed dealing includes cultivation of the GM wheat lines and retention of all
wheat seed for storage, laboratory research or further planting in the following season. The
applicant proposes that wheat seeds would be harvested by hand at the end of the growing
season, double-bagged and transported in containers to secure storage facilities, thus limiting
the potential for accidental dispersal of the GM wheat seed beyond the trial sites. Wheat seed
produced in the proposed field trials would not be used for human food, nor would wheat
seed, straw or other wheat products be used for animal feed.

184. Conditions of heavy rainfall could result in seed from this site being washed away.
However, the applicant has stated that the trial site is flat and not subject to flooding. It is a

APPENDIX 3 WEEDINESS 43

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